bluuebunny/crossref_metadata_2025 huggingface dataset:
Social deprivation and traumatic dental injuries among 14‐year‐old schoolchildren in Newham, London
Authors: Marcenes W. , Murray S.
Published: March 2003

Abstract – Aims: To assess the epidemiology of dental injuries among schoolchildren in Newham, London. Material and methods: This study was carried out as part of the annual Community Dental Service’s School Dental Inspection (SDI) programme in Newham. All schoolchildren aged 14 years were invited to participate in the study. Dental examinations were carried out by one trained examiner (SM) and included presence of clinical evidence of dental injuries, treatment provided and needed in the upper and lower permanent incisors, size of incisal overjet and type of lip coverage. The postcode was recorded to assess the levels of social deprivation. Findings: 2242 out of 2684 children were examined. The prevalence of dental injuries was 23.7%. Treatment of dental injuries was neglected. Being male, having an incisal overjet greater than 5 mm and living in an overcrowded household increased the risk of having dental injuries. Conclusions: The prevalence of dental injuries in Newham (23.7%), which is a deprived area of London, was much higher than the overall prevalence in the UK (17%). Traumatic dental injuries seem to be a serious dental public health problem among children in deprived areas. Thus, there is an urgent need to collect local data on dental injuries in order to obtain a more comprehensive picture of the dental health.
Comparison of a new dental trauma splint device (TTS) with three commonly used splinting techniques
Authors: Von Arx Thomas , Filippi Andreas , Lussi Adrian
Published: March 2003

Abstract – Splinting is the standard of care for stabilization of replanted or repositioned permanent teeth following trauma. The present experimental study compared four dental trauma splints in 10 volunteers. The evaluated splints included a wire‐composite splint (WCS), a button‐bracket splint (BS), a resin splint (RS), and a new device (TTS=Titanium Trauma Splint) specifically developed for splinting traumatized teeth. All splints were bonded to the labial surfaces of the maxillary lateral and central incisors. Splints were left in place for 1 week. After splint removal, the next splint was placed after a 1‐week rest period. The sequence of splint application was randomized for each individual. The following parameters were assessed: tooth mobility with horizontal and vertical Periotest values (PTV) before and after splint application and splint removal, respectively; probing depths, plaque and bleeding on probing indices before splint application and removal, and chair time needed for splint application and removal. After splint application, horizontal PTV were significantly lower in central incisors for BS compared to TTS (P=0.04), and for RS compared to TTS (P=0.005) and to WCS (P=0.006). Reduction of lateral tooth mobility (=splint effect) expressed by the difference between horizontal pre‐ and postoperative PTV was significantly greater in RS compared to TTS and WCS (P<0.05) for central as well as for lateral incisors. However, changes of vertical tooth mobility were not significant across the splinting techniques. Periodontal parameters remained unchanged, reflecting the excellent oral hygiene by the study subjects. The chair time needed for splint application was significantly shorter for TTS (P<0.01). In conclusion, all tested splints appeared to maintain physiologic vertical and horizontal tooth mobility. However, the latter was critically reduced in RS splints.
Internal root resorption studied by radiography, stereomicroscope, scanning electron microscope and computerized 3D reconstructive method
Authors: Lyroudia Kleoniki M. , Dourou Vasiliki I. , Pantelidou Ourania CH. , Labrianidis Theodoros , Pitas Ioannis K.
Published: March 2003

Abstract – Aim and methology: Two cases of internal tooth resorption were examined. A mandibular premolar and a mandibular canine were studied after they were extracted using radiographs, a stereomicroscope (SM) and a scanning electron microscope (SEM). Lastly, 3D images of the sectioned teeth were obtained(3D).

Results: Radiographically, internal root resorption was shown as a uniform radiolucency. By SM examination, an extensive destruction of dentin was seen, while, by SEM examination, a disappearance of dentinal tubules was clear. The 3D reconstructive method revealed a circumscribed, oval‐shaped defect that did not perforate the cemental layer.

Conclusions: Internal root resorption is a rare remodeling process that can be studied using different experimental methods.
Delayed removal of a fully intruded primary incisor through the nasal cavity: a case report
Authors: Thor Andreas L. I.
Published: March 2003

Abstract – Complete intrusion of a primary anterior tooth may initially be diagnosed as an avulsion injury. The importance of a correct first clinical and radiological examination of a young patient who has sustained trauma to the anterior primary teeth is stressed in this article. This issue is illustrated by a case presentation where a 3‐year‐old girl had a delayed treatment of 5 days after a primary incisor was fully impacted by a fall trauma into the nasal cavity. This was due to an incomprehensible clinical and radiological examination by the first examining dentist. The impacted tooth was later removed through the right nostril under general anesthesia. Two‐year follow‐up showed no adverse clinical or radiological problems even though eruption of the permanent successor had not taken place at that time.
The effect of fish oil on bone resorption following pulp exposure in rats
Authors: Indahyani Didin E. , Pudyani Pinandi S. , Santoso AL. Supartinah , Jonarta Alma L. , Sosroseno Wihaskoro
Published: March 2003

Abstract  – The aim of this study was to determine whether treatment with fish oil may alter the periapical bone resorption following pulp exposure in rats. Untreated and pulp‐exposed animals served as the negative and positive control, respectively. Other pulp‐exposed animals were orally treated with fish oil at different concentrations and frequencies. Periapical tissue sections were stained with tartrate‐resistant acid phosphatase (TRAP), and then the numbers of both periapical osteoclasts and preosteoclasts were determined. The levels of bone resorption were assessed using the osteoclast–bone interface (OBI) index. The results showed that no differences between the numbers of both osteoclasts and preosteoclasts in the fish oil‐treated animals and the negative control at day 14 could be found. Similarly, the periapical bone resorption in the animals treated with fish oil for 14 days significantly reduced to the levels of that in the negative control. The results of the present study, therefore, suggest that oral treatment with fish oil may inhibit bone resorption following pulp exposure in rats and hence, may have a therapeutic modality for inflamed periradicular tissue.
Fractured dens evaginatus and unusual periapical radiolucency
Authors: Chu Frederick C. S. , Sham Arthur S. K. , Yip Kevin H. K.
Published: March 2003

Abstract   –  Dens evaginatus (DE) is not uncommon. It can affect the premolars of people of oriental ethnicity, and it can lead to different clinical problems. This article describes the diagnosis and management of a patient presenting with a fractured tubercle of DE and an unusual buccal abscess in relation to periapical radiolucency extending around the mandibular second premolar and first molar.
Assessment of post‐traumatic PDL cells viability by a novel collagenase assay
Authors: Pileggi Roberta , Dumsha Thomas C. , Nor Jacques E.
Published: March 2003

Abstract – Both length of extra‐alveolar time and type of storage media are significant factors that can affect the long‐term prognosis for replanted teeth. Numerous studies have examined various media in an attempt to determine the ideal material for storage of the avulsed tooth. The purpose of this study was to compare the number of viable periodontium ligament (PDL) cells in different storage media using a collagenase assay. Thirty‐three freshly extracted human teeth were divided into four experimental and two control groups. The positive and negative controls corresponded to 0 min and an 8‐h dry time, respectively. The experimental teeth were stored dry for 30 min and then immersed in one of four media (Hank's balanced salt solution (HBSS), milk, saline, water) for 45 min. The teeth were then treated with dispase grade II and collagenase for 30 min. The number of viable and nonviable PDL cells was counted with a hemocytometer and analyzed. An anova demonstrated no statistically significant differences in the viability of PDL cells among saline, HBSS and milk. Within the parameters of this study, it appears that milk or saline is an equally viable alternative to HBSS for storage of avulsed teeth.
Traumatic injuries in the primary dentition
Authors: Flores Marie Therese
Published: March 2003

Abstract   –  As a support for the guidelines published in 2001, a review of the literature was carried out using the evidence‐based approach in order to update the state of the art regarding epidemiology and treatment of traumatic dental injuries in the primary dentition. An online search in Medline, and a review of expert literature, lead to the conclusion that most luxation injuries heal spontaneously, and unless more conclusive evidence is available, conservative treatment of acute dental trauma for preschool children is suggested. Pain control, the dentist's ability to cope with the child's anxiety, and follow‐up instructions in oral hygiene techniques will allow many primary teeth that are currently lost at the time of injury to be saved.
The use of MTA in teeth with necrotic pulps and open apices1
Authors: Giuliani Valentina , Baccetti Tiziano , Pace Riccardo , Pagavino Gabriella
Published: March 2003

Abstract  – Three clinical cases have been treated with the use of an apical plug of MTA for apexification. All three cases were central incisors that had suffered premature interruption of root development as a consequence of trauma. According to the treatment protocol, the root canals were rinsed with 5% NaOCl; then, calcium hydroxide paste was placed in the canals for 1 week. Consequently, the apical portion of the canal (4 mm) was filled with MTA. The remaining portion of the root canals was then closed with thermoplastic gutta‐percha. At 6‐month and 1‐year follow‐up period the clinical and radiographic appearance of the teeth showed resolution of the periapical lesions. MTA appears to be a valid option for apexification with its main advantage being the speed at which the treatment can be completed.
Age‐related changes in the human pulpal blood flow measured by laser Doppler flowmetry
Authors: Ikawa Motohide , Komatsu Hideji , Ikawa Kyoko , Mayanagi Hideaki , Shimauchi Hidetoshi
Published: March 2003

Abstract  – The purpose of this study was to examine the age‐related changes in human pulpal blood flow (PBF). Recordings were taken from 22 clinically healthy upper central incisors in 22 healthy participants (age: 8–75 years). A Moor blood flow monitor (type MBF3D) was used to measure PBF; and subjects' electrocardiogram (ECG), mean blood pressure (BP) and heart rate (HR) were also recorded. The resting PBF was significantly decreased with the increased age of the participants (Pearson's correlation coefficient, P < 0.001). The examined tooth crown was briefly(1 s) cooled using a dental coolant, and significant reductions in PBF were induced (one‐way repeated measures anova, P < 0.05). The magnitude of the reduction (%) was significantly decreased with the increased age of the participants (Pearson's correlation coefficient, P < 0.05). The findings indicate that the hemodynamics in the human pulp is reduced with age.
Remifentanil and the tunnelling phase of paediatric ventriculoperitoneal shunt insertion. A double‐blind, randomised, prospective study
Authors: Chambers N. , Lopez T. , Thomas J. , James M. F. M.
Published: March 2003

Summary Sixty‐two children were randomly allocated to receive, during inhalational anaesthesia with isoflurane and nitrous oxide, either 1.0 µg.kg−1 remifentanil (n = 33) or saline (n = 29) just before the tunnelling phase of ventriculoperitoneal shunt insertion, in a double‐blind study. The remifentanil group showed little stress response to tunnelling as indicated by median (interquartile range [range]) change in heart rate −5.2 (−11.4 to 9.8 [−19.4 to 30.4])%, mean arterial pressure −5.0 (−20.8 to 15.5 [−40.9 to 42.9])% or plasma norepinephrine −13.5 (−38.1 to −2.5 [−77.7 to 81.5])% compared with the saline group, in which the changes were 20.1 (11.5–36.1 [2.1–83.1])%, 42.7 (27.1–56.8 [3.2–73.5])% and 13.3 (0.8–70.0 [−45.2 to 337.5])%, respectively (p < 0.001 for all comparisons). These changes were consistent across most different age categories. The cardiovascular response in the saline group lasted for 8 (4–15 [0–39]) min. Tracheal extubation occurred after 3 (2–4 [1–8]) min in the remifentanil group and 3 (2–6␣[0–15]) min in the saline group (p = 0.29), with transfer to the recovery area and discharge to␣the ward, respectively, 4 (4–5 [1–10]) min and 9 (7–13 [2–32]) min in the remifentanil group and 7␣(4–8 [2–18]) min and 14 (10–19 [7–44]) min in the saline group (p = 0.06 and 0.01, respectively). Postoperatively there was some evidence of respiratory depression and increased oxygen requirements in all age categories, but this was similar in both groups. Overall, the maximum increase from baseline in transcutaneous carbon dioxide tension was 41.2 (11.3–66.7 [−2.0 to 141.7])% in the remifentanil group compared with 30.7 (20.5–55.1 [1.7–159])% in the saline group (p = 0.8), and the time taken for transcutaneous carbon dioxide tension to decrease to < 6.0 kPa was 4 (0–13 [0–60]) min compared with 7 (0–13 [0–60]) min, respectively (p = 0.75). There was no difference between the two groups in postoperative analgesic requirements or in blood loss and there were no significant side‐effects. We conclude that remifentanil is an appropriate and safe analgesic to provide balanced␣anaesthesia to cover the tunnelling phase of paediatric ventriculoperitoneal shunt insertion.
Orotracheal fibreoptic intubation for rapid sequence induction of anaesthesia*
Authors: Pandit J. J. , Dravid R. M. , Iyer R. , Popat M. T.
Published: March 2003

Summary We assessed whether flexible fibreoptic‐guided orotracheal intubation could be rapidly and successfully achieved during a simulated rapid sequence induction in 30 anaesthetised and paralysed patients. Rapid sequence induction was simulated by applying practised cricoid pressure. Using a␣flexible fibreoptic laryngoscope with camera and closed circuit television, an anaesthetist experienced with the technique performed orotracheal endoscopy and intubation with a cuffed 7.0‐mm Portex tracheal tube through a VBM Bronchoscope Airway. Fibreoptic intubation was successful at the first attempt in 28 patients (93%); two patients required two attempts. Mean (SD) time from removal of the facemask from the patient's face to the appearance of carbon dioxide in the expired breath after intubation was 111 (46) s (median 100 s; range 54–195 s). There were one or more difficulties in 13 patients (43%). These difficulties were largely avoidable and included problems with fibreoptic equipment, the Bronchoscope Airway, copious secretions, cricoid pressure or railroading of the tracheal tube. Flexible fibreoptic‐guided orotracheal intubation may have a place in the management of failed intubation during a rapid sequence induction.
Retracted: The effect of ulinastatin pre‐treatment on vecuronium‐induced neuromuscular block in patients with hepatic cirrhosis
Authors: Saitoh Y. , Kaneda K. , Murakawa M.
Published: March 2003

The aim of this study was to investigate the effect of ulinastatin, a protease inhibitor, on the neuromuscular block produced by vecuronium in patients with hepatic cirrhosis. Thirty adult patients with hepatic cirrhosis were randomly allocated to receive ulinastatin (cirrhosis/ulinastatin group, n = 15) or saline (cirrhosis/saline group, n = 15). Fifteen healthy adult patients without hepatic cirrhosis comprised a control group. Patients were given a standardised anaesthetic that␣included nitrous oxide and isoflurane in oxygen, and fentanyl. A bolus dose of ulinastatin 5000 unit.kg−1 was given to members of the cirrhosis/ulinastatin group. The same volume of normal saline was given to the other two groups. Two minutes later, vecuronium 0.1 mg.kg−1 was given. The onset of neuromuscular block was significantly slower in the cirrhosis/ulinastatin group than in the cirrhosis/saline and control groups (p < 0.05). Spontaneous recovery of neuromuscular function was significantly quicker in the cirrhosis/ulinastatin and control groups than in the cirrhosis/saline group (p < 0.05). The time course of recovery in the cirrhosis/ulinastatin and control groups was similar. We conclude that in cirrhotic patients, ulinastatin delays the onset of neuromuscular block produced by vecuronium. After pretreatment with ulinastatin, the speed of recovery from neuromuscular block in patients with cirrhosis becomes similar to that seen in healthy patients.
Severe hyperphosphataemia and associated electrolyte and metabolic derangement following the administration of sodium phosphate for bowel preparation
Authors: Tan H. L. , Liew Q. Y. , Loo S. , Hawkins R.
Published: March 2003

Sodium phosphate is widely used as an effective bowel preparation agent. It is used in smaller volumes, leading to improved patient tolerance. Although it is generally safe, cases of severe hyperphosphataemia following sodium phosphate administration have been reported in the literature. The common risk factors identified are advanced age, impaired renal function, impaired colonic motility and multiple doses. However, many doctors remain unaware of the dangers associated with this agent. We report six cases of severe electrolyte and metabolic derangement due to sodium phosphate bowel preparation: two patients had delayed awakening from general anaesthesia, and four patients suffered life‐threatening consequences.
Negative selection of peripheral blood stem cells  to support a tandem autologous transplantation  programme in multiple myeloma
Authors: Barbui Anna Maria , Galli Monica , Dotti Gianpietro , Belli Nadia , Borleri Gianmaria , Gritti Giovanna , Bellavita Piermario , Viero Piera , Comotti Benedetto , Barbui Tiziano , Rambaldi Alessandro
Published: March 2003

Summary.We recently described a two‐step negative selection procedure whereby peripheral blood stem cells (PBSCs) were efficiently purged of contaminating neoplastic cells by a combination of monoclonal antibodies. Here, we report 60 newly diagnosed multiple myeloma (MM) patients treated with a double transplant programme and randomized to receive either unmanipulated orin vitropurged PBSCs. We demonstrated that this technique is feasible and safe without significant loss of either CD34+ or CD3+ cells. Haematological engraftment and immunological reconstitution were rapid without treatment‐related mortality. Using polymerase chain reaction (PCR), we compared the level of minimal residual disease (MRD) in PBSC before and afterin vitropurging andin vivoafter transplant. A median of one tumour cell per 102normal cells (range 101−105) was seen in the unmanipulated aphereses with a 3–4 log reduction after manipulationin vitro. However, despite this tumour debulking, all patients remained PCR positivein vivo.At 3 years, the estimated event‐free survival was 40% in the control arm and 72% in the experimental arm (P = 0·05), whereas the estimated overall survival was 83% in both arms. This suggests that autologous transplantation using efficiently purged PBSCs can be performed safely, but confirms the need for innovative protocols for MRD eradicationin vivo.
Transfusion‐related acute lung injury caused by human leucocyte antigen class II antibody
Authors: Flesch Brigitte K. , Neppert Jürgen
Published: March 2003

Summary.  Fcγ receptor (FcγR)‐mediated destruction of immunoglobulin‐coated red blood cells (RBCs) is the underlying mechanism of haemolytic disease of the newborn. Human leucocyte antigen (HLA) antibodies in vitro are able to block monocyte FcγRs and prevent phagocytosis. The intention was to demonstrate this effect in vivo upon a volunteer. Plasma containing a non‐cytotoxic HLA‐DR alloantibody was infused into the subject. The FcγR blockade was achieved and persisted for about 2·5 d, but, unexpectedly, a mild transfusion‐related acute lung injury (TRALI) was also caused. Monocytes disappeared completely from the peripheral blood within the first hour after infusion and a mild pulmonary oedema was observed within 3–4 h. The subject recovered within 2 d.
Hypermethylation of the P15INK4b and P16INK4a in agnogenic myeloid metaplasia (AMM) and AMM in leukaemic transformation
Authors: Wang Jen C. , Chen Wilson , Nallusamy Selvarany , Chen Chi , Novetsky Allan D.
Published: March 2003

Summary. Hypermethylation of p15 and p16 genes was determined in 32 patients with agnogenic myeloid metaplasia(AMM), also known as idiopathic myelofibrosis (MF). These included 10 patients in leukaemic transformation phase. Using polymerase chain reaction‐based methylation analysis assay methods, with substantiation using Southern blot analysis, the study showed no hypermethylation of p15 or p16 genes in the chronic phase of AMM, but p15 gene hypermethylation was found in four patients (40%) and p16 gene hypermethylation in two patients (20%) when they were in leukaemic transformation stage. Furthermore, two of the patients in leukaemic transformation were found to have both p15 and p16 gene hypermethylation, demonstrating possible multiple gene hypermethylation in the same patient. Thus, hypomethylation agents for treating patients with AMM in leukaemic transformation may be appropriate for future trials.
Interleukin 10, produced in abundance by human newborn T cells, may be the regulator of increased tolerance associated with cord blood stem cell transplantation
Authors: Rainsford Eleanor , Reen Denis J.
Published: March 2003

Summary. The use of human umbilical cord blood as an alternative source of stem cells to bone marrow for the reconstitution of the immune system is associated with less frequent and less severe incidence of graft‐versus‐host disease (GVHD). This study focuses on aspects of cord blood T‐cell biology that may contribute to a perceived increased tolerance associated with the neonatal immune response. A skewing of the T‐helper (Th)1/Th2 phenotype of cord blood T cells towards a Th2 response has frequently been cited as a possible cause. In this study, primary and repeated stimulation via the T‐cell receptor (TCR) complex induced a Th0‐type cytokine response, with both adult and cord blood‐derived naïve T cells producing interferon γ (IFN‐γ), interleukin 4 (IL‐4) and IL‐5. IL‐10 was induced in cord blood T‐cell cultures during primary stimulation, while adult T cells began to secrete IL‐10 only after repeated stimulation. The presence of the antigen‐presenting cell (APC)‐derived cytokine IL‐1β inhibited IL‐10 production by cord blood cells. The effects of IL‐12 and IL‐4 on T‐cell cytokine responses were also examined. In addition to their differential Th1/Th2 skewing effects on cord and adult T cells, both cytokines augmented IL‐10 production in both T‐cell populations. These findings demonstrate that cord blood T cells may secrete large amounts of the anti‐inflammatory cytokine IL‐10 and that the presence of IL‐1β or Th1/Th2 skewing cytokines can regulate its production. This data provides support for the recognized tolerant nature of the newborn immune response that may contribute to the reduced incidence of GVHD associated with cord blood transplantation.
Melphalan treatment in patients with myelofibrosis with myeloid metaplasia
Authors: Petti M. C. , Latagliata R. , Spadea T. , Spadea A. , Montefusco E. , Aloe Spiriti M. A. , Avvisati G. , Breccia M. , Pescarmona E. , Mandelli F.
Published: March 2003

Summary.Between January 1985 and December 1992, 104 consecutive patients with symptomatic myelofibrosis with myeloid metaplasia (MMM) [splenic enlargement > 5 cm and/or transfusional requirement or Hb < 10 g/dl and/or white blood cell (WBC) count > 20 × 109/l and/or platelets > 1·0 × 109/l] received low‐dose Melphalan (2·5 mg/3 times/week) to evaluate the efficacy and toxicity of this approach. Among 99 evaluable patients, 66 (66·7%) achieved a response after a median time of 6·7 months: 26 (26·3%) had a normalization of all clinical and haematological parameters (complete response, CR) and 40 (40·4%) showed an improvement > 50% (partial response, PR). Thirty‐three patients (33·3%) were resistant. Reversible haematological toxicity was the most common complication. Median durations of CR and PR were 28·4 and 26 months respectively: median survival of CR + PR patients was 71·2 months (95%CI: 33·8–108·7) versus 36·5 months (95%CI: 24·5–48·5) for the non‐responders (log‐rank test,P= 0·002). In the multivariate analysis, the following variables were significantly associated with a shorter survival: anaemia [hazard risk (HR) = 2·7], WBC count > 20 × 109/l (HR = 2·4) and not achieving any type of response, either partial or complete (HR = 3·9). In conclusion, Melphalan could be a promising first‐line option for MMM patients with clinical or haematological symptoms requiring treatment.
Clinical evaluation of a polymerase chain reaction assay to detect Aspergillus species in bronchoalveolar lavage samples of neutropenic patients
Authors: Buchheidt Dieter , Baust Corinna , Skladny Heyko , Baldus Michael , Bräuninger Susanne , Hehlmann Rüdiger
Published: March 2003

Summary. The increasing incidence of invasive aspergillosis, a life‐threatening infection in immunocompromised patients, emphasizes the need to improve the currently limited diagnostic tools. Using a recently developed two‐step polymerase chain reaction (PCR) assay to detect 10 fg of Aspergillus DNA, corresponding to 1–5 colony‐forming units (CFU)/ml of spiked samples in vitro, we prospectively examined 197 bronchoalveolar lavage (BAL) samples from 176 subjects, including 141 neutropenic, febrile patients with lung infiltrates, at risk for invasive fungal disease. Underlying diseases of these patients were haematological malignancies; 93 patients suffered from acute leukaemias. Thirty‐one of these immunocompromised patients (17·6%) were PCR positive, correlating with positive BAL culture, positive histology from lung surgery or from autopsy, positive computerized tomography scans or positive galactomannan enzyme‐linked immunosorbent assay. Six patients (4·3%) of this group had positive PCR results without any correlation to clinical or other diagnostic data, probably owing to contamination of the samples by ubiquitous Aspergillus spores. The samples of two patients (1·4%) with a subsequent histologically proven mould infection were PCR negative. All 102 immunocompromised patients (72·3%) with a negative PCR showed no evidence of invasive fungal disease. From 35 patients without immunodeficiency, four (11·4%) showed positive results, without evidence of invasive or non‐invasive pulmonary aspergillosis. In this haematological population, the sensitivity and specificity values of the test reached 93·9% and 94·4%, the positive predictive value 83·8%, the negative predictive value 98·1%. Our data support the considerable clinical value of this PCR assay for confirming and improving diagnosis of pulmonary aspergillosis in high‐risk patients.
Botulinum toxin A in the treatment of patients with Frey syndrome
Authors: Beerens A J F , Snow G B
Published: March 2003

Background

This was a prospective study of the treatment of Frey syndrome, also known as gustatory sweating, with botulinum toxin A.

Methods

Thirteen patients with a mean involved skin area of 53 (range 36–80) cm2, as assessed with the Minor starch–iodine test, were treated with 0·1 ml toxin (75 units/ml) injected intracutaneously into every 4 cm2 of involved skin. The mean total dose was 100 (range 67·5–150) units. Treatment results were assessed every 3 months with the Minor test. The Frey Questionnaire Card (FQC) was used for subjective assessment. The mean follow-up after primary treatment was 20 (range 9–24) months. Treatment was repeated if the symptoms recurred.

Results

After 3 months 11 of the 13 patients showed a decrease of gustatory sweating of more than 90 per cent. All but one patient with a follow-up of 2 years suffered recurrent gustatory sweating. The mean recurrence-free period after primary treatment was 11 months and that after secondary treatment was 15 months. FQC score and objective assessment correlated well. Treatments were well tolerated, although two patients developed a temporary perioral muscle paresis.

Conclusion

Botulinum toxin A produces good results in the treatment of Frey syndrome. Repeated treatment improves on the results of primary treatment.
Ultrasonography is superior to plain radiography in the diagnosis of pneumoperitoneum
Authors: Chen S-C , Yen Z-S , Wang H-P , Lin F-Y , Hsu C-Y , Chen W-J
Published: March 2003

Background

The aim of this study was to compare plain radiography with abdominal ultrasonography in the detection of pneumoperitoneum.

Methods

A total of 188 patients with suspected hollow organ perforation were studied. All patients had abdominal ultrasonography, upright chest radiography and left lateral decubitus abdominal radiography examinations. The sensitivity, specificity, positive and negative predictive value, and accuracy of chest and abdominal radiography were compared with that of abdominal ultrasonography.

Results

One hundred and seventy-eight patients underwent laparotomy; 170 patients had hollow organ perforation, five patients had perforated appendicitis and three had acute cholecystitis. In the diagnosis of pneumoperitoneum, ultrasonography had improved sensitivity (92 versus 78 per cent), negative predictive value (39 versus 20 per cent) and accuracy (88 versus 76 per cent), and similar specificity (both 53 per cent) and positive predictive value (95 versus 94 per cent) compared with plain radiography.

Conclusion

Ultrasonography is more sensitive than plain radiography in the diagnosis of pneumoperitoneum.
The AUG start codon of the Saccharomyces cerevisiae NFS1 gene can be substituted for by UUG without increased initiation of translation at downstream codons
Authors: Nett Jürgen H. , Kessl Jacques , Wenz Tina , Trumpower Bernard L.
Published: March 2003

The selection of the site for initiation of translation for the Saccharomyces cerevisiae NFS1 gene was examined using mutated AUG1, AUG2 and AUG3 codons. When AUG1 of the yeast NFS1 gene was mutated to UUG and the resulting mRNA was translated in vitro using a reticulocyte system, initiation from the mutated codon was abolished and occurred instead at downstream codons at increased rates. When the same mRNA was translated using a yeast extract, translation initiated at the mutated codon, albeit at a reduced rate, and there was no increased translation at downstream AUG codons. The NFS1 gene in which AUG1 was replaced by UUG was also able to substitute for the wild‐type gene in vivo in yeast. Western blots confirmed that the encoded protein was the same size as that encoded by the wild‐type gene and that both the wild‐type and mutated proteins localized to mitochondria. This is apparently the first example of a yeast protein where mutagenesis of AUG1 does not lead to alternate use of a downstream AUG.
Molecular dynamics simulations of solvated UDP–glucose in interaction with Mg2+ cations
Authors: Petrová Pavla , Koča Jaroslav , Imberty Anne
Published: March 2003

Glycosyltransferases are key enzymes involved in biosynthesis of oligosaccharides. Nucleotide‐sugars, the glycosyltransferase substrates, serve as activated donors of sugar residues during the enzymatic reaction Although very little is known about the catalytic mechanism of these enzymes, it appears that the catalytic activity in most glycosyltransferases is dependent upon the presence of a divalent cation, for example Mn2+ or Mg2+. It is not known whether the ion is bound to the enzyme before its interaction with the substrate, or if it binds the substrate before the enzymatic reaction to modify its conformation to fit better the active site of the enzyme. We have inspected the latter possibility by running four 2‐ns molecular dynamics trajectories on fully solvated UDP‐glucose in the presence of Mg2+ ions. Our results indicate that the divalent cation interacts strongly with the nucleotide‐sugar in solution, and that it can alter its conformational behavior. It is also shown that a conformation of the pyrophosphate moiety that results in an eclipsed or almost eclipsed orientation of two of the oxygen atoms, and which is found in protein interacting with a nucleotide di‐ or tri‐phosphate X‐ray data, is energetically favored. The results are also discussed in light of existing NMR data, and are found to be in a good agreement with them.
Supermolecular organization of photosystem II and its associated light‐harvesting antenna in Arabidopsis thaliana
Authors: Yakushevska Alevtyna E. , Jensen Poul E. , Keegstra Wilko , van Roon Henny , Scheller Henrik V. , Boekema Egbert J. , Dekker Jan P.
Published: March 2003

The organization of Arabidopsis thaliana photosystem II (PSII) and its associated light‐harvesting antenna (LHCII) was studied in isolated PSII–LHCII supercomplexes and native membrane‐bound crystals by transmission electron microscopy and image analysis. Over 4000 single‐particle projections of PSII–LHCII supercomplexes were analyzed. In comparison to spinach supercomplexes [Boekema, E.J., van Roon, H., van Breemen, J.F.L. & Dekker, J.P. (1999) Eur. J. Biochem. 266, 444–452] some striking differences were revealed: a much larger number of supercomplexes from Arabidopsis contain copies of M‐type LHCII trimers. M‐type trimers can also bind in the absence of the more common S‐type trimers. No binding of l‐type trimers could be detected. Analysis of native membrane‐bound PSII crystals revealed a novel type of crystal with a unit cell of 25.6 × 21.4 nm (angle 77°), which is larger than any of the PSII lattices observed before. The data show that the unit cell is built up from C2S2M2 supercomplexes, rather than from C2S2M supercomplexes observed in native membrane crystals from spinach [Boekema, E.J., Van Breemen, J.F.L., Van Roon, H. & Dekker, J.P. (2000) J. Mol. Biol.301, 1123–1133]. It is concluded from both the single particle analysis and the crystal analysis that the M‐type trimers bind more strongly to PSII core complexes in Arabidopsis than in spinach.
Characterization of a major form of human isatin reductase and the reduced metabolite
Authors: Usami Noriyuki , Kitahara Kouei , Ishikura Shuhei , Nagano Makoto , Sakai Syunsuke , Hara Akira
Published: March 2003

Isatin, an endogenous indole, has been shown to inhibit monoamine oxidase, and exhibit various pharmacological actions. However, the metabolism of isatin in humans remains unknown. We have found high isatin reductase activity in the 105 000 g supernatants of human liver and kidney homogenates, and have purified and characterized a major form of the enzyme in the two tissues. The hepatic and renal enzymes showed the same properties, including an Mr of 31 kDa, substrate specificity for carbonyl compounds and inhibitor sensitivity, which were also identical to those of recombinant human carbonyl reductase. The identity of the isatin reductase with carbonyl reductase was immunologically demonstrated with an antibody against the recombinant carbonyl reductase. About 90% of the soluble isatin reductase activity in the liver and kidney was immunoprecipitated by the antibody. The Km (10 µm) and kcat/Km (1.7 s−1·µm−1) values for isatin at pH 7.0 were comparable to those for phenanthrenequinone, the best xenobiotic substrate of carbonyl reductase. The reduced product of isatin was chemically identified with 3‐hydroxy‐2‐oxoindole, which is also excreted in human urine. The inhibitory potency of the reduced product for monoamine oxidase A and B was significantly lower than that of isatin. The results indicate that the novel metabolic pathway of isatin in humans is mediated mainly by carbonyl reductase, which may play a critical role in controlling the biological activity of isatin.
Antiproliferative effect of nitric oxide on rat glomerular mesangial cells via inhibition of mitogen‐activated protein kinase
Authors: Chin Ting‐Yu , Lin Yin‐Shan , Chueh Sheau‐Huei
Published: March 2003

The effect of nitric oxide (NO) donors and lipopolysaccharide (LPS) on the proliferation of rat glomerular mesangial cells was characterized. Exogenous application of a NO donor inhibited serum‐induced proliferation in a time‐ and dose‐dependent manner. S‐Nitrosoglutathione (GSNO) also increased cGMP generation and arachidonic acid release, but it did not cause any measurable increase in the cytosolic Ca2+ concentration. Chelation of cytosolic Ca2+ or inhibition of mitogen‐activated protein kinase (MAPK) kinase had an inhibitory effect on proliferation, but neither enhanced the antiproliferative effect of GSNO. In contrast, inhibition of guanylate cyclase or phospholipase A2 had no effect on proliferation, but partially reversed GSNO‐induced antiproliferation by ≈ 98 and 65%, respectively. GSNO did not cause cell death. Incubation of cells with LPS induced endogenous NO generation and had an antiproliferative effect. LPS‐induced antiproliferation was reversed completely by inhibition of nitric oxide synthase and partially by inhibition of guanylate cyclase or phospholipase A2. GSNO or LPS inhibited serum‐induced MAPK activation, and both effects were partially reversed by inhibition of guanylate cyclase or phospholipase A2. Inclusion of 8‐bromo‐cGMP or arachidonic acid in the growth medium resulted in a similar antiproliferative effect. In conclusion, in rat glomerular mesangial cells, MAPK inhibition and an antiproliferative effect could be induced by either an increase in the cellular concentration of NO or exposure of the cells to LPS. Part of the effect of NO was attributable to the increased cellular cGMP generation and arachidonic acid release.
Nucleotide binding induces global and local structural changes of myosin head in muscle fibres
Authors: Lörinczy Dénes , Belagyi Joseph
Published: March 2003

Thermal stability and internal dynamics of myosin heads in fiber bundles from rabbit psoas muscle has been studied by electron paramagnetic resonance (EPR) spectroscopy and differential scanning calorimetry (DSC). Using ADP, ATP and orthovanadate (Vi), three intermediate states of the ATP hydrolysis cycle were simulated in glycerinated muscle fibers. DSC transitions contained three overlapping endotherms in each state. Deconvolution showed that the transition temperature of 58.4 °C was almost independent of the intermediate state of myosin, while nucleotide binding shifted the melting temperatures of 54.0 and 62.3 °C, and changed the enthalpies. These changes suggest global rearrangements of the internal structure in myosin head. In the presence of ADP and ADP plus Vi, the conventional EPR spectra showed changes in the ordering of the probe molecules, suggesting local conformational and motional changes in the internal structure of myosin heads. Saturation transfer EPR measurements reported increased rotational mobility of spin labels in the presence of ATP plus orthovanadate corresponding to a weakly binding state of myosin to actin.
Identification of an element within the promoter of human selenoprotein P responsive to transforming growth factor‐β
Authors: Mostert Volker , Wolff Sandra , Dreher Ingeborg , Köhrle Josef , Abel Josef
Published: March 2003

Selenoprotein P (SeP) is a plasma protein that contains up to 10 selenocysteine residues and accounts for about 50% of total selenium in human plasma. We have previously shown that SeP expression in the human liver cell line HepG2 is inhibited by transforming growth factor (TGF)‐β1 on a transcriptional level. Smad proteins are the transcriptional mediators of TGF‐β signalling and putative Smad‐binding elements (SBE) comprising the core sequence CAGACA are present at two positions in the SeP promoter. The aim of our study was to investigate whether Smad molecules are involved in inhibition of SeP expression by TGF‐β1 and to locate the promoter region critical for this effect. As seen in electrophoretic‐mobility‐shift assays, TGF‐β1 treatment led to enhanced binding of nuclear proteins to a putative SBE from the SeP promoter. Overexpression of Smad 3 and 4, but not of Smad 2, resulted in a marked down‐regulation of SeP mRNA expression. Similar effects were observed for luciferase expression under control of a human SeP‐promoter construct. Deletion as well as point‐mutation of putative SBEs led to a loss of promoter sensitivity towards TGF‐β1 treatment. Hence, we demonstrated an involvement of Smad 3 and 4 in transcriptional regulation of SeP by TGF‐β1 and we were able to identify the TGF‐β‐responsive element in the SeP promoter.
Toxicity of novel C‐terminal prion protein fragments and peptides harbouring disease‐related C‐terminal mutations
Authors: Daniels Maki , Cereghetti Grazia Maria , Brown David R.
Published: March 2003

Mice expressing a C‐terminal fragment of the prion protein instead of wild‐type prion protein die from massive neuronal degeneration within weeks of birth. The C‐terminal region of PrPc (PrP121–231) expressed in these mice has an intrinsic neurotoxicity to cultured neurones. Unlike PrPSc, which is not neurotoxic to neurones lacking PrPc expression, PrP121–231 was more neurotoxic to PrPc‐deficient cells. Human mutations E200K and F198S were found to enhance toxicity of PrP121–231 to PrP‐knockout neurones and E200K enhanced toxicity to wild‐type neurones. The normal metabolic cleavage point of PrPc is approximately amino‐acid residue 113. A fragment of PrPc corresponding to the whole C‐terminus of PrPc (PrP113–231), which is eight amino acids longer than PrP121–231, lacked any toxicity. This suggests the first eight amino residues of PrP113–121 suppress toxicity of the toxic domain in PrP121–231. Addition to cultures of a peptide (PrP112–125) corresponding to this region, in parallel with PrP121–231, suppressed the toxicity of PrP121–231. These results suggest that the prion protein contains two domains that are toxic on their own but which neutralize each other's toxicity in the intact protein. Point mutations in the inherited forms of disease might have their effects by diminishing this inhibition.
Inhibition of the SERCA Ca2+ pumps by curcumin
Authors: Bilmen Jonathan G. , Khan Shahla Zafar , Javed Masood‐ul‐Hassan , Michelangeli Francesco
Published: March 2003

Curcumin is a compound derived from the spice, tumeric. It is a potent inhibitor of the SERCA Ca2+ pumps (all isoforms), inhibiting Ca2+‐dependent ATPase activity with IC50 values of between 7 and 15 µm. It also inhibits ATP‐dependent Ca2+‐uptake in a variety of microsomal membranes, although for cerebellar and platelet microsomes, a stimulation in Ca2+ uptake is observed at low curcumin concentrations (<10 µm). For the skeletal muscle isoform of the Ca2+ pump (SERCA1), the inhibition of curcumin is noncompetitive with respect to Ca2+, and competitive with respect to ATP at high curcumin concentrations (≈10–25 µm). This was confirmed by ATP binding studies that showed inhibition in the presence of curcumin: ATP‐dependent phosphorylation was also reduced. Experiments with fluorescein 5′‐isothiocyanate (FITC)‐labelled ATPase also suggest that curcumin stabilizes the E1 conformational state. The fact that FITC labels the nucleotide binding site of the ATPase (precluding ATP from binding), and the fact that curcumin affects FITC fluorescence indicate that curcumin must be binding to another site within the ATPase that induces a conformational change to prevent ATP from binding. This observation is interpreted, with the aid of recent structural information, as curcumin stabilizing the interaction between the nucleotide‐binding and phosphorylation domains, precluding ATP binding.
Cys359 of GrdD is the active‐site thiol that catalyses the final step of acetyl phosphate formation by glycine reductase from Eubacterium acidaminophilum
Authors: Kohlstock Ulf‐Martin , Rücknagel Karl Peter , Reuter Michael , Schierhorn Angelika , Andreesen Jan R. , Söhling Brigitte
Published: March 2003

In the amino‐acid‐fermenting anaerobe Eubacterium acidaminophilum, acetyl phosphate is synthesized by protein C of glycine reductase from a selenoprotein A‐bound carboxymethyl‐selenoether. We investigated specific thiols present in protein C for responsibility for acetyl phosphate liberation. After cloning of the genes encoding the large and the small subunit (grdC1, grdD1), they were expressed separately in Escherichia coli and purified as Strep‐tag proteins. GrdD was the only subunit that catalysed arsenate‐dependent hydrolysis of acetyl phosphate (up to 274 U·mg−1), whereas GrdC was completely inactive. GrdD contained two cysteine residues that were exchanged by site‐directed mutagenesis. The GrdD(C98S) mutant enzyme still catalysed the hydrolysis of acetyl phosphate, but the GrdD(C359A) mutant enzyme was completely inactive. Next, these thiols were analysed further by chemical modification. After iodoacetate treatment of GrdD, the enzyme activity was lost, but in the presence of acetyl phosphate enzyme activity was protected. Subsequently, the inactivated carboxymethylated enzyme and the protected enzyme were both denatured, and the remaining thiols were pyridylethylated. Peptides generated by proteolytic cleavage were separated and subjected to mass spectrometry. Cys98 was not accessible to carboxymethylation by iodoacetate in the native enzyme in the presence or absence of the substrate, but could be alkylated after denaturation. Cys359, in contrast, was protected from carboxymethylation in the presence of acetyl phosphate, but became accessible to pyridylethylation upon prior denaturation of the protein. This clearly confirmed the catalytic role of Cys359 as the active site thiol of GrdD responsible for liberation of acetyl phosphate.
The cytochrome cbb3 from Pseudomonas stutzeri displays nitric oxide reductase activity
Authors: Forte Elena , Urbani Andrea , Saraste Matti , Sarti Paolo , Brunori Maurizio , Giuffrè Alessandro
Published: March 2003

The cytochrome cbb3 is an isoenzyme in the family of cytochrome c oxidases. This protein purified from Pseudomonas stutzeri displays a cyanide‐sensitive nitric oxide reductase activity (Vmax=100±9 mol NO·mol

·min−1 and Km=12±2.5 µm), which is lost upon denaturation. This enzyme is only partially reduced by ascorbate, and readily re‐oxidized by NO under anaerobic conditions at a rate consistent with the turnover number for NO consumption. As shown by transient spectroscopy experiments and singular value decomposition (SVD) analysis, these results suggest that the cbb3‐type cytochromes, sharing structural features with bacterial nitric oxide reductases, are the enzymes retaining the highest NO reductase activity within the heme‐copper oxidase superfamily.