Twice daily allied health rehabilitation is feasible in acutely hospitalised older people: an observational study.

ObjectiveTo evaluate the feasibility of twice daily rehabilitation in older patients admitted to an acute care of the elderly (ACE) hospital ward.MethodThis was a prospective single-site, cohort study of twice daily interventions provided by ACE physiotherapists, occupational therapists and/or allied health assistants in an ACE hospital ward. The feasibility of twice daily therapy was evaluated using a range of outcomes including satisfaction, fidelity and limited efficacy.ResultsA total of 220 patients were included (median age 86 [IQR 81-91]years, 54% female, with a median length of hospital stay of 7 [IQR 5-10] days). Twice daily therapy was delivered on 71% (n=757) of patient admitted weekdays (fidelity). Moderate-to-large effect sizes were observed in patient functional and mobility measures during their hospital stay and most patients (74%) were able to be successfully discharged home (limited efficacy). Both staff and patients reported high levels of satisfaction with physiotherapy and occupational therapy while on the ACE ward.ConclusionTwice daily therapy with acutely hospitalised elderly patients is feasible, facilitated discharge home, and is associated with high patient and staff satisfaction.

Atmospheric drying as the main driver of dramatic glacier wastage in the southern Indian Ocean.

The ongoing retreat of glaciers at southern sub-polar latitudes is particularly rapid and widespread. Akin to northern sub-polar latitudes, this retreat is generally assumed to be linked to warming. However, no long-term and well-constrained glacier modeling has ever been performed to confirm this hypothesis. Here, we model the Cook Ice Cap mass balance on the Kerguelen Islands (Southern Indian Ocean, 49°S) since the 1850s. We show that glacier wastage during the 2000s in the Kerguelen was among the most dramatic on Earth. We attribute 77% of the increasingly negative mass balance since the 1960s to atmospheric drying associated with a poleward shift of the mid-latitude storm track. Because precipitation modeling is very challenging for the current generation of climate models over the study area, models incorrectly simulate the climate drivers behind the recent glacier wastage in the Kerguelen. This suggests that future glacier wastage projections should be considered cautiously where changes in atmospheric circulation are expected.

Defective initiation on natural messenger RNA by cell free systems from Krebs ascites cells incubated at elevated temperatures.

1. Cell-free systems prepared from Krebs II ascites cells incubated at 45 degrees C have a much lower endogenous activity than those from cells incubated at 37 degrees C. The endogenous activity is mainly due to completion of polypeptide chains initiated in the intact cell. The low activity of the 45 degrees C system is due to a lesion in initiation in cells incubated at 45 degrees C. 2. Cell-free systems from cells incubated at 45 degrees C can translate efficiently poly (U) at 8 mM Mg2+. However, they initiate poorly on globin mRNA, indicating that these systems reflect the situation in the intact cell. 3. The lesion in globin mRNA translation in 45 degrees C systems can be overcome by addition of reticulocyte initiation factors. At saturation concentrations of factors, the response of a 45 degrees C system is restored to almost normal. 4. 45 degrees C systems from 40-S initiation complexes with methionyl tRNAfmet almost as efficiently as normal, but their ability for form 80-S complexes with globin mRNA is impaired, unless they are supplied with exogenous initiation factors.

The mechanism of inhibition of initiation of protein synthesis in reticulocyte lysates by pyrocatechol violet.

1. Pyrocatechol violet inhibits the initiation of protein synthesis in reticulocyte lysates. It prevents the formation of 80-S initiation complexes by inhibiting the binding of mRNA to 40-S initiation complexes. 2. Pyrocatechol violet inhibits the hydrolysis of methionyl-tRNAmetf by reticulocyte lysates or reticulocyte cell sap. 3. Concentrations of pyrocatechol violet sufficient to inhibit initiation have little effect on amino acid activation. Higher concentrations of pyrocatechol violet were strongly inhibitory.

Post-transcriptional regulation of cytoskeletal actin and T lymphocyte receptor beta chain mRNA by phorbol ester.

The protein kinase C activator phorbol 12-myristate 13-acetate (TPA) augments the level of cytoplasmic mRNA for cytoskeletal actin and the beta chain of the T cell antigen receptor (TCR beta) in peripheral blood lymphocytes. Calcium ionophore A23187 has an antagonistic effect on these increases. The immunosuppressive drug cyclosporin A blocked both positive and negative effects of A23187 without affecting those of TPA. TPA-mediated increases in cytoplasmic RNA for both genes can occur when transcription is blocked with actinomycin D and no increase is observed in the corresponding total cellular RNA. Activation of protein kinase C can thus mediate the accumulation of these mRNAs by a post-transcriptional mechanism.

Initiation reactions in the mRNA-dependent reticulocyte lysate.

Reticulocyte lysates depleted of mRNA by digestion with micrococcal nuclease still show an unexpectedly high rate of formation of 80 S initiation complexes. Formation of these complexes is sensitive to all inhibitors of the normal protein synthesis initiation process tested. Such lysates contain high concentrations of mRNA fragments which can be utilized for initiation, with which exogenous mRNA must compete. As a consequence of this competition, mRNAs that are weak initiators may be translated poorly by this system even at low exogenous mRNA concentrations.

The control of protein synthesis during the stimulation of lymphocytes by phytohaemagglutinin. III. Poly(U) translation and the rate of polypeptide chain elongation.

1. Cell-free systems from phytohaemagglutinin-stimulated pig lymphocytes are much more active in both endogenous protein synthesis and the translation of poly(U) than those from unstimulated lymphocytes. 2. Addition of tRNA stimulates the translation of poly(U) and greatly reduces the difference between systems from stimulated and unstimulated lymphocytes. Endogenous protein synthesis is not increased by added tRNA. 3. Systems from stimulated lymphocytes have an increased capacity to form aminoacyl-tRNA with several different amino acids. This reaction is limited by the amount of tRNA present in the cell-free system. 4. The rate of polypeptide elongation is not affected by lymphocyte stimulation. The increased rate of protein synthesis must therefore be due to an increase in the frequency of initiation of the synthesis of protein molecules.

Amino acid transport in pig lymphocytes. Enhanced activity of transport system asc following mitogenic stimulation.

Changes in neutral amino acid transport activity caused by addition of phytohaemagglutinin-P to quiescent peripheral pig lymphocytes have been evaluated by measurements of 14C-labelled neutral and analogue amino acids under conditions approaching initial entry rates. Utilizing methylaminoisobutyric acid, the best model substrate of System A, we confirmed our previous report (Borghetti, A.F., Kay, J.E. and Wheeler, K.P. (1979) Biochem. J. 182, 27-32) on the absence of this transport system in quiescent cells and its emergence following stimulation. Furthermore, we demonstrated the presence in quiescent cells of an Na+-dependent transport system for neutral amino acids that has been characterized as System ASC by several criteria including intolerance to methylaminoisobutyric acid, strict Na+-dependence, the property of transtimulation and specificity for pertinent substrates such as alanine, serine, cysteine and threonine. Analysis of the relationship between influx and substrate concentration revealed that two independent saturable components contribute to entry of alanine in quiescent cells: a low affinity (Km = approximately 4 mM) and a high affinity (Km = approximately 0.2 mM) component. The high affinity component could be inhibited in a competitive way by serine, cysteine and threonine, but methylaminoisobutyric acid did not change appreciably its constants. The enhanced activity of alanine transport through the ASC system observed in activated cells resulted from a large increase in the capacity (V) of the high affinity component without any substantial change in the apparent affinity constant (Km).

Cyclolinopeptide A (CLA) mediates its immunosuppressive activity through cyclophilin-dependent calcineurin inactivation.

The immunosuppressive cyclic nonapeptide cyclolinopeptide A inhibits calcium-dependent, but not calcium-independent, activation of T lymphocytes comparably to the actions of cyclosporin A and FK506. The concentration required for complete inhibition, however, is 10 times higher than that of cyclosporin A. In addition, we demonstrate that calcineurin, a phosphatase which plays an important role in T lymphocyte signalling, is inhibited in vitro by cyclolinopeptide A by a mechanism dependent on the peptidyl-prolyl cis-trans isomerase (PPIase) cyclophilin A but not FKBP12. Direct binding of cyclolinopeptide A to cyclophilin A was confirmed using tryptophan fluorescence studies and PPIase assays. These results represent a third example of the production of a natural product that neutralises calcineurin by a mechanism dependent on the primary binding to a PPIase.

Binding of a putative and a known chaperone protein revealed by immunogold labeling transmission electron microscopy: A suggested use of chaperones as probes for the distribution of their target proteins.

Parvulins are a distinct family within the peptidyl-prolyl cis-trans isomerase group of proteins that catalyse the cis-trans isomerization of proline-containing peptides. The intracellular distribution of a novel human parvulin homologue (hEPVH) has been investigated in a human kidney cell line (HEK 293) by immunogold labeling transmission electron microscopy (TEM). This showed hEPVH to be distributed throughout HEK 293 cells but in highest concentration within mitochondria. Unexpectedly, preabsorption of anti-hEPVH antiserum with recombinant hEPVH exaggerated the observed immunolabel density in a pattern that mirrored that of the endogenous hEPVH. The hEPVH protein itself was then used to label sections, and the specificity of its binding was confirmed with the use of polyclonal and monoclonal antibodies in conjunction with homologous and irrelevant protein controls. The pattern of hEPVH binding also mirrored that of endogenous hEPVH. A known chaperone protein, Pin1, was also found to bind to cells in a pattern mirroring that of the endogenous protein. This lends considerable weight to our hypothesis that hEPVH is binding to its target protein(s) within the cell, reflecting its postulated chaperone function. Finally, we suggest that chaperone proteins in general might be used as TEM probes for their target (or substrate) proteins. (J Histochem Cytochem 47:1633-1640, 1999)

Mechanisms of T lymphocyte activation.

The proliferation of T lymphocytes in response to an antigenic stimulus requires the successive ligation of a range of T lymphocyte receptors by MHC-presented antigen, molecules expressed on the surfaces of accessory cells and lymphokines. A range of intracellular signalling systems are involved, with the possibility of alternative activation pathways utilising different intracellular signalling systems, though protein kinase C activation appears to play a key role. Appreciation of the complexity of the response may allow more selective clinical modulation of T lymphocyte-dependent immunological responses.

The role of the transferrin receptor in lymphocyte activation.

Blockade of the transferrin receptors whose expression is induced in lymphocytes incubated with the mitogenic lectin phytohaemagglutinin (PHA) does not affect the initial stimulation of protein synthesis but does strongly and progressively inhibit the subsequent induction of DNA synthesis. When the effects of transferrin receptor blockade on the induction of the enzymes uridine kinase (whose induction begins early in G1 phase of the cell cycle) and thymidine kinase (whose induction is closely associated with DNA synthesis) were examined, both enzymes were found to be induced normally. This indicates that the function of the transferrin receptor is directly to provide a component essential for DNA synthesis itself (probably iron) rather than to act as the receptor for a general signal required to initiate entry into S-phase.

T lymphocyte activation through the C28 pathway is insensitive to inhibition by the immunosuppressive drug FK-506.

Nanomolar concentrations of the novel immunosuppressive drug FK-506 inhibit the proliferation of human T lymphocytes in vitro induced by mitogenic lectins or by monoclonal antibodies directed against the CD3 or CD2 surface antigens. However, the alternative pathway of T lymphocyte proliferation induced by monoclonal antibodies specific for CD28 together with phorbol esters is unaffected by FK-506.

The effects of indomethacin on T lymphocyte stimulation.

Immunomodulatory effects of indomethacin on mitogen-induced [3H]thymidine incorporation in cultured pig peripheral blood lymphocytes were demonstrated. In monocyte-containing lymphocyte cultures, indomethacin enhanced lymphocyte stimulation, whereas in monocyte-depleted cultures indomethacin inhibited the response. Indomethacin effects were most marked at suboptimal to optimal concentrations of mitogen and decreased at superoptimal concentrations in both types of culture. The effects of prostaglandin E2 at superoptimal mitogen concentrations were found paradoxically to be stimulatory. The results indicate that arachidonic acid metabolites may have both positive and negative effects on lymphocyte activation.

Activation of T lymphocytes by 12-O-tetradecanoylphorbol-13-acetate is resistant to inhibition by cyclosporin A.

Cultured T lymphocytes from pig blood can be activated by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). Activation is additive with that induced by the mitogenic lectin phytohaemagglutinin (PHA). Activation by TPA differs from that induced by PHA or other mitogenic lectins in that it is not inhibited even by high concentrations of the immunosuppressive drug cyclosporin A (CS-A). Neither co-culture of lymphocytes with PHA and TPA nor addition of culture supernatants from TPA-stimulated cultures affected the sensitivity to CS-A of the response to PHA.

Stability of polyadenylated mRNA in pig lymphocytes.

The stability of cytoplasmic poly(A)-containing mRNA in unstimulated pig lymphocytes has been determined by [3H]poly(U) hybridization after actinomycin D treatment. This approach has shown that the majority of the mRNA is much more unstable than has been suggested by earlier pulse-chase experiments, and has a half-life of between 1 and 2 h.