Nationwide Acute Care Physical Therapist Practice Analysis Identifies Knowledge, Skills, and Behaviors That Reflect Acute Care Practice

Background and Purpose. Acute care physical therapists have experienced the effects of dramatic changes in health care reimbursement systems and population demographics. Acute care hospitals now serve a patient population of much older, chronically ill patients who are hospitalized for shorter periods of time in a practice environment in which physical therapy staffing resources are often inadequate. The purposes of this study were to document common experiences in the practice of acute care physical therapy and to identify differences in the perceptions of physical therapists with varying levels of experience and in various sizes of acute care facilities. Subjects and Methods. A survey questionnaire was mailed to 500 randomly selected physical therapists employed in acute care facilities. The therapists answered questions regarding the frequency of various physical therapy evaluation and treatment practices, problems encountered in delivering physical therapy services, coordination of the discharge planning process, and perceptions of staffing trends in the acute care setting. The responses of 188 physical therapists who completed the survey were compared by their experience levels and the size of the institutions in which they practiced. Results. Subjects reported that patient factors, such as medical complications and cooperation; organizational factors, such as staffing shortages and large caseloads; and health care system constraints, such as difficulty changing orders and limited time in which to work with the patient interfered with patients reaching physical therapy goals. Conclusion and Discussion. Inadequate skills for successful acute care practice and maladaptive therapist beliefs about acute care career possibilities may adversely affect physical therapist career longevity in the acute care setting

Nationwide Acute Care Physical Therapist Practice Analysis Identifies Knowledge, Skills, and Behaviors That Reflect Acute Care Practice

Background and Purpose. Acute care physical therapists have experienced the effects of dramatic changes in health care reimbursement systems and population demographics. Acute care hospitals now serve a patient population of much older, chronically ill patients who are hospitalized for shorter periods of time in a practice environment in which physical therapy staffing resources are often inadequate. The purposes of this study were to document common experiences in the practice of acute care physical therapy and to identify differences in the perceptions of physical therapists with varying levels of experience and in various sizes of acute care facilities. Subjects and Methods. A survey questionnaire was mailed to 500 randomly selected physical therapists employed in acute care facilities. The therapists answered questions regarding the frequency of various physical therapy evaluation and treatment practices, problems encountered in delivering physical therapy services, coordination of the discharge planning process, and perceptions of staffing trends in the acute care setting. The responses of 188 physical therapists who completed the survey were compared by their experience levels and the size of the institutions in which they practiced. Results. Subjects reported that patient factors, such as medical complications and cooperation; organizational factors, such as staffing shortages and large caseloads; and health care system constraints, such as difficulty changing orders and limited time in which to work with the patient interfered with patients reaching physical therapy goals. Conclusion and Discussion. Inadequate skills for successful acute care practice and maladaptive therapist beliefs about acute care career possibilities may adversely affect physical therapist career longevity in the acute care setting

Interfering with Glycolysis Causes Sir2-Dependent Hyper-Recombination of Saccharomyces cerevisiae Plasmids

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key metabolic regulator implicated in a variety of cellular processes. It functions as a glycolytic enzyme, a protein kinase, and a metabolic switch under oxidative stress. Its enzymatic inactivation causes a major shift in the primary carbohydrate flux. Furthermore, the protein is implicated in regulating transcription, ER-to-Golgi transport, and apoptosis. We found that Saccharomyces cerevisiae cells null for all GAPDH paralogues (Tdh1, Tdh2, and Tdh3) survived the counter-selection of a GAPDH–encoding plasmid when the NAD+ metabolizing deacetylase Sir2 was overexpressed. This phenotype required a fully functional copy of SIR2 and resulted from hyper-recombination between S. cerevisiae plasmids. In the wild-type background, GAPDH overexpression increased the plasmid recombination rate in a growth-condition dependent manner. We conclude that GAPDH influences yeast episome stability via Sir2 and propose a model for the interplay of Sir2, GAPDH, and the glycolytic flux

Regulation of plant cytosolic glyceraldehyde 3-phosphate dehydrogenase isoforms by thiol modifications.

Cytosolic NAD-dependent glyceraldehyde 3-P dehydrogenase (GAPDH; GapC; EC 1.2.1.12) catalyzes the oxidation of triose phosphates during glycolysis in all organisms, but additional functions of the protein has been put forward. Because of its reactive cysteine residue in the active site, it is susceptible to protein modification and oxidation. The addition of GSSG, and much more efficiently of S-nitrosoglutathione, was shown to inactivate the enzymes from Arabidopsis thaliana (isoforms GapC1 and 2), spinach, yeast and rabbit muscle. Inactivation was fully or at least partially reversible upon addition of DTT. The incorporation of glutathione upon formation of a mixed disulfide could be shown using biotinylated glutathione ethyl ester. Furthermore, using the biotin-switch assay, nitrosylated thiol groups could be shown to occur after treatment with nitric oxide donors. Using mass spectrometry and mutant proteins with one cysteine lacking, both cysteines (Cys-155 and Cys-159) were found to occur as glutathionylated and as nitrosylated forms. In preliminary experiments, it was shown that both GapC1 and GapC2 can bind to a partial gene sequence of the NADP-dependent malate dehydrogenase (EC 1.2.1.37; At5g58330). Transiently expressed GapC-green fluorescent protein fusion proteins were localized to the nucleus in A. thaliana protoplasts. As nuclear localization and DNA binding of GAPDH had been shown in numerous systems to occur upon stress, we assume that such mechanism might be part of the signaling pathway to induce increased malate-valve capacity and possibly other protective systems upon overreduction and initial formation of reactive oxygen and nitrogen species as well as to decrease and protect metabolism at the same time by modification of essential cysteine residues

Influence of the photoperiod on redox regulation and stress responses in Arabidopsis thaliana L. (Heynh.) plants under long- and short-day conditions

Becker B, Holtgrefe S, Jung S, et al. Influence of the photoperiod on redox regulation and stress responses in Arabidopsis thaliana L. (Heynh.) plants under long- and short-day conditions. PLANTA. 2006;224(2):380-393.Arabidopsis thaliana L. (Heynh.) plants were grown in low light (150 mu mol photons m(-2) s(-1) and 20 degrees C) either in short days (7.5 h photoperiod) or long days ( 16 h photoperiod), and then transferred into high light and low temperature (350-800 mu mol photons m(-2) s(-1) at 12 degrees C). Plants grown in short days responded with a rapid increase in NADP-malate dehydrogenase (EC 1.1.1.82) activation state. However, persisting overreduction revealed a new level of regulation of the malate valve. Activity measurements and Northern-blot analyses indicated that NADP-malate dehydrogenase transcript and protein levels increased within a few hours. Using macroarrays, additional changes in gene expression were identified. Transcript levels for several enzymes of glutathione metabolism and of some photosynthetic genes increased. The cellular glutathione level increased, but its redox state remained unchanged. A different situation was observed in plants grown in long-day conditions. Neither NADP-malate dehydrogenase nor glutathione content changed, but the expression of several antioxidative enzymes increased strongly. We conclude that the endogenous systems that measure day length interact with redox regulation, and override the interpretation of the signals, i.e. they redirect redox-mediated acclimation signals to allow for more efficient light usage and redox poising in short days to systems for the prevention of oxidative damages when grown under long-day conditions

Regulation of plant cytosolic aldolase functions by redox-modifications.

From the five genes which code for cytosolic fructose 1,6-bisphosphate aldolases in Arabidopsis thaliana L., the cDNA clone of cAld2 (At2g36460), was heterologously expressed in E. coli and incubated under various oxidizing and reducing conditions. Covalent binding of a GSH moiety to the enzyme was shown by incorporation of biotinylated GSH (BioGEE) and by immunodetection with monoclonal anti-GSH serum. Nitrosylation after incubation with GSNO or SNP was demonstrated using the biotin-switch assay. Mass-spectrometry analysis showed glutathionylation and/or nitrosylation at two different cysteine residues: GSH was found to be attached to C68 and C173, while the nitroso-group was incorporated only into C173. Non-reducing SDS-PAGE conducted with purified wild-type and various Cys-mutant proteins revealed the presence of disulfide bridges in the oxidized enzyme, as described for rabbit muscle aldolase. Incubation of the purified enzyme with GSSG (up to 25 mM) led to partial and reversible inactivation of enzyme activity; NADPH, in the presence of the components of the cytosolic NADP-dependent thioredoxin system, could reactivate the aldolase as did DTT. Total and irreversible inactivation occurred with low concentrations (0.1 mM) of nitrosoglutathione (GSNO). Inactivation was prevented by co-incubation of cAld2 with fructose-1,6-bisphosphate (FBP). Nuclear localization of cAld2 and interaction with thioredoxins was shown by transient expression of fusion constructs with fluorescent proteins in isolated protoplasts

Detection of S-nitrosated nuclear proteins in pathogen-treated <em>Arabidopsis</em> cell cultures using biotin switch technique.

Nitric oxide (NO) is an important signaling molecule involved in various plant physiological processes. The main effect of NO arises from its reaction with proteins. S-Nitrosation is the most studied NO-mediated protein posttranslational modification in plants. Via S-nitrosation, NO derivatives react with thiol groups (SHs) of protein cysteine residues and produce nitrosothiol groups (SNOs). From the time of discovering the biological function of NO in plants, an interesting case of study has been the detection of the endogenous S-nitrosated proteins in different plants, tissues, organelles, and various conditions. Maps of S-nitrosated proteins provide hints for deeper studies on the function of this modification in specific proteins, biochemical pathways, and physiological processes. Many functions of NO have been found to be related to plant defense; on the other hand the involvement of nuclear proteins in regulation of plant defense reactions is well studied. Here, an approach is described in which the Arabidopsis cell cultures first are treated with P. syringae, afterward their bioactive nuclear proteins are extracted, then the nuclear proteins are subjected to biotin switch assay in which S-nitrosated proteins are specifically converted to S-biotinylated proteins. The biotin switch technique (BST) which was introduced by Jaffrey et al. (Nat Cell Biol 3:193-197, 2001) solves the instability issue of SNOs. Additionally, it provides detection and purification of biotinylated proteins by anti-biotin antibody and affinity chromatography, respectively