bluuebunny/crossref_metadata_2025 huggingface dataset:
Developing interprofessional assessment
Authors: Morison Susan L. , Stewart Moira C.
Published: November 2005

Undergraduate medical and nursing education should enable the development of communication and teamworking skills and of reflective practice, which should be assessed and continued into professional practice. This study aimed to examine appropriate methods for the assessment of interprofessional learning of clinical, teamwork and communication skills for undergraduate students in Paediatrics and Children's Nursing and to involve Senior House Officers (SHOs) in this process. During the 2002/3 academic year, 31 students, four nurse educators, two medical educators and six SHOs were involved in the development and implementation of a programme of interprofessional learning and assessment. Objective structured clinical skills examination (OSCE) and role‐play assessment methods were used to explore the potential to develop common activities and standards for both professions. This study found that common learning activities and assessment methods acceptable to both professions could be developed and common standards set. It also concluded that development and delivery by an interprofessional team is particularly important and that formative feedback has a vital role to play in the process. Involving SHOs in the planning and implementation of interprofessional education (IPE) has the potential to allow its extension into different clinical areas and to meet training requirements at the postgraduate level.
Learning bioscience in nursing education: perceptions of the intended and the prescribed curriculum
Authors: Friedel June M. , Treagust David F.
Published: November 2005

This study used a curriculum inquiry framework to investigate the perceptions of 184 nursing students and nurse educators in relation to bioscience in the nursing curriculum. These participants’ attitudes to science and bioscience, self‐efficacy in bioscience and perceptions of bioscience were investigated using scales based on previous research. Nursing students were found to have significantly more positive attitudes to bioscience in nursing education than nurse educators, and nurse educators were not found to have significantly better self‐efficacy in bioscience than the students, although this might have been expected. The results of focus group discussions, used to investigate this in more depth, suggested that some nurse educators and clinical preceptors may not have sufficient science background or bioscience knowledge, to help nursing students apply bioscience knowledge to practice. As a result of this, it is suggested that the aims of the intended and prescribed nursing curricula are not being fulfilled in the implemented curriculum. Because of the increasing acuity of illness in hospitals, the fact that nurses are becoming more autonomous in their practice and the increasing expectation that nurses, in some cases, will be able to prescribe as well as administer medications, bioscience knowledge is becoming even more important. The teaching of bioscience and its application to clinical practice needs to improve if nurses are to be credible members of the multi‐disciplinary team.
Oral lesions as indicators of HIV infection among routine dental patients in Lagos, Nigeria
Authors: Agbelusi GA , Wright AA
Published: November 2005

Objectives:  To document the incidental oral lesions of human immunodeficiency virus (HIV) infection, the pattern and frequency of the lesions based on clinical presentation and oral manifestations in routine dental patients who tested positive in Nigeria.

Subjects and methods:  The study was conducted at the Oral Diagnosis/Oral Medicine clinic of the Lagos University Teaching Hospital, Lagos, Nigeria between May 2002 and April 2003. During this period, all patients with oral lesions suggestive of HIV/acquired immunodeficiency syndrome (AIDS) as described in the EEC‐WHO Classification and diagnostic criteria of oral lesions of HIV were counseled and offered voluntary HIV testing. All the 35 patients who consented and tested positive were included in this study.

Results:  Of a total of 700 patients 53 patients with oral lesions suggestive of HIV/AIDS were seen, thirty‐eight (72%) consented to HIV screening, 15 patients (28%) refused. Thirty‐five patients (92%), mean age 36 ± 13 years were confirmed positive for HIV.

Oral candidiasis was the commonest lesion seen (43%) the second common being Herpes zoster (23%). Other lesions seen included erythema multiforme in two (6%), facial palsy in two (6%) and oral hairy leukoplakia in one (3%).

Conclusion:  An oral mucosal lesion may be the presenting lesion of HIV/AIDS in routine patients attending the dental clinic. Oral health care workers should practice optimal infection control based on the Centers for Disease Control ‘Standard Precautions’ guidelines on infection control for all patients to minimize occupational transmission of HIV.
Norwegian LongoVital® and recurrent aphthous ulceration: a randomized, double‐blind, placebo‐controlled study
Authors: Kolseth I , Herlofson BB , Pedersen A
Published: November 2005

Objective:  LongoVital® (LV) is a herbal‐based tablet enriched with the recommended daily doses of vitamins. The present study was undertaken to investigate possible prevention of recurrent aphthous ulceration (RAU) during 4 months daily intake of the Norwegian LV.

Design:  The study was a placebo‐controlled, double‐blind, randomized, clinical trial.

Subjects:  Sixty otherwise healthy patients with at least one attack of minor RAU per 2 months were included in the study.

Methods:  After an introduction period (IP) of 60 days, the patients were randomly divided into three groups and given either LV, the herbs of LV only, or placebo. Three test tablets were taken every day together with breakfast for 4 months [tablet period (TP)] and the patients followed up for another 4 months (F‐UP). The number of new ulcers (NU) and ulcer‐free days (UFD) were observed.

Results:  Fifty‐two patients completed the study. Neither NU nor UFD showed any statistical significant differences between any of the groups in any of the periods. All three groups, however, showed a significant increase in UFD during the first 2 months of TP compared to IP. Within the LV group only, there was a further increase in UFD after 2 months intake of the tablets. The number of NU and UFD decreased significantly in both the LV and the herbal group in F‐UP compared with TP.

Conclusion:  Neither the Norwegian LV nor the herbal component alone was superior to placebo in the prevention of RAU. The results, however, indicate that neither the LV nor the herbal group benefited from the treatment.
Clinical features of hypodontia and associated dental anomalies: a retrospective study
Authors: Kırzıoğlu Z , Köseler Şentut T , Özay Ertürk MS , Karayılmaz H
Published: November 2005

Objective:  Hypodontia is the most commonly known developmental dental anomaly in man. This paper aims to investigate the characteristics of tooth absence and associated dental anomalies among the patients attending our paediatric dentistry clinic.

Subjects and methods:  For this study, 192 patients aged between 1 and 18 years with congenitally missing teeth were selected among the patients who attended the Department of Paediatric Dentistry of Süleyman Demirel University between January 2000 and December 2003. Clinical and radiographic examinations were performed by two dentists in order to detect the missing teeth.

Results:  We determined 503 congenitally missing teeth in 192 patients (male=93, female=99), excluding third molars. Twelve of these teeth were deciduous and 491 were permanent. A higher incidence of missing teeth was observed in girls than boys, in mandibular arch than maxillary and on the right side than left side. However the differences between arches and sides were not statistically significant in both sexes.

Conclusion:  The diagnosis of the hypodontia should be performed as early as possible, thus alternative treatment modalities can be planned and performed in a multidisciplinary team approach, in order to establish an aesthetic and functional dentition in the future.
Tendon and Muscle Imaging in Sports
Authors: Kainberger F. , Weidekamm C.
Published: November 2005

Tendon and muscle lesions appear in manifold forms clinically as well as on magnetic resonance or ultrasound images. Tendinous abnormalities may be classified and staged with respect to their biomechanical role in a kinetic chain by using the concept of the tendon overuse syndrome (TOS). In the first phase, painful functional impairment of movements occurs without any morphological changes. In the second stage, abnormalities of the gliding tissue in the form of bursitis, tendovaginitis or peritendinitis are observed. In the third stage, such lesions are followed by degenerative changes of the tendon itself. Often, they present more clearly than during the early forms of TOS and three types of tendon degeneration can be differentiated: tendinosis at distinct points along the course of the tendon, fibroostosis at the tendon insertion and compression or impingement syndromes. Rupture of fibres following tendinosis may be considered as the last or fourth stage of TOS.
Novel Developments in Sports Injuries: The Ankle
Authors: Faletti C.
Published: November 2005

Sports injuries may cause traumatic and degenerative injuries at both an amatuer and professional level and are caused by direct athletic gestures or biomechanical modifications triggered by repetitive movements. Because of its particular morphology, the ankle is often involved in these injuries, particularly in soccer, athletics and basketball. The most common pathologies involve sprains, but there may also be osteochondral fractures, impingement syndrome, stress fractures or tendon injuries. There is a complex diagnostic cycle which must include an evaluation of the clinical history, specific information from specialists and diagnostic imaging. Nowadays, there are many and varied methodologies available that vary in sensibility and specificity. The correct diagnostic imaging cycle must be followed, integrating the methodologies in such a way as to obtain the most reliable diagnostic data as possible.
Professional issues in emergency medicine: UK perspective
Authors: Hughes Geoffrey
Published: November 2005

The present paper describes the history, the professional development and current state of the speciality of emergency medicine in the UK, and outlines the role of parent bodies that administer the speciality.
Factors associated with walkout of patients from New South Wales hospital emergency departments, Australia
Authors: Mohsin Mohammed , Young Lis , Ieraci Sue , Bauman Adrian E
Published: November 2005

Objective To explore the association between daily patient numbers, sociodemographic, clinical and system characteristics and walkout rates of patients from New South Wales (NSW) public hospital ED (without seeing a medical officer).

Methods This was a secondary analysis of the NSW Emergency Department Data Collection, between 1 January 1999 and 31 December 2001, with a total of 4 356 323 ED attendances. Bivariate and multiple logistic regression analyses were performed to explore the relationship of walkout from ED and explanatory variables.

Results About 5.7% of the patients attending in NSW public hospital ED left without seeing a doctor. The results from bivariate and multivariate analyses revealed that walkout rates significantly varied by sociodemographic and clinical characteristics of the patients. Patient groups that more likely to walk out were those aged 15–44 years, from a non‐English‐speaking background, Aboriginal, with lower socioeconomic status, with no private health insurance coverage and longer waiting times for triage. Patient volume in the ED showed a significant positive association with walkout rates.

Conclusions After adjusting for triage category (urgency of presentation) and triage time, patients from lower socioeconomic backgrounds and those without private health insurance coverage were more likely to leave ED without treatment. These results have important implications for Health Services. Future strategies aimed at minimizing walkouts from public hospital ED should prioritize and target factors identified in the present study.
Non‐urgent presentations to a paediatric emergency department: Parental behaviours, expectations and outcomes
Authors: Ryan Mary , Spicer Maureen , Hyett Carlie , Barnett Peter
Published: November 2005

Objectives:  To identify factors that influence the attendance of children with non‐urgent medical problems to a tertiary paediatric ED and to define parental expectations of the hospital visit.

Methods:  A questionnaire administered to a convenience sample of 200 parents/carers who attended the paediatric ED and whose children were categorized as having non‐urgent problems that had been present for at least 4 weeks. Subsequent outpatient attendances were then analysed retrospectively.

Results:  Most patients had visited a general practitioner but remained unhappy with the outcome of the consultation. The hospital was expected to provide immediate diagnosis and initiate appropriate treatment by the vast majority of parent. Only four patients required admission.

Conclusions:  The expectations of parents who attend the paediatric ED with children who have non‐urgent chronic complaints are high. Rapid access to outpatient clinics might provide an acceptable alternative for these patients, and free valuable ED resources.
Techniques for reduction of anteroinferior shoulder dislocation
Authors: Cunningham Neil J
Published: November 2005

Dislocation of the shoulder joint is common. The shoulder is affected in up to 60% of all major joint dislocations, one study citing an incidence of 1.7% in the general population. The most common form is anteroinferior dislocation. A variety of techniques to reduce shoulder dislocation has been described. The key to successful relocation is a thorough understanding of the anatomy of both the enlocated and the dislocated shoulder joint.
Definitive management of acute cardiac tamponade secondary to blunt trauma
Authors: Fitzgerald Mark , Spencer Jack , Johnson Fiona , Marasco Silvana , Atkin Chris , Kossmann Thomas
Published: November 2005

Blunt cardiac injuries are a leading cause of fatalities following motor‐vehicle accidents. Injury to the heart is involved in 20% of road traffic deaths. Structural cardiac injuries (i.e. chamber rupture or perforation) carry a high mortality rate and patients rarely survive long enough to reach hospital. Chamber rupture is present at autopsy in 36–65% of death from blunt cardiac trauma, whereas in clinical series it is present in 0.3–0.9% of cases and is an uncommon clinical finding. Patients with large ruptures or perforations usually die at the scene or in transit – the rupture of a cardiac cavity, coronary artery or intrapericardial portion of a major vein or artery is usually instantly fatal because of acute tamponade. The small, rare, remaining group of patients who survive to hospital presentation usually have tears in a cavity under low pressure and prompt diagnosis and surgery can now lead to a survival rate of 70–80% in experienced trauma centres. As regional trauma systems evolve, patients with severe, but potentially survivable cardiac injury are surviving to ED. Two distinct syndromes are apparent – haemorrhagic shock and cardiac tamponade. Any patient with severe chest trauma, hypotension disproportionate to estimated loss of blood or with an inadequate response to fluid administration should be suspected of having a cardiac cause of shock. For patients with severe hypotension or in extremis, the treatment of choice is resuscitative thoracotomy with pericardotomy. Closed chest cardiopulmonary resuscitation is ineffective in these circumstances. Blunt traumatic cardiac injury presenting with shock is associated with a poor prognosis. The majority of survivors of blunt or penetrating cardiac injury present to the ED/trauma centre with vital signs. The main pathophysiologic determinant for most survivors is acute pericardial tamponade. The presence of normal clinical signs or normal ECG studies does not exclude tamponade. In recent years the widespread availability and use of ultrasound for the initial assessment of severely injured patients has facilitated the early diagnosis of cardiac tamponade and associated cardiac injuries. Two cases of survival from blunt traumatic cardiac trauma are described in the present paper to demonstrate survivability in the context of rapid assessment and intervention.
Quality of poisoning management advice in the Monthly Index of Medical Specialties Annual
Authors: Mallows James , Chan Betty , Graudins Andis
Published: November 2005

Background:  The Monthly Index of Medical Specialties (MIMS) contains Therapeutic Goods Administration‐approved product information supplied by manufacturers. It is widely used by health‐care professionals but is not specifically designed as a toxicology reference.

Objectives:  To determine how widespread the use of MIMS is as a toxicology reference. To evaluate the quality of poisoning management advice it contains.

Methods:  First, a survey of 500 consecutive calls to the NSW Poison Information Centre (PIC) was undertaken asking health‐care workers which toxicology references were consulted prior to calling and which references they would use if the PIC were not available. Second, a consensus opinion for poisoning management was obtained, for 25 medications which are either commonly involved in poisoning or potentially life‐threatening in overdose, by review of 5 current toxicology references for contraindicated treatments, ineffective treatments and specific recommended treatments and antidotes. MIMS poisoning management advice was then compared with this toxicology consensus opinion.

Results:  In total, 276 doctors and 222 nurses were surveyed. Prior to calling the PIC 22.8% of doctors and 6.8% of nurses consulted MIMS. In total, 25.7% of doctors and 39.6% nurses stated they would use the MIMS for poisoning management advice if the PIC were not available. For the 25 drugs assessed, 14 contained inaccurate poisoning management: 1 recommended ineffective treatments and 14 omitted specific treatments or antidotes.

Conclusion:  The MIMS is often used as a toxicology reference by physicians prior to calling the PIC. It contains a number of significant inaccuracies pertaining to management of poisonings and should not be used as a primary reference for poisoning advice.
Unusual cause of sudden onset headache: Spontaneous intracranial hypotension
Authors: Broadley Simon A , Park Nick , Renowden Shelley , Ferguson Iain T
Published: November 2005

Spontaneous intracranial hypotension is a very distinctive but unusual cause of acute headache. The postural nature of the headache can be easily overlooked in the celerity to exclude subarachnoid haemorrhage. We describe the clinical and radiological features of a case that emphasizes some of the diagnostic difficulties. An approach to management and treatment for this condition is outlined.
THREADS: A Life in Geochemistry
Authors: Turekian Karl K.
Published: November 2005

I returned and saw under the sun, that the race is not to the swift, nor the battle to the strong, neither yet bread to the wise, nor yet riches to men of understanding, nor yet favour to men of skill; but time and chance happeneth to them all. Ecclesiastes 9:11

(King James version)
The Rotavirus Vaccine Saga
Authors: Shaw Alan R.
Published: November 2005

Rotavirus is the single most common cause of acute, dehydrating gastroenteritis worldwide. This is a highly contagious and highly democratic disease. The attack rate in infants and young children is similar regardless of sanitation, socioeconomics or geography. Rotavirus vaccine development began in the early 1980s using a “Jennerian” approach based on rotaviruses that normally infect animals. Although these vaccines were found to be generally safe, protection from disease was inconsistent. The second generation of vaccines was based on the same animal viruses configured to carry the relevant coat proteins of human rotaviruses. An attenuated human rotavirus vaccine has also been developed. After close to 20 years of laboratory and clinical studies, safe and effective rotavirus vaccines are approaching regulatory approval.
REGULATION OF NORMAL AND CYSTIC FIBROSIS AIRWAY SURFACE LIQUID VOLUME BY PHASIC SHEAR STRESS
Authors: Tarran Robert , Button Brian , Boucher Richard C.
Published: November 2005

▪ Abstract  The physical removal of viruses and bacteria on the mucociliary escalator is an important aspect of the mammalian lung's innate defense mechanism. The volume of airway surface liquid (ASL) present in the respiratory tract is a critical determinant of both mucus hydration and the rate of mucus clearance from the lung. ASL volume is maintained by the predominantly ciliated epithelium via coordinated regulation of (a) absorption, by the epithelial Na+channel, and (b) secretion, by the Ca2+-activated Cl−channel (CaCC) and CFTR. This review provides an update on our current understanding of how shear stress regulates ASL volume height in normal and cystic fibrosis (CF) airway epithelia through extracellular ATP- and adenosine (ADO)-mediated pathways that modulate ion transport and ASL volume homeostasis. We also discuss (a) how derangement of the ADO-CFTR pathway renders CF airways vulnerable to viral infections that deplete ASL volume and produce mucus stasis, and (b) potential shear stress–dependent therapies for CF.
Effect of exercise intensity and hypoxia on skeletal muscle AMPK signaling and substrate metabolism in humans
Authors: Wadley G. D. , Lee-Young R. S. , Canny B. J. , Wasuntarawat C. , Chen Z. P. , Hargreaves M. , Kemp B. E. , McConell G. K.
Published: November 2005

We compared in human skeletal muscle the effect of absolute vs. relative exercise intensity on AMP-activated protein kinase (AMPK) signaling and substrate metabolism under normoxic and hypoxic conditions. Eight untrained males cycled for 30 min under hypoxic conditions (11.5% O2, 111 ± 12 W, 72 ± 3% hypoxia V̇o2 peak; 72% Hypoxia) or under normoxic conditions (20.9% O2) matched to the same absolute (111 ± 12 W, 51 ± 1% normoxia V̇o2 peak; 51% Normoxia) or relative (to V̇o2 peak) intensity (171 ± 18 W, 73 ± 1% normoxia V̇o2 peak; 73% Normoxia). Increases ( P < 0.05) in AMPK activity, AMPKα Thr172 phosphorylation, ACCβ Ser221 phosphorylation, free AMP content, and glucose clearance were more influenced by the absolute than by the relative exercise intensity, being greatest in 73% Normoxia with no difference between 51% Normoxia and 72% Hypoxia. In contrast to this, increases in muscle glycogen use, muscle lactate content, and plasma catecholamine concentration were more influenced by the relative than by the absolute exercise intensity, being similar in 72% Hypoxia and 73% Normoxia, with both trials higher than in 51% Normoxia. In conclusion, increases in muscle AMPK signaling, free AMP content, and glucose disposal during exercise are largely determined by the absolute exercise intensity, whereas increases in plasma catecholamine levels, muscle glycogen use, and muscle lactate levels are more closely associated with the relative exercise intensity.
Isolated Fetal Hyperechogenic Bowel Associated with Intra-Uterine Parvovirus B19 Infection
Authors: Jouannic Jean-Marie , Gavard Laurent , Créquat Joël , Muller Françoise , Serero Stéphane , Bénifla Jean-Louis , Costa Jean-Marc
Published: November 2005

We report a case of fetal hyperechogenic bowel diagnosed at midgestation that was associated with fetal parvovirus B19 infection. Isolated hyperechogenic bowel was detected at 25 weeks. Cystic fibrosis, chromosomal abnormalities and cytomegalovirus infection were excluded, whereas polymerase chain reaction DNA for parvovirus B 19 was found positive on amniotic fluid. The hyperechogenic bowel decreased with complete resolution by 32 weeks of gestation. No other signs of fetal B19 infection were detected prenatally and the baby had normal postnatal outcome. This case provides additional arguments in favor of a possible intestinal tropism of parvovirus B19 during fetal life. Fetal B19 infection should be systematically incorporated in the prenatal evaluation of isolated fetal hyperechogenic bowel.
Reduced Capacity of Whole Blood Leucocytes to Express Tumour Necrosis Factor-α and Interleukin-10 following Major Orthopaedic Surgery
Authors: Reikeraas O. , Sun J. , Krohn C.D. , Wang J.E. , Aasen A.O.
Published: November 2005

Background: Severe trauma is a challenge to the immune response and may cause reduced immune capacity. As a marker of decreased cellular activity, studies with ex vivo lipopolysaccharide (LPS) stimulation of whole blood or isolated mononuclear cells from injured patients have revealed reduced production of inflammatory cytokines. To gain further insight into immune alterations in orthopaedic surgery, we studied LPS-induced tumour necrosis factor (TNF)-α and interleukin (IL)-10 in whole blood of patients during peri- and postoperative phases of total hip replacement. Methods: Four females and 3 males undergoing elective total hip replacement were included in the study. Ex vivo LPS-induced TNF-α and IL-10 were measured in a whole blood assay before, during and at 1 and 6 days after operation. In addition, the counts of white blood cells were determined. Results: During the operation, there were significant reductions in the number of monocytes, but at day 1 and 6 after surgery, there were significant increases as compared to the levels before surgery. The capacity of whole blood to express TNF-α and IL-10 did not change significantly during the operation and the following postoperative day. At day 6, however, there were significant reductions in expression of both TNF-α and IL-10 as compared to the levels before the operation. In relation to the values of monocytes, there was a significant reduction in the expression of TNF-α also at day 1 after operation. Conclusion: Our data indicate that in the course of at least 6 days after a major orthopaedic trauma, there is suppression of the whole blood capacity to express the inflammatory cytokine TNF-α and the anti-inflammatory cytokine IL-10 when exposed to LPS. During this time, then, the patient is particular susceptible to septic complications.
Interleukin-10 Gene Transfection of Donor Pancreas Grafts Protects against Rejection after Heterotopic Pancreas Transplantation in a Rat Model
Authors: Özden H. , Kabay B. , Guven G. , Acikalin M.F. , Erbis H. , Alatas O.
Published: November 2005

Objective: The aim of this study was to assess the effect of immunoregulatory cytokine interleukin-10 (IL-10) gene therapy on pancreas tissue rejection in a heterotopic pancreas transplantation model. Background: Modulation of inflammatory responses by anti-inflammatory cytokines (e.g., IL-10) has been suggested to minimize organ rejection. In this context, modulation of cytokines using gene therapy could be a new therapeutic modality in preventing organ rejection. Methods: The study was performed using male inbred Wistar rats as recipients and Sprague-Dawley rats as donors. 24 h before transplantation, groups of rats, named IL-10 (n = 20) and green fluorescent protein (GFP, n = 20), were injected with viral vectors Ad5CMVhIL10 or Ad5CMVGFP. Sham-operated rats (n = 20) underwent saline injection only before transplantation. The pancreatic tissue from each of these donor rats was subsequently transplanted into the corresponding groups of streptozotocin-induced diabetic recipient rats. Recipients were thus transfected with either IL-10 (n = 20), GFP-only carrying viral vectors (n = 20) or no viral vectors (normal saline, n = 20). A selected number of animals from each recipient group (n = 5) was sacrificed at weekly intervals for 3 weeks and some were further followed up to 12 weeks before sacrifice. Histological assessment of the pancreatic tissue was made based on rejection and GFP expression. Blood glucose levels were checked daily in all groups until sacrifice. Upon sacrifice, serum cytokine and insulin levels were measured. Histopathological correlations between blood glucose levels, serum insulin levels and serum IL-10 levels were made. Results: IL-10 gene therapy significantly attenuated pancreas rejection compared to controls, provided more normal blood glucose levels and elevated plasma insulin levels. Upon assumed natural deactivation of transferred viruses after 4 weeks, differences between groups in terms of rejection, blood glucose and insulin levels disappeared. Conclusion: These findings suggest that IL-10 gene therapy significantly reduced pancreas rejection.
Irradiated Pancreatic Cancer Cells Undergo both Apoptosis and Necrosis, and Could Be Phagocytized by Dendritic Cells
Authors: Shimamura H. , Sunamura M. , Tsuchihara K. , Egawa S. , Takeda K. , Matsuno S.
Published: November 2005

The interaction of immature dendritic cells (DC) with irradiated pancreatic cancer cells was examined. Flow cytometric analysis using annexin V and propidium iodide revealed that ionizing radiation (25–35 Gy X-ray) induced both apoptosis and necrosis in pancreatic cancer cell lines. After irradiation, PK-1 and Panc-1 cells were likely to undergo necrosis, whereas MIAPaCa-2 cells underwent apoptosis. When DiO-stained immature DCs were co-incubated with DiI-stained irradiated MIAPaCa-2, it was observed under fluorescent microscopy that DCs phagocytized dead tumor cells as early as 4 h after co-incubation. The DCs’ phagocytosis of irradiated tumor cells was also confirmed by flow cytometry. These results suggest that irradiated pancreatic cancer cells, which undergo both apoptosis and necrosis, could be a good source of tumor-associated antigens for cross-presentation by DCs.
Effect of Povidone-Iodine Liposome Hydrogel on Colonic Anastomosis
Authors: Ustek S. , Kismet K. , Akkus M.A. , Ozcan A.H. , Aydogan A. , Renda N.
Published: November 2005

Background: To evaluate the effectof povidone-iodine liposome hydrogel on colonic anastomosis. Methods: 70 Wistar-Albino male rats were randomly divided into seven groups. The left colon was transected and end-to-end anastomosis was performed. PVP-I liposome hydrogel was applied around the anastomoses in groups 2 and 5. Colonic bursting pressures and tissue hydroxyproline contents were measured on postoperative days 3 and 7. Results: PVP-I application did not cause any difference in the bursting pressures on postoperative day 3, but anastomotic strength was significantly increased by the use of PVP-I on postoperative day 7. The use of PVP-I liposome hydrogel had no effect on the level of perianastomotic hydroxyproline on postoperative day 3 but had positive effects on postoperativeday 7. Conclusions: We concluded that the beneficial effects of PVP-I liposome hydrogel might be due to the combination of broad-spectrum antimicrobial activity of PVP-I with the drug delivery properties and moisturizing molecular film effects of the liposome hydrogel.
Effect of Pantoprazole Administered Subcutaneously on the Healing of Sutured Gastric Incisions in Rats
Authors: Bonin E.A. , Campos A.C.L. , Coelho J.C.U. , Matias J.E.F. , Malafaia O. , Jonasson T.H.
Published: November 2005

Background: The properties of proton pump inhibitors most investigated are related to peptic diseases and upper gastrointestinal bleeding, but their influence on the healing of sutured gastric incisions has not been assessed. In the present study we evaluated the effect of subcutaneously administered pantoprazole on the healing of sutured gastric incisions in rats. Methods: Sixty rats were divided into a pantoprazole group and a control group. All rats were submitted to gastric suture in the gastric body and in the gastric fundus and had their gastric pH measured. The pantoprazole group received 20 mg/kg pantoprazole and the control group received 0.9% isotonic NaCl, both subcutaneously t.i.d. Healing analysis was carried out using collagen picrosirius red F3BA staining, and breaking strength was measured on the 4th and 7th postoperative days in all groups. Results: Gastric pH was higher in the pantoprazole group. In the fundus, the pantoprazole group had a higher measurement of breaking strength and a higher proportion of type-I over type-III collagen on the 7th postoperative day. In the body, the pantoprazole group had a higher proportion of type-I over type-III collagen on the 4th and 7th postoperative days. Conclusions: Pantoprazole given subcutaneously promoted a reduction in gastric acid secretion and was associated with improved healing of the sutured gastric incision in the fundus (squamous epithelium) of rats. These findings suggest that pantoprazole has healing properties in sutured gastric incisions with potential benefits in gastric surgery.
Complete Inventory of ABC Proteins in Human Pathogenic Yeast, Candida albicans
Authors: Gaur Manisha , Choudhury Devapriya , Prasad Rajendra
Published: November 2005

The recent completion of the sequencing project of the opportunistic human pathogenic yeast, Candida albicans (http://www.ncbi.nlm.nih.gov/), led us to analyze and classify its ATP-binding cassette (ABC) proteins, which constitute one of the largest superfamilies of proteins. Some of its members are multidrug transporters responsible for the commonly encountered problem of antifungal resistance. TBLASTN searches together with domain analysis identified 81 nucleotide-binding domains, which belong to 51 different putative open reading frames. Considering that each allelic pair represents a single ABC protein of the Candida genome, the total number of putative members of this superfamily is 28. Domain organization, sequence-based analysis and self-organizing map-based clustering led to the classification of Candida ABC proteins into 6 distinct subfamilies. Each subfamily from C. albicans has an equivalent in Saccharomyces cerevisiae suggesting a close evolutionary relationship between the two yeasts. Our searches also led to the identification of a new motif to each subfamily in Candida that could be used to identify sequences from the corresponding subfamily in other organisms. It is hoped that the inventory of Candida ABC transporters thus created will provide new insights into the role of ABC proteins in antifungal resistance as well as help in the functional characterization of the superfamily of these proteins.
Molecular Cloning and Biochemical Characterization of L-N-Carbamoylase from Sinorhizobium meliloti CECT4114
Authors: Martínez-Rodríguez Sergio , Clemente-Jiménez Josefa María , Rodríguez-Vico Felipe , Las Heras-Vázquez Francisco Javier
Published: November 2005

An N-carbamoyl-L-amino acid amidohydrolase (L-N-carbamoylase) from Sinorhizobium meliloti CECT 4114 was cloned and expressed in Escherichia coli. The recombinant enzyme catalyzed the hydrolysis of N-carbamoyl α-amino acid to the corresponding free amino acid, and its purification has shown it to be strictly L-specific. The enzyme showed broad substrate specificity, and it is the first L-N-carbamoylase that hydrolyses N-carbamoyl-L-tryptophan as well as N-carbamoyl L-amino acids with aliphatic substituents. The apparent Km values for N-carbamoyl-L-methionine and tryptophan were very similar (0.65 ± 0.09 and 0.69 ± 0.08 mM, respectively), although the rate constant was clearly higher for the L-methionine precursor (14.46 ± 0.30 s–1) than the L-tryptophan one (0.15 ± 0.01 s–1). The enzyme also hydrolyzed N-formyl-L-methionine (kcat/Km = 7.10 ± 2.52 s–1·mM–1) and N-acetyl-L-methionine (kcat/Km = 12.16 ± 1.93 s–1·mM–1), but the rate of hydrolysis was lower than for N-carbamoyl-L-methionine (kcat/Km = 21.09 ± 2.85). This is the first L-N-carbamoylase involved in the ‘hydantoinase process’ that has hydrolyzed N-carbamoyl-L-cysteine, though less efficiently than N-carbamoyl-L-methionine. The enzyme did not hydrolyze ureidosuccinic acid or 3-ureidopropionic acid. The native form of the enzyme was a homodimer with a molecular mass of 90 kDa. The optimum conditions for the enzyme were 60°C and pH 8.0. Enzyme activity required the presence of divalent metal ions such as Ni2+, Mn2+, Co2+ and Fe2+, and five amino acids putatively involved in the metal binding were found in the amino acid sequence.
Increasing PCR Fragment Stability and Protein Yields in a Cell-Free System with Genetically Modified Escherichia coli Extracts
Authors: Michel-Reydellet Nathalie , Woodrow Kim , Swartz James
Published: November 2005

Escherichia coli cell-free protein synthesis is a highly productive system that can be applied to high throughput expression from polymerase chain reaction (PCR) products in 96-well plates for proteomic studies as well as protein evolution. However, linear DNA instability appears to be a major limitation of the system. We modified the genome of the E. coli strain A19 by removing the endA gene encoding the endonuclease I and replacing the recCBD operon (in which recD encodes the exonuclease V) by the λ phage recombination system. Using the cell extract from this new strain increased the stability of PCR products amplified from a plasmid containing the cat gene. This resulted in CAT (chloramphenicol acetyltransferase) production from PCR products comparable to that from plasmids (500–600 µg/ml) in a batch reaction. We show that cell-free protein synthesis reactions using PCR products amplified from genomic DNA and extended with the T7 promoter and the T7 terminator give the same high yields of proteins (550 µg/ml) in 96-well plates. With this system, it was possible to rapidly express a range of cytoplasmic and periplasmic proteins.
Quantitative Effects of Carbohydrates and Aromatic Amino Acids on Clostridium botulinum Toxin Gene Expression Using a Rapid Competitive RT/PCR Assay
Authors: Sharkey Freddie H. , Dooley James S. , Haylock Richard W.
Published: November 2005

A rapid competitive RT/PCR assay was developed to determine the effects of nutrients on Clostridium botulinum type E toxin gene expression. The type E strain (EVH) was grown in a nutrient-rich broth containing 1% glucose (base medium). Toxin gene expression was quantified at both mid and late exponential phases of growth. It was found that toxin encoding mRNA levels were highly growth phase dependent with elevated levels found in late exponential phase compared to mid exponential phase. Changing the carbohydrate source had a smaller effect on toxin encoding mRNA levels but as earlier results have suggested, toxin encoding mRNA levels show a strong correlation with type E growth rate. The results have important implications for the food industry whereby risk of type E botulism could be correlated to the nutrient composition of the contaminated food or assessed from C. botulinum growth rates in challenged foodstuffs.
Heterologous Aquaporin (AQY2–1) Expression Strongly Enhances Freeze Tolerance of Schizosaccharomyces pombe
Authors: Tanghe An , Kayingo Gerald , Prior Bernard A. , Thevelein Johan M. , van Dijck Patrick
Published: November 2005

Aquaporin membrane proteins enable the transport of water across membranes in various organisms. In yeast their expression has been shown to correlate strongly with freeze tolerance. When we analyzed the freeze tolerance of Schizosaccharomyces pombe, an organism whose genome sequence has revealed no genes encoding a bona fide water channel, we found very low intrinsic freeze tolerance compared to other yeast species with aquaporin-encoding genes. Deletion of Spac977.17, which encodes a putative glycerol facilitator, resulted in no significant differences in freeze tolerance with its corresponding wild-type strain in all growth conditions tested. However, when we expressed the Saccharomyces cerevisiae aquaporin-encoding gene AQY2–1 in S. pombe cells, we found that the relatively low freeze tolerance of S. pombe could be significantly enhanced. Therefore, (i) the absence of a bona fide water channel in S. pombe might provide in part an explanation for its overall low freeze tolerance compared to other yeast species, and (ii) aquaporin overexpression might be a tool to improve cryopreservation of many other cell types as well, as has recently been shown for mouse oocytes and fish embryos.
The Streptomyces lividans Cytoplasmic Signal Recognition Particle Receptor FtsY Is Involved in Protein Secretion
Authors: Palomino Carmen , Mellado Rafael P.
Published: November 2005

The bacterial version of the mammalian signal recognition particle (SRP) and its receptor α-subunit (FtsY) is well conserved and essential to all known bacteria. In Gram-negative bacteria, the SRP pathway mediates a co-translational targeting of most inner membrane proteins. Additionally, in Streptomyces lividans, a Gram-positive bacterium, SRP also targets secretory proteins to the translocon. The role of S. lividans FtsY has been assessed in this work. Co-immunoprecipitation studies confirmed that FtsY is associated with the S. lividans SRP in the cytoplasm and that this complex also co-immunoprecipitated with pre-agarase, suggesting that the SRP receptor is involved in SRP-mediated targeting of secretory proteins in S. lividans. Furthermore, the SRP remains attached for the most part to the cellular membrane when the cleavage of pre-secretory proteins is severely reduced in a strain lacking the gene coding for the major type-I signal peptidase.
Clonal Populations of Chondrocytes with Progenitor Properties Identified within Human Articular Cartilage
Authors: Thornemo M. , Tallheden T. , Sjögren Jansson E. , Larsson A. , Lövstedt K. , Nannmark U. , Brittberg M. , Lindahl A.
Published: November 2005

The aim of the present study was to identify and characterize progenitor properties of human articular chondrocytes selected by using agarose suspension culture. In this chondrogenic selective culture condition, about 3.6% of seeded surplus chondrocytes from patients undergoing articular chondrocyte transplantation proliferated and formed cell clusters after 6 weeks. Phase-contrast microscopy and transmission electron microscopy revealed four different types of cell clusters differing in cellular content and matrix production. Based on their morphological features, they were named the homogenous (H), the homogenous matrix (HM), the differentiated matrix (DM) and the differentiated (D) cell clusters. All cell clusters showed positive safranin O staining, and matrix was positive for antibodies detecting type II collagen and aggrecan. The clusters were further demonstrated to express the genes for fibroblast growth factor receptor 3, type IIA collagen and type IIB collagen, while type X collagen was not expressed. After subcloning, the H and HM clusters demonstrated the best proliferative capacity. Chondrocytes from these two cell clusters also showed phenotypic plasticity in chondrogenic, adipogenic as well as osteogenic assays. This study demonstrates that existing subpopulations of cells with chondroprogenitor properties can be isolated from human adult articular cartilage using agarose suspension cultures.
Rhodopsin, Violet and Blue Opsin Expressions in the Chick Are Highly Dependent on Tissue and Serum Conditions
Authors: Jacob Vanessa , Rothermel Andrée , Wolf Peter , Layer Paul G.
Published: November 2005

The molecular, cellular or tissue environment can influence the expression of genes and thereby regulate processes of tissue formation. Here we determined the tissue and serum dependence of the expression of all photopigments in the chick by a series of distinct retinal cell cultures, analyzed by RT-PCR using specific primers for all four opsins and rhodopsin followed by quantitative scanning of the respective gel bands. For comparison, we first determined expression of all opsins during normal chick retinogenesis, which began with red and violet opsins at E12, shortly followed by blue and green opsins and finally rhodopsin at E14. This period corresponds to the time of synaptogenesis in the inner retina. All cultures were started with 6-day-old dissociated retinal cells. Cells were kept at low or high cell density (called LoDens or HiDens), or they were reaggregated as retinal spheres, whereby all of them were raised at low (2%) or high serum (12%) levels (called LoSer or HiSer). In LoDens/HiSer cultures, expression of all opsins was weak. At HiDens/LoSer red and green opsin expression was strong, while rhodopsin and violet/blue remained low. In HiDens/HiSer cultures the expression of red and green was strong; rhodopsin was almost normal, while violet and green were low. In reaggregates at high serum the expression came closest to a normal retina, but violet and blue opsins were still below normal. At low serum, however, violet and blue were negligible and rhodopsin was low. This in vitro study shows that rhodopsin, followed by violet and blue opsin expressions is highly dependent on serum, cell density and tissue conditions, while red and green opsins are more autonomous.
Age-Related Morphological Changes in the Human Hyoid Bone
Authors: Shimizu Yoshinaka , Kanetaka Hiroyasu , Kim Yeon-Hwan , Okayama Keisuke , Kano Mitsuhiro , Kikuchi Masayoshi
Published: November 2005

Object: Age-related morphological changes in the human hyoid bone were investigated radiographically and histologically. Materials and Methods: Thirty-two measurements were performed on radiographs of 238 hyoid bones from autopsy cases of known age and sex. Thirty-one hyoid bones that were studied by radiography were also examined histologically in horizontal sections. Results: Analysis of the length and width of the hyoid bone revealed significant increases in the body and the anterior part of the greater cornu and a significant decrease in the posterior part of the greater cornu with aging. Most measurements of the body and the greater cornu revealed differences between male and female hyoid bones. The outer margins of the body and the greater cornu were situated further outside in older males compared with females. The breadth of the joint space showed a significant age-related decrease, and the degree of fusion showed a significant age-related increase. Histological findings showed ossified or calcified fusion, with osteoclasts in the marginal area of the joint space. Conclusions: Increasing age induces fusion of the body and the greater cornu. The morphometric changes in the shape of the hyoid bone may represent functional adaptation to articulation fixation.
The Current Status of Gene Therapy in Autologous Transplantation
Authors: Becker Pamela S.
Published: November 2005

Autologous hematopoietic cells have been used as targets of gene transfer, with applications in inherited disorders, cell therapy, and acquired immunodeficiency. The types of cells include hematopoietic progenitor cells, lymphocytes, and mesenchymal stem cells. The inherited disorders thus far approached in clinical trials include severe combined immunodeficiency, common variable γ-chain immunodeficiency, chronic granulomatous disease, and Gaucher disease. Preclinical studies are vigorously under way in thalassemia, sickle cell anemia, Wiskott-Aldrich syndrome and Fanconi anemia. Clinical trials of immunological therapy with gene-modified lymphocytes are under study in the treatment of malignancies. Clinical trials using anti-viral strategies for HIV infection in combination with autologous transplantation have begun, with additional approaches being developed. Gene therapy vectors are being developed to eliminate tumor cells contaminating autologous stem cell products. However, the risk of insertional mutagenesis and the potential for development of leukemia was highlighted by the first gene therapy trials in inherited immunodeficiency syndromes that achieved a therapeutic effect. Despite the slow progress of the field to date, there is extraordinary promise for gene therapy in the future.