bluuebunny/crossref_metadata_2025 huggingface dataset:
The Herbal Medicine Unkei‐to Stimulates the Secretion of a Cytokine‐induced Neutrophil Chemoattractant, CINC/gro, in the Rat Ovarian Cell Culture
Authors: Yasui Toshiyuki , Matsuzaki Toshiya , Ogata Rie , Kiyokawa Machiko , Ushigoe Kenjiro , Uemura Hirokazu , Kuwahara Akira , Ikawa Hiroshi , Maegawa Masahiko , Furumoto Hiroyuki , Aono Toshihiro , Irahara Minoru
Published: March 2003

Yasui T, Matsuzaki T, Ogata R, Kiyokawa M, Ushigoe K, Uemura H, Kuwahara A, Ikawa H, Maegawa M, Furumoto H, Aono T, Irahara M. The herbal medicine Unkei‐to stimulates the secretion of a cytokine‐induced neutrophil chemoattractant, CINC/gro, in the rat ovarian cell culture. AJRI 2003; 49:–© Blackwell Munksgaard, 2003

PROBLEM AND METHOD OF STUDY: We investigated the ovulation‐inducing effects of Unkei‐to, a Japanese herbal medicine, in relation to the production of sex steroid hormones (17β‐estradiol and progesterone), cytokine‐induced neutrophil chemoattractant (CINC/gro), interleukin‐1β (IL‐1β) and tumor necrosis factor α (TNF‐α) in the rat ovarian cell culture.

RESULTS: Unkei‐to at a concentration of 100 μg/mL significantly stimulated the secretions of 17β‐estradiol and progesterone (P < 0.01) in cultured whole ovarian dispersates. Unkei‐to also enhanced the secretion of CINC/gro in a dose‐dependent manner, and the secretions of CINC/gro increased significantly at concentrations of 10 and 100 μg/mL (P < 0.01). These stimulatory effects of Unkei‐to on steroidgenesis and CINC/gro production are very similar to those of another Japanese herbal medicine, Toki‐Shakuyaku‐san. In addition, Unkei‐to significantly (P < 0.01) enhanced the secretions of both IL‐1β and TNF‐α, which are known to stimulate the secretion of CINC/gro in the ovulatory process, at concentrations of 10 and 100 μg/mL. The stimulatory effect of Unkei‐to at a concentration of 100 μg/mL on IL‐1β/was significantly (P < 0.01) lower than that of Toki‐Shakuyaku‐san, while the stimulatory effects of these two herbal medicines at a concentration of 100 μg/mL on TNF‐α were similar.

CONCLUSIONS: These results show that Unkei‐to can stimulate ovarian steroidgenesis and the ovulatory process by inducing the secretion of CINC/gro with IL‐1 β and TNF‐αin vitro. Unkei‐to has stimulatory effects on both steroidgenesis and the ovulatory process in the ovary as well as a stimulatory effect on the hypothalamus‐pituitary axis, and it may be useful for treating patients with ovulatory disorders.
Progesterone Regulates IL12 Expression in Pregnancy Lymphocytes by Inhibiting Phospholipase A2
Authors: Par G. , Geli J. , Kozma N. , Varga P. , Szekeres‐Bartho J.
Published: March 2003

Par G, Geli J, Kozma N, Varga P, Szekeres‐Bartho J. Progesterone regulates IL12 expression in pregnancy lymphocytes by inhibiting phospholipase A2. AJRI 2003; 49:1–5 © Blackwell Munksgaard, 2003

PROBLEM: Progesterone‐induced blocking factor (PIBF) is one of the pathways that mediate the immunological effects of progesterone. PIBF inhibits natural killer (NK) cytotoxic activity. Recently we showed that neutralization of PIBF results in an increased interleukin (IL)‐12 expression, which is corrected by cyclooxygenase inhibitors. As exogenous arachidonic acid (AA) voids the NK blocking effect of PIBF, it is likely that PIBF acts before the level of the cyclooxygenase enzyme. Therefore in this study we investigated the effect of PIBF neutralizing antibody and simultaneous phospholipase A2 inhibitor quinacrine (Q) treatment on IL‐12 production.

METHODS: Pregnancy lymphocytes were treated with anti‐PIBF antibody or lipopolysaccharide (LPS) as a positive control, in the presence or absence of Q. IL‐12 expression by PBMC was detected by immunocytochemistry.

RESULTS: Neutralization of PIBF as well as LPS treatment resulted in an increased IL‐12 expression, which was corrected by simultaneous Q treatment. Pre‐treatment of lymphocytes with progesterone prevented the stimulating effect of LPS on IL‐12 production.

CONCLUSION: Progesterone binding of the lymphocytes is followed by the release of PIBF that inhibits AA release. The subsequent block of prostaglandin synthesis reduces IL‐12 production and results in a lowered cytotoxic NK activity, which may contribute to a normal pregnancy outcome.
In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in four media at room temperature
Authors: Ashkenazi M. , Marouni M. , Sarnat H.
Published: March 2003

Abstract – The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objective of this study was to compare the effectiveness of four recommended storage media (Hank's balanced salt solution [HBSS], culture medium, α minimal essential medium [α‐MEM], and ViaSpan) to preserve cultured periodontal ligament fibroblasts (PDLF) at room temperature (22°C). PDLF were obtained from explants of extracted healthy human teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at room temperature. A control group was incubated with culture medium at 37°C. After incubation, viability of the cells was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Viability of PDLF stored up to 24 h was comparable in all tested media, and the differences were limited to 1%–3%. PDLF stored for up to 24 h in various media had statistically comparable mitogenicity to the control group. After 8 h of storage, the differences were limited to 2%–9%, except for the α‐MEM group which had 23%–29% lower mitogenic capacity compared to the control group. Increasing the storage time up to 24 h further decreased the mitogenicity of the cells by 22%–47%. The highest mitogenicity after 24 h of storage was found in PDLF stored in culture medium or HBSS, and the lowest in α‐MEM. PDLF stored for 2–8 h in various media had a comparable clonogenic capacity to the control group. However, after 24 h, the cells' clonogenic ability dropped by 14%–66%. A similar trend of reduction was noted in the mitogenic and clonogenic capacity, although it was statistically significant only in the clonogenic capacity. Culture medium and ViaSpan, followed by HBSS, were the most effective in preserving the clonogenic capacity of PDLF after 24 h of storage. The lowest clonogenic capacity after 24 h of storage was in the α‐MEM group (66%, P<0.0025). In conclusion, culture medium, followed by HBSS and ViaSpan, was the most effective media for preserving the viability, mitogenicity and clonogenic capacity of PDLF stored for up to 24 h at room temperature. The lowest functional abilities were found in PDLF stored in α‐MEM.
Utilization of gutta‐percha for retrograde root fillings
Authors: Sauveur G. , Sobel M. , Boucher Y.
Published: March 2003

Abstract – Just as gutta‐percha used with a root canal sealer is a recommended material for orthograde root fillings, it could similarly be the material of choice for retrograde fillings. Unfortunately, clinical accessibility and visibility do not always facilitate such a technique. The aim of this article is to present a new technique which enables retrograde fillings to be achieved with gutta‐percha and a sealer. After the apex had been resected, a hole was drilled perpendicular to the plane of section of the apex about 1 mm coronally. The bucco‐lingual depth required to reach the main canal was calculated. The cavity was then dried, coated with the sealer, and obturated with gutta‐percha in accordance with thermo‐mechanical compaction techniques. After excess filling material had been removed, the gutta‐percha was cold burnished and the angles of the root were smoothed. Clinical cases illustrating healing of the periapical tissues are shown.
Malformations in maxillary incisors: case report of radicular palatal groove
Authors: Schäfer E. , Cankay R. , Ott K.
Published: March 2003

Abstract – This report describes malformations of anterior teeth and a case of a palatal radicular groove in an upper lateral incisor with periodontal complications. The patient was first diagnosed with an endodontic problem. Endodontic treatment alone failed to resolve the pain. Therefore, after the palatal groove had been properly diagnosed, a palatal flap procedure was carried out including removal of the granulation tissue and careful scaling and root planing of the area with the groove. No odontoplasty was done. Endodontic therapy of the tooth was completed and periapical healing was evident. The treatment effectively reduced the gingival probing depth and led to an asymptomatic tooth.
Bacterial leakage in obturated root canals following the use of different intracanal medicaments
Authors: Barthel C. R. , Zimmer S. , West G. , Roulet J.‐F.
Published: March 2003

Abstract – The aim of this study was to examine whether intracanal medication prior to root canal obturation has an inhibitory effect on corono‐apical penetration of bacteria. 93 single rooted teeth were instrumented and sterilized with ethylene oxide. They were assigned to three control groups and four test groups with n=20 each. For one week, they were dressed with different medicaments: The first group with a 5% chlorhexidine gel, the second with Ledermix, the third with a fresh mix of calcium hydroxide and water and the fourth without any medication. After obturation (lateral condensation, AH26) the roots were fixed between a top and a bottom chamber. The top chamber contained 3 mL trypticase soy broth with 108Staphylococcus epidermidis CFU's/mL, whereas the bottom chamber contained sterile trypticase soy broth. For one year, the mounts were incubated at 37°C. They were checked on a regular basis for turbidity in their bottom chambers indicating bacterial growth. None of the test samples leaked for three months. After one year, the calcium hydroxide group had only 6 leaking samples whereas the chlorhexidine group had 14, the Ledermix group 15, and the unmedicated group had 13 leaking samples. It may be concluded that under the conditions of this study, calcium hydroxide was the medicament of choice to avoid bacterial penetration of the root canal. Ledermix did not perform better than no pre‐medication. Chlorhexidine was superior to Ledermix in the second third of the observation period.
Dental trauma management awareness of Singapore pre‐school teachers
Authors: Sae‐Lim Varawan , Lim Lum Peng
Published: March 2003

Abstract – The purpose of this study was to assess the awareness of pre‐school teachers concerning the management of traumatized teeth. A questionnaire survey of teachers’ backgrounds, attitudes/practices and knowledge was distributed to all teachers who attended a dental education program organized by the Singapore Dental Health Foundation. Of a total of 291 teachers surveyed, 29% had received tertiary education, while the rest (71%) had received secondary education. The mean teaching experience was 6.8±6.9 years. About a quarter had previous experience in handling dental trauma. Sixty‐three percent admitted having no knowledge of dental trauma; 79% were unsatisfied with their level of knowledge in this area; 95% were keen to have further education in dental trauma; 65% thought dental trauma emergency should be dealt with as soon as possible. Concerning knowledge, during‐office hour emergency services were more familiar (84%) than after‐office hour emergency services (15%), as was the concept of management of avulsed teeth (71%) compared to that of fractured teeth (51%). Knowledge about optimal storage media for avulsed permanent teeth was especially poor – being as low as 15%. Using multiple logistic regression analysis, it was found that teaching experience significantly influenced the respondents’ self‐assessed knowledge and their level of satisfaction with their knowledge (P=0.012). Teachers with more teaching experience had better knowledge about the replantation of permanent teeth (P=0.003). It is recommended that public education targeted at teachers should be carried out to increase dental trauma management awareness.
Prevalence and correlates of traumatic injuries to the permanent teeth of school‐children aged 9–14 years in Belo Horizonte, Brazil
Authors: Cortes M. I. S. , Marcenes W. , Sheiham A.
Published: March 2003

Abstract – A cross‐sectional survey was carried out on 3702 boys and girls aged 9–14 years, attending public and private primary schools in Belo Horizonte, Brazil. A multi‐stage sampling technique using an equal probability scheme was adopted to select the children. The response rate for the total sample was 97%. Dental examinations were carried out by one dentist (MISC). Intra‐examiner agreement was very good. The prevalence of dental injuries increased from 8% at the age of 9 years to 13.6% at 12 and 16.1% at 14 years. Adjusted results showed that children from high socio‐economic backgrounds were 1.4 (95% CI=1.15–1.79) times more likely to present with a dental injury than children with low SES. Boys were 1.7 times (95% CI=1.41–2.16) more likely to have dental injuries than girls. Children with an overjet size greater than 5.0 mm were 1.37 times (95% CI=1.06–1.80) more likely to have a dental injury than children with an overjet size equal or lower than 5.0 mm. Finally, children with an adequate lip coverage were 0.56 times (95% CI=0.44–0.72) less likely to have a traumatic dental injury than those with inadequate lip coverage.
Prevalence of dental trauma in 5–6‐ and 12–14‐year‐old boys in Riyadh, Saudi Arabia
Authors: Al‐Majed Ibrahim , Murray John J. , Maguire Anne
Published: March 2003

Abstract – This study involved 354 boys aged 5–6 years and 862 boys aged 12–14 years, attending 40 schools in Riyadh. All children were examined at school by a single dental examiner, using criteria similar to those employed in the survey of children’s dental health in the United Kingdom. The prevalence of dental trauma in 354 Saudi boys aged 5–6 years was 33%. The most common type of dental trauma was fracture of enamel (71%) followed by loss of tooth due to trauma (13%), fracture into enamel and dentine (7%), discolouration (5%), pulp involvement (4%). No relationship between the degree of overjet and the occurrence of dental trauma in the primary dentition was observed. The prevalence of dental trauma in 862 12–14‐year‐old boys was 34%. The commonest dental trauma was fracture of enamel (74%) followed by fracture into enamel and dentine (15%), fracture into enamel‐dentine and pulp (5%), loss of tooth due to trauma (3%), and discolouration (0.4%). A significant relationship (P=0.02) between the increased overjet (≥ 6 mm) and the occurrence of dental trauma in the permanent dentition was reported. The present study found no evidence of dental care provided for traumatised primary incisors in 5–6‐year‐old boys. The treatment of dental trauma in 12–14‐year‐old boys was negligible (2.4%). The present Saudi Arabian study showed higher prevalence of dental trauma in 5–6‐ and 12–14‐year‐old boys than the reported results of the United Kingdom Children’s Dental Health Survey of the same age groups.
Response of the pulp of dogs to capping with mineral trioxide aggregate or a calcium hydroxide cement
Authors: Faraco Italo Medeiros , Holland Roberto
Published: March 2003

Abstract – This study was conducted to observe the response of dogs’ dental pulp to mineral trioxide aggregate (MTA) and a calcium hydroxide cement when used as pulp capping materials. After the pulps of 30 teeth were exposed, they were capped with either MTA or a calcium hydroxide cement. Histological analysis was performed 2 months after treatment. Results showed a healing process with complete tubular dentin bridge formation and no inflammation in any of the pulps capped with MTA. On the other hand, only five specimens from the calcium hydroxide cement group formed a complete dentin bridge. In this experimental group, pulp inflammation was observed in all but three cases. In conclusion, MTA exhibited better results than the calcium hydroxide cement for the capping of the pulp in dogs.
Periodontal healing after bonding treatment of vertical root fracture
Authors: Sugaya Tsutomu , Kawanami Masamitsu , Noguchi Hiroshi , Kato Hiroshi , Masaka Nobuo
Published: March 2003

Abstract – Vertical root fractures lead to advanced periodontal breakdown with deep periodontal pockets and vertical bone defects. The purpose of this study is to evaluate clinically the periodontal healing of root fracture treatment using adhesive resin cement. In 22 patients, 23 teeth with vertical root fractures were treated with 4‐META/MMA‐TBB resin cement. Eleven fractured roots were bonded through the root canal (group A) and 12 fractured roots were bonded extra‐orally and replanted (group B). All teeth were then restored with full cast crowns (n=20) or coping (n=3). Mean probing depth was 6.6 mm at pre‐treatment and 4.4 mm 6 months after the treatment in group A, and 7.4 mm and 4.6 mm, respectively, in group B. Bleeding scores were 100% at pre‐treatment and 36.4% after 6 months in group A and 91.7% and 8.3%, respectively in group B. Radiographic bone level was 56.8% at pre‐treatment and 59.1% after 6 months in group A, and 18.8% and 29.2%, respectively, in group B. Two roots of group A and three roots of group B were extracted due to refracture, deterioration of periodontal inflammation, mobility, and luxation. The remaining roots (n=18) presented no discomfort to the patients and there was no deterioration of periodontal conditions over a mean period of 33 months (range 14–74 months) in group A and over a mean period of 22 months (range 6–48 months) in group B. There was no ankylosed teeth nor was any root resorption detected. The results suggested that the treatment of vertical root fracture using 4‐META/MMA‐TBB resin has good prognostic possibilities.
Traumatized permanent teeth in Brazilian children assisted at the Federal University of Santa Catarina, Brazil
Authors: De Carvalho Rocha Maria José , Cardoso Mariane
Published: March 2003

Abstract – The goal of this study was to determine factors related to the occurrence of dental trauma in permanent teeth of children assisted at the Pediatric Dentistry Clinic of the Federal University of Santa Catarina (UFSC) Florianópolis, Brazil. During a period of 18 months, 36 children between 7 and 12 years of age had 72 traumatized teeth treated. The children were all assisted by one professional, a dentist working as a trainee of the Pediatric Dentistry Clinic. The occurrence of trauma was higher in male patients (61.3%) and in children between 8 and 9 years old with an average age of x̄=9.4 years. In the group assisted, 63.9% of the children had more than one traumatized tooth and trauma reoccurred 19.4% of the time. The maxillary anterior teeth represented 96.1% of the cases and the central incisor teeth were the most affected. Both sides of the mouth had approximately the same number of traumas. Fractures were more frequent (51.4%) than luxations (48.6%). Enamel/dentin crown fractures represented 51.4% of the total traumatized teeth. Falls were the main cause of trauma (83.3%). A dental professional assisted 36.1% of the children in some way during the first 24 h after the incident. The study concluded that the permanent dentition is mostly affected by crown fractures that occur especially on maxillary central incisor teeth in patients between 8 and 9 years of age. The major etiological factor is falls that affected more than one tooth. Re‐occurrence of trauma is fairly common.
Dental trauma and level of information: mouthguard use in different contact sports
Authors: Ferrari Carlos Henrique , De Medeiros João Marcelo Ferreira
Published: March 2003

Abstract  – The objective of the present work was to study the occurrence of dental trauma in different sports, as well as to check if athletes used mouthguard during sport activities, and knew the siginificance of its utilization. The professional and semi‐professional athletes who practised contact sports were interviewed, and were analyzed to determine the occurrence of dental trauma during sport practice, as well as if the athletes used a mouthguard and knew its indication. Results showed a high rate of dental trauma in athletes compared to the general population. The majority of athletes also demonstrated little utilization of mouthguard, in spite of the information about usage during sports practices and a general knowledge of its uses.
Autotransplantation of immature third molars: influence of different splinting methods and fixation periods
Authors: Bauss Oskar , Schilke Reinhard , Fenske Christian , Engelke Werner , Kiliaridis Stavros
Published: March 2003

Abstract  
–
 Different data were published documenting the influence of fixation methods and periods on the outcome of autotransplantations of teeth. Besides studies reporting increased ankylosis and disturbances of pulpal revascularization following rigid or extended fixation, there are studies revealing no connection in this matter. The clinical and radiological results of 76 transplanted germs of third molars were to be assessed after a rigid fixation for 4 weeks or after a suture splinting for 7 days and compared with each other. The choice of fixation method depended on the initial stability of the transplant. After a mean observation period of 3.4 years (range 1.0–6.1 years), 92.9% of the teeth stabilized with a suture, but only 73.5% fixed with an acid‐etch composite and wire splint could be classified as successful (P = 0.029). The significant increases in ankylosis (P = 0.036) and pulp necrosis (P = 0.041) were the factors for the less favorable results of the rigidly fixed teeth. Our data support the results of other experimental and clinical studies with regard to the negative influences of an extended and rigid fixation on the success of tooth transplantation. However, apart from a correlation with the method and period of fixation, we also suspect the influence of an incongruity between the transplant and recipient site.
The effect of age of refrigerated conditioned medium on cell survivability in vitro
Authors: Vajrabhaya La‐Ongthong , Vongphan Nattaya , Hongskul Pranee , Suwannawong Suwanna K.
Published: March 2003

Abstract  – Samples of a conditioned medium (CM), obtained from human ginvigal fibroblasts culture, were kept at 4°C each for 2 weeks, 1, 2, 3 and 6 months. Their effectiveness in maintaining cell survival for their contact with L929 cells in 96 well tissue culture plates for 6 and 10 h at 4°C was compared with Hank's Balanced Salt Solution (HBSS). Surviving cells were stained with neutral red dye and the dye intensity was measured spectrophotometrically at 540 nm. The target cells responded to all the above mentioned CM and the HBSS in the same manner at 6‐h contact time. However, by 10‐h contact time, the percentage of the target cells' survival was higher in the 2‐week, 1‐ and 2‐month CM than in HBSS, 3‐ and 6‐month CM.
Quality of handover to the postanaesthesia care unit nurse
Authors: Anwari J. S.
Published: March 2003

Summary I surveyed 276 patients admitted to the postanaesthetic care unit (PACU). The quality of handover to the PACU nurse was assessed by scoring four indicators: the quality of verbal information about the patient, the condition of the patient on admission, the professional behaviour of the anaesthetist and the nurse's satisfaction with the handover. Maximum scores were attained by 32.6% of anaesthetists for the quality of ‘verbal information’ about the patient given to the PACU nurse, and by 62% of anaesthetists for the ‘condition of the patient’. ‘Professional behaviour’ was judged satisfactory or good in 87% of anaesthetists and almost half of the anaesthetists (48%) scored full marks for the nurse's satisfaction with the handover. The scores of all four indicators were aggregated to give an overall impression of quality of handover; this was good in fewer than half the number of patients brought to the PACU.
A new, simple, safe, effective and cost‐effective procedure for genuine stress incontinence: Midurethral polypropylene sling
Authors: Demirci Fuat , Ozdemir Ismail , Yucel Oguz , Alhan Ali
Published: March 2003

We developed a cost‐effective procedure for genuine stress incontinence (GSI) that has the advantages of the tension‐free vaginal tape (TVT). The midurethral polypropylene sling procedure (MPS) is carried out under local anaesthesia. A self‐fashioned sling (7.5 × 1 cm) was created from a polypropylene mesh with two lengthening polypropylene sutures at the ends. The sutures are carried through the rectus fascia using a needle and the sling is placed around the urethra. Ten patients underwent the MPS and were followed up for a mean of 6.2 months. All patients were cured. The short‐term results of the MPS were comparable to those of the TVT. The procedure costs approximately US$9. We conclude that the MPS can be considered as an alternative to the TVT procedure.
Radical surgery with omental flap for uncontrolled locally recurrent breast cancer
Authors: Henderson Michael A. , Burt James D. , Jenner David , Crookes Peter , Bennett Richard C.
Published: March 2003

Background:  Uncontrolled chest wall disease due to breast cancer is a highly morbid condition causing pain, ulceration, malodour and the need for frequent dressings. Aggressive surgical approaches are rarely justified because most patients will succumb to metastatic breast cancer within a short period. A highly selected group of patients with minimal or no evidence of metastatic disease and good performance status may benefit from radical chest wall surgery. Omental transposition flaps are ideal for reconstructing extensive surgical defects following chest wall surgery.

Methods:  A retrospective review was carried out of 61 female patients treated consecutively between 1980 and 1995. The surgical technique is described herein.

Results:  All patients were symptomatic preoperatively. Symptoms included ulceration (80%), pain (44%) and malodour (40%). Twenty‐nine patients had uncontrolled local recurrence following initial treatment for locally advanced breast cancer and 32 patients developed uncontrolled recurrence after treatment for operable breast cancer by mastectomy or conservation surgery. Median survival following chest wall surgery was 21 months and the median local recurrence‐free interval was 20 months. Morbidity was ­limited. There were no cases of major flap loss. Twenty‐nine patients (48%) had no further local disease. Eighteen patients (30%) developed soft‐tissue recurrence at the edge of the omental flap or in surrounding skin and 14 (23%) developed recurrence beneath the flap.

Conclusion:  In a highly selected group of patients with symptomatic uncontrolled chest wall recurrence who are fit and have an expectation of at least moderate‐term survival, radical chest wall surgery and omental flap transposition offers excellent palliation and local control in the majority of patients
Why do some patients with > 80% stenosis of the internal carotid artery not undergo surgery? A retrospective review
Authors: Gunawardena Indunil D. , Burnett Alison , Makeham Virginia , Harris John P.
Published: March 2003

Background: Carotid endarterectomy is known to benefit both symptomatic and asymptomatic patients with high‐grade internal carotid artery stenosis. Duplex scanning is the ‘gold standard’ for non‐invasive preoperative investigation of carotid artery stenosis. The aim of the present study was to analyse the indications for duplex scanning and to identify other factors that influenced the management of patients with high‐grade stenosis who did not undergo carotid endarterectomy.

Methods: A total of 271 patients was observed to have > 80% stenosis of the internal carotid artery on duplex scanning during the period of review. Of these patients, 85 did not undergo carotid endarterectomy. The vascular laboratory database and hospital records of these patients were retrospectively reviewed.

Results: The indications for requesting a carotid duplex scan in the 85 patients were transient ischaemic attack (22%), stroke (25%), symptomatic bruit (7%), asymptomatic bruit (12%), and stroke and symptomatic bruit combined (7%). Falls and preoperative carotid assessment prior to coronary surgery were the commonest indications in the remaining patients. The main risk factors were cardiac (68%), hypertension (60%), respiratory (21%), diabetes (25%), peripheral vascular disease (19%), neoplasm (16%) and renal disease (16%). Twenty‐five per cent of the patients were over 80 years of age.

Conclusion: In the present study risk factors associated with increased perioperative morbidity and mortality were the commonest explanation for patients with high‐grade stenosis of the internal carotid artery not undergoing surgery. These patients would generally not meet the inclusion criteria for the major carotid endarterectomy trials.
Massive pseudotumour resection with recombinant  factor VIIa (NovoSeven) cover
Authors: O'Connell N. , Chen J. , Byrne M. , O'Shea E. , Smyth H. , Smith O. P.
Published: March 2003

Summary.  Surgical resection is the only definitive treatment for haemophilic pseudotumour. For patients with high‐ responding factor (F)VIII inhibitors, the lack of reliable by‐passing agents in the past has meant that surgery has been avoided in favour of conservative measures. The development of recombinant factor VIIa (rFVIIa) has revolutionized the management of surgery in these patients. We document the first successful report of the surgical resection of a massive pseudotumour in a patient with high responding FVIII inhibitors using rFVIIa to achieve haemostasis. Serial post‐operative measurements indicated that FVII coagulation activity (FVII:C) levels were more sensitive to rFVIIa dose changes than the prothrombin time.
A novel deletion of the l‐ferritin iron‐responsive element responsible for severe hereditary hyperferritinaemia–cataract syndrome
Authors: Cazzola Mario , Foglieni Barbara , Bergamaschi Gaetano , Levi Sonia , Lazzarino Mario , Arosio Paolo
Published: March 2003

Summary. In the last few years, mutations that cause disease through increased efficiency of mRNA translation have been discovered. Hereditary hyperferritinaemia–cataract syndrome (HHCS) arises from various point mutations or deletions within the iron‐responsive element (IRE) in the 5′‐UTR of the l‐ferritin mRNA. Each unique mutation confers a characteristic degree of hyperferritinaemia and severity of cataract in affected individuals. We report a novel six‐nucleotide deletion identified in an Italian family presenting with elevated serum ferritin and early onset bilateral cataract. This deletion involves a sequence with a TCT repetition and may have occurred through a mechanism of slippage mispairing. Because of the above repetition, the observed mutation can be interpreted as deletion 22–27, 23–28, 24–29 or 25–30. Structural modelling predicted an IRE stem modification that is expected to markedly reduce the binding to iron‐regulatory proteins. A double‐gradient denaturing gradient gel electrophoresis (DG‐DGGE) method easily detected the above deletion.
High multidrug resistance protein activity in acute myeloid leukaemias is associated with poor response to chemotherapy and reduced patient survival
Authors: Laupeze B. , Amiot L. , Drenou B. , Bernard M. , Branger B. , Grosset J. M. , Lamy T. , Fauchet R. , Fardel O.
Published: March 2003

Summary.  Multidrug resistance protein (MRP) activity was investigated in 44 newly diagnosed acute myeloid leukaemia (AML) patients using a functional assay based on efflux of carboxy‐2′,7′‐dichlorofluorescein, an anionic dye handled by both MRP1 and MRP2. Elevated MRP transport was detected in 29% of cases, but was not significantly correlated with sex, age, white blood cell count at diagnosis or karyotype. In contrast, it was associated with secondary AML (P = 0·002), CD34 positivity (P = 0·041) and P‐glycoprotein activity (P = 0·01). There was a lower rate of complete remission in MRP‐positive patients versus MRP‐negative patients (23% versus 81%; P = 0·001); overall survival was also better for MRP‐negative patients (P = 0·004). These data indicate a probable role for MRP activity in the clinical outcome of AML.
Randomized double-blind controlled trial of roxithromycin for prevention of abdominal aortic aneurysm expansion
Authors: Vammen S , Lindholt J S , Østergaard L , Fasting H , Henneberg E W
Published: March 2003

Background

Macrolide treatment has been reported to lower the risk of recurrent ischaemic heart disease. The influence of macrolides on the expansion rate of abdominal aortic aneurysms (AAAs) remains unknown. The aim was to investigate the effect of roxithromycin on the expansion rate of small AAAs.

Methods

A total of 92 subjects with a small AAA were recruited from two populations. One population consisted of 6339 men aged 65–73 years who were offered a hospital-based mass screening programme for AAA. From this population 66 subjects were recruited. The remaining 26 men were recruited from among 49 subjects diagnosed at interval screening for an initial aortic diameter between 25 and 29 mm. Subjects were randomized to receive either oral roxithromycin 300 mg once daily for 28 days or matching placebo, and followed for a mean of 1·5 years.

Results

The mean annual expansion rate of AAAs was reduced by 43 per cent in the intervention group (1·56 mm per year), compared with 2·75 mm per year following placebo (P = 0·02). Multiple linear regression analysis showed that roxithromycin treatment and initial AAA size were significantly related to AAA expansion when adjusted for smoking, diastolic blood pressure and immunoglobulin A level of 20 enzyme immunounits or more. Logistic regression analysis confirmed a significant difference in expansion rates above 2 mm annually between the intervention and placebo groups: odds ratio = 0·09 (95 per cent confidence interval 0·01–0·75).

Conclusion

In comparison to placebo, roxithromycin 300 mg daily for 4 weeks reduced the expansion rate of AAAs.
Fate of the rectum in patients undergoing total colectomy for Crohn's disease
Authors: Cattan P , Bonhomme N , Panis Y , Lémann M , Coffin B , Bouhnik Y , Allez M , Sarfati E , Valleur P
Published: March 2003

Background

This study aimed to analyse disease recurrence and fate of the rectum in patients who had a total colectomy for Crohn's disease.

Methods

One hundred and forty-four patients who had a total colectomy for Crohn's colitis were reviewed retrospectively. Ileorectal anastomosis (IRA) was performed in 118 patients, while 26 never had an IRA after colectomy because of severe anorectal lesions. Factors associated with recurrence and rectal preservation failure were studied.

Results

The probability of clinical recurrence after IRA was 58 and 83 per cent at 5 and 10 years respectively. The probability of rectal preservation at 5 and 10 years was 70 and 63 per cent after colectomy, and 86 and 86 per cent after IRA, respectively. Patients with extraintestinal manifestation had a higher risk of recurrence and of rectal preservation failure. Previous ileal involvement was associated with a higher rate of ileal recurrence after IRA. After IRA, prophylactic treatment with 5-aminosalicylic acid was associated with a lower rate of recurrence and of failure to preserve the rectum.

Conclusion

Overall, 63 per cent of patients had a functioning IRA 10 years after total colectomy. Absence of extraintestinal manifestation and prophylactic treatment with 5-aminosalicylates after IRA were the main factors associated with long-term rectal preservation.
Identification of novel splice variants of the human catalytic subunit cβ of cAMP‐dependent protein kinase
Authors: Ørstavik Sigurd , Reinton Nils , Frengen Eirik , Langeland Bjørn Tore , Jahnsen Tore , Skålhegg Bjørn S.
Published: March 2003

Four different isoforms of the catalytic subunit of cAMP‐dependent protein kinase, termed Cα, Cβ, Cγ and PrKX have been identified. Here we demonstrate that the human Cβ gene encodes six splice variants, designated Cβ1, Cβ2, Cβ3, Cβ4, Cβ4ab and Cβ4abc. The Cβ splice variants differ in their N‐terminal ends due to differential splicing of four different forms of exon 1 designated exon 1‐1, 1‐2, 1‐3, 1‐4 and three exons designated a, b and c. All these exons are located upstream of exon 2 in the Cβ gene. The previously identified human Cβ variant has been termed Cβ1, and is similar to the Cβ isoform identified in the mouse, ox, pig and several other mammals. Human Cβ2, which is the homologue of bovine Cβ2, has no homologue in the mouse. Human Cβ3 and Cβ4 are homologous to the murine Cβ3 and Cβ2 splice variants, whereas human Cβ4ab and Cβ4abc represent novel isofoms previously not identified in any other species. At the mRNA level, the Cβ splice variants reveal tissue specific expression. Cβ1 was most abundantly expressed in the brain, with low‐level expression in several other tissues. The Cβ3 and Cβ4 splice variants were uniquely expressed in human brain in contrast to Cβ2, which was most abundantly expressed in tissues of the immune system, with no detectable expression in brain.

We suggest that the various Cβ splice variants when complexed with regulatory subunits may give rise to novel holoenzymes of protein kinase A that may be important for mediating specific effects of cAMP.
Phenylcoumaran benzylic ether and isoflavonoid reductases are a new class of cross‐reactive allergens in birch pollen, fruits and vegetables
Authors: Karamloo Fariba , Wangorsch Andrea , Kasahara Hiroyuki , Davin Laurence B. , Haustein Dieter , Lewis Norman G. , Vieths Stefan
Published: March 2003

We investigated the biochemical function of the birch pollen allergen Bet v 6 and its role in the IgE‐cross‐reactivity between birch pollen and plant foods, and characterized Pyr c 5, a Bet v 6‐related food allergen, from pear; the proteins were expressed as His‐Tag fusion proteins in Eschershia coli and purified by Ni‐chelate affinity chromatography under native conditions. Nonfusion proteins were obtained by factor Xa protease treatment. The highest degree of amino‐acid sequence identity of Pyr c 5 and Bet v 6 was found with a plant protein related to a defense mechanism, which we have named phenylcoumaran benzylic ether reductase (PCBER) based on its ability to catalyze the NADPH‐dependent reduction of 8–5′ linked lignans such as dehydrodiconiferyl alcohol to give isodihydrodehydrodiconiferyl alcohol. Enzymatic assays with recombinant Pyr c 5 and Bet v 6 showed PCBER catalytic activity for both recombinant allergens. Both Pyr c 5 and Bet v 6 allergens had similar IgE binding characteristics in immunoblotting and enzyme allergosorbent tests (EAST), and bound IgE from 10 sera of birch‐pollen‐allergic patients including six pear‐allergic subjects. EAST inhibition experiments with Pyr c 5 as the solid phase antigen suggested that homologous allergens may be present in many vegetable foods such as apple, peach, orange, lychee fruit, strawberry, persimmon, zucchini (courgette), and carrot. In extracts of pear, apple, orange, and persimmon, the presence of proteins of approximately 30–35 kDa containing Bet v 6 cross‐reactive epitopes was demonstrated with two Bet v 6‐specific monoclonal antibodies. Recombinant Pyr c 5 triggered a strong, dose‐dependent mediator release from basophils of a pear‐allergic subject, suggesting that Pyr c 5 has the potential to elicit type I allergic reactions.
New antigenic regions of streptokinase are identified by affinity‐directed mass spectrometry
Authors: Coffey Jonathan A. , Jennings Keith R. , Dalton Howard
Published: March 2003

Streptokinase (SK) is a bacterial protein used for the treatment of myocardial infarction, which is immunogenic in humans. Here we report the use of an affinity‐directed MS approach to determine the minimal epitopes involved in the binding between SK and patient antibodies. Using this method we have identified two novel epitopes and mapped these to the minimal recognition regions formed by the amino acids D96–E99 and F323–D328. We have also located three previously identified antigenic regions and have now mapped them and shown that they can be defined more precisely as residues P4–L8, P171–P177 and K334–N338.
Biosynthesis of the cancer‐related sialyl‐α2,6‐lactosaminyl epitope in colon cancer cell lines expressing β‐galactoside α2,6‐sialyltransferase under a constitutive promoter
Authors: Dall'Olio Fabio , Chiricolo Mariella , Mariani Erminia , Facchini Andrea
Published: March 2003

An elevation of β‐galactoside α2,6‐sialyltransferase (ST6Gal.I) enzyme activity and an increased α2,6‐sialylation of cell membranes are among the most prominent glycosylation changes associated with colon cancer; both modifications correlate with a worse prognosis. In our previous studies, we have frequently observed a discrepancy between the ST6Gal.I level within a colon cancer sample or cell line and the respective level of reactivity with the α2,6‐sialyl‐specific lectin from Sambucus nigra (SNA). In this study, we have investigated quantitatively the biosynthesis of the sialyl‐α2,6‐lactosaminyl epitope in two colon cancer cell types expressing the ST6Gal.I cDNA under the control of a constitutive promoter. By measuring the amount of ST6Gal.I mRNA using competitive RT‐PCR, the expression of α2,6‐sialylated lactosaminic structures with SNA and anti‐CDw75 Ig, and the presence of unsubstituted lactosaminic termini by Erythrina cristagalli lectin, we reached the following conclusions: (a) a high proportion of the cell surface lactosaminic termini remains unsubstituted, even in the presence of a very high ST6Gal.I activity. This proportion is strongly dependent on the cell type; (b) ST6Gal.I‐transfected colon cancer cells do not express the CDw75 epitope; (c) the level of ST6Gal.I enzyme activity only partially correlates with the mRNA level; (d) despite the control by a constitutive promoter, the ST6Gal.I mRNA is not constantly expressed over time; and (e) a very large portion of the enzyme molecules is secreted in the extracellular milieu. These results indicate that post‐transcriptional and post‐translational mechanisms play a pivotal role in the control of α2,6‐sialylation in colon cancer cells.
Importance of the amino‐acid composition of the shutter region of plasminogen activator inhibitor‐1 for its transitions to latent and substrate forms
Authors: Hansen Martin , Busse Marta N. , Andreasen Peter A
Published: March 2003

The serpins are of general protein chemical interest due to their ability to undergo a large conformational change consisting of the insertion of the reactive centre loop (RCL), which becomes strand 4, into the central β sheet A. To make space for the incoming RCL, the ‘shutter region’ opens by the β strands 3A and 5A sliding apart over the underlying α helix B. Loop insertion occurs during the formation of complexes of serpins with their target serine proteinases and during latency transition. This type of loop insertion is unique to plasminogen activator inhibitor‐1 (PAI‐1). We report here that amino‐acid substitutions in a buried cluster of three residues forming a hydrogen bonding network in the shutter region drastically accelerate PAI‐1 latency transition; that the rate was in all cases normalized by the PAI‐1 binding protein vitronectin; and that substitution of an adjacent β strand 5A Lys residue, believed to anchor β strand 5A to other secondary structural elements, had differential effects on the rates of latency transition in the absence and the presence of vitronectin, respectively. An overlapping, but not identical set of substitutions resulted in an increased tendency to substrate behaviour of PAI‐1 at reaction with its target proteinases. These findings show that vitronectin regulates the movements of the RCL through conformational changes of the shutter region and β strand 5A, are in agreement with RCL insertion proceeding by different routes during latency transition and complex formation, and contribute to the biochemical basis for the potential use of PAI‐1 as a therapeutic target in cancer and cardiovascular diseases.
l‐Threonine aldolase, serine hydroxymethyltransferase and fungal alanine racemase
Authors: Contestabile Roberto , Paiardini Alessandro , Pascarella Stefano , di Salvo Martino L. , D'Aguanno Simona , Bossa Francesco
Published: March 2003

Serine hydroxymethyltransferase (SHMT) is a member of the fold type I family of vitamin B6‐dependent enzymes, a group of evolutionarily related proteins that share the same overall fold. The reaction catalysed by SHMT, the transfer of Cβ of serine to tetrahydropteroylglutamate (H4PteGlu), represents in the cell an important link between the breakdown of amino acids and the metabolism of folates. In the absence of H4PteGlu and when presented with appropriate substrate analogues, SHMT shows a broad range of reaction specificity, being able to catalyse at appreciable rates retroaldol cleavage, racemase, aminotransferase and decarboxylase reactions. This apparent lack of specificity is probably a consequence of the particular catalytic apparatus evolved by SHMT. An interesting question is whether other fold type I members that normally catalyse the reactions which for SHMT could be considered as ‘forced errors’, may be close relatives of this enzyme and have a catalytic apparatus with the same basic features. As shown in this study, l‐threonine aldolase from Escherichia coli is able to catalyse the same range of reactions catalysed by SHMT, with the exception of the serine hydroxymethyltransferase reaction. This observation strongly suggests that SHMT and l‐threonine aldolase are closely related enzymes specialized for different functions. An evolutionary analysis of the fold type I enzymes revealed that SHMT and l‐threonine aldolase may actually belong to a subgroup of closely related proteins; fungal alanine racemase, an extremely close relative of l‐threonine aldolase, also appears to be a member of the same subgroup. The construction of three‐dimensional homology models of l‐threonine aldolase from E. coli and alanine racemase from Cochliobolus carbonum, and their comparison with the SHMT crystal structure, indicated how the tetrahydrofolate binding site might have evolved and offered a starting point for further investigations.
Evaluation of two biosynthetic pathways to δ‐aminolevulinic acid in Euglena gracilis
Authors: Iida Katsumi , Mimura Ippei , Kajiwara Masahiro
Published: March 2003

δ‐Aminolevulinic acid (ALA), which is an intermediate in the biosynthesis of chlorophyll a, can be biosynthesized via the C5 pathway and the Shemin pathway in Euglena gracilis. Analysis of the 13C‐NMR spectrum of 13C‐labeled methyl pheophorbide a, derived from 13C‐labeled chlorophyll a biosynthesized from d‐[1‐13C]glucose by E. gracilis, provided evidence suggesting that ALA incorporated in the 13C‐labeled chlorophyll a was synthesized via both the C5 pathway and the Shemin pathway in a ratio of between 1.5 and 1.7 to one. The methoxyl carbon of the methoxycarbonyl group at C‐132 of chlorophyll a was labeled with 13C. The phytyl moiety of chlorophyll a was labeled on C‐P2, C‐P31, C‐P4, C‐P6, C‐P71, C‐P8, C‐P10, C‐P111, C‐P12, C‐P14, C‐P151 and C‐P16.