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Abstract: Background: Pain and anxiety are a common problem in all recovery phases after a burn. The Burns Specific Pain Anxiety Scale (BSPAS) was proposed to assess anxiety in burn patients related to painful procedures. Objectives: To assess internal consistency, discriminative construct validity, dimensionality and convergent construct validity of the Brazilian-Portuguese version of the Burns Specific Pain Anxiety Scale. Design: In this cross-sectional study, the original version of the BSPAS, adapted into Brazilian Portuguese, was tested for internal consistency (Cronbach''s Alpha), discriminative validity (related to total body surface area burned and sex), dimensionality (through factor analysis), and convergent construct validity (applying the Visual Analogue Scale for pain and State-Anxiety—STAI) in a group of 91 adult burn patients. Results: The adapted version of the BSPAS displayed a moderate and positive correlation with pain assessments: immediately before baths and dressings (r =0.32; p <0.001), immediately after baths and dressings (r =0.31; p <0.001) and during the relaxation period (r =0.31; p <0.001) and with anxiety assessments (r =0.34; p <0.001). No statistically significant differences were observed when comparing the mean of the adapted version of the BSPAS scores with sex (p =0.194) and total body surface area burned (p =0.162) (discriminative validity). The principal components analysis applied to our sample seems to confirm anxiety as one single domain of the Brazilian-Portuguese version of the BSPAS. Cronbach''s Alpha showed high internal consistency of the adapted version of the scale (0.90). Conclusion: The Brazilian-Portuguese version of the BSPAS 9-items has shown statically acceptable levels of reliability and validity for pain-related anxiety evaluation in burn patients. This scale can be used to assess nursing interventions aimed at decreasing pain and anxiety related to the performance of painful procedures. [ABSTRACT FROM AUTHOR]

The current study explored whether immersive virtual reality (VR) continues to reduce pain (via distraction) during more than one wound care session per patient. Thirty-six patients aged 8 to 57 years (mean age, 27.7 years), with an average of 8.4% TBSA burned (range, 0.25-25.5 TBSA) received bandage changes, and wound cleaning. Each patient received one baseline wound cleaning/debridement session with no-VR (control condition) followed by one or more (up to seven) subsequent wound care sessions during VR. After each wound care session (one session per day), worst pain intensity was measured using a visual analog thermometer, the dependent variable. Using a within-subjects design, worst pain intensity during wound care with no-VR (baseline, day 0) was compared with pain during wound care while using immersive VR (up to 7 days of wound care during VR). Compared with pain during no-VR baseline (day 0), pain ratings during wound debridement were statistically lower when patients were in VR on days 1, 2, and 3, and although not significant beyond day 3, the pattern of results from days 4, 5, and 6 are consistent with the notion that VR continues to reduce pain when used repeatedly. Results from the present study suggest that VR continues to be effective when used for three (or possibly more) treatments during severe burn wound debridement.

Background: The Burn Specific Health Scale Brief (BSHS-B), which is the only multidimensional measure to evaluate burn-specific aspects of health status, has previously been validated in several languages across the world. However, the stability of the underlying construct was not cross-culturally evaluated. The current study reports on measurement invariance across two samples of Swedish- and Dutch- speaking patients with burns., Methods: In a prospective study, 231 and 275 Swedish and Dutch-Belgian patients with burns, completed the BSHS-B at 9 or 12 months, respectively, after burn. Using a multigroup confirmatory factor analysis, measurement invariance across languages (Swedish and Dutch) was tested., Results: The results of the confirmatory factor analysis in the total sample revealed that the scale structure for the earlier reported three-factor structure and the original nine-factor structure was adequate. However, an eight-factor structure in which hand function and simple abilities were merged provided the best fit. This structure was used to test measurement invariance across the two language groups. The two-group outcomes testing measurement invariance across Swedish- and Dutch-speaking patients indicated a stable, configural invariance., Conclusion: The BSHS-B seems to function uniformly across both language groups. The BSHS-B can be used to compare cross-cultural results in both countries.

This study investigated traumatic stress symptoms in severely burned survivors of two fire disasters and two comparison groups of patients with "non-disaster" burn injuries, as well as risk factors associated with acute and chronic stress symptoms. Patients were admitted to one out of eight burn centers in The Netherlands or Belgium. The Impact of Event Scale (IES) was administered to 61 and 33 survivors respectively of two fire disasters and 54 and 57 patients with "non-disaster" burn etiologies at 2 weeks, 3, 6, 12 and 24 months after the event. We used latent growth modeling (LGM) analyses to investigate the stress trajectories and predictors in the two disaster and two comparison groups. The results showed that initial traumatic stress reactions in disaster survivors with severe burns are more intense and prolonged during several months relative to survivors of "non-disaster" burn injuries. Excluding the industrial fire group, all participants' symptoms on average decreased over the two year period. Burn severity, peritraumatic anxiety and dissociation predicted the long-term negative outcomes only in the industrial fire group. In conclusion, fire disaster survivors appear to experience higher levels of traumatic stress symptoms on the short term, but the long-term outcome appears dependent on factors different from the first response. Likely, the younger age, and several beneficial post-disaster factors such as psychosocial aftercare and social support, along with swift judicial procedures, contributed to the positive outcome in one disaster cohort.

Background: Methylation of histone H3 lysine 79 (H3K79) by Dot1 is highly conserved among species and has been associated with both gene repression and activation. To eliminate indirect effects and examine the direct consequences of Dot1 binding and H3K79 methylation, we investigated the effects of targeting Dot1 to different positions in the yeast genome., Results: Targeting Dot1 did not activate transcription at a euchromatic locus. However, chromatin-bound Dot1 derepressed heterochromatin-mediated gene silencing over a considerable distance. Unexpectedly, Dot1-mediated derepression was established by both a H3K79 methylation-dependent and a methylation-independent mechanism; the latter required the histone acetyltransferase Gcn5. By monitoring the localization of a fluorescently tagged telomere in living cells, we found that the targeting of Dot1, but not its methylation activity, led to the release of a telomere from the repressive environment at the nuclear periphery. This probably contributes to the activity-independent derepression effect of Dot1., Conclusions: Targeting of Dot1 promoted gene expression by antagonizing gene repression through both histone methylation and chromatin relocalization. Our findings show that binding of Dot1 to chromatin can positively affect local gene expression by chromatin rearrangements over a considerable distance.

Background: Methylation of lysine 79 on histone H3 by Dot1 is required for maintenance of heterochromatin structure in yeast and humans. However, this histone modification occurs predominantly in euchromatin. Thus, Dot1 affects silencing by indirect mechanisms and does not act by the recruitment model commonly proposed for histone modifications. To better understand the role of H3K79 methylation gene silencing, we investigated the silencing function of Dot1 by genetic suppressor and enhancer analysis and examined the relationship between Dot1 and other global euchromatic histone modifiers., Result: We determined that loss of H3K79 methylation results in a partial silencing defect that could be bypassed by conditions that promote targeting of Sir proteins to heterochromatin. Furthermore, the silencing defect in strains lacking Dot1 was dependent on methylation of H3K4 by Set1 and histone acetylation by Gcn5, Elp3, and Sas2 in euchromatin. Our study shows that multiple histone modifications associated with euchromatin positively modulate the function of heterochromatin by distinct mechanisms. Genetic interactions between Set1 and Set2 suggested that the H3K36 methyltransferase Set2, unlike most other euchromatic modifiers, negatively affects gene silencing., Conclusion: Our genetic dissection of Dot1's role in silencing in budding yeast showed that heterochromatin formation is modulated by multiple euchromatic histone modifiers that act by non-overlapping mechanisms. We discuss how euchromatic histone modifiers can make negative as well as positive contributions to gene silencing by competing with heterochromatin proteins within heterochromatin, within euchromatin, and at the boundary between euchromatin and heterochromatin.

An assessment carried out on adolescents aged 11-18 who had suffered severe burns in early childhood 10-11 years previously indicated that the levels of problem behavior were similar and the levels of depression lower than those reported by adolescents in comparable reference populations. Adolescents' self-reports revealed higher levels of the personality traits emotional stability, agreeableness and extraversion. Self-reports further revealed lower levels of passive coping than are found in a normative reference population. Regression analyses showed that less emotional stability, less agreeableness and more passive coping styles were related to increased behavioral problems and symptoms of depression. The authors argue that future studies should focus on resilience and within-group differences.

Dot1 methylates histone H3 lysine 79 (H3K79) on the nucleosome core and is involved in Sir protein-mediated silencing. Previous studies suggested that H3K79 methylation within euchromatin prevents nonspecific binding of the Sir proteins, which in turn facilitates binding of the Sir proteins in unmethylated silent chromatin. However, the mechanism by which the Sir protein binding is influenced by this modification is unclear. We performed genome-wide synthetic genetic array (SGA) analysis and identified interactions of DOT1 with SIR1 and POL32. The synthetic growth defects found by SGA analysis were attributed to the loss of mating type identity caused by a synthetic silencing defect. By using epistasis analysis, DOT1, SIR1, and POL32 could be placed in different pathways of silencing. Dot1 shared its silencing phenotypes with the NatA N-terminal acetyltransferase complex and the conserved N-terminal bromo adjacent homology (BAH) domain of Sir3 (a substrate of NatA). We classified all of these as affecting a common silencing process, and we show that mutations in this process lead to nonspecific binding of Sir3 to chromatin. Our results suggest that the BAH domain of Sir3 binds to histone H3K79 and that acetylation of the BAH domain is required for the binding specificity of Sir3 for nucleosomes unmethylated at H3K79.

Treatment of people with burn injuries includes recovery of optimal function for survivors to fully participate in society, psychologically and physically. Increased likelihood of physical survival has led to greater concern for potential psychological morbidity for the burn survivor. Based on research and on many years of clinical experience in providing psychosocial care to burned children and adults, the authors outline their approach to assisting burn survivors and their families through the arduous process of recovery from admission through critical care, inpatient recuperation and reintegration upon hospital discharge. A philosophy of rehabilitation, a process that may occur for many months or years after patients' discharge from their acute hospitalization, is presented in the form of seven guidelines for working with burn survivors.

Background: Itching (pruritus) following burns is a well-known clinical problem. However, there are no long-term prospective studies that document the course and the extent of the problem. Studies on risk factors are anecdotal., Objectives: To study self-reported itching in a multicentre cohort among adults with burns at 3, 12 and 24 months postburn. Further, to examine psychological and injury characteristics in relation to itching at these three points in time., Methods: Itching was assessed as part of a self-report scar complaint list in a prospective longitudinal cohort study. Injury characteristics, demographics and self-reported post-traumatic stress symptoms were examined as possible risk factors in three linear regression models., Results: A total of 510 persons participated. The reported prevalence rates of mild to severe itching were as high as 87%, 70% and 67% at the three respective points in time. Significant predictors of itching at all three points in time were deep dermal injury and early post-traumatic stress symptoms. Along with these, total burned surface area and female gender were predictors at 3 months postburn., Conclusions: Itching remains a significant problem over a 2-year period. Individuals having undergone surgical procedures and experiencing early post-traumatic distress are more likely to suffer from long-term and persistent itching. Implications regarding practice and research are discussed.

Adult burn patients experience pain during wound care despite pharmacological interventions. Additional nursing interventions are needed to improve pain management. A systematic review was undertaken in order to examine the implications of previous research for evidence based decisions concerning the use of non-pharmacological nursing interventions and for future research. Twenty-six studies met the inclusion criteria and were discussed. The majority of the included studies concerned behavioural nursing interventions and focussed on promotion of psychological comfort. Although 17 studies showed that the intervention had a positive effect on pain outcomes and no adverse effects of the reviewed interventions were reported, the best available evidence was found for active hypnosis, rapid induction analgesia and distraction relaxation. However, in order to reduce methodological limitations, further research is needed before well-founded evidence based decisions for nursing practice can be made. Aspects that seem important for future research, like the type of the intervention, theoretical framework, manner of giving instruction and guidance, cost, outcomes, measurement instruments and data collection points are considered.

Like most eukaryotes, Saccharomyces cerevisiae cells contain a minor 5.8SL rRNA that, relative to the major 5.8SS species, carries several extra nucleotides at the 5'-end. The two species are produced by alternative pathways that differ in the events removing the 3'-terminal region of Internal Transcribed Spacer 1 from the 27SA2 pre-rRNA. Whereas the pathway leading to 5.8SS rRNA is well established, that producing the 5'-end of 5.8SL (called B1L) is poorly understood. Northern analysis of two different mutants of S. cerevisiae that overproduce 5.8SL rRNA revealed the presence of a fragment corresponding to the 3'-terminal region of Internal Transcribed Spacer 1 (ITS1) directly upstream from site B1L. Immunoprecipitation experiments showed this fragment to be associated with the trans-acting factor Rrp5p required for processing at the early sites A0-A3. Together these data clearly support that the 5'-end of 5.8SL rRNA is an endonucleolytic event. In vivo mutational analysis demonstrated the lack of any cis-acting sequence elements directing this cleavage within ITS1.

The Health Outcomes Burn Questionnaire (HOBQ) is a self-administered questionnaire to monitor outcome after burns in young children. This study aimed to assess feasibility, reliability and validity of the Dutch version of the HOBQ. The HOBQ was adapted into Dutch and tested in a population of children aged 0-4 years with a primary admission to a Dutch burn centre in March 2001-February 2004. Parents of 413 children were sent a questionnaire. To assess validity, a generic outcome instrument was included, the Infant Toddler Quality of Life Questionnaire (ITQOL). The response rate was 50.0% (n=196). Mean self-reported completion time was 16.7 min. The internal consistency of all the HOBQ-scales was good (Cronbach's alpha's>0.69). Test-retest results showed no differences in 7 out of 10 scales. High correlations between HOBQ-scales and conceptually equivalent ITQOL and scales were found in 5 out of 7 comparisons. The majority of the HOBQ-scales (7 out of 10) showed significant differences in the expected direction between children with a long versus short length of stay. Our data support the reliability and validity of the Dutch HOBQ. The HOBQ can be used as a research tool, to monitor functional outcome after burns in young children. Further research in other samples is recommend to fully establish the reliability and validity of the HOBQ.

Rrp5p is a trans-acting factor important for biogenesis of both the 40S and 60S subunit of the Saccharomyces cerevisiae ribosome. The protein contains 12 tandemly repeated S1 RNA binding motifs in its N-terminal region, suggesting the ability to interact directly with the pre-rRNA. In vitro binding studies, using immunopurified Rrp5p and in vitro transcribed, 32P-UTP-labeled RNA fragments, revealed that Rrp5p is a general RNA-binding protein with a strong preference for single-stranded sequences rich in uridines. Co-immunoprecipitation studies in yeast cells expressing ProtA-tagged Rrp5p showed that the protein is still associated with pre-ribosomal particles containing 27SA2 pre-rRNA but not with particles containing the 27SB precursor. Thus, Rrp5p appears to dissociate from the 66S pre-ribosome upon or immediately after further processing of 27SA2 pre-rRNA, suggesting the presence of (an) important binding site(s) within the 3'-terminal portion of ITS1. The location of these possible binding site(s) was further delimited using rrp2-1 mutant cells, which accumulate the 5'-extended 5.8S pre-rRNA species. The results indicate that association of Rrp5p with the pre-ribosome is abolished upon removal of a 30-nt region downstream from site A2, which contains two short, single-stranded U stretches. Sequence comparison shows that only the most 5' of these two U-rich stretches is conserved among yeast species whose ITS1 can functionally replace the S. cerevisiae spacer. The implications for the role of Rrp5p in yeast ribosome biogenesis are discussed.

The aim of this study was to assess if the pain observation scale for young children (POCIS) and the visual analogue scale (VAS) are reliable and valid instruments to measure procedural and background pain in burned children aged 0-4 years. Burn care nurses (n=73) rated pain from 24 fragments of videotaped children during wound care procedures and during periods of rest using the POCIS and the VAS. Intraclass correlations were used to assess inter-rater and intra-rater reliability for the POCIS and the VAS. Internal consistency for POCIS was assessed by Cronbach's alpha. The POCIS has shown poor to moderate inter-rater reliability, moderate to good intra-rater reliability and an acceptable internal consistency. The VAS turned out to have poor inter-rater reliability and poor to moderate intra-rater reliability. Due to poor results of inter-rater reliability in both scales, construct validation is left undone until more acceptable results are obtained. Factors explaining the results are the large number of raters, the manner they were trained and a lack of variation between pain classes in video fragments. Although not all results were satisfying, an easy to use scale as POCIS has promising qualities and deserves further reliability research.

Eukaryotes have two types of ribosomes containing either 5.8SL or 5.8SS rRNA that are produced by alternative pre-rRNA processing. The exact processing pathway for the minor 5.8SL rRNA species is poorly documented. We have previously shown that the trans-acting factor Rrp5p and the RNA exonuclease Rex4p genetically interact to influence the ratio between the two forms of 5.8S rRNA in the yeast Saccharomyces cerevisiae. Here we report a further analysis of ITS1 processing in various yeast mutants that reveals genetic interactions between, on the one hand, Rrp5p and RNase MRP, the endonuclease required for 5.8SS rRNA synthesis, and, on the other, Rex4p, the RNase III homolog Rnt1p, and the debranching enzyme Dbr1p. Yeast cells carrying a temperature-sensitive mutation in RNase MRP (rrp2-1) exhibit a pre-rRNA processing phenotype very similar to that of the previously studied rrp5-33 mutant: ITS2 processing precedes ITS1 processing, 5.8SL rRNA becomes the major species, and ITS1 is processed at the recently reported novel site A4 located midway between sites A2 and A3. As in the rrp5-Delta3 mutant, all of these phenotypical processing features disappear upon inactivation of the REX4 gene. Moreover, inactivation of the DBR1 gene in rrp2-1, or the RNT1 gene in rrp5-Delta3 mutant cells also negates the effects of the original mutation on pre-rRNA processing. These data link a total of three RNA catabolic enzymes, Rex4p, Rnt1p, and Dbr1p, to ITS1 processing and the relative production of 5.8SS and 5.8SL rRNA. A possible model for the indirect involvement of the three enzymes in yeast pre-rRNA processing is discussed.

Yeast Rrp5p, one of the few trans-acting proteins required for the biogenesis of both ribosomal subunits, has a remarkable two-domain structure. Its C-terminal region consists of seven tetratricopeptide motifs, several of which are crucial for cleavages at sites A(0) to A(2) and thus for the formation of 18S rRNA. The N-terminal region, on the other hand, contains 12 S1 RNA-binding motifs, most of which are required for processing at site A(3) and thus for the production of the short form of 5.8S rRNA. Yeast cells expressing a mutant Rrp5p protein that lacks S1 motifs 10 to 12 (mutant rrp5Delta6) have a normal growth rate and wild-type steady-state levels of the mature rRNA species, suggesting that these motifs are irrelevant for ribosome biogenesis. Here we show that, nevertheless, in the rrp5Delta6 mutant, pre-rRNA processing follows an alternative pathway that does not include the cleavage of 32S pre-rRNA at site A(2). Instead, the 32S precursor is processed directly at site A(3), producing exclusively 21S rather than 20S pre-rRNA. This is the first evidence that the 21S precursor, which was observed previously only in cells showing a substantial growth defect or as a minor species in addition to the normal 20S precursor, is an efficient substrate for 18S rRNA synthesis. Maturation of the 21S precursor occurs via the same endonucleolytic cleavage at site D as that used for 20S pre-rRNA maturation. The resulting D-A(3) fragment, however, is degraded by both 5'-->3' and 3'-->5' exonuclease digestions, the latter involving the exosome, in contrast to the exclusively 5'-->3' exonucleolytic digestion of the D-A(2) fragment. We also show that rrp5Delta6 cells are hypersensitive to both hygromycin B and cycloheximide, suggesting that, despite their wild-type growth rate, their preribosomes or ribosomes may be structurally abnormal.

Biogenesis of eukaryotic ribosomal subunits proceeds via a series of precursor ribonucleoprotein particles that correspond to different stages in the maturation pathway. The different pre-ribosomal particles each contain a distinct complement of non-ribosomal, trans-acting factors that are crucial for correct and efficient progress of the maturation process. Although in recent years we have gained considerable insight into the composition of the pre-ribosomal particles, our knowledge how the ordered association with and their dissociation from the pre-ribosome of these trans-acting factors is controlled is still quite limited. Here, we have studied the mutual dependence between three of these factors, Rrp5p, U3 snoRNP and Rok1p, all essential for the early stages of pre-rRNA processing/assembly, for association with the 35S pre-rRNA in Saccharomyces cerevisiae. Using co-immunoprecipitation assays, we show that Rrp5p and U3 snoRNP associate independently of each other and that the two factors do not detectably interact prior to incorporation into the pre-ribosome. In contrast, association of the putative RNA helicase Rok1p, which is known to genetically interact with Rrp5p, is absolutely dependent on the presence of the latter protein but does not require U3.

The authors' goal was to examine the course and predictors of posttraumatic stress symptoms among persons hospitalized for burns. A total of 301 participants completed self-report measures assessing peritraumatic mental state, anxiety related to pain, and posttraumatic stress symptoms. Twenty-six percent of the participants were suffering from posttraumatic stress symptoms at 2-3 weeks postburn and 15% of them at 12 months postburns. In general, a decrease in symptoms was observed over time, although a substantial part of the participants with acute stress symptoms suffers from chronic posttraumatic stress symptoms 1-year postburn. Symptoms were predicted by anxiety measures and objective factors, such as female gender, locus, and severity of injury.

Saccharomyces cerevisiae Rio2p (encoded by open reading frame Ynl207w) is an essential protein of unknown function that displays significant sequence similarity to Rio1p/Rrp10p. The latter was recently shown to be an evolutionarily conserved, predominantly cytoplasmic serine/threonine kinase whose presence is required for the final cleavage at site D that converts 20 S pre-rRNA into mature 18 S rRNA. A data base search identified homologs of Rio2p in a wide variety of eukaryotes and Archaea. Detailed sequence comparison and in vitro kinase assays using recombinant protein demonstrated that Rio2p defines a subfamily of protein kinases related to, but both structurally and functionally distinct from, the one defined by Rio1p. Failure to deplete Rio2p in cells containing a GAL-rio2 gene and direct analysis of Rio2p levels by Western blotting indicated the protein to be low abundant. Using a GAL-rio2 gene carrying a point mutation that reduces the kinase activity, we found that depletion of this mutant protein blocked production of 18 S rRNA due to inhibition of the cleavage of cytoplasmic 20 S pre-rRNA at site D. Production of the large subunit rRNAs was not affected. Thus, Rio2p is the second protein kinase that is essential for cleavage at site D and the first in which the processing defect can be linked to its enzymatic activity. Contrary to Rio1p/Rrp10p, however, Rio2p appears to be localized predominantly in the nucleus.

Rrp5p is the only protein so far known to be required for the processing of yeast pre-rRNA at both the early sites A0, A1 and A2 leading to 18S rRNA and at site A3, the first step specific for the pathway leading to 5.8S/25S rRNA. Previous in vivo mutational analysis of Rrp5p demonstrated that the first 8 of its 12 S1 RNA-binding motifs are involved in the formation of the 'short' form of 5.8S rRNA (5.8S(S)), which is the predominant species under normal conditions. We have constructed two strains in which the genomic RRP5 gene has been replaced by an rrp5 deletion mutant lacking either S1 motifs 3-5 (rrp5-Delta3) or 5-8 (rrp5-Delta4). The first mutant synthesizes almost exclusively 5.8S(L) rRNA, whereas the second one still produces a considerable amount of the 5.8S(S) species. Nevertheless, both mutations were found to block cleavage at site A3 completely. Instead, a novel processing event occurs at a site in a conserved stem-loop structure located between sites A2 and A3, which we have named A4. A synthetic lethality screen using the rrp5-Delta3 and rrp-Delta4 mutations identified the REX4 gene, which encodes a non-essential protein belonging to a class of related yeast proteins that includes several known 3'-->5' exonucleases. Inactivation of the REX4 gene in rrp5-Delta3 or rrp-Delta4 cells abolished cleavage at A4, restored cleavage at A3 and returned the 5.8S(S):5.8S(L) ratio to the wild-type value. The sl phenotype of the rrp5Delta/rex4(-) double mutants appears to be due to a severe disturbance in ribosomal subunit assembly, rather than pre-rRNA processing. The data provide direct evidence for a crucial role of the multiple S1 motifs of Rrp5p in ensuring the correct assembly and action of the processing complex responsible for cleavage at site A3. Furthermore, they clearly implicate Rex4p in both pre-rRNA processing and ribosome assembly, even though this protein is not essential for yeast.

Saccharomyces cerevisiae contains three nonessential genes (NGL1, NGL2, and NGL3) that encode proteins containing a domain with similarity to a Mg(2+)-dependent endonuclease motif present in the mRNA deadenylase Ccr4p. We have investigated a possible role of these proteins in rRNA processing, because for many of the pre-rRNA processing steps, the identity of the responsible nuclease remains elusive. Analysis of RNA isolated from cells in which the NGL2 gene has been inactivated (ngl2delta) demonstrates that correct 3'-end formation of 5.8S rRNA at site E is strictly dependent on Ngl2p. No role in pre-rRNA processing could be assigned to Ngl1p and Ngl3p. The 3'-extended 5.8S rRNA formed in the ngl2delta mutant is slightly shorter than the 6S precursor previously shown to accumulate upon combined deletion of the 3' --> 5' exonuclease-encoding REX1 and REX2 genes or upon depletion of the exosomal subunits Rrp40p or Rrp45p. Thus, our data add a further component to the set of nucleases required for correct 3'-end formation of yeast 5.8S rRNA.

Mutational analysis has shown that the integrity of the region in domain III of 25S rRNA that is involved in binding of ribosomal protein L25 is essential for the production of mature 25S rRNA in the yeast Saccharomyces cerevisiae. However, even structural alterations that do not noticeably affect recognition by L25, as measured by an in vitro assay, strongly reduced 25S rRNA formation by inhibiting the removal of ITS2 from the 27S(B) precursor. In order to analyze the role of L25 in yeast pre-rRNA processing further we studied the effect of genetic depletion of the protein or mutation of each of its three previously identified functional domains, involved in nuclear import (N-terminal), RNA binding (central) and 60S subunit assembly (C-terminal), respectively. Depletion of L25 or mutating its (pre-)rRNA-binding domain blocked conversion of the 27S(B) precursor to 5.8S/25S rRNA, confirming that assembly of L25 is essential for ITS2 processing. However, mutations in either the N- or the C-terminal domain of L25, which only marginally affect its ability to bind to (pre-)rRNA, also resulted in defective ITS2 processing. Furthermore, in all cases there was a notable reduction in the efficiency of processing at the early cleavage sites A0, A1 and A2. We conclude that the assembly of L25 is necessary but not sufficient for removal of ITS2, as well as for fully efficient cleavage at the early sites. Additional elements located in the N- as well as C-terminal domains of L25 are required for both aspects of pre-rRNA processing.

In a cross-sectional study of patients 12-24 months after a burn injury, the need for a multidisciplinary burn specific outpatient clinic was examined in relation to aftercare consumption, physical and psychological problems. Four hundred and twenty nine patients were assessed by means of three self-report questionnaires: IES, SCL(PTSD-SL) and BSHS-SV-S. Results indicated that the current aftercare providers are almost exclusively medical doctors and that a quarter of the patients are dissatisfied with received aftercare. However, dissatisfaction about current aftercare was not the only criterion to determine whether patients wanted burn-specific aftercare. The severity of psychological and physical problems predicted interest in a multi-disciplinary outpatient clinic. Self-reported psychological and physical problems were found to be related to one another. Univariate logistic regression outcomes suggested that patients with serious psychological and physical problems are more likely to express interest in a burn-specific outpatient clinic, but that, in a multivariate regression analysis, physical problems and psychological problems measured on the SCL(PTSD-SLEEP) do not contribute to the prediction of the interest in a burn-specific outpatient clinic anymore once symptoms of PTSD are taken care of. Results suggest that psychological aftercare for burn patients needs to be improved. Possible steps to improve aftercare to meet patients' needs are discussed.

Analogous to a hospital survey in North America (the US and Canada) a survey in Europe was performed to determine the extent of psychological services offered in burn units. The questionnaire used to acquire this information was exactly the same in both continents. One hundred and seventy questionnaires and self-addressed stamped envelopes were sent to the medical manager of the European burn units with the request to pass the questionnaire to the person who provides psychological care to patients in the burn units. The questionnaire was translated into four other languages. Response was 103 (62.8%). The questions posed inquired after what percentage of patients received psychological testing, psychological therapy, displayed symptoms of depression and post-traumatic stress, what percentage is given referral and if emotional follow-up is arranged. Also questions about whether staff members of the burn unit displayed symptoms of depression and post-traumatic stress were enclosed. Results indicated that few burn units in Europe provide formal psychological testing for their patients and in half of the European hospitals, less than 20% of the patients receive psychological therapy or counselling during their stay in the hospital. Aftercare services are also capable of improvement. Compared with North America, Europe provided less psychological services. No statistical differences were found between countries within Europe.

Pre-rRNA processing in eukaryotic cells requires participation of several snoRNPs. These include the highly conserved and abundant U3 snoRNP, which is essential for synthesis of 18S rRNA. Here we report the characterization of Rrp9p, a novel yeast U3 protein, identified via its homology to the human U3-55k protein. Epitope-tagged Rrp9p specifically precipitates U3 snoRNA, but Rrp9p is not required for the stable accumulation of this snoRNA. Genetic depletion of Rrp9p inhibits the early cleavages of the primary pre-rRNA transcript at A0, A1, and A2 and, consequently, production of 18S, but not 25S and 5.8S, rRNA. The hU3-55k protein can partially complement a yeast rrp9 null mutant, indicating that the function of this protein has been conserved. Immunoprecipitation of extracts from cells that coexpress epitope-tagged Rrp9p and various mutant forms of U3 snoRNA limits the region required for association of Rrp9p to the U3-specific box B/C motif. Box C is essential, whereas box B plays a supportive role.

The authors examined ratings on a scale of pain-related anxiety in 173 burn patients in three groups: patients with small burns, patients with moderate burns and patients with extensive burns. The data suggest a greater degree of anxiety during procedures and before procedures in the burn patients with extensive burns than in burn patients with small and moderate burns. This study introduces a novel measure of pain-related anxiety in clinical burn patients, the abbreviated Burn Specific Pain Anxiety Scale (BSPAS), which showed a high degree of reliability. The alpha coefficients were high for the BSPAS subscales.

The purpose of this study was to assess the reaction to burn trauma, the hospitalisation characteristics and psychosocial adjustment in 174 burn victims who agreed to participate in a structured interview. It was postulated that once patients sustain a burn, the level of loneliness feelings and feelings of shame is a function of the visibility of the scars. Contrary to expectation, the visibility of the burn scars was not a predictor of pathological feelings of shame. By contrast, social introversion was a factor significantly associated with the development of burn related pathological feelings of shame.

Burn patients were assessed 14-24 months following their injury to estimate the prevalence of chronic posttraumatic stress in the adult burn population in The Netherlands. Among 428 patients, 33 per cent suffered severe posttraumatic stress symptomatology. Burn related health problems and shame were strongly associated with posttraumatic stress. Bootstrapping was used to estimate standardized regression weights in a structural equation model.

This article describes the relationship between post-traumatic morbidity, anxious anticipation of pain and pain perception in 33 adult burn patients. Burn patients were assessed, on average, 7 days after admission to the hospital. Five times a day nurses asked the patients to provide pain ratings. The more patients suffered from post-traumatic stress, the more their anxiety state was elevated. The association between post-traumatic stress and pain perception was, controlling for the effects of anxious anticipation, spurious.

Forty-six burned adults were seen, on average 7 days after admission to hospital. Eighty per cent suffered from one or more dissociative symptoms during trauma and three or more dissociative phenomena were experienced by 19.5 per cent. Blunting or absence of emotion was the symptom most frequently reported. The number of dissociative phenomena during trauma was statistically significant and associated with extreme anxiety. The intensity of the dissociation-anxiety reaction during trauma was a predictor for psychopathology measured at the moment of discharge.

Forty-three consecutive patients were assessed for post-traumatic stress 7 days after admission to hospital. Patients were asked to complete the Impact-of-Event Scale. Findings indicated that among the 43 patients 52.1 per cent experienced serious levels of stress symptomatology. A Visual Analog Thermometer was employed to measure the pain experienced at rest and during therapeutic procedures. High levels of post-traumatic stress were associated with higher pain scores during therapeutic procedures and more pain when at rest.

The burn specific pain anxiety scale (BSPAS) is a nine-item self-report scale for the assessment of pain-related and anticipatory anxiety in burned patients. This paper describes a study designed to explore the psychometric properties of the scale. The study used 35 burned patients hospitalized in Rotterdam and Groningen, The Netherlands, to confirm the internal consistency of the instrument and provide an assessment of its validity. The alpha coefficient was high: 0.94. The BSPAS correlated statistically significantly with the STAI-S, procedural pain, non-procedural pain, and nurses' visual analog observation ratings of tension.

We have developed a system for mutational analysis of Saccharomyces cerevisiae ribosomal RNA in vivo in which yeast cells can be made completely dependent on mutant rRNA and ribosomes by a simple switch in carbon source. The system is based on a yeast strain defective in RNA polymerase I (Pol I) transcription [Nogi et al. (1991) Proc. Natl. Acad. Sci. USA 88, 3962-3966]. This normally inviable strain was rescued by integration of multiple copies of the complete 37S pre-rRNA operon under control of the inducible, Pol II-transcribed GAL7 promoter into the rDNA repeat on chromosome XII. The resulting YJV100 strain can only grow on medium containing galactose as the carbon source. A second, episomal vector was constructed in which the rDNA unit was placed under control of the constitutive PGK1 promoter. YJV100 cells transformed with this vector are now also able to grow on glucose-based medium making the cells completely dependent on plasmid-encoded rRNA. We show that the Pol II-transcribed pre-rRNA is processed and assembled similarly to authentic Pol I-synthesised pre-rRNA, making this 'in vivo Pol II system' suitable for the detailed analysis of rRNA mutations, even highly deleterious ones, affecting ribosome biogenesis or function. A clear demonstration of this is our finding that an insertion into variable region V8 in 17S rRNA, previously judged to be neutral with respect to processing of 17S rRNA, its assembly into 40S subunits and the polysomal distribution of these subunits [Musters et al. (1989), Mol. Cell. Biol. 9, 551-559], is in fact a lethal mutation.

Approximately 18 months after discharge we were able to screen 42 burned adult patients for psychological and social problems. Using independent semi-standardized interviews, validated psychological questionnaires and standardized self-reports of the patients, we obtained three separate evaluations. The interreliability between the three different 'judges' varied from poor to moderate. Research suggests that 40% of burned adult patients have psychological problems. The cycle of recovery is dynamic. Only 21% need psychological help 18 months after discharge, but half of these are in danger of becoming 'drop-outs' because they do not receive the necessary treatment. A follow-up consisting of psychological questionnaires six months post-discharge appears to be a good and efficient instrument in limiting the drop-out risk.