Brazilian version of the Care Transitions Measure: translation and validation.

AIM: To translate, adapt and evaluate psychometric properties of the complete (15 items) and reduced (three items) versions of the Care Transitions Measure into Brazilian Portuguese. INTRODUCTION: The Care Transitions Measure assesses the quality of care transitions, from the perspective of patients. As accomplishing effective transitions is a challenge to healthcare systems, the instrument provides an opportunity to assess care transitions and improve quality initiatives. METHODS: A three-phased design was used for cross-cultural adaptation, pre-testing and evaluation of psychometric properties of the measurement in a Brazilian hospital. After forward translation, back translation and expert committee review, patients evaluated the instrument in a pre-test. Psychometric testing included face and content validity, reliability, stability and factorial analysis. RESULTS: Cross-cultural adaptation was completed successfully with a high clarity rate. Internal consistency was good in the 15-item version and was moderate in the three-item version. Test-retest reliability showed good stability of the two versions over time. The three-item version had satisfactory criterion validity. Four factors were extracted for the 15-item measure. LIMITATIONS: Samples were restricted to a group of patients from one hospital in southern Brazil. Future studies should test the measurement's construct and predictive validity. CONCLUSIONS: The translated version of the Care Transitions Measure has good face and content validity, reliability and stability. It has shown to be a valid measurement for evaluating the quality of care transitions in Brazil. IMPLICATIONS FOR NURSING AND HEALTH POLICY: Results are beneficial for nurses, managers and policy makers for evaluating care transitions and support the need for changes in policies and practices.

Heat shock resistance conferred by expression of the human HSP27 gene in rodent cells.

Heat shock induces in cells the synthesis of specific proteins called heat shock proteins (HSPs) and a transient state of thermotolerance. The putative role of one of the HSPs, HSP27, as a protective molecule during thermal stress has been directly assessed by measuring the resistance to hyperthermia of Chinese hamster and mouse cells transfected with the human HSP27 gene contained in plasmid pHS2711. One- and two-dimensional gel electrophoresis of [3H]leucine- and [32P]orthophosphate-labeled proteins, coupled with immunological analysis using Ha27Ab and Hu27Ab, two rabbit antisera that specifically recognize the hamster and the human HSP27 protein respectively, were used to monitor expression and inducibility of the transfected and endogenous proteins. The human HSP27 gene cloned in pHS2711 is constitutively expressed in rodent cells, resulting in accumulation of the human HSP27 and all phosphorylated derivatives. No modification of the basal or heat-induced expression of endogenous HSPs is detected. The presence of additional HSP27 protein provides immediate protection against heat shock administered 48 h after transfection and confers a permanent thermoresistant phenotype to stable transfectant Chinese hamster and mouse cell lines. Mild heat treatment of the transfected cells results in an induction of the full complement of the endogenous heat shock proteins and a small increase in thermoresistance, but the level attained did not surpass that of heat-induced thermotolerant control cells. These results indicate that elevated levels of HSP27 is sufficient to give protection from thermal killing. It is concluded that HSP27 plays a major role in the increased thermal resistance acquired by cells after exposure to HSP inducers.

Sequence and regulation of a gene encoding a human 89-kilodalton heat shock protein.

Vertebrate cells synthesize two forms of the 82- to 90-kilodalton heat shock protein that are encoded by distinct gene families. In HeLa cells, both proteins (hsp89 alpha and hsp89 beta) are abundant under normal growth conditions and are synthesized at increased rates in response to heat stress. Only the larger form, hsp89 alpha, is induced by the adenovirus E1A gene product (M. C. Simon, K. Kitchener, H. T. Kao, E. Hickey, L. Weber, R. Voellmy, N. Heintz, and J. R. Nevins, Mol. Cell. Biol. 7:2884-2890, 1987). We have isolated a human hsp89 alpha gene that shows complete sequence identity with heat- and E1A-inducible cDNA used as a hybridization probe. The 5'-flanking region contained overlapping and inverted consensus heat shock control elements that can confer heat-inducible expression on a beta-globin reporter gene. The gene contained 10 intervening sequences. The first intron was located adjacent to the translation start codon, an arrangement also found in the Drosophila hsp82 gene. The spliced mRNA sequence contained a single open reading frame encoding an 84,564-dalton polypeptide showing high homology with the hsp82 to hsp90 proteins of other organisms. The deduced hsp89 alpha protein sequence differed from the human hsp89 beta sequence reported elsewhere (N. F. Rebbe, J. Ware, R. M. Bertina, P. Modrich, and D. W. Stafford (Gene 53:235-245, 1987) in at least 99 out of the 732 amino acids. Transcription of the hsp89 alpha gene was induced by serum during normal cell growth, but expression did not appear to be restricted to a particular stage of the cell cycle. hsp89 alpha mRNA was considerably more stable than the mRNA encoding hsp70, which can account for the higher constitutive rate of hsp89 synthesis in unstressed cells.

Modulation of cellular thermoresistance and actin filament stability accompanies phosphorylation-induced changes in the oligomeric structure of heat shock protein 27.

Phosphorylation of heat shock protein 27 (HSP27) can modulate actin filament dynamics in response to growth factors. During heat shock, HSP27 is phosphorylated at the same sites and by the same protein kinase as during mitogenic stimulation. This suggests that the same function of the protein may be activated during growth factor stimulation and the stress response. To determine the role of HSP27 phosphorylation in the heat shock response, several stable Chinese hamster cell lines that constitutively express various levels of the wild-type HSP27 (HU27 cells) or a nonphosphorylatable form of human HSP27 (HU27pm3 cells) were developed. In contrast to HU27 cells, which showed increased survival after heat shock, HU27pm3 cells showed only slightly enhanced survival. Evidence is presented that stabilization of microfilaments is a major target of the protective function of HSP27. In the HU27pm3 cells, the microfilaments were thermosensitized compared with those in the control cells, whereas wild-type HSP27 caused an increased stability of these structures in HU27 cells. HU27 but not HU27pm3 cells were highly resistant to cytochalasin D treatment compared with control cells. Moreover, in cells treated with cytochalasin D, wild-type HSP27 but not the phosphorylated form of HSP27 accelerated the reappearance of actin filaments. The mutations in human HSP27 had no effect on heat shock-induced change in solubility and cellular localization of the protein, indicating that phosphorylation was not involved in these processes. However, induction of HSP27 phosphorylation by stressing agents or mitogens caused a reduction in the multimeric size of the wild-type protein, an effect which was not observed with the mutant protein. We propose that early during stress, phosphorylation-induced conformational changes in the HSP27 oligomers regulate the activity of the protein at the level of microfilament dynamics, resulting in both enhanced stability and accelerated recovery of the filaments. The level of protection provided by HSP27 during heat shock may thus represent the contribution of better maintenance of actin filament integrity to overall cell survival.

Changes in the stability of a human H3 histone mRNA during the HeLa cell cycle.

A major component of the regulation of histone protein synthesis during the cell cycle is the modulation of the half-life of histone mRNA. We have uncoupled transcriptional and posttranscriptional regulation by using a Drosophila hsp70-human H3 histone fusion gene that produces a marked human H3 histone mRNA upon heat induction. Transcription of this gene can be switched on and off by raising and lowering cell culture temperatures, respectively. HeLa cell lines containing stably integrated copies of the fusion gene were synchronized by double thymidine block. Distinct populations of H3 histone mRNA were produced by heat induction in early S-phase, late S-phase, or G2-phase cells, and the stability of the induced H3 histone mRNA was measured. The H3 histone mRNA induced during early S phase decayed with a half-life of 110 min, whereas the same transcript induced during late S phase had a half-life of 10 to 15 min. The H3 histone mRNA induced in non-S-phase cells is more stable than that induced in late S phase, with a half-life of 40 min. Thus, the stability of histone mRNA is actively regulated throughout the cell cycle. Our results are consistent with an autoregulatory model in which the stability of histone mRNA is determined by the level of free histone protein in the cytoplasm.

In vivo chaperone activity of heat shock protein 70 and thermotolerance.

Heat shock protein 70 (Hsp70) is thought to play a critical role in the thermotolerance of mammalian cells, presumably due to its chaperone activity. We examined the chaperone activity and cellular heat resistance of a clonal cell line in which overexpression of Hsp70 was transiently induced by means of the tetracycline-regulated gene expression system. This single-cell-line approach circumvents problems associated with clonal variation and indirect effects resulting from constitutive overexpression of Hsp70. The in vivo chaperone function of Hsp70 was quantitatively investigated by using firefly luciferase as a reporter protein. Chaperone activity was found to strictly correlate to the level of Hsp70 expression. In addition, we observed an Hsp70 concentration dependent increase in the cellular heat resistance. In order to study the contribution of the Hsp70 chaperone activity, heat resistance of cells that expressed tetracycline-regulated Hsp70 was compared to thermotolerant cells expressing the same level of Hsp70 plus all of the other heat shock proteins. Overexpression of Hsp70 alone was sufficient to induce a similar recovery of cytoplasmic luciferase activity, as does expression of all Hsps in thermotolerant cells. However, when the luciferase reporter protein was directed to the nucleus, expression of Hsp70 alone was not sufficient to yield the level of recovery observed in thermotolerant cells. In addition, cells expressing the same level of Hsp70 found in heat-induced thermotolerant cells containing additional Hsps showed increased resistance to thermal killing but were more sensitive than thermotolerant cells. These results suggest that the inducible form of Hsp70 contributes to the stress-tolerant state by increasing the chaperone activity in the cytoplasm. However, its expression alone is apparently insufficient for protection of other subcellular compartments to yield clonal heat resistance to the level observed in thermotolerant cells.

Stress response protein (srp-27) determination in primary human breast carcinomas: clinical, histologic, and prognostic correlations.

Expression of an estrogen-regulated protein known as the 27,000-d heat-shock or stress-response protein (srp-27) was evaluated in human breast carcinomas and established breast cancer cell lines. Results obtained by Northern and Western blot analyses and immunohistochemical methods were concordant. Immunohistochemical assessment of srp-27 expression in 300 breast carcinomas (with median patient follow-up of 8 years) was performed. Twenty-six percent of lymph node-negative and 45% of lymph node-positive tumors were overexpressors. Univariate analysis demonstrated significant correlations between srp-27 overexpression and estrogen receptor (ER) content, pS2 protein expression, nodal metastases, advanced T stage, lymphatic/vascular invasion, and a shorter disease-free survival period (but not a shorter overall survival) for the study population as a whole. Regression tree analysis showed that srp-27 expression was an independent prognostic indicator for disease-free survival only in patients with one to three positive lymph nodes. The Cox proportional hazards model confirmed the independent prognostic significance of nodal involvement, T stage, and ER content but failed to recognize srp-27 overexpression as a significant independent parameter predictive of patient outcome in the patient population as a whole. The observed associations between srp-27 overexpression and more aggressive tumors suggest a biologic role for srp-27 in human breast carcinomas.

Cloning and sequencing of a cDNA encoding the canine HSP27 protein.

The nucleotide (nt) sequence encoding a 27-kDa heat-shock protein (HSP27) was determined from cDNAs cloned from a canine smooth muscle library. The primary structure deduced from the nt sequence reveals a 209-amino-acid protein having 86-89% identity with human, mouse, rat and hamster small HSP. Similar to human HSP27, the canine protein contains three Ser residues that are potential MAPKAP kinase II substrates.

Molecular cloning of sequences encoding the human heat-shock proteins and their expression during hyperthermia.

Plasmids containing cDNA copies of mRNAs induced in HeLa cells by heat shock have been isolated and characterized. In vitro translation of RNAs selected by hybridization to plasmid DNAs identified sequences representing the three major classes (89, 70 and 27-kDa) of heat-shock proteins (hsp) and a 60-kDa minor hsp. Plasmids with inserts specific for the 27, 60, and 70-kDa hsp each hybridize with a single discrete size class of heat-inducible mRNA. Plasmids specific for the 89-kDa protein, however, hybridize with either a 2.7- or 2.95-kb mRNA species. Both mRNAs are coordinately induced during heat shock. We show that the characteristic pattern of induction and repression of each class of hsp during sustained hyperthermia is the result of changes in the steady state level of each mRNA.

Influence of phosphorylation and oligomerization on the protective role of the small heat shock protein 27 in rat adult cardiomyocytes.

Recent reports have demonstrated that the heat shock proteins (hsp) and in particular the hsp70 confer protection against cardiac ischemic damage. More recently, we have shown that increased expression of another heat shock protein, the hsp27, through an adenovirus vector system protects adult cardiomyocytes against ischemic injury. This small heat shock protein undergoes phosphorylation when the cell is under stress. This has led many to speculate that phosphorylation of hsp27 is required for the protective role this protein plays in the cell. In order to investigate this possibility, we have mutated the serines that are the sites of phosphorylation on the hsp27, to glycines or alanines. These nonphosphorylatable mutants of hsp27 were cloned into adenoviral vectors and used to infect adult rat cardiomyocytes to assess their ability in protecting against ischemic injury. In addition, we used a specific inhibitor of p38 MAP kinase that is a key member of the kinase pathway responsible for phosphorylating the hsp27. Our present results show that the nonphosphorylated hsp27 forms larger oligomeric complexes than the phosphorylated hsp27. Interestingly, phosphorylation of hsp27 seems not to play a role in its ability to protect adult rat cardiomyocytes against ischemic damage.

The surgical tray.

This article provides an overview of the most commonly used instruments for excisional surgery. It has also discussed some of the specialty instruments that may be of value. It may serve as a starting point for the outfitting of a dermatologic surgery practice. However, it is no substitute for seeing, touching, and handling the instruments. At many of the larger specialty meetings, the instrument vendors will be displaying their wares. These displays offer an excellent opportunity to evaluate the many instruments. All of these fancy shiny instruments are merely tools to be used as an extension of the surgeon's hand and brain. They should work for the surgeon in a manner that is comfortable and efficient. Only the surgeon can decide what works best.

Drug therapy of fungal and parasitic diseases.

This article on therapy provides an updated review, including pharmacology, indications, doses, and side effects, of the major antifungal and antiparasitic drugs.

Reticulate pigmented anomaly of the flexures (Dowling-Degos disease): a case report associated with hidradenitis suppurativa and squamous cell carcinoma.

We report the first case of reticulate pigmented anomaly of the flexures (Dowling-Degos disease) associated with hidradenitis suppurativa. Our patient also showed a squamous cell carcinoma of the groin, which is a rare complication of hidradenitis suppurativa. The relationship between Dowling-Degos disease and hidradenitis is discussed and the literature on squamous cell carcinoma and hidradenitis suppurativa is reviewed.

P-V-T, Thermodynamic and Related Properties of Oxygen from the Triple Point to 300 K at Pressures to 33 MN/m2.

The results of new experimental pressure-volume-temperature measurements on oxygen are presented. The data range in temperature from 54 to 300 K and in pressure from 0.1 to about 33 MN/m2. The following properties are tabulated for selected isobars: molar volume, (∂P/∂ρ) T , (∂P/∂T)ρ, internal energy, enthalpy, entropy, specific heats at constant volume and at constant pressure, and the velocity of sound. Additional tables present the above properties for saturated liquid and vapor, the freezing liquid P-V-T relationship, and the derived Joule-Thomson inversion curve. New values for the critical density and triple point density are presented, and the second and third virial coefficients are tabulated.