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2.
PLoS One ; 10(3): e0119641, 2015.
Article in English | MEDLINE | ID: mdl-25811607

ABSTRACT

Phosphoglucose isomerase (PGI) catalyzes the reversible isomerization of glucose-6-phosphate and fructose-6-phosphate. It is involved in glycolysis and in the regeneration of glucose-6-P molecules in the oxidative pentose phosphate pathway (OPPP). In chloroplasts of illuminated mesophyll cells PGI also connects the Calvin-Benson cycle with the starch biosynthetic pathway. In this work we isolated pgi1-3, a mutant totally lacking pPGI activity as a consequence of aberrant intron splicing of the pPGI encoding gene, PGI1. Starch content in pgi1-3 source leaves was ca. 10-15% of that of wild type (WT) leaves, which was similar to that of leaves of pgi1-2, a T-DNA insertion pPGI null mutant. Starch deficiency of pgi1 leaves could be reverted by the introduction of a sex1 null mutation impeding ß-amylolytic starch breakdown. Although previous studies showed that starch granules of pgi1-2 leaves are restricted to both bundle sheath cells adjacent to the mesophyll and stomata guard cells, microscopy analyses carried out in this work revealed the presence of starch granules in the chloroplasts of pgi1-2 and pgi1-3 mesophyll cells. RT-PCR analyses showed high expression levels of plastidic and extra-plastidic ß-amylase encoding genes in pgi1 leaves, which was accompanied by increased ß-amylase activity. Both pgi1-2 and pgi1-3 mutants displayed slow growth and reduced photosynthetic capacity phenotypes even under continuous light conditions. Metabolic analyses revealed that the adenylate energy charge and the NAD(P)H/NAD(P) ratios in pgi1 leaves were lower than those of WT leaves. These analyses also revealed that the content of plastidic 2-C-methyl-D-erythritol 4-phosphate (MEP)-pathway derived cytokinins (CKs) in pgi1 leaves were exceedingly lower than in WT leaves. Noteworthy, exogenous application of CKs largely reverted the low starch content phenotype of pgi1 leaves. The overall data show that pPGI is an important determinant of photosynthesis, energy status, growth and starch accumulation in mesophyll cells likely as a consequence of its involvement in the production of OPPP/glycolysis intermediates necessary for the synthesis of plastidic MEP-pathway derived hormones such as CKs.


Subject(s)
Arabidopsis/metabolism , Cytokinins/metabolism , Glucose-6-Phosphate Isomerase/metabolism , Mesophyll Cells/metabolism , Photosynthesis , Starch/metabolism , Alleles , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Erythritol/analogs & derivatives , Erythritol/metabolism , Genetic Loci , Glucose-6-Phosphate Isomerase/chemistry , Glucose-6-Phosphate Isomerase/genetics , Metabolic Networks and Pathways , Mutation , Phenotype , Plant Leaves/metabolism , Sugar Phosphates/metabolism
3.
Blood ; 121(1): 197-206, 2013 Jan 03.
Article in English | MEDLINE | ID: mdl-23109243

ABSTRACT

Although peripheral blood stem cells (PBSCs) have replaced bone marrow (BM) as the most common unrelated donor progenitor cell product collected, a direct comparison of concurrent PBSC versus BM donation experiences has not been performed. We report a prospective study of 2726 BM and 6768 PBSC donors who underwent collection from 2004 to 2009. Pain and toxicities were assessed at baseline, during G-CSF administration, on the day of collection, within 48 hours of donation, and weekly until full recovery. Peak levels of pain and toxicities did not differ between the 2 donation processes for most donors. Among obese donors, PBSC donors were at increased risk of grade 2 to 4 pain as well as grade 2 to 4 toxicities during the pericollection period. In contrast, BM donors were more likely to experience grade 2 to 4 toxicities at 1 week and pain at 1 week and 1 month after the procedure. BM donors experienced slower recovery, with 3% still not fully recovered at 24 weeks, whereas 100% of PBSC donors had recovered. Other factors associated with toxicity included obesity, increasing age, and female sex. In summary, this study provides extensive detail regarding individualized risk patterns of PBSC versus BM donation toxicity, suggesting donor profiles that can be targeted with interventions to minimize toxicity.


Subject(s)
Blood Component Removal/adverse effects , Blood Donors , Bone Marrow Transplantation , Fatigue/etiology , Fever/etiology , Granulocyte Colony-Stimulating Factor/adverse effects , Hematopoietic Stem Cell Mobilization/adverse effects , Pain/etiology , Peripheral Blood Stem Cell Transplantation , Tissue Donors , Tissue and Organ Harvesting/adverse effects , Adolescent , Adult , Anesthesia/adverse effects , Blood Cell Count , Convalescence , Exanthema/epidemiology , Exanthema/etiology , Fatigue/epidemiology , Female , Fever/epidemiology , Filgrastim , Gastrointestinal Diseases/epidemiology , Gastrointestinal Diseases/etiology , Granulocyte Colony-Stimulating Factor/pharmacology , Hematopoietic Stem Cell Mobilization/methods , Hospitalization/statistics & numerical data , Humans , Male , Middle Aged , Obesity/epidemiology , Pain/epidemiology , Prospective Studies , Recombinant Proteins/adverse effects , Recombinant Proteins/pharmacology , Syncope/epidemiology , Syncope/etiology , Tissue and Organ Harvesting/methods , United States , Young Adult
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