The effect of group or individualised pelvic floor exercises with or without ultrasonography guidance for urinary incontinence in elderly women - A pilot study.

INTRODUCTION: Pelvic floor exercises combined with patient education has shown to be a promising intervention for women suffering from urinary incontinence. This pilot study investigated the effect of patient education combined with group or individualised pelvic floor exercises, or individualised pelvic floor exercises with ultrasonography guidance. METHODS: Thirty-three elderly women with urinary incontinency completed a block-randomised, assessor-blinded study combining patient education with 12-weeks of pelvic floor exercises either group-based or individual with or without ultrasonography guidance. Urinary incontinence symptoms were assessed using the Incontinence Impact Questionnaire-7 (IQ-7) and Urogenital Distress Inventory-6 (UDI-6). Furthermore, daily fluid intake and number of bathroom visits were recorded. Pelvic floor muscle strength was assessed using a manual squeeze test (Oxford Scale, 6-point). RESULTS: An increase in pelvic floor strength was observed after 12 weeks for both the individual (P = 0.038) and the individual ultrasonography-guided (P = 0.01) exercise groups. However, only the latter group maintained an increased strength at the 24-week follow-up (P = 0.008). Across all groups, the intervention led to a decrease in bathroom visits (P = 0.002) that was maintained at the 24-week follow-up (P < 0.001). The interventions led to a decrease in UDI-6 both after the 12-week intervention (P = 0.009) and at the 24-week follow-up (P = 0.032). CONCLUSIONS: These findings indicate that pelvic floor exercises together with patient education can reduce urogenital distress and bathroom visits without change in fluid intake. Furthermore, when pelvic floor exercises were conducted individually, pelvic floor strength increased, but pelvic floor strength was maintained over time only for individualised pelvic floor exercises with ultrasonography guidance.

A nuclear-localized protein, KOLD SENSITIV-1, affects the expression of cold-responsive genes during prolonged chilling in Arabidopsis.

Plants respond to cold by transcriptional and metabolic responses which underlie tolerance and acclimation mechanisms, but details at the molecular level are incomplete. Here we describe KOLD SENSITIV-1 (KOS1), a new gene required for responses to cold. KOS1 protein is predicted to have coiled-coil, Structural Maintenance of Chromosomes and nuclear-targeting domains. GFP-labeled KOS1 localizes to the nucleus. Null mutants could not be isolated but two independent knockdown T-DNA mutants were obtained. Growth and development of kos1 knockdown mutant plants was comparable to wild type when grown at 21°C. However, when grown at 4°C these mutants exhibited accelerated leaf yellowing and smaller rosette size than wild type. Quantitative RT-PCR revealed that in the cold kos1 mutants had reduced expression of cold-responsive transcripts COR15A, COR15B, BAM3 and AMY3. Metabolite profiling revealed that ascorbate levels were lower in the mutants in the cold relative to wild type. KOS1 therefore represents a new gene that influences the regulation of transcript and metabolite levels in response to prolonged chilling temperatures.

Similar protein phosphatases control starch metabolism in plants and glycogen metabolism in mammals.

We report that protein phosphorylation is involved in the control of starch metabolism in Arabidopsis leaves at night. sex4 (starch excess 4) mutants, which have strongly reduced rates of starch metabolism, lack a protein predicted to be a dual specificity protein phosphatase. We have shown that this protein is chloroplastic and can bind to glucans and have presented evidence that it acts to regulate the initial steps of starch degradation at the granule surface. Remarkably, the most closely related protein to SEX4 outside the plant kingdom is laforin, a glucan-binding protein phosphatase required for the metabolism of the mammalian storage carbohydrate glycogen and implicated in a severe form of epilepsy (Lafora disease) in humans.

Osmotic stress changes carbohydrate partitioning and fructose-2,6-bisphosphate metabolism in barley leaves.

Carbohydrate metabolism was investigated in barley leaves subjected to drought or osmotic stress induced by sorbitol incubation. Both drought and osmotic stress resulted in accumulation of hexoses, depletion of sucrose and starch, and 5-10-fold increase in the level of the regulatory metabolite fructose-2,6-bisphosphate (Fru-2,6-P2). These changes were paralleled by an increased activity ratio of fructose-6-phosphate,2-kinase / fructose-2,6-bisphosphatase (F2KP). The drought-induced changes in carbohydrate content and Fru-2,6-P2 metabolism were reversed upon re-watering. This reveals a reversible mechanism for modification of the F2KP enzyme activity. This suggests that F2KP might be involved in altering carbohydrate metabolism during osmotic stress. However, labelling with [14C]-CO2 showed that sucrose synthesis was not inhibited, despite the increased Fru-2,6-P2 levels, and demonstrated that increased flux into the hexose pools probably derived from sucrose hydrolysis. Similar effects of osmotic stress were observed in leaf sections incubated in the dark, showing that the changes did not result from altered rates of photosynthesis. Metabolism of [14C]-sucrose in the dark also revealed increased flux into hexoses and reduced flux into starch in response to osmotic stress. The activities of a range of enzymes catalysing reactions of carbohydrate metabolism in general showed only a marginal decrease during osmotic stress. Therefore, the observed changes in metabolic flux do not rely on a change in the activity of the analysed enzymes. Fructose-2,6-bisphosphate metabolism responds strongly to drought stress and this response involves modification of the F2KP activity. However, the data also suggests that the sugar accumulation observed during osmotic stress is mainly regulated by another, as yet unidentified mechanism.

Identification of more than 200 glucose-responsive Arabidopsis genes none of which responds to 3-O-methylglucose or 6-deoxyglucose.

The response of some plant genes to glucose analogues 3-O-methylglucose (3OMG) or 6-deoxyglucose (6DOG) has been cited as evidence for metabolism-independent glucose signalling. To analyse such signalling using a genetic approach, we sought to identify Arabidopsis glucose-responsive genes which also respond to 3OMG and 6DOG in seedlings. Microarray analysis of gene expression in glucose-treated seedlings and RT-PCR analysis of glucose-treated leaf sections identified more than 200 glucose-responsive genes, but none responded to 3OMG or 6DOG. These data together with other published data on individual genes fail to identify any Arabidopsis sugar-responsive genes which also respond to 3OMG or 6DOG.

Fructose-2,6-bisphosphate: a traffic signal in plant metabolism.

Fructose-2,6-bisphosphate (Fru-2,6-P(2)) regulates key reactions of the primary carbohydrate metabolism in all eukaryotes. In plants, Fru-2,6-P(2) coordinates the photosynthetic carbon flux into sucrose and starch biosynthesis. The use of transgenic plants has allowed the regulatory models to be tested by modifying the Fru-2,6-P(2) levels and the enzymes regulated by Fru-2,6-P(2). Genes for the bifunctional plant enzyme that synthesizes and degrades Fru-2,6-P(2) have been isolated and molecular characterization has provided new insight into structure and molecular regulation of the enzyme. Advances in Fru-2,6-P(2) physiology and molecular biology are discussed. These advances have not only enlightened in vivo operation of Fru-2,6-P(2) but also revealed that the Fru-2,6-P(2) regulatory system is highly complex and interacts with other regulatory mechanisms.

Phosphorylation and 14-3-3 binding of Arabidopsis 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase.

Fructose 2,6-bisphosphate (fru-2,6-P2) is a signalling metabolite that regulates photosynthetic carbon partitioning in plants. The content of fru-2,6-P2 in Arabidopsis leaves varied in response to photosynthetic activity with an abrupt decrease at the start of the photoperiod, gradual increase through the day, and modest decrease at the start of the dark period. In Arabidopsis suspension cells, fru-2,6-P2 content increased in response to an unknown signal upon transfer to fresh culture medium. This increase was blocked by either 2-deoxyglucose or the protein phosphatase inhibitor, calyculin A, and the effects of calyculin A were counteracted by the general protein kinase inhibitor K252a. The changes in fru-2,6-P2 at the start of dark period in leaves and in the cell experiments generally paralleled changes in nitrate reductase (NR) activity. NR is inhibited by protein phosphorylation and binding to 14-3-3 proteins, raising the question of whether fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase protein from Arabidopsis thaliana (AtF2KP), which both generates and hydrolyses fru-2,6-P2, is also regulated by phosphorylation and 14-3-3s. Consistent with this hypothesis, AtF2KP and NR from Arabidopsis cell extracts bound to a 14-3-3 column, and were eluted specifically by a synthetic 14-3-3-binding phosphopeptide (ARAApSAPA). 14-3-3s co-precipitated with recombinant glutathione S-transferase (GST)-AtF2KP that had been incubated with Arabidopsis cell extracts in the presence of Mg-ATP. 14-3-3s bound directly to GST-AtF2KP that had been phosphorylated on Ser220 (SLSASGpSFR) and Ser303 (RLVKSLpSASSF) by recombinant Arabidopsis calcium-dependent protein kinase isoform 3 (CPK3), or on Ser303 by rat liver mammalian AMP-activated protein kinase (AMPK; homologue of plant SNF-1 related protein kinases (SnRKs)) or an Arabidopsis cell extract. We have failed to find any direct effect of 14-3-3s on the F2KP activity in vitro to date. Nevertheless, our findings indicate the possibility that 14-3-3 binding to SnRK1-phosphorylated sites on NR and F2KP may regulate both nitrate assimilation and sucrose/starch partitioning in leaves.