bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of opium use by students of medical sciences
Authors: Ahmadi Jamshid , Fallahzadeh Hossein , Salimi Alireza , Rahimian Mohsen , Salehi Vahid , Khaghani Maryam , Babaeebeigi Mohammadali
Published: March 2006

Aims and objectives.  To investigate the prevalence of opium use in university students.

Background.  University health professionals and authorities are very concerned about substance use among university students.

Design.  A survey with a representative sample of 2519 (1126 men and 1393 women) university students and opium use disorders assessed by means of DSM‐IV criteria (Diagnostic Statistical Manual‐IV Axis I during the year 2003).

Findings.  Mean age of the sample was 23·8 year and SD was 3·9. Of the students, 110 (4·4%) admitted using of opium once or more during their lives (9.1% of men and 0·6% of women; P < 0·01). Fifty (2%) were occasional opium user (4·2% of men and 0·2% of women; P < 0·01). Nineteen (0·8%) were current opium user (1·4% of men and 0·2% of women; P = 0·001). Mean age of opium users was higher than the remainder. Opium use was significantly related to gender (P = 0·001), and life stress (P = 0·04).

Conclusion.  These findings can be considered for preventive and therapeutic programmes, because early intervention during the formative university years may present an opportunity to reduce the risk of long‐term problems, to decrease social and individual harm and also to promote public health of society.

Relevance to clinical practice.  These findings can be considered in clinical practice for detection and treatment of opioid abuse.
Why don't trained community nurse prescribers prescribe?
Authors: Hall Jason , Cantrill Judith , Noyce Peter
Published: March 2006

Aims and objectives.  (i) To identify barriers that could either prevent community nurses from prescribing altogether or reduce the number of times that a nurse might prescribe. (ii) To determine how wide spread the barriers identified above were.

Background.  Many community nurses who are trained to prescribe are either not prescribing at all or not routinely doing so.

Method.  (i) Qualitative semi‐structured interviews with nurses prescribers and Trust prescribing leads. (ii) Postal questionnaires sent to the nurse prescribing leads in each Primary Care Trust across three strategic health authorities in England.

Results.  Several barriers were identified in this study. Those that (i) prevented prescribing included roles with no patient contact, prescription pads not issued, opposition from general practitioners and lack of confidence; (ii) prevented some prescribing included lack of time in clinics, inability to prescribe for patients registered with another Trust, security concerns, lack of access to patient medical records and the use of alternative methods of supply; (iii) made prescribing more difficult included keeping records, informing general practitioner of items prescribed, delivering prescribed items to housebound patients and situations requiring items from more than one prescriber.

Conclusion.  Many barriers have been identified that have the potential to reduce the impact of nurse prescribing on patient care. Trusts should monitor the prescribing of their nurse prescribers, target appropriate support and encouragement to address problems with prescriber confidence, develop strategies to integrate their nurse prescribers into the healthcare team to improve access to patient's medical notes, and improve the efficiency of the prescribing process.

Relevance to clinical practice.  There is a danger that the anticipated benefits to patient care resulting from the introduction of nurse prescribing may not materialize if the barriers identified in this study are not addressed.
The gap between saying and doing in postoperative pain management
Authors: Dihle Alfhild , Bjølseth Gunnar , Helseth Sølvi
Published: March 2006

Aim.  To understand how nurses contribute to postoperative pain management in a surgical setting and to identify barriers to achieving optimal postoperative pain alleviation.

Background.  Postoperative pain is inadequately managed. Nurses play an important role in assessment, treatment and evaluation of postoperative pain in surgical wards, but combined observational and interview studies about how they approach these activities have rarely been undertaken.

Design and methods.  The study design is descriptive. Observations and in‐depth interviews were conducted with nine nurses on three surgical wards at two hospitals. Each nurse was observed during five shifts, day and night, and interviewed after the final observation. The collection and analysis of data followed principles of qualitative research.

Findings.  One main theme emerged about the nurses’ approach to postoperative pain management: a discrepancy between what the nurses said they did and what they actually did.

Conclusion.  The study revealed a gap between what nurses said and did in postoperative pain management, and this gap was smaller when the nurses took an active approach. An active approach towards patients about postoperative pain seemed to enhance pain alleviation.

Relevance to clinical practice.  Nursing education and practice both need to promote knowledge of pain and pain management, as well as empathy and empathic communication in relation to pain. They need to collaborate in guiding nurses to act in accord with theoretical knowledge and so enhance competence in nursing actions related to postoperative pain management.
Establishing nurses’ preferences in wound assessment: a concept evaluation
Authors: Maylor Miles E
Published: March 2006

Aims and objectives.  The aim of the study was to contribute to increasing reliability of wound assessment. The objectives were to capture terminology used and to assess whether these words reflected personality preferences for information.

Background.  Wound assessment forms are expected to be used reliably by non‐experts. This assumes that they mean the same thing to all practitioners. But many wound assessment forms are too complicated and do not necessarily show whether a wound is healing or not. Literature indicates that people have different information preferences that they use to aid judgement.

Design.  A correlational study investigating the relationship between word‐preferences and assessor's personality type.

Method.  Postregistration students (n = 45) and Tissue Viability Nurses (n = 16) suggested words or phrases to be used in an hypothetical wound assessment. Analysis was performed of how they preferred to describe healing, stasis and deterioration. They ranked the three most important signs they would look for in the three phases of wounds. Words were classified into categories (e.g. subjective, objective, value judgement), then matched with preferences seen in a personality type indicator.

Results.  Healing and deteriorating wounds were assessed relative to different signs or symptoms and static wounds were the most difficult to describe (mostly by absence of signs of healing). Personal types of approach to assessment could be identified: including subjectifiers and quantifiers.

Conclusions.  In this preliminary study, statistically significant results support the view that wound assessment involves assessor preferences. Assessors judge by the absence rather than the presence of some signs and symptoms.

Relevance to clinical practice.  Wound assessment forms need to take account of assessor preferences for subjectivity or quantification if reliability is to be increased, also different wound assessment forms could suit different wound phases. Potentially, assessor types could seek agreement more on outcomes of assessment rather than input.
Hip protectors: a pilot study of older people in Taiwan
Authors: Huang Hui‐Chi , Lee Chian‐Her , Wu Shang‐Liang
Published: March 2006

Aims and objectives.  This research explores the compliance by older people in Taiwan with wearing hip protectors and whether protectors can prevent fractures or decrease injuries after falls.

Background.  The ageing process causes degeneration of nerves, joints, muscles and then the incidence of falls in the older people is higher than in the young. Therefore, fractures caused by falls, particularly hip fractures of older people, need to be studied. Hip protectors are designed to reduce hip fractures and hip injuries when falls occur, by distributing the impact load and absorbing the impact force generated during a fall.

Design.  This research was designed as a longitudinal study using purposive sampling and intervention for six months.

Methods.  The target population were older people at high risk for falls and the study was carried out during a six‐month period. Self‐reported data were collected by interview and questionnaires. The expert validity of the questionnaires, including demographics, high‐risk group, compliance and correct wearing of hip protectors and fall injuries, as assessed by three experts, was 0·94 and the test–retest correlation coefficient was 0·95. Individuals were excluded from the research sample if they were 24‐hour bedridden, non‐mobile or had an existing hip injury.

Results.  Ten female subjects participated in this study. Their mean age was 76·1 years. During a six‐month follow‐up, there were 12 falls while wearing hip protectors, however, no fractures occurred because of the falls.

Conclusions.  This is the first research into hip protectors in Taiwan. The reasons for low compliance by older people were (1) unaccustomed to use, (2) discomfort, (3) wearing only during the daytime, (4) lack of assistance from family caregivers.

Relevance to clinical practice.  Fall prevention, exercise and the use of hip protectors can significantly decrease the incidence of hip fractures. Although hip protectors are effective in reducing fall fracture risk, the compliance rate remains low. In the future, the researchers will develop a hip protector program related to teaching the correct method for wearing protectors. Helping older people learn how to overcome the discomfort of wearing hip protectors and encouraging a willingness to wear them is imperative.
Health‐related quality of life among haemodialysis patients – relationship with clinical indicators, morbidity and mortality
Authors: Morsch Cássia Maria , Gonçalves Luiz Felipe , Barros Elvino
Published: March 2006

Aim.  To verify the association between quality of life and morbidity, mortality and clinical indicators in haemodialysis patients.

Background.  While a number of therapies have been reported to increase quality of life in end‐stage renal disease, patients report that they remain substantially burdened by limited physical functioning and by dialysis‐related symptoms. Indeed, quality of life may be the most critical outcome for those undergoing haemodialysis. Furthermore, quality of life has been associated with clinical indicators, morbidity and survival in haemodialysis patients.

Design.  Descriptive cohort study of patients undergoing haemodialysis at the Nephrology Hemodialysis Unit of the Hospital de Clínicas in Porto Alegre, Brazil.

Methods.  Forty haemodialysis patients were followed for 12 months and evaluated for demographics, time on dialysis, diabetes mellitus, clinical indicators (dose of dialysis – Kt/V, haematocrit and serum albumin) and comorbidities. The comorbidities were evaluated with the end‐stage renal disease severity index and health‐related quality of life with The Medical Outcomes Study 36 (SF‐36).

Results.  Men present higher health‐related quality of life scores in the energy and fatigue component (P = 0·04). Patients treated for over one year at the beginning of follow up and patients with less schooling had better results in General Health Perception (P < 0·05). The health‐related quality of life evaluation of patients who later died showed that they already had a worse perception of physical functioning as compared to the survivors (P = 0·05). Patients with diagnosed diabetes perceived their physical functioning more negatively compared with those with other etiologies of end‐stage renal disease (P = 0·045). We found a correlation between physical functioning and serum albumin (r = 0·341, P < 0·05) and between physical functioning and haematocrit (r = 0·317, P < 0·05). The end‐stage renal disease severity index was more strongly related to physical functioning (r = −0·538, P < 0·001). Comparing the patients’ results to the indicators above and below the established targets, we observed a trend to worse health‐related quality of life in patients with Kt/V above target. However, in the case of albumin, patients with results above target tended to have better results.

Conclusion.  A close relationship was observed between quality of life and morbidity and mortality. Among the clinical indicators, albumin and haematocrit have the greatest influence on quality of life.

Relevance to clinical practice.  Haemodialysis patients experience various problems that may adversely influence their quality of life. Special care must be given to those who have diabetes mellitus, high morbidity scores, low serum albumin and low haematocrits.
The research assessment exercise in nursing: learning from the past, looking to the future
Authors: Cecil Rosanne , Thompson Kathryn , Parahoo Kader
Published: March 2006

Aims and objectives.  The aim of the study was to enable those planning to make submissions to the 2008 Research Assessment Exercise to make informed decisions regarding their research strategies. The objective of the study was to identify the factors that distinguish those units of assessment (Universities and Higher Education Institutions) that were highly rated from those that received a low rating for nursing in the Research Assessment Exercise of 2001.

Background.  Nursing research differs in kind from other types of biomedical research. There is a tendency for research in nursing to be characterized by an inward‐looking focus and dominated by a concern with the profession itself.

Design.  The examination of the research output of nursing.

Methods.  Data from the abstracts of journal articles submitted to the 2001 Research Assessment Exercise for nursing were extracted, classified, collated and analysed.

Results.  The publications submitted by those in the higher‐rated units showed a greater tendency than those submitted by those in the lower‐rated units to report on a study involving the collection of primary data; to be multi‐authored; to use either qualitative methodology or randomized‐controlled trials; and to focus upon clinical issues and have patients as the subjects of the research.

Conclusion.  Nursing research is in a process of change and growth and is still of variable quality. The development of research towards patient care and clinical issues, and away from issues relating to the profession itself, is most evident among those units of assessment rated highly in the 2001 Research Assessment Exercise.

Relevance to clinical practice.  The study indicates that the best of current nursing research is focused upon clinical issues and patient care.
Patients undergoing total hip arthroplasty: a perioperative pain experience
Authors: Stomberg Margareta Warrén , Öman Ulla‐Britt
Published: March 2006

Aim.  The aim was to evaluate patient's perioperative pain experience after total hip replacement and patients’ satisfaction with pain management.

Background.  Total hip arthroplasty is a common surgical procedure intended to reduce pain and return patients to better function. Realistic expectations about total hip arthroplasty is important for optimal postoperative recovery and the information must be adapted to fit the individual patient.

Methods.  A descriptive design was used comparing patients outcome data. Pitman's test was used for statistical analyses. Adult patients (n = 112) undergoing surgical hip replacement procedures answered a 17‐item questionnaire on the fourth postoperative day. The questionnaire included given alternatives and visual analogue scales (0–100 mm) for the pain assessment.

Result.  The patients’ postoperative pain experience after hip replacement surgery was in average low, 33·1 mm on a 100 mm visual analogue scale. Patient's pain experience was reported to be highest on the first postoperative day for most of the patients. The preoperative pain experience tends to be higher than the postoperative pain experience. Older patients reported less average pain level postoperatively. Satisfaction with pain management was high.

Conclusion.  The pain experience tends to be higher preoperatively than postoperatively. Patients who reported a higher pain experience postoperatively reported that their pain experience was significant higher than preoperative expected.

Relevance to practice.  It is important for the postoperative outcome measure that the patients have a realistic expectation of pain experiences after total hip arthroplasty. The nurse is one of the staff members responsible for information to the individual patient.
Wound measurement comparing the use of acetate tracings and VisitrakTM digital planimetry
Authors: Gethin Georgina , Cowman Seamus
Published: March 2006

Aims.  This study aimed to compare acetate tracing with the new digital planimetry system VisitrakTM (Smith + Nephew). It also aimed to compare the percentage change in wound size after four weeks using both methods.

Background.  There is a need for development and more widespread use of objective tools in wound management. Wound measurement is an integral part of wound assessment and provides possibly the most objective tool available. To achieve this, practitioners use a variety of methods including acetate tracing and digital planimetry. Studies have demonstrated good inter‐ and intrarater reliability using both methods.

Methods.  A retrospective, random selection of wound tracings of superficial leg ulcers was completed until 25 tracings of <10 cm2 and 25 tracings of >10 cm2 were obtained. Wound area was calculated firstly by manually counting squares completely within the wound border and those half or more within the border as 1 cm2. The area was then calculated using the VisitrakTM system.

Results.  The difference between the acetate and VisitrakTM for wounds <10 cm2 not significant (P = 0·330). For wounds >10 cm2 the difference between both methods was significant (P = 0·008). After four weeks the number of tracings available was too small for any conclusions to be made.

Conclusion.  Wound measurement should become part of routine practice. Acetate offers a good indicator of wound size and digital planimetry can enhance the precision of the measurement.

Relevance to clinical practice.  Accurate and objective wound measurement is a vital component of wound management. It is essential that nurses have adequate training and skills in wound measuring to assess wounds correctly and objectively. Findings from this study can be used when deciding which method should be used in a particular setting when outcomes and cost have been considered. Accurate on‐going wound measuring contributes to improved quality of patient care, more effective communication between professionals, as a predictor to wound healing, to document progress and assess treatment efficacy.
Massage to prevent pressure ulcers: knowledge, beliefs and practice. A cross‐sectional study among nurses in the Netherlands in 1991 and 2003
Authors: Duimel‐Peeters Inge GP , Hulsenboom Mirjam A , Berger Martijn PF , Snoeckx Luc HEH , Halfens Ruud JG
Published: March 2006

Abstract Background.  Pressure ulcers are a major problem in all areas of health care in the Netherlands. National guidelines for the prevention and treatment of pressure ulcers were originally developed in 1985 and revised in 1991 and 2002. The value of these guidelines can be questioned because it seems they are not in line with the beliefs and practice of the caregivers and only 5% of them are evidence‐based.

Aims and objectives.  To get a better insight into nurses’ current knowledge, beliefs and performed practices relating to massage, a study was designed to assess changes in these three aspects after the publication of the 2002 Dutch national guidelines. The outcome was compared with the situation in 1991, the year in which the previous guidelines were published.

Design and method.  A cross‐sectional comparative study was designed using written questionnaires. Questions were formulated regarding knowledge and beliefs about prevention methods and the actual use of these methods in the prevention of pressure ulcers. The 2003‐survey population consisted of nurses working in the Netherlands and was approached at random via subscriptions to Nursing News (i.e. Verpleegkunde Nieuws), a Dutch professional journal or via affiliations to an institution participating in the 2003 National Prevalence Survey of Pressure Ulcers.

Results and conclusions.  Compared with the 1991 findings, the 2003 results show an improvement for the topic of pressure ulcers, but a deterioration regarding dehydration. It is obvious that the knowledge of the current CBO‐guidelines on massage is still not widely distributed as it should be. Our overall conclusion is that the differences in responses between 1991 and 2003 are significant and suggest that the nurses were better informed in 2003.

Relevance to clinical practice.  For the improvement of health care in the domain of pressure ulcer preventions, we need more than an accurate implementation of new or existing guidelines. Guidelines should also be based on qualitative, methodological well‐designed studies to be evidence‐based.
Psychiatric wards with locked doors – advantages and disadvantages according to nurses and mental health nurse assistants
Authors: Haglund K , Von Knorring L , Von Essen L
Published: March 2006

Aims and objective.  To describe nurses’ and mental health nurse assistants’ perceptions of advantages and disadvantages about working on a psychiatric ward with a locked entrance door.

Background.  Psychiatric staff sometimes needs to protect patients from harming themselves or others. To keep the entrance door locked may help staff to achieve this goal. How locked entrance doors at psychiatric wards are experienced by staff, working on these wards, has been investigated to a very limited extent.

Design.  The study was explorative and descriptive.

Method.  Audio taped, semi‐structured interviews with open‐ended questions about advantages and disadvantages about working on a psychiatric ward with a locked entrance door, were conducted with 20 nurses and 20 mental health nurse assistants. Data were analyzed with content analysis.

Results.  A content analysis revealed eight categories of advantages and 18 categories of disadvantages. Most advantages mentioned by nurses and mental health nurse assistants were categorized as providing staff with control over patients, providing patients with a secure and efficient care and protecting patients and staff against ‘the outside’. Most disadvantages mentioned by nurses were categorized as causing extra work for staff, making patients feel confined, making patients feel dependent and creating a non‐caring environment. Most disadvantages mentioned by mental health nurse assistants were categorized as causing extra work for staff, making patients feel confined, causing emotional problems for patients, making staff's power obvious and forcing patients to adapt to other patients’ needs. Nurses and mental health nurse assistants mentioned more disadvantages than advantages and nurses mentioned more disadvantages than mental health nurse assistants.

Conclusion.  Nurses and mental health nurse assistants perceive a number of advantages and disadvantages for themselves, patients and significant others with a locked door at a psychiatric ward. Most of these concern patients’ experiences.

Relevance to clinical practice.  It is important for staff working within psychiatric care to reflect upon the fact that a locked entrance door is connected with a range of negative as well as positive perceptions and to minimize patient and own concerns connected to the locked door.
The RNA polymerase‐binding protein RbpA confers basal levels of rifampicin resistance on Streptomyces coelicolor
Authors: Newell Katy V. , Thomas Derek P. , Brekasis Dimitris , Paget Mark S. B.
Published: March 2006

RbpA is an RNA polymerase‐binding protein that occurs in the actinomycete family of bacteria and is regulated by the disulphide stress‐response sigma factor, σR, in Streptomyces coelicolor. Here we demonstrate that rbpA null mutants exhibit a slow‐growth phenotype and are particularly sensitive to the transcription inhibitor rifampicin. Strikingly, transcription mapping experiments revealed that rbpA expression is induced upon exposure of S. coelicolor to rifampicin and that this, in part, involves an increase in the activity of σR. In contrast, the ribosomal RNA operon promoter rrnDp3, which is recognized by the vegetative sigma factor σHrdB, was strongly inhibited by rifampicin. Reconstitution of RNAP from an rbpA null mutant with purified RbpA revealed that RbpA stimulates transcription from rrnDp3, even in the presence of rifampicin. The data presented suggest that RbpA confers basal levels of rifampicin resistance and is a novel regulator of rRNA synthesis in S. coelicolor.
Modulation of Mycobacterium tuberculosis proliferation by MtrA, an essential two‐component response regulator
Authors: Fol Marek , Chauhan Ashwini , Nair Naveen K. , Maloney Erin , Moomey Meredith , Jagannath Chinnaswamy , Madiraju Murty V. V. S. , Rajagopalan Malini
Published: March 2006

Paired two‐component regulatory systems consisting of a sensor kinase and a response regulator are the major means by which bacteria sense and respond to different stimuli. The role of essential response regulator, MtrA, in Mycobacterium tuberculosis proliferation is unknown. We showed that elevating the intracellular levels of MtrA prevented M. tuberculosis from multiplying in macrophages, mice lungs and spleens, but did not affect its growth in broth. Intracellular trafficking analysis revealed that a vast majority of MtrA overproducing merodiploids were associated with lysosomal associated membrane protein (LAMP‐1) positive vacuoles, indicating that intracellular growth attenuation is, in part, due to an impaired ability to block phagosome–lysosome fusion. A merodiploid strain producing elevated levels of phosphorylation‐defective MtrA (MtrAD53N) was partially replicative in macrophages, but was attenuated in mice. Quantitative real‐time PCR analyses revealed that expression of dnaA, an essential replication gene, was sharply upregulated during intramacrophage growth in the MtrA overproducer in a phosphorylation‐dependent manner. Chromatin immunoprecipitation using anti‐MtrA antibodies provided direct evidence that MtrA regulator binds to dnaA promoter in vivo indicating that dnaA promoter is a MtrA target. Simultaneous overexpression of mtrA regulator and its cognate mtrB kinase neither inhibited growth nor sharply increased the expression levels of dnaA in macrophages. We propose that proliferation of M. tuberculosis in vivo depends, in part, on the optimal ratio of phosphorylated to non‐phosphorylated MtrA response regulator.
P159 is a proteolytically processed, surface adhesin ofMycoplasma hyopneumoniae: defined domains of P159 bind heparin and promote adherence to eukaryote cells
Authors: Burnett Tracey A. , Dinkla Katrin , Rohde Manfred , Chhatwal Gursharan S. , Uphoff Cord , Srivastava Mukesh , Cordwell Stuart J. , Geary Steven , Liao Xiaofen , Minion F. Chris , Walker Mark J. , Djordjevic Steven P.
Published: March 2006

Mycoplasma hyopneumoniae, the causative agent of porcine enzootic pneumonia, colonizes the respiratory cilia of affected swine causing significant economic losses to swine production worldwide. Heparin is known to inhibit adherence ofM. hyopneumoniaeto porcine respiratory epithelial cilia.M. hyopneumoniaecells bind heparin but the identity of the heparin‐binding proteins is limited. Proteomic analysis ofM. hyopneumoniaelysates identified 27 kDa (P27), 110 kDa (P110) and 52 kDa (P52) proteins representing different regions of a 159 kDa (P159) protein derived from mhp494. These cleavage fragments were surface located and present at all growth stages. Following purification of four recombinant proteins spanning P159 (F1P159, F2P159, F3P159and F4P159), only F3P159and F4P159bound heparin in a dose‐dependent manner (Kdvalues 142.37 ± 22.01 nM; 75.37 ± 7.34 nM respectively). Scanning electron microscopic studies showedM. hyopneumoniaebound intimately to porcine kidney epithelial‐like cells (PK15 cells) but these processes were inhibited by excess heparin and F4P159. Similarly, latex beads coated with F2P159and F4P159adhered to and entered PK15 cells, but heparin, F2P159and F4P159was inhibitory. These findings indicate that P159 is a post‐translationally cleaved, glycosaminoglycan‐binding adhesin ofM. hyopneumoniae.
The chromosome partitioning proteins Soj (ParA) and Spo0J (ParB) contribute to accurate chromosome partitioning, separation of replicated sister origins, and regulation of replication initiation in Bacillus subtilis
Authors: Lee Philina S. , Grossman Alan D.
Published: March 2006

Soj (ParA) and Spo0J (ParB) of Bacillus subtilis belong to a conserved family of proteins required for efficient plasmid and chromosome partitioning in many bacterial species. Unlike most Par systems, for which intact copies of both parA and parB are required for the Par system to function, inactivating soj does not cause a detectable chromosome partitioning phenotype whereas inactivating spo0J leads to a 100‐fold increase in the production of anucleate cells. This suggested either that Soj does not function like other ParA homologues, or that a cellular factor might compensate for the absence of soj. We found that inactivating smc, the gene encoding the structural maintenance of chromosomes (SMC) protein, unmasked a role for Soj in chromosome partitioning. A soj null mutation dramatically enhanced production of anucleate cells in an smc null mutant. To look for effects of a soj null on other phenotypes perturbed in a spo0J null mutant, we analysed replication initiation and origin positioning in (soj‐spo0J)+, Δsoj, Δspo0J and Δ(soj‐spo0J) cells. All of the mutations caused increased initiation of replication and, to varying extents, affected origin positioning. Using a new assay to measure separation of the chromosomal origins, we found that inactivating soj, spo0J or both led to a significant defect in separating replicated sister origins, such that the origins remain too close to be spatially resolved. Separation of a region outside the origin was not affected. These results indicate that there are probably factors helping to pair sister origin regions for part of the replication cycle, and that Soj and Spo0J may antagonize this pairing to contribute to timely separation of replicated origins. The effects of Δsoj, Δspo0J and Δ(soj‐spo0J) mutations on origin positioning, chromosome partitioning and replication initiation may be a secondary consequence of a defect in separating replicated origins.
Quantification of Alzheimer pathology in ageing and dementia: age‐related accumulation of amyloid‐β(42) peptide in vascular dementia
Authors: Lewis H. , Beher D. , Cookson N. , Oakley A. , Piggott M. , Morris C. M. , Jaros E. , Perry R. , Ince P. , Kenny R. A. , Ballard C. G. , Shearman M. S. , Kalaria R. N.
Published: March 2006

Clinicopathological observations suggest there is considerable overlap between vascular dementia (VaD) and Alzheimer's disease (AD). We used immunochemical methods to compare quantities of amyloid‐β (Aβ) peptides in post mortem brain samples from VaD, AD subjects and nondemented ageing controls. Total Aβ peptides extracted from temporal and frontal cortices were quantified using a previously characterized sensitive homogenous time‐resolved fluorescence (HTRF) assay. The HTRF assays and immunocapture mass spectrometric analyses revealed that the Aβ(42) species were by far the predominant form of extractable peptide compared with Aβ(40) peptide in VaD brains. The strong signal intensity for the peak representing Aβ(4–42) peptide confirmed that these N‐terminally truncated species are relatively abundant. Absolute quantification by HTRF assay showed that the mean amount of total Aβ(42) recovered from VaD samples was approximately 50% of that in AD, and twice that in the age‐matched controls. Linear correlation analysis further revealed an increased accumulation with age of both Aβ peptides in brains of VaD subjects and controls. Interestingly, VaD patients surviving beyond 80 years of age exhibited comparable Aβ(42) concentrations with those in AD in the temporal cortex. Our findings suggest that brain Aβ accumulates increasingly with age in VaD subjects more so than in elderly without cerebrovascular disease and support the notion that they acquire Alzheimer‐like pathology in older age.
Behavioural and histopathological alterations in mice with cerebral malaria
Authors: Lackner P. , Beer R. , Heussler V. , Goebel G. , Rudzki D. , Helbok R. , Tannich E. , Schmutzhard E.
Published: March 2006

Different features of sensorimotor function and behaviour were studied in murine cerebral malaria (CM) and malaria without cerebral involvement (non‐CM) applying the primary screen of the SHIRPA protocol. Histopathological analysis of distinct brain regions was performed and the relative size of haemorrhages and plugging of blood cells to brain vasculature was analysed. Animals suffering from CM develop a wide range of behavioural and functional alterations in the progressive course of the disease with a statistically significant impairment in all functional categories assessed 36 h prior to death when compared with control animals. Early functional indicators of cerebral phenotype are impairments in reflex and sensory system and in neuropsychiatric state. Deterioration in function is paralleled by the degree of histopathological changes with a statistically significant correlation between the SHIRPA score of CM animals and the mean size of brain haemorrhage. Furthermore, image analysis yielded that the relative area of the brain lesions was significantly larger in the forebrain and brainstem compared with the other regions of interest. Our results indicate that assessment of sensory and motor tasks by the SHIRPA primary screen is appropriate for the early in vivo discrimination of cerebral involvement in experimental murine malaria. Our findings also suggest a correlation between the degree of functional impairment and the size of the brain lesions as indicated by parenchymal haemorrhage. Applying the SHIRPA protocol in the functional characterization of animals suffering from CM might prove useful in the preclinical assessment of new antimalarial and potential neuroprotective therapies.
Iron overload and antioxidative role of perivascular astrocytes in aceruloplasminemia
Authors: Oide T. , Yoshida K. , Kaneko K. , Ohta M. , Arima K.
Published: March 2006

Aceruloplasminemia (ACP) is an inherited disorder of iron metabolism caused by the lack of ceruloplasmin activity; the neuropathological hallmarks are excessive iron deposition, neuronal loss, bizarrely deformed astrocytes, and numerous ‘grumose or foamy spheroid bodies (GFSBs)’. We histopathologically examined two autopsied ACP brains, and observed for the first time that GFSBs form in clusters at the ends of perivascular astrocytic foot processes. Both the deformed astrocytes and the GFSBs contained ferric iron and were intensely immunolabelled with antibodies against the antioxidant proteins ferritin and manganese superoxide dismutase (Mn SOD). Ceruloplasmin is largely produced by perivascular astrocytes in the central nervous system and exhibits a ferroxidase activity that inhibits iron‐associated lipid peroxidation and hydroxyl radical formation; therefore, the lack of ceruloplasmin causes direct oxidative stress on astrocytes. The intense immunolabelling of ferritin and Mn SOD most likely reflects a defensive response to iron‐mediated oxidative stress. This study suggests that astrocytes play key roles in iron trafficking and the detoxification of iron‐mediated free radicals at the blood–brain barrier and in the parenchyma in ACP brain. The antioxidative ability of astrocytes is one of their essential neuroprotective effects, and the decompensation of this ability may lead to secondary neuronal cell death in ACP.
Rarity of JC virus DNA sequences and early proteins in human gliomas and medulloblastomas: the controversial role of JC virus in human neurooncogenesis
Authors: Muñoz‐Mármol A. M. , Mola G. , Ruiz‐Larroya T. , Fernández‐Vasalo A. , Vela E. , Mate J. L. , Ariza A.
Published: March 2006

JC virus (JCV), the agent of progressive multifocal leucoencephalopathy (PML), exerts an oncogenic effect in several laboratory animal models. Moreover, JCV genomic DNA and early viral protein T‐antigen have been detected in various types of human central nervous system (CNS) neoplasms. To further explore this association we have studied paraffin‐embedded brain biopsy tissue from 60 neoplasms (55 gliomas and five medulloblastomas) and 15 reactive gliosis cases for the presence of JCV DNA sequences and proteins. Four post mortem cases of HIV‐associated PML were used as positive controls. Samples were assessed by polymerase chain reaction (PCR) amplification of early (large T antigen) and late (virion protein 3) sequences and immunohistochemistry (IHC) with both PAb 2024 and anti‐SV40 large T antigen monoclonal antibodies. Five cases (three neoplasms and two reactive gliosis instances) showed low viral DNA levels when PCR‐tested for VP3 or large T, while no case was immunoreactive for any of the two antibodies used. The four PML cases yielded positive results with both PCR and IHC. Additionally, IHC with both antibodies was applied to a tissue micro‐array including 109 CNS tumours and 21 reactive gliosis samples. No immunoreactivity was detected in any of these tissue micro‐array samples. The rarity of JCV DNA sequences and early proteins in our brain tumours enriches the controversy over the role of JCV in human neurooncogenesis, whose clarification is in need of further molecular and epidemiologic studies.
Serine proteases purified from sera of patients with amyotrophic lateral sclerosis (ALS) induce contrasting cytopathology in murine motoneurones to IgG
Authors: Demestre M. , Howard R. S. , Orrell R. W. , Pullen A. H.
Published: March 2006

Affinity purified IgG from sera of patients with amyotrophic lateral sclerosis (ALS) is claimed to enhance transmitter release, induce apoptotic death of cultured motoneurones, and elicit a distinctive cytopathology with raised Ca2+ in mouse motoneurones. An alternative hypothesis attributes these events to serine proteases in ALS sera. To test this, motoneurones in BALB/c mice injected intraperitoneally with plasminogen affinity purified from sera of ALS patients and healthy controls were analysed using immunochemical and ultrastructural morphometric methods. The responses were validated in motoneurones of mice injected with commercially purified plasminogen, tissue plasminogen activator (tPA), or plasmin. Motoneurones in non‐injected mice had normal morphology and ultrastructure without evidence of electron‐dense degeneration. Purified plasminogen from both ALS patients and healthy controls, evoked electron‐dense motoneurone degeneration, as did commercially purified plasminogen and tPA. The common cytopathology comprised disruption and distension of Nissl body rough endoplasmic reticulum, cytoplasmic polyribosomal proliferation, and significant Ca2+ enhancement in mitochondria. By contrast, using affinity purified serum immunoglobulins, ALS‐IgG but not IgG from healthy or disease controls, elicited necrosis, with 30% of ALS‐IgGs tested evoking electron‐dense degeneration in 40% of motoneurones. The primary cytopathology was extensive swelling of Golgi endoplasmic reticulum and mitochondria, with enhancement of Ca2+ in Golgi endoplasmic reticulum and presynaptic boutons. We conclude that serine proteases purified from sera of ALS patients elicits a distinctive cytopathology and pattern of Ca2+ enhancement in motoneurones different from that found on passive transfer of affinity purified ALS‐IgG.
Changed distribution pattern of the constitutive rather than the inducible HSP70 chaperone in neuromelanin‐containing neurones of the Parkinsonian midbrain
Authors: Andringa G. , Bol J. G. J. M. , Wang X. , Boekel A. , Bennett M. C. , Chase T. N. , Drukarch B.
Published: March 2006

Aberrant protein aggregation has been recognized as an important factor in the degeneration of melanized dopaminergic neurones in Parkinson's disease (PD). The constitutive (HSP73) and (heat)‐inducible (HSP72) proteins of the heat shock 70 family form a major defence system against pathological protein aggregation. However, the distribution patterns of these chaperones in nigral neuromelanin‐laden neurones are largely unknown. The present study determined the distribution of HSP72 and HSP73 in control and Parkinsonian substantia nigra, using immunohistochemistry. In the neuromelanin‐laden neurones of controls, HSP72 was nondetectable, whereas HSP73 was weakly expressed in both the cytosol and the nucleus. Surprisingly, in PD subjects, marked nuclear HSP73, but not HSP72 immunoreactivity was observed, while cytosolic immunoreactivity of the two chaperones resembled the labelling pattern observed in controls. Furthermore, HSP73 immunoreactivity was observed in a subset of the Lewy bodies (LBs) detected in the substantia nigra of PD subjects, whereas only few of these LBs were labelled with HSP72. Interestingly, HSP72 and to a lesser extent HSP73 immunoreactivity was much stronger in nonmelanized neurones as compared with melanized neurones in this area. Thus, we conclude that the distribution pattern of HSP73 rather than HSP72 is changed in the nigral neuromelanin‐laden neurones of PD subjects as compared with control subjects. The impaired ability of aged, dopaminergic neurones to express high levels of chaperones, may contribute to the preferential vulnerability of the latter cells in PD.
In vitro identification and functional characterization of glial precursor cells in human gliomas
Authors: Colin C. , Baeza N. , Tong S. , Bouvier C. , Quilichini B. , Durbec P. , Figarella‐Branger D.
Published: March 2006

Human gliomas including astrocytomas and oligodendrogliomas are defined as being composed of neoplastic astrocytes and oligodendrocytes respectively. Here, on the basis of in vitro functional assays, we show that gliomas contain a mixture of glial progenitor cells and their progeny. We have set up explant cultures from pilocytic astrocytomas, glioblastomas and oligodendrogliomas and studied antigens that characterize glial lineage, from the precursor cells (glial restricted precursors and oligodendrocyte‐type2‐astrocyte/oligodendrocyte precursor cells expressing the A2B5 ganglioside) to the differentiated cells (oligodendrocyte and type‐1 and type‐2 astrocytes). All tumoral explants contain A2B5+ cells and can generate migrating cells with distinctive functional properties according to glioma subtypes. In pilocytic astrocytomas, very few migrating cells are dividing and can differentiate in type‐2 astrocytes or towards the oligodendrocyte lineage. In glioblastomas, most migrating cells are dividing, express A2B5 or glial fibrillary acid protein (GFAP) and can generate oligodendrocytes and type‐1 and type‐2 astrocytes in appropriate medium. Oligodendroglioma explants are made by actively dividing glial precursor cells expressing A2B5 or PSA‐NCAM. Only few cells can migrate and differentiation towards oligodendrocyte lineage does not occur. Isolated A2B5+ cells from both glioblastomas and oligodendrogliomas showed similar genetic alterations as the whole tumour. Therefore, pilocytic astrocytomas contain slowly dividing oligodendrocyte‐type2‐astrocyte/oligodendrocyte precursor cells in keeping with their benign behaviour whereas both glioblastomas and oligodendrogliomas contain neoplastic glial restricted precursor cells. In oligodendrogliomas, these cells are trapped in undifferentiated and proliferating state. The precursor cells properties present in gliomas give new insight into their histogenesis and open up new avenues for research in the field of gliomagenesis.
The characteristics of Ca++‐activated Cl–channels of the salt‐tolerant charophyteLamprothamnium
Authors: BEILBY MARY J. , SHEPHERD VIRGINIA A.
Published: March 2006

The dependence of the Ca++‐activated Cl–channels on potential difference (PD) was extracted from current–voltage (I/V) profiles recorded at the time of hypotonic regulation while the large conductance (G) K+channels were blocked by tetraethylammonium (TEA). The total clamp current (I) was dominated by the Cl–I,iCl, with small contribution from the backgroundI(ibackground). TheiClwas fitted by the Goldman–Hodgkin–Katz (GHK) model with enhanced PD dependence simulated by Boltzmann probability distributions. Theibackgroundwas modelled by an empirical equation. TheiClresponded to PD changes within tens of milliseconds. TheGmaxima were located between −20 and −150 mV. The Cl–channel number and channel permeability parameter,NClPCl, decreased as a function of time in a hypotonic medium (from 0.45 × 10−7to 0.17 × 10−7 ms−1in 19 min), with the positive half activation PD,V50+, shifting from +35 to −65 mV, and the negative half activation PD,V50–, shifting from −134 to −310 mV. The fitted Cl–concentration [Cl–]cytat the time of hypotonic regulation indicated rapid equalization of vacuolar and cytoplasmic concentrations. Excellent data obtained under similar experimental conditions in a previous study enabled us to infer [Ca++]cytinfluences on the Cl–channel characteristics. Thick sulphated polysaccharide mucilage, found onLamprothamniumcells acclimated to more saline media, eliminated the activation of theiClat the time of the hypotonic regulation. This effect was reversed by the application of the enzyme heparinase. The characteristics of theiClwere found to be consistent with a component of the excitationIs at the time of the action potential (AP). The short duration of the excitation transients was contrasted with that of the hypotonic regulation. The mechanisms for Cl–channel activation (and hence the Ca++channel activation) were considered.
Evidence for separate origins of the two Pseudomonas avellanae lineages
Authors: Scortichini M. , Natalini E. , Marchesi U.
Published: March 2006

Pseudomonas avellanae is the causal agent of hazelnut (Corylus avellana) decline, both in northern Greece and central Italy, and two lineages related to the geographical origins of the pathogen have previously been identified. Forty strains, obtained from all the areas where the disease has so far been observed, and representing six different subpopulations of the two lineages, were further assessed using insertion‐sequence PCR genomic fingerprinting. The data previously obtained from repetitive‐sequence PCR using ERIC and BOX primer sets and insertion‐sequence PCR (IS50) were analysed using statistical methods, enabling genetic diversity and gene flow among the populations to be elucidated, as well as verifying the possible correlation between genetic diversity and geographical origin. The Mantel test performed with ERIC, BOX and IS50‐PCR data revealed that the P. avellanae populations that are spatially distant from each other are also genetically dissimilar: gene flow estimates confirmed this. The present study supports the hypothesis that P. avellanae originated separately in Greece and Italy, and that the two lineages of the pathogen underwent separate local evolution.