bluuebunny/crossref_metadata_2025 huggingface dataset:
Anticancer activity of RecQL1 helicase siRNA in mouse xenograft models
Authors: Futami Kazunobu , Kumagai Emi , Makino Hiroshi , Sato Ayumi , Takagi Motoki , Shimamoto Akira , Furuichi Yasuhiro
Published: April 2008

Small interfering RNAs (siRNAs) are expected to have a medical application in human therapy as drugs with a high specificity for their molecular target mRNAs. RecQL1 DNA helicase in the human RecQ helicase family participates in DNA repair and recombination pathways in the cell cycle of replication. Silencing the RecQL1 expression by RecQL1‐siRNA induces mitotic death in vitro specifically in growing cancer cells. By contrast, the same RecQL1 silencing does not affect the growth of normal cells, emphasizing that RecQL1 helicase is an ideal molecular target for cancer therapy. In this study, we show that local and systemic administration of RecQL1‐siRNA mixed with polyethyleneimine polymer or cationic liposomes prevented cancer cell proliferation in vivo in mouse models of cancer without noticeable adverse effects. The results indicate that RecQL1‐siRNA in a complex with a cationic polymer is a very promising anticancer drug candidate, and that in particular, RecQL1‐siRNA formulated with a cationic liposome has an enormous potential to be used by intravenous injection for therapy specific for liver cancers, including metastasized cancers from the colon and pancreas. (Cancer Sci 2008; 99: 1227–1236)
THIS ARTICLE HAS BEEN RETRACTED  Epigenetic regulator polycomb group protein complexes control cell fate and cancer
Authors: Kanno Rieko , Janakiraman Harinarayanan , Kanno Masamoto
Published: April 2008

The chromatin‐associated Polycomb group (PcG) proteins were first identified in genetic screens for homeotic transformations in Drosophila melanogaster. Besides body patterning, members of the PcG are now known to regulate epigenetic cellular memory, stem cell self‐renewal, and cancer development. Here, we discuss the multifarious functions of the PcG family, isoforms of protein complexes, and its enzymatic activities, for example histone methylation, links to DNA methylation, its phosphorylation status, H2A mono‐ubiquitination, SUMOylation, and links to non‐coding RNA. We also discuss the function of cytosolic PcG complexes as a regulator of receptor‐induced actin polymerization and proliferation in a methylation‐dependent manner. We propose that the functional versatility of PcG protein complexes contributed significantly to the complexity of heritable gene repression mechanisms, signal transduction, and cell proliferation in cancer development. (Cancer Sci 2008; 99: 1077–1084)
Role of human T‐cell leukemia virus type I Tax in expression of the human telomerase reverse transcriptase (hTERT) gene in human T‐cells
Authors: Hara Toshifumi , Matsumura‐Arioka Yuuki , Ohtani Kiyoshi , Nakamura Masataka
Published: April 2008

The viral product Tax encoded by human T‐cell leukemia virus type I (HTLV‐I) is thought to play a central role in leukemogenesis. Clonal expansion of HTLV‐I‐infected cells requires the extension of cell division with telomere maintenance, which is regulated by the ribonucleoprotein enzyme telomerase. However, the roles of Tax in the expression of telomerase activity in T‐cells remains controversial. Our previous study indicated that expression of the human telomerase reverse transcriptase subunit (hTERT) gene, which determines telomerase activity, is tightly regulated in human T‐cells. In the present study, we investigated Tax‐mediated regulation of hTERT gene expression by Tax in human T‐cells. HTLV‐I Tax induced expression of the hTERT gene in human peripheral blood leukocytes. Reporter assays revealed that Tax activated the hTERT promoter in quiescent Kit 225 cells, while the promoter activity was repressed by Tax in proliferating Jurkat cells. Both up‐regulation and down‐regulation by Tax were mediated through the 43‐bp sequences in the promoter, which carried at least two elements that independently functioned as repressors. The two elements bound distinct factors. G1 to S phase transition induced by introduction of either cyclin D2 with cdk4 or p130‐specific shRNA also activated the hTERT promoter, implying that activation of the hTERT promoter in quiescent Kit 225 cells is associated with cell cycle progression. Our findings suggest that the cell cycle state critically influences Tax‐mediated regulation of hTERT expression. (Cancer Sci 2008; 99: 1155–1163)
Overexpression of calcineurin B subunit (CnB) enhances the oncogenic potential of HEK293 cells
Authors: Wang Yan‐Li , Wang Ying , Tong Li , Wei Qun
Published: April 2008

Calcineurin (CaN) is a Ca2+/calmodulin (CaM)‐stimulated protein phosphatase. It is a heterodimeric enzyme consisting of a catalytic A subunit (CnA) and a Ca2+‐binding regulatory B subunit (CnB). CaN's key role in vivo is well known, while the function of CnB keeps unclear except that it acts as a regulator of CaN. The present paper shows that CnB overexpression promotes proliferation of human embryonic kidney HEK293 cells by comparing with vector control cells in the complete or serum reduced medium. Furthermore, stable CnB transfectants showed dramatically improved growth in soft agar. And the migration ability of CnB overexpressors also was enhanced remarkably. But in the progress of transformation, the activity of CaN remained unchanged between CnB overexpressors and controls. Therefore, CnB, rather than CaN, is involved in the proliferation promotion of HEK293 cells. Subsequently, 11 proteins with different expression levels between CnB transfectants and controls were identified using two‐dimensional gel electrophoresis and electrospray ionization time‐of‐fight mass spectrometry. Therein, the expression of heat shock protein 27 (HSP27) and protein DJ‐1 increased along with CnB overexpression. The important role of CnB in cell neoplastic transformation was found and the possible mechanism was analyzed. (Cancer Sci 2008; 99: 1100–1108)
Plasma matrix metalloproteinase‐7 as a metastatic marker and survival predictor in patients with renal cell carcinomas
Authors: Ramankulov Azizbek , Lein Michael , Johannsen Manfred , Schrader Mark , Miller Kurt , Jung Klaus
Published: April 2008

We evaluated the clinical usefulness of plasma matrix metalloproteinase‐7 (MMP‐7) as a diagnostic and prognostic biomarker in patients with renal cell carcinoma (RCC). MMP‐7 was quantified in plasma of 50 healthy subjects and 97 RCC patients using a Fluorokine MultiAnalyte Profiling assay. RCC patients were stratified into the following groups: without metastases (N0M0; n = 39), with lymph nodes (N1M0; n = 13), and with distant metastases (M1; n = 45). Diagnostic performance of MMP‐7 was analyzed by the receiver operating characteristics (ROC) curve. Kaplan–Meier analysis and the Cox regression model were used to estimate the impact of MMP‐7 on the cancer‐specific survival outcome of RCC patients. MMP‐7 was significantly higher in both metastatic groups N1M0 and M1 (medians, 3.82 and 3.34 µg/L) compared to N0M0 group or controls (medians, 1.85 and 1.64 µg/L; all P < 0.001). In ROC analysis, the area under the ROC curve of MMP‐7 was 0.80 in the detection of metastases in RCC (P < 0.0001). In the Kaplan–Meier analysis, patients with MMP‐7 above the 95th percentile of controls showed less favorable survival rates compared to those with normal MMP‐7 (log‐rank test, 15.7; P < 0.0001). High MMP‐7 was associated with cancer‐related mortality estimated by univariate Cox regression (risk ratio, 4.34, 95% CI, 1.12–10.6; P = 0.032). The multivariate Cox regression model determined MMP‐7 (risk ratio, 2.70, 95% CI, 1.39–5.24; P = 0.003) and metastases (risk ratio, 5.81, 95% CI, 2.77–12.2; P < 0.0001) as independent determinants of cancer‐related survival outcomes. In conclusion, increased plasma MMP‐7 could be related to metastatic disease and poor prognosis in patients with RCC. (Cancer Sci 2008; 99: 1188–1194)
Functional analysis of single amino‐acid mutations in the thrombopoietin‐receptor Mpl underlying congenital amegakaryocytic thrombocytopenia
Authors: Tijssen Marloes R. , Di Summa Franca , Van Den Oudenrijn Sonja , Zwaginga Jaap Jan , Van Der Schoot C. Ellen , Voermans Carlijn , De Haas Masja
Published: April 2008

Congenital amegakaryocytic thrombocytopenia (CAMT) is a rare disorder that presents with severe thrombocytopenia and absence of megakaryocytes in the bone marrow. The disease may develop into bone marrow aplasia. Genetic defects in the gene encoding the thrombopoietin (Tpo) receptor, MPL, are the cause of this disease. In a previous study, we identified four missense mutations in CAMT patients, predicting Arg102Pro, Pro136His, Arg257Cys and Pro635Leu. To investigate whether these mutations result in defective Tpo‐binding and/or signalling, full‐length wildtype and mutant MPL were transduced into K562 cells. Expression levels and the ability to activate the mitogen‐activated protein kinase, Janus kinase‐signal transducer and activator of transcription and phosphoinositide‐3 kinase pathways upon Tpo‐binding were studied. The results predicted that MPL carrying the P136H or P635L mutation was not properly expressed, whereas the R102P and R257C mutations resulted in impaired signal transduction. Our results indicate that a severe clinical course may be expected when these mutations lead to absent Mpl expression or signalling in CAMT patients with missense mutations.
Developing idiotype vaccines for lymphoma: from preclinical studies to phase III clinical trials
Authors: Park Hyun Jun , Neelapu Sattva S.
Published: April 2008

Therapeutic vaccines for B‐cell non‐Hodgkin lymphoma (NHL) using the clonal tumour immunoglobulin idiotype (Id) have been under development for more than three decades. A major obstacle for rapid progress in the field has been that the Id vaccine is patient‐specific and required the generation of a custom‐made product. The manufacturing issues were recently overcome by advances in hybridoma and recombinant DNA technology which facilitated the completion of several phase I and II clinical trials. The strong immunogenicity and apparent clinical benefit observed on the early phase studies led to the initiation of three randomized phase III clinical trials that are also nearing completion. This review will focus on the development of Id vaccines before and after the introduction of rituximab for the treatment of B‐cell NHL and also discuss potential strategies to enhance the efficacy of active immunotherapy in the future.
Results of bone marrow examination in 275 patients with histological features that suggest an indolent type of cutaneous B‐cell lymphoma
Authors: Senff Nancy J. , Kluin‐Nelemans Hanneke C. , Willemze Rein
Published: April 2008

Whether or not bone marrow biopsies should be performed routinely in patients with skin lesions that show histological features consistent with an indolent B‐cell lymphoma [marginal zone lymphoma (MZL) or follicle centre lymphoma (FCL)] has been debated. As no studies have addressed this question for this group of lymphomas, we evaluated the results of bone marrow biopsy examination in 275 patients with histological features consistent with MZL (n = 82) or FCL (n = 193) first presenting in the skin. In the MZL group, two of 82 patients (2%) showed bone marrow involvement, which was the only extracutaneous localization in one of these patients. In the FCL group, 22 of 193 patients (11%) had bone marrow involvement and was the only extracutaneous localization in nine of them. FCL patients with skin lesions and a positive bone marrow had a significantly worse prognosis when compared with patients with only skin lesions (5‐year disease‐specific survival 63% vs. 95%; P = 0·001). These results indicate that bone marrow investigation is essential for staging patients with a FCL first presenting in the skin. Bone marrow examination appears to have limited value in patients with MZL presenting in the skin.
Triiodothyronine stimulates hepatocyte proliferation in two models of impaired liver regeneration
Authors: Columbano A. , Simbula M. , Pibiri M. , Perra A. , Deidda M. , Locker J. , Pisanu A. , Uccheddu A. , Ledda‐Columbano G. M.
Published: April 2008

Abstract.  Objectives: Liver regeneration is attenuated in old age and is substantially slower after 90% than after 70% partial hepatectomy (PH). We have previously demonstrated that the proliferative response to a primary mitogen is intact in aged mice, indicating that impaired liver regeneration is not due to loss of proliferative capacity. Here, we have investigated whether mitogenic effects of triiodothyronine (T3) could reverse the impaired regeneration of ageing or 90% hepatectomy, in the rat. Materials and methods: T3 (20 µg/100 g body weight) was administered to 14‐month‐old rats subjected to 70% PH or to young rats subjected to 90% PH. Cell‐proliferative capacity was determined by bromodeoxyuridine incorporation and microscopy and changes of cell cycle‐related proteins were analysed by Western blot analysis. Results: Treatment of old intact rats with T3 increased cyclin D1 expression that was followed by an enhanced proliferative response, the labelling index (LI), being 7.8% versus 1.3% of controls. T3 given before 70% PH stimulated regenerative response (LI was 10.8% versus 2.28%), and expression of cyclin D1 and proliferating cell nuclear antigen (PCNA) 24 h after PH. Pre‐treatment with T3 also improved the regenerative response of the liver after 90% hepatectomy (LI was 27.9% versus 14.2%). Conclusions: These findings show in principle that mitogen‐induced hyperplasia could be applied to human therapy in patients with reduced regenerative capacity or massive loss of hepatocytes.
Allergenicity and cross‐reactivity of Senecio pollen: identification of novel allergens using the immunoproteomics approach
Authors: Luengo O. , Mollá R. , Gámez C. , Cardona V. , López E. , Sastre B. , Waisel Y. , Belmonte J. , Cadahia Á. , Lahoz C. , Del Pozo V.
Published: April 2008

Background
The genus Senecio is the largest genus of the family Asteraceae (Compositae).

The allergenicity of Senecio has not been assessed previously.

Objective
The aim of this study was to investigate the allergens of Senecio jacobea pollen and to determine their immunological characteristics and clinical relevance.

Methods
Fifty patients with rhinoconjunctivitis and a positive skin prick test (SPT) to Senecio were recruited. The clinical relevance of this pollen was assessed by means of a nasal provocation test (NPT). Allergens were characterized by one‐dimensional electrophoresis (SDS‐PAGE) and two‐dimensional gel electrophoresis and immunoblotting. Furthermore, characterization and identification of the allergens were performed by mass spectrometry (MS). In vitro inhibition tests were performed to evaluate cross‐reactivity with other pollen.

Results
Three predominant allergens, both in the intensity of reaction and the frequency of recognition by human‐allergic sera, were 59 (60%), 42 (50%) and 31 kDa (50%). The two‐dimensional analysis allowed the identification of several allergens. One spot around 42 kDa was identified as a protein homologous to pectate lyase and three other spots were homologous to malate dehydrogenase by MS. S. jacobea proteins showed cross‐reactivity with other proteins of the Asteraceae family and also with Parietaria judaica. This was demonstrated by immunoblotting and ELISA inhibition studies.

Conclusion
S. jacobea constitute a newly discovered allergenic source. It shows cross‐reactivity with other members of the Asteraceae plant family as well as with P. judaica.
Cyclooxygenase‐2 gene polymorphisms in an Australian population: association of the −1195G > A promoter polymorphism with mild asthma
Authors: Shi J. , Misso N. L. , Kedda M.‐A. , Horn J. , Welch M. D. , Duffy D. L. , Williams C. , Thompson P. J.
Published: April 2008

Background
Cyclooxygenase (COX)‐2 is an inducible enzyme responsible for catalysing the formation of prostaglandins (PGs) in settings of inflammation. Single nucleotide polymorphisms (SNPs) of the COX‐2 gene may influence gene transcription and PG production in the asthmatic airway.

Objective
To evaluate the frequencies of COX‐2 SNPs in an Australian Caucasian population, and determine potential associations between common COX‐2 promoter SNPs and asthma, asthma severity and aspirin‐intolerant asthma (AIA).

Methods
The frequencies of 25 COX‐2 SNPs were determined in a random population (n=176). The SNPs with a minor allele frequency of >10% were then studied in asthmatic (n=663), non‐asthmatic controls (n=513) and AIA subjects (n=58). Genotype, allele and haplotype associations were assessed. Functional assessment of SNPs was performed by transfection into HeLa cells measured using the luciferase dual‐reporter assay system.

Results
Eighteen COX‐2 SNPs were not detected, five were rare and two promoter SNPs, −1195G>A (rs689465), and −1290A>G (rs689466), were further studied. The A allele of the −1195 SNP was present at a significantly higher frequency among all asthmatic subjects (P=0.012). Over 60% of the asthmatic individuals were −1195A homozygotes compared with 54.6% of the control subjects (odds ratio, 1.35; 95% CI, 1.06–1.72, P=0.03). After classifying for severity, the mild asthmatics represented 64.6% of −1195AA individuals, the highest of all the asthma groups compared with 54.6% of the control subjects (odds ratio, 1.5; 95% CI, 1.12–2.02, P=0.02). The −1290A/−1195G/−765G haplotype was associated with a reduced incidence of asthma (odds ratio, 0.76; 95% CI, 0.61–0.95, P=0.017).

Conclusion
The −1195G>A polymorphism appears to be associated with asthma, and in particular with mild asthma.
Secondary hyperparathyroidism in primary osteoporosis and osteopenia: optimizing calcium and vitamin D intakes to levels recommended by expert panels may not be sufficient for correction
Authors: Yendt Edmund R. , Kovacs Katherine A. , Jones Glenville
Published: April 2008

Objective  To compare biochemical variables, renal function and calcium and vitamin D intakes in euparathyroid and hyperparathyroid patients with primary osteoporosis and osteopenia and describe the measures necessary to normalize serum PTH in the patients with secondary hyperparathyroidism.

Design and patients  We reviewed the charts of normocalcemic patients with primary osteoporosis and osteopenia first seen during the years 1991–2003 and identified 75 with elevated serum PTH levels at baseline. These patients were compared to all the 143 euparathyroid patients first seen in 1998 and 1999. Patients were restudied after 1 year and we attempted to follow patients with secondary hyperparathyroidism until PTH levels became normal.

Measurements  At baseline serum PTH, ionized calcium, inorganic phosphate, alkaline phosphatase, creatinine, a complete blood count and serum 25 hydroxy vitamin D were measured in the early morning fasting state. These tests were repeated at follow up.

Results  In one‐third of the hyperparathyroid patients, the standard baseline treatment failed to correct the secondary hyperparathyroidism necessitating extraordinary measures including unusually large doses of vitamin D (i.e. 50 000 IU vitamin D2 twice weekly) or the substitution of calcium citrate for calcium carbonate as a calcium supplement.

Conclusion  Large doses of vitamin D are frequently necessary to suppress secondary hyperparathyroidism in patients with primary osteoporosis and osteopenia. This suggests that vitamin D metabolism may be altered in some of these patients.
Relationship between calcium absorption and plasma dehydroepiandrosterone sulphate (DHEAS) in healthy males
Authors: Chen R. Y. T. , Nordin B. E. C. , Need A. G. , Scopacasa F. , Wishart J. , Morris H. A. , Horowitz M.
Published: April 2008

Context  Impaired gut sensitivity to 1,25‐dihydroxyvitamin D (1,25(OH)2D), leading to reduced intestinal calcium absorption, has been reported in older men and women. While this phenomenon in postmenopausal women has been attributed to oestrogen deficiency, it is unclear whether the same observation in older men correlates with the age‐related decline in androgen concentrations.

Objective  To examine the relationship between androgens and intestinal calcium absorption in older men.

Design  Cross‐sectional study on 55 healthy male volunteers, divided into younger (n = 27) and older (n = 28) groups separated according to the median age of 59 years.

Main outcome measures  Calcium absorption, total and free (calculated) testosterone, dehydroepiandrosterone sulphate (DHEAS), SHBG, and 1,25(OH)2D, among others, were measured.

Results  Calcium absorption, free testosterone and DHEAS, but not 1,25(OH)2D, declined significantly with age. After adjusting for age and body mass index, stepwise regression showed that 1,25(OH)2D and serum albumin were the only significant determinants of calcium absorption in younger men, while the sole determinant in older men was DHEAS, not testosterone. Residual deviations from the regression of calcium absorption on 1,25(OH)2D, reflecting the efficiency of 1,25(OH)2D‐induced calcium absorption, was positively correlated with DHEAS (r = 0·27, P = 0·027).

Conclusions  DHEAS is an independent determinant of calcium absorption in older men, although its manner of influence is, as yet, undefined. The age‐related decline of DHEAS may, partly, account for the observed ‘intestinal resistance to 1,25(OH)2D’ in older men.
Clinical manifestations of familial paraganglioma and phaeochromocytomas in succinate dehydrogenase B (SDH‐B) gene mutation carriers
Authors: Srirangalingam Umasuthan , Walker Lisa , Khoo Bernard , MacDonald Fiona , Gardner Daphne , Wilkin Terence J. , Skelly Robert H. , George Emad , Spooner David , Monson John P. , Grossman Ashley B. , Akker Scott A. , Pollard Patrick J. , Plowman Nick , Avril Norbert , Berney Daniel M. , Burrin Jacky M. , Reznek Rodney H. , Kumar V.K. Ajith , Maher Eamonn R. , Chew Shern L.
Published: April 2008

Objective  Phaeochromocytomas and paragangliomas are familial in up to 25% of cases and can result from succinate dehydrogenase (SDH) gene mutations. The aim of this study was to describe the clinical manifestations of subjects with SDH‐B gene mutations.

Design  Retrospective case‐series.

Patients  Thirty‐two subjects with SDH‐B gene mutations followed up between 1975 and 2007. Mean follow‐up of 5·8 years (SD 7·4, range 0–31). Patients seen at St Bartholomew's Hospital, London and other UK centres.

Measurements  Features of clinical presentation, genetic mutations, tumour location, catecholamine secretion, clinical course and management.

Results  Sixteen of 32 subjects (50%) were affected by disease. Two previously undescribed mutations in the SDH‐B gene were noted. A family history of disease was apparent in only 18% of index subjects. Mean age at diagnosis was 34 years (SD 15·4, range 10–62). 50% of affected subjects had disease by the age of 26 years. 69% (11 of 16) were hypertensive and 80% (12 of 15) had elevated secretions of catecholamines/metabolites. 24% (6 of 25) of tumours were located in the adrenal and 76% (19 of 25) were extra‐adrenal. 19% (3 of 16) had multifocal disease. Metastatic paragangliomas developed in 31% (5 of 16). One subject developed a metastatic type II papillary renal cell carcinoma. The cohort malignancy rate was 19% (6 of 32). Macrovascular disease was noted in two subjects without hypertension.

Conclusion  SDH‐B mutation carriers develop disease early and predominantly in extra‐adrenal locations. Disease penetrance is incomplete. Metastatic disease is prominent but levels are less than previously reported. Clinical manifestations may include papillary renal cell carcinoma and macrovascular disease.
Incidence of radiation thyroiditis and thyroid remnant ablation success rates following 1110 MBq (30 mCi) and 3700 MBq (100 mCi) post‐surgical 131I ablation therapy for differentiated thyroid carcinoma
Authors: Cherk Martin H. , Kalff Victor , Yap Kenneth S. K. , Bailey Michael , Topliss Duncan , Kelly Michael J.
Published: April 2008

Objective  To evaluate the relationships between thyroid remnant 131I uptake, radiation thyroiditis and remnant ablation success rate between lower (1110 MBq) and higher (3700 MBq) initial ablative 131I dose for post‐surgical ablation therapy for differentiated thyroid cancer.

Design  Patients having post‐surgical administration of 1110 MBq (68 patients) or 3700 MBq (115 patients) 131I were retrospectively reviewed. Thyroid remnant 131I uptake on a 48 h post‐administration scan was correlated with neck symptoms experienced. Patients were classified as having insignificant, mild or severe thyroiditis based on symptoms. Absent thyroid bed 131I uptake on a follow‐up 74 MBq 131I study was considered successful ablation.

Results  183 patients were included. Median 131I remnant uptake was 37 MBq. 21% (39/183) of patients developed thyroiditis. Incidence and severity of thyroiditis increased with increasing remnant 131I activity (P ≤ 0·0001). Risk of thyroiditis increased by 64% for every 37 MBq increase in remnant 131I uptake. OR = 1·64 [1·36–1·99]. Severe thyoriditis was only seen with remnant 131I uptake > 73 MBq. For patients treated with 1110 MBq and 3700 MBq, incidence of thyroiditis was 12% and 27% (P = 0·02) and remnant ablation success rate was 76% and 84% (P = NS), respectively. Occurrence of thyroiditis did not correlate with successful ablation.

Conclusions  Incidence and severity of radiation thyroiditis following 131I remnant ablation therapy is directly related to thyroid remnant 131I uptake. As 1110 MBq 131I is associated with a significantly lower frequency of thyroiditis but similar remnant ablation rate to 3700 MBq, it warrants consideration for thyroid remnant ablation particularly in patients with low risk disease.
Experimental haemophilic arthropathy in a mouse model of a massive haemarthrosis: gross, radiological and histological changes
Authors: HAKOBYAN N. , ENOCKSON C. , COLE A. A. , SUMNER D. R. , VALENTINO L. A.
Published: April 2008

Summary.  Recurrent haemarthrosis results in chronic synovitis and destructive arthropathy. The long‐term effect of a single haemorrhage is not known. To investigate the histopathological changes following a single, but major joint haemorrhage, an animal model of massive haemarthrosis without mechanical trauma was developed and is described in this manuscript. The knee joint capsule of mice deficient in coagulation factor VIII or IX and non‐haemophilic wild type mice was punctured to induce a one time, but massive haemorrhage. The single joint puncture resulted in acute haemarthrosis in both types of haemophilic mice but not in wild type mice. Subsequent to injury, the changes in the knee joints were analysed using gross, histological and radiographic assessments and compared with the uninjured knee. In addition, a novel imaging modality, micro‐computed tomography, was used to document the structural damage to the joint. Our results indicate that the long‐term changes classically observed in patients with advanced haemophilic arthropathy are evident following a single massive haemarthrosis. This model will allow a thorough investigation of the pathobiology of blood‐induced joint disease and will be useful to test the efficacy of innovative therapeutic strategies to prevent haemophilic synovitis and arthropathy.
Guideline on the selection and use of therapeutic products to treat haemophilia and other hereditary bleeding disorders
Authors: KEELING D. , TAIT C. , MAKRIS M.
Published: April 2008

Summary.  Evidence‐based guidelines are presented on the selection and use of therapeutic products to treat haemophilia and other hereditary bleeding disorders. They include details of therapeutic products available in the UK and they update and replace previous United Kingdom Haemophilia Centre Doctors’ Organisation guidelines.
Apoptotic cells induce dendritic cell‐mediated suppression via interferon‐γ‐induced IDO
Authors: Williams Charlotte A. , Harry Rachel A. , McLeod Julie D.
Published: April 2008

Dendritic cells (DC) are sensitive to their local environment and are affected by proximal cell death. This study investigated the modulatory effect of cell death on DC function. Monocyte‐derived DC exposed to apoptotic Jurkat or primary T cells failed to induce phenotypic maturation of the DC and were unable to support CD4+ allogeneic T‐cell proliferation compared with DC exposed to lipopolysaccharide (LPS) or necrotic cells. Apoptotic cells coincubated with LPS‐ or necrotic cell‐induced mature DC significantly suppressed CD80, CD86 and CD83 and attenuated LPS‐induced CD4+ T‐cell proliferation. Reduced levels of interleukin‐12 (IL‐12), IL‐10, IL‐6, tumour necrosis factor‐α and interferon‐γ (IFN‐γ) were found to be concomitant with the suppressive activity of apoptotic cells upon DC. Furthermore, intracellular staining confirmed IFN‐γ expression by DC in association with apoptotic environments. The specific generation of IFN‐γ by DC within apoptotic environments is suggestive of an anti‐inflammatory role by the induction of indoleamine 2,3‐dioxygenase (IDO). Both neutralization of IFN‐γ and IDO blockade demonstrated a role for IFN‐γ and IDO in the suppression of CD4+ T cells. Moreover, we demonstrate that IDO expression within the DC was found to be IFN‐γ‐dependent. Blocking transforming growth factor‐β (TGF‐β) also produced a partial release in T‐cell proliferation. Our study strongly suggests that apoptosis‐induced DC suppression is not an immunological null event and two prime mediators underpinning these functional effects are IFN‐γ‐induced IDO and TGF‐β.
Differential contribution of TAP and tapasin to HLA class I antigen expression
Authors: Belicha‐Villanueva Alan , McEvoy Sarah , Cycon Kelly , Ferrone Soldano , Gollnick Sandra O. , Bangia Naveen
Published: April 2008

Expression of class I human leucocyte antigens (HLA) on the surface of malignant cells is critical for their recognition and destruction by cytotoxic T lymphocytes. Surface expression requires assembly and folding of HLA class I molecules in the endoplasmic reticulum with the assistance of proteins such as Transporter associated with Antigen Processing (TAP) and tapasin. Interferon‐γ induces both TAP and tapasin so dissection of which protein contributes more to HLA class I expression has not been possible previously. In this study, we take advantage of a human melanoma cell line in which TAP can be induced, but tapasin cannot. Interferon‐γ increases TAP protein levels dramatically but HLA class I expression at the cell surface does not increase substantially, indicating that a large increase in peptide supply is not sufficient to increase HLA class I expression. On the other hand, transfection of either allelic form of tapasin (R240 or T240) enhances HLA‐B*5001 and HLA‐B*5701 antigen expression considerably with only a modest increase in TAP. Together, these data indicate that in the presence of minimal TAP activity, tapasin can promote substantial HLA class I expression at the cell surface.
Importance of CD80/CD86–CD28 interactions in the recognition of target cells by CD8+CD122+ regulatory T cells
Authors: Shi Zhe , Rifa’i Muhaimin , Lee Young Ho , Shiku Hiroshi , Isobe Ken‐ichi , Suzuki Haruhiko
Published: April 2008

CD8+CD122+ regulatory T cells are a newly identified, naturally occurring type of regulatory T cell that produce interleukin‐10 (IL‐10) and effectively suppress interferon‐γ (IFN‐γ) production from both CD8+ and CD4+ target cells. Molecular mechanisms responsible for the recognition of target cells by CD8+CD122+ regulatory T cells were investigated in this study by using an in vitro culture system that reconstitutes the regulatory action of these cells. CD8+CD122+ regulatory T cells did not produce IL‐10 and did not suppress the IFN‐γ production of allogeneic target T cells when they were stimulated by immobilized anti‐CD3 antibody alone, but they clearly produced IL‐10 and suppressed the IFN‐γ production of target cells when stimulated by anti‐CD3 plus anti‐CD28‐coated beads. IFN‐γ production by major histocompatibility complex‐class I‐deficient T cells was also suppressed by CD8+CD122+ regulatory T cells stimulated with anti‐CD3 plus anti‐CD28 antibody but was not suppressed by cells stimulated by anti‐CD3 alone. Experiments examining the blockade of cell surface molecules expressed on either the regulatory cells or the target cells by adding specific neutralizing antibodies in the culture indicated that CD80, CD86, and CD28 molecules were involved in the regulatory action, but cytotoxic T lymphocyte antigen‐4, inducible costimulatory molecule (ICOS) and programmed death‐1 (PD‐1) molecules were not. Finally, CD8+CD122+ cells isolated from CD28‐knockout (CD28−/−) mice showed no regulatory activity. These results indicate that CD8+CD122+ regulatory T cells recognize target T cells via the interaction of CD80/CD86–CD28 molecules to become active regulatory cells that produce suppressive factors such as IL‐10.
CD40 Ligand‐activated, antigen‐specific B cells are comparable to mature dendritic cells in presenting protein antigens and major histocompatibility complex class I‐ and class II‐binding peptides
Authors: Ahmadi Tahamtan , Flies Amanda , Efebera Yvonne , Sherr David H.
Published: April 2008

Dendritic cells (DC) are increasingly exploited for cell‐based immunotherapy. However, limitations in ex vivo DC growth and DC functional heterogeneity have motivated development of complementary antigen‐presenting cell sources. Here, the ability of CD40 ligand (CD40L)‐activated B cells to fulfil that role was investigated. We demonstrate for the first time that non‐specific or antigen‐specific murine B cells can be grown for extended periods of time by stimulation with CD40L. These cells rapidly up‐regulate and maintain high levels of co‐stimulatory molecules. In a head‐to‐head comparison with DC, CD40L‐expanded B cells were comparable to DC in the presentation of peptides to CD4+ and CD8+ T cells. While DC were superior to antigen non‐specific CD40L‐activated B cells with regard to whole protein (NP‐BSA) processing and presentation, CD40L‐expanded B cells from NP‐BSA‐immunized mice were comparable to DC when presenting BSA or NP‐BSA to primed primary T cells or when presenting NP linked to an unrelated carrier, CGG, to naïve T cells. Thus, the combination of CD40L activation, which supports B‐cell growth and augments intracellular protein processing, and antigen uptake via the B‐cell receptor, allows for efficient uptake, processing, and presentation of whole protein antigens in a fashion comparable to that observed with mature DC. Like DC, CD40L‐activated B cells efficiently home to secondary lymphoid organs in vivo. This system represents a unique tool for studying primary antigen‐specific B cells and the results suggest that the outgrowth of large numbers of highly activated B cells represents a viable and practical complement to DC for cell‐based immunotherapy.
Pulp stones: a review
Authors: Goga R. , Chandler N. P. , Oginni A. O.
Published: April 2008

Pulp stones are a frequent finding on bitewing and periapical radiographs but receive relatively little attention in textbooks. A review of the literature was therefore performed, initially using the PubMed database and beginning the search with ‘pulp calcifications’ and ‘pulp stones’. Each term provided more than 400 references, many of which related to pulp calcification in general rather than pulp stones, and focussed largely on the problems these changes presented to clinicians. A manual search using references from this source was carried out. Contemporary textbooks in endodontology were also consulted, and an historic perspective gained from a number of older books and references. The factors involved in the development of the pulp stones are largely unknown. Further research may determine the reasons for their formation, but with current endodontic instruments and techniques this is unlikely to alter their relevance to clinicians.
Conventional and surgical endodontic retreatment of a maxillary lateral incisor with unusual anatomy
Authors: Karabucak B. , Ishii H. , Kratchman S. I.
Published: April 2008

Aim  To describe conventional and surgical endodontic treatment of a maxillary right lateral incisor with an unusual root dilaceration and to discuss the treatment outcome.

Summary  Root dilacerations refer to severe curvature or angulations that may occur anywhere along the length of a root. This anatomical deviation may present an unfavourable outcome if clinicians fail to recognize the difficulties. The following case report describes conventional and surgical endodontic treatment of a maxillary right lateral incisor with an unusual root dilaceration.

Key learning points 

Root morphology may present a challenge for clinicians during root canal treatment.

Clinicians should be aware of the importance of surgical endodontics and be familiar with microsurgery principals and techniques.
Effects of post system and length on the strain and fracture resistance of root filled bovine teeth
Authors: Santos‐Filho P. C. F. , Castro C. G. , Silva G. R. , Campos R. E. , Soares C. J.
Published: April 2008

Aim  To investigate ex vivo the effects of different post systems and lengths on the strain and fracture resistance of root‐filled teeth.

Methodology  One hundred and thirty‐five bovine incisors were sectioned 15 mm from their apices, root‐filled and divided into three groups (n = 45): fibreglass post; prefabricated steel post; cast post and core. Each group was divided into three subgroups (n = 15) according to the post length: 5.0 mm; 7.5 mm; 10.0 mm. All teeth were restored with metal crowns. For strain‐gauge measurement, two strain gauges per sample were used. The fracture resistance (N) was measured and the data were analysed with two‐way analyses of variance, followed by the Tukey’s HSD test (α = 05).

Results  For all posts, decreased lengths resulted in increased microstrain values. However, the fibreglass posts were associated with lower increases when compared with cast post and cores and prefabricated steel posts, which showed microstrain values two times higher when the post length was 5.0 mm. The two‐way analyses of fracture resistance values revealed that post length was statistically significant for the metal posts and not significant for the fibreglass post. The fracture mode analysis indicated that all groups tended to demonstrate root fractures in groups restored with metal posts and resin core fractures in groups restored with fibreglass posts.

Conclusions  The cast post and core when the length was 10.0 mm had the highest fracture resistance; however, the fibreglass post was effective with the three post lengths, with higher fracture resistance than metal posts when the length was 5.0 mm.
Microbiological evaluation of primary endodontic infections in teeth with and without sinus tract
Authors: Sassone L. M. , Fidel R. , Faveri M. , Fidel S. , Figueiredo L. , Feres M.
Published: April 2008

Aim  To examine the microbiological status of primary endodontic infections in teeth with and without a sinus tract.

Methodology  Samples were collected by means of a size 15 H‐type file and two sterile paper points from 30 cases of primary endodontic infections with (n = 15) or without (n = 15) a sinus tract. The presence of 40 bacterial species was determined by the checkerboard DNA‐DNA hybridization method.

Results  The species found at the highest levels and prevalence were Fusobacterium nucleatum sp. vincentii, Porphyromonas gingivalis, Veillonella parvula, Enterococcus faecalis, Campylobacter gracilis and Neisseria mucosa. Total bacterial counts were similar between teeth with (44 × 105) and without (50 × 105) a sinus tract (t‐test: P > 0.05). E. faecalis, Streptococcus anginosus, Capnocytophaga sputigena and Capnocytophaga gingivalis had significantly higher counts in the absence of sinus tract (Mann–Whitney test, P < 0.05). Higher levels of P. gingivalis and Fusobacterium nucleatum sp. nucleatum were observed in cases with a sinus tract. Leptotrichia buccalis (OR = 1.83; CI 95%) and Porphyromonas endodontalis (OR = 2.15; CI 95%) were associated with an increased chance of subjects having a sinus tract.

Conclusions  Primary endodontic infections were associated with a large variety of bacterial species. Specific differences between the composition of the microbiota of primary root canal infections were observed in cases with or without a sinus tract.
Spatial and temporal patterns of phenotypic variation in a Neotropical frog
Authors: Robertson J. M. , Robertson A. D.
Published: April 2008

Aim  Studies of the spatial and temporal patterns of phenotypic diversity help to elucidate the fine‐scale evolutionary and ecological mechanisms underlying geographical differentiation. The red‐eyed tree frog, Agalychnis callidryas, is a widespread Neotropical frog that exhibits a broad range of polymorphism of coloration and flank‐stripe pattern. The goal of this study was two‐fold: first, to investigate the stability of polymorphisms over a 38‐year period; and second, to evaluate biogeographical hypotheses of diversification between lower Central American populations through quantification of phenotypic diversity on a fine geographical scale.

Location  This study was conducted at 12 sites across four biogeographical regions in Costa Rica and Panama.

Methods  We quantified colour, categorized flank‐stripe pattern from digital photos taken during field sampling, and measured body size for each individual. We compared the regional frequency of each flank‐stripe pattern in 2005 with the frequency distribution from a previous study of the same sites in 1967 using logistic regression analyses. We determined the geographical signal of leg coloration by employing linear discriminant function analyses to generate a classification matrix based on covariance similarities, and by comparison of the average hue values within and between regions.

Results  We found a temporal shift in the frequency of flank‐stripe patterns in three of four regions over 38 years. Based on measures of leg coloration, the frequency distribution of flank‐stripe patterns and body size, we conclude that A. callidryas populations are easily distinguishable at a regional scale.

Main conclusions  Agalychnis callidryas exhibits regional differentiation in all phenotypic traits measured in this study, supporting the role of three major biogeographical barriers to gene exchange. We found evidence of a putative contact zone between polytypic regions in Costa Rica. In addition, we report temporal instability of the relative frequency of stripe patterns located on the flanks. The ecological and evolutionary mechanisms that may underlie this variation include sexual selection and avoidance of predators.
Post‐glacial range fragmentation is responsible for the current distribution of Potentilla matsumurae Th. Wolf (Rosaceae) in the Japanese archipelago
Authors: Ikeda Hajime , Senni Kei , Fujii Noriyuki , Setoguchi Hiroaki
Published: April 2008

Aim  We aimed to elucidate how the current geographic distribution of alpine plants in the Japanese archipelago was shaped during Quaternary climatic oscillations, using Potentilla matsumurae as a case study. According to previous phylogeographic studies, post‐glacial range fragmentation (vicariance scenario) and stepwise migration (dispersal scenario) are both possible. We thus aimed to assess which scenario is more probable for the distribution changes of alpine plants in the Japanese archipelago.

Location  The alpine zone in the Japanese archipelago.

Methods  Using amplified fragment length polymorphism we determined the genotype of 161 individuals of P. matsumurae from 22 populations. Relationships among individuals and populations were examined using principal coordinates analysis and a neighbour‐joining (NJ) tree, respectively. To examine the genetic population structure, we performed analysis of molecular variance (amova) and structure analysis.

Results  Differentiation between central Honshu and northern Japan was not very strong based on the principal coordinates analysis among individuals, the NJ tree of populations (59% bootstrap support), or amova (12% of genetic variation). Moreover, structure analysis did not detect clear geographic differentiation across populations. Although the populations in central Honshu were structured geographically (Mantel test: r = 0.45, P < 0.005; NJ tree), those in northern Japan did not exhibit geographic structure regardless of geographic distance (Mantel test: r = 0.26, P = 0.03; NJ tree). Population relationships in the NJ tree did not always reflect the geographic location.

Main conclusions  The current geographic structure of P. matsumurae could not be explained by stepwise migration. This suggests that a single continuous distribution during the last glacial period was later fragmented, perhaps by recovering forest, during the post‐glacial period, resulting in the current distribution and phylogeographic structure of P. matsumurae. Our data support the vicariance scenario.
The ‘oriental’ component of the Balkan flora: evidence of presence on the Thracian Plain during the Weichselian late‐glacial
Authors: Magyari E. K. , Chapman J. C. , Gaydarska B. , Marinova E. , Deli T. , Huntley J. P. , Allen J. R. M. , Huntley B.
Published: April 2008

Aim  To obtain palaeobotanical evidence enabling evaluation of the viability of the hypothesis that the ‘oriental’ element of the Balkan flora reached south‐east Europe from Turkey prior to the Holocene, probably via the Thracian Plain during a late Quaternary glacial stage but no later than the late Weichselian.

Location  Ezero wetland, northern Thracian Plain, Bulgaria.

Methods  We undertook analyses of pollen and microspores, plant macrofossils, wood fragments and molluscs recovered from sediments deposited in the Ezero wetland during the late Weichselian and Weichselian late‐glacial. Sediment chronology was determined using radiocarbon age estimates.

Results  Six metres of sediments were recovered from the basin, of which the lower 3 m, extending from c. 15,450 cal yr bp to the early Allerød, was analysed. A major hiatus occurred after c. 13,900 cal yr bp, the overlying sediments being of late Holocene age. Palaeobotanical evidence indicates predominantly open vegetation during the Weichselian late‐glacial, although macrofossil remains of woody taxa demonstrate the local presence of patches of wooded steppe and gallery forest. Changes in the composition of the steppe vegetation, and in the nature of the sediments deposited in the basin, indicate changes in climatic conditions, especially in the hydrological regime and in the moisture available to vegetation. After an initially relatively moister phase, the final centuries of the late Weichselian were drier, as was a short interval that may correlate with the Older Dryas. Moister conditions characterize intervals corresponding to the Bølling and Allerød sub‐units of the Weichselian late‐glacial interstadial. Although the pollen evidence is thus consistent with that from previous studies of this period in south‐east Europe and south‐west Asia, indicating predominantly open steppe vegetation, the macrofossil evidence indicates the persistent local presence of woody taxa. The woody taxa recorded include Celtis tournefortii‐type and Juniperus cf. J. excelsa, two taxa today characteristic of the wooded steppes of Anatolia and members of the ‘oriental’ element of the southern Balkan flora, as well as Rosaceae Subfams. Maloideae and Prunoideae, Alnus and Fraxinus.

Main conclusions  The late Weichselian vegetation of the northern Thracian Plain included patches of wooded steppe that supported members of the ‘oriental’ element of the modern Balkan flora. The presence of such taxa renders viable the hypothesis that they could have reached south‐east Europe from Turkey via the Thracian Plain during glacial times. Such hypotheses in historical biogeography can be evaluated critically using the evidence obtained from plant macrofossil analyses in combination with that from pollen analysis.