bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel PTEN mutation in a Japanese patient with Cowden disease
Authors: Kubo Y. , Urano Y. , Hida Y. , Ikeuchi T. , Nomoto M. , Kunitomo K. , Arase S. 
Published: March 2003

Abstract
Cowden disease (CD) is an autosomal dominant syndrome characterized by multiple hamartomatous lesions and an increased risk for malignancies. Recent evidence has indicated that the PTEN gene, encoding a protein tyrosine phosphatase, is the CD susceptibility gene. However, another line of evidence has suggested that CD might be genetically heterogeneous. Clinical features of CD are variable, and there are interfamilial differences in the expression of skin lesions . Therefore, information on PTEN mutations in CD patients should be accumulated to clarify the genotype–phenotype correlation. In the present study, we found heterozygous germline mutations of PTEN in all of three Japanese patients with CD examined, indicating no genetic heterogeneity among our patients. The mutations included two non-sense mutations of R335X and R130X, and a mis-sense mutation of C136R. To the best of our knowledge, the C136R mutation has not previously been reported in CD patients. This novel mutation was located outside the core motif of the phosphatase domain of PTEN protein, where most of the missense mutations previously reported in CD patients were clustered. Mucocutaneous manifestations were far fewer in the patient with this mutation than in the patients with nonsense mutations. Whether the phenotypic difference in mucocutaneous features was due to the different mutations remains unclear.
Role for tumour necrosis factor-α receptors in ultraviolet-induced skin tumours
Authors: Starcher B. 
Published: March 2003

Abstract
The biological effects of tumour necrosis factor (TNF)-α are mediated through either the TNFR1 or the TNFR2 receptor. In the present study, the effects of ultraviolet (UV) irradiation on skin pathology and tumour promotion were studied in hairless mice deficient in either the TNFR1 or the TNFR2 receptor. SKH-1 hairless mice were crossed with either TNFR1 knockout (KO) mice or TNFR2 KO mice to develop a strain of hairless mice deficient in either of these receptors. Elastosis and other pathological indications of UVB irradiation were not affected by the loss of either receptor. The absence of either receptor, however, resulted in a highly significant reduction in skin tumours in response to UVB irradiation. Inflammatory cell influx following chronic UV irradiation was virtually eliminated in the TNFR1 KO mice, while the TNFR2 KO mice responded to UV irradiation with the normal increase in inflammatory cells throughout the lower and upper dermis. Contact hypersensitivity responses were eliminated in the TNFR2 KO mice, whereas the TNFR1 KO mice retained normal contact hypersensitivity reactions. These studies suggest that TNF-α plays no part in the accumulation of excessive elastin in the skin during chronic UVB exposure. However, there appears to be an important role for TNF-α in mediating tumorigenesis, distinct from its role in initiating cutaneous immune responses.
Two cases of vulval pigmented extramammary Paget’s disease: histochemical and immunohistochemical studies
Authors: Chiba H. , Kazama T. , Takenouchi T. , Nomoto S. , Yamada S. , Tago O. , Ito M. 
Published: March 2003

Abstract
We describe two Japanese female patients with pigmented extramammary Paget’s disease (EMPD); one patient had a dark brown plaque and the other had a reddish patch with a pigmented area, both affecting the vulval region. Histochemical and immunohistochemical examinations confirmed EMPD with melanocyte colonization; plump tumour cells with a large nucleus and pale cytoplasm that were positive for CAM 5.2 and CEA proliferated singly or in nests in the epidermis, and scattered among the tumour cells were many dendritic cells with a large amount of melanin that were positive for HMB-45 and S-100 protein. Fontana–Masson (FM) stain showed many positive cells with well-developed dendritic processes within and around tumour nests. Histochemical and immunohistochemical studies of non-pigmented EMPD cases on the same region showed that HMB-45 positive cells were sparse or not detected at all, and that also FM staining-positive cells were decreased or not detected, and their dendritic processes were poorly formed. The present study suggests that there might be heterogeneity in EMPD in terms of relationships between Paget’s cells and melanocytes.
A phase II trial of interleukin‐2 in myelodysplastic syndromes
Authors: Nand Sucha , Stock Wendy , Stiff Patrick , Sosman Jeffrey , Martone Brenda , Radvany Ruta 
Published: March 2003

Patients with myelodysplastic syndromes (MDS) show a decrease in the number and function of natural killer (NK) cells, including lymphokine activated killer (LAK) cell activity. Interleukin‐2 (IL‐2) stimulates the proliferation and activity of these lymphocytes. Anecdotal clinical experience has shown haematological and cytogenetic improvement in myelodysplasia by low‐dose IL‐2 treatment.A total of 10 patients with MDS were treated with 1 million units of IL‐2 subcutaneously daily for 12 weeks. Even though improvement in CD16+/CD56+ cell numbers was seen in a majority of the patients, the haematological status and transfusion requirements remained unchanged. There was minimal toxicity from this therapy.
A comparison of artificially‐depleted, lyophilized coumarin and fresh coumarin plasmas in thromboplastin calibration
Authors: Poller  , Van Den Besselaar  , Jespersen  , Tripodi  , Houghton  
Published: March 2003

Artificially‐depleted lyophilized plasmas and lyophilized coumarin plasmas were prepared and compared with fresh coumarin plasmas to assess their comparative reliability in local thromboplastin calibration using the manual prothrombin time (PT) technique. Their certified PT values were inserted in turn on the vertical axis in place of the PT obtained with fresh coumarin plasmas. PT results were obtained at eight ECAA national laboratories (‘test centres’) and inserted on the horizontal axis. The resulting thromboplastin calibration slopes were compared with conventional fresh coumarin plasma calibration slopes at the same ‘test centres’. When 60 artificially‐depleted plasmas were substituted for 60 fresh plasmas, the mean calibration slopes with the human plain International Reference Preparation (IRP) were 4.2% higher. For comparison with 20 lyophilized coumarins, three sets of 20 artificially‐depleted plasmas were selected in sequential order from the 60. The lyophilized coumarin plasmas gave a mean deviation of 9.6% from the fresh plasma calibration slopes compared with values of 2.0%, 6.1% and 11.7% for the three sets of 20 depleted plasmas.Although both types of lyophilized plasma calibration slopes give measurable differences from conventional fresh plasmas, these may be regarded as acceptable in clinical terms.
Paediatric Hodgkin's disease in Spain: association with Epstein‐Barr virus strains carrying latent membrane protein‐1 oncogene deletions and high frequency of dual infections
Authors: SantÓn Almudena , MartĹn Carmen , Manzanal Ana Isabel , Preciado MarĹa Victoria , Bellas Carmen 
Published: March 2003

The present report analyses the distribution of 30‐base pair (bp) latent membrane protein‐1 (LMP‐1) oncogene deletions in 24 cases of Epstein‐Barr virus (EBV)‐positive paediatric Hodgkin's disease (HD) and 39 normal controls.The 30 bp deletion was identified in 19/24 paediatric HD cases (79.2%), of which seven (29.2%) showed the deleted fragment alone, whereas in the remaining 12 (50%) it was accompanied by the nondeleted fragment. Conversely, the deletion was found in 8/22 (36.4%) EBV‐positive healthy children, in two (9.1%) of whom the deleted fragment was alone, and was coinfecting with the nondeleted fragment in the other six (27.3%). The LMP‐1 deletion was significantly associated with paediatric HD, both including dual infections (P = 0.006) or excluding them (P = 0.01).Type 2 EBV was carried by 25% of HD children, whereas all controls harboured type 1 EBV. The 30 bp deletion was present in all the paediatric HD specimens that contained type 2 EBV, suggesting that a deleted type 2 EBV strain may be more tumourigenic than a nondeleted type 2 EBV strain.These findings indicate that EBV strains carrying a 30 bp deletion in the third exon of the LMP‐1 oncogene may have a more important role in the pathogenesis of paediatric HD than full‐length EBV strains. Dual infection by LMP‐1 deleted and nondeleted EBV strains is a frequent event both in healthy children and in the paediatric HD population.
Molecular, haematological and clinical studies of a silent β‐gene C → G mutation at 6 bp 3′ to the termination codon (+1480 C → G) in twelve Greek families
Authors: Maragoudaki  , Vrettou  , Kanavakis  , Traeger‐Synodinos  , Metaxotou‐Mavrommati  , Kattamis  
Published: March 2003

We report the clinical, haematological, biosynthetic and molecular data of 12 β‐thalassaemia intermedia patients and their heterozygous parents, all of whom carried a rare C → G mutation at nucleotide position 6 3′ to the termination codon (term. cd +6 C → G) in the 3′ untranslated region (3′ UTR) of the β‐globin gene (+1480 C → G). This mutation has been reported previously in a single β‐thalassaemia intermedia patient of Greek origin. The 12 patients of the present study had the clinical phenotype of mild non‐transfusion‐dependent thalassaemia intermedia, preserving haemoglobin levels around 9 g/dl and haemoglobin F levels < 25%. All were compound heterozygotes for the +1480 C → G mutation and common severe β‐thalassaemia mutations. The haematological parameters of heterozygotes with this mutation were within the normal range with the exception of a slightly raised α/non‐α‐globin chain synthesis (1.2–1.9).mRNA analysis demonstrated a 20–34% reduction in mRNA levels associated with the +1480 C → G mutation compared to normal β‐globin alleles. These findings confirm that the C → G mutation at position 6 3′ to the termination codon is a mild β‐thalassaemia mutation causing slight reduction in β‐globin mRNA levels and β‐globin chain synthesis. It becomes clinically relevant when co‐inherited with a severe β‐thalassaemia mutation in trans.
Expression of p210 and p190 BCR‐ABL due to alternative splicing in chronic myelogenous leukaemia
Authors: Lichty Brian D. , Keating Armand , Callum Jeannie , Yee Karen , Croxford Ruth , Corpus George , Nwachukwu Bevoline , Kim Peter , Guo Joyce , Kamel‐Reid Suzanne 
Published: March 2003

The hallmark of chronic myelogenous leukaemia (CML) is the presence of the Philadelphia chromosome and its resultant fusion message, BCR‐ABL, and fusion protein, p210. Patients with CML in blast crisis, or with Philadelphia positive acute lymphoblastic leukaemia (ALL), can have a smaller BCR‐ABL fusion transcript possessing only the first exon of BCR fused to ABL. This smaller transcript encodes a 190 kD protein which is more strongly transforming than the p210 protein derived from the larger CML‐associated transcript. We performed RT‐PCR on samples from CML patients in chronic phase to determine the frequency and mechanism of p190 and p210 co‐expression and to see if this correlated with clinical indices.We examined the peripheral blood or marrow of 67 patients with CML and found that 35 of them expressed both transcripts whereas the remainder expressed the p210‐encoding transcript exclusively. Additional PCR products of an intermediate size were also frequently detected and have been isolated and sequenced. Data from two of these products indicate that they are the result of alternative splicing and include variable combinations of BCR exons. We believe that the expression of the p190‐encoding transcript in the chronic phase of CML is also due to alternative splicing. A comparison of patients co‐expressing the p190‐ and p210‐encoding transcripts with those patients who expressed only the p210‐encoding transcript detected significantly higher white blood cell (WBC) counts and blast cell counts at time of testing as well as significantly higher white blood cell counts at diagnosis.
Stable recombinant expression and characterization of the two haemophilic factor VIII variants C329S (CRM−) and G1948D (CRMr)
Authors: David Dezsö , Saenko Evgueni L. , Santos I. M. Ana , Johnson Daniel J. D. , Tuddenham Edward G. D. , McVey John H. , Kemball‐Cook Geoffrey 
Published: March 2003

In haemophilia A, the functional defect at the molecular level of most factor VIII (FVIII) missense mutations remains unknown. Site‐directed mutagenesis of B domain‐deleted FVIII cDNA (FVIIISQ) was used to introduce two mutations associated with severe cross‐reacting material (CRM)‐negative (FVIII‐C329S) or mild/moderate CRM‐reduced (FVIII‐G1948D) haemophilia A. Wild‐type (FVIIISQ‐WT) and variant FVIIISQ proteins were successfully expressed after stable transfection in Chinese hamster ovary (CHO) cells, and partially characterized at the intracellular, molecular and functional levels. Reverse transcription polymerase chain reaction analysis confirmed that both transcription and mRNA processing appeared normal in CHO cells transfected with both the wild‐type and two variant constructs. In contrast to FVIIISQ‐WT, immunofluorescence analysis of both CRM− and CRMr variants showed intracellular FVIII accumulation within the rough endoplasmic reticulum, suggesting secretion defects in transfected CHO cells. Immunoblot analysis of the FVIIISQ variant proteins that were secreted showed that they were expressed as mixed populations of uncleaved 170 kDa polypeptides, processed 90 kDa heavy chains and 80 kDa light chains, similar to FVIIISQ‐WT. Phenotypic analysis of the B domain‐deleted FVIIISQ variants expressed in CHO cells correlated well with the patients' reduced FVIII activity and, in addition, surface plasmon resonance studies demonstrated that both missense mutations were associated with increased rates of A2 domain dissociation following thrombin activation. We conclude that the mutations found are responsible for the haemophilia A phenotype, through intracellular retention and decreased stability of the active cofactor FVIIIa.
High levels of reticulated platelets and thrombopoietin characterize fetal thrombopoiesis
Authors: Jilma‐Stohlawetz P. , Homoncik M. , Jilma B. , Folman C. C. , Von Dem Borne A. E. G. KR. , Bernaschek G. , Deutinger J. , Ulm B. , Eppel W. , Panzer S. 
Published: March 2003

To characterize fetal thrombopoiesis, we determined plasma thrombopoietin (TPO) and glycocalicin levels, platelet counts and reticulated platelets (RP) of fetuses and compared them with the respective values of their mothers. Percutaneous umbilical vein sampling in abnormal pregnancies revealed twofold higher thrombopoietin levels and 20‐fold higher reticulated platelet counts, but lower levels of glycocalicin in fetuses compared with their mothers (P < 0·05). Neither the expression of platelet glycoprotein Ib and IIb on platelets nor the platelet counts were different between mothers and their fetuses. These data indicate enhanced thrombopoiesis and/or increased platelet turnover in fetuses.
Age as the major predictive factor of long‐term response to splenectomy in immune thrombocytopenic purpura
Authors: Fabris F. , Tassan T. , Ramon R. , Carraro G. , Randi M. L. , Luzzatto G. , Moschino P. , Girolami A. 
Published: March 2003

Sixty‐one consecutive patients undergoing splenectomy for chronic immune thrombocytopenia were retrospectively evaluated. Platelet response was considered as complete (CR) when platelet count rose to > 100 × 109/l, partial (PR) when 30–100 × 109/l or absent (NR) if otherwise. Follow‐up (mean time 7·6 years) was possible in 54 patients. Forty‐eight patients (88%) had an immediate response to splenectomy (39 CR, 9 PR) whereas six (12%) were NR. Thirty‐six responders (67%) had sustained remission (31 CR; 5 PR) without further treatment; thrombocytopenia recurred in 12 patients (33%). The probability curve of continued remission showed a constant relapse‐rate during the first 36 months; a further step of relapse was observed beginning 70 months after surgery. The only positive predictive factor for the long‐term response to splenectomy was age < 40 (P < 0·005). Neither duration of thrombocytopenia nor previous response to medical treatment (steroids and/or intravenous immunoglobulins) were related to splenectomy response.
Influence of highly active anti‐retroviral therapy on response to treatment and survival in patients with acquired immunodeficiency syndrome‐related non‐Hodgkin's lymphoma treated with cyclophosphamide, hydroxydoxorubicin, vincristine and prednisone
Authors: Navarro José‐Tomás , Ribera Josep‐Maria , Oriol Albert , Vaquero Manuel , Romeu Joan , Batlle Montserrat , Flores Alonso , Millá Fuensanta , Feliu Evarist 
Published: March 2003

Combined highly active anti‐retroviral therapy (HAART) with protease and reverse transcriptase inhibitors has modified the natural history of opportunistic infections and neoplasms in human immunodeficiency virus (HIV)‐infected patients. We analysed the influence of HAART on the response to treatment and survival in a series of 58 patients with acquired immune deficiency syndrome (AIDS)‐related non‐Hodgkin's lymphoma (NHL) treated with CHOP (cyclophosphamide, hydroxydoxorubicin, vincristine and prednisone). Two groups of patients were included: (i) forty‐one patients diagnosed with NHL between 1988 and 1996 who were not treated with HAART; (ii) seventeen patients diagnosed since 1996, who were receiving or commenced HAART when NHL was diagnosed. The response rate to CHOP was higher in group 2 (13 out of 17 cases; 75%) than in group 1 (14 out of 41 cases; 34%) (P = 0·003). The 2‐year probability of event‐free survival (EFS) [95% confidence interval (CI)] for group 1 was 0·5 (0·24–0·74), whereas for group 2 it was 0·85 (0·61–0·90) (P = 0·024). The lymphoma‐free survival (LFS) was also significantly different for both groups (2‐year LFS probability 0·53 vs. 1·0,P = 0·04). The median (95% CI) overall survival (OS) for group 1 was 7 months (range, 3–10·8 months), whereas it was not reached in group 2 (P = 0·0015). In the multivariate analysis for remission attainment, the only variables with a higher probability to achieve complete remission (CR) were HAART (P = 0·01) and International Prognostic Index score 1 (P = 0·02). The only statistically significant variable in the multivariate analysis for EFS was HAART (P = 0·049) and the variables with prognostic value for OS in the multivariate analysis were B symptoms (P = 0·01) and HAART (P = 0·003). Patients with AIDS‐related NHL treated with CHOP and HAART had a higher CR rate than those treated only with CHOP. In this study, HAART was an independent prognostic factor for CR, OS and EFS in patients with AIDS‐related NHL.
Protein S levels are lower in women receiving desogestrel‐containing combined oral contraceptives (COCs) than in women receiving levonorgestrel‐containing COCs at steady state and on cross‐over
Authors: Mackie Ian J. , Piegsa Karin , Furs Sally‐ann , Johnson Juliet , Bounds Walli , Machin Samuel J. , Guillebaud John 
Published: March 2003

This study aimed to identify specific haemostatic changes that might account for previous observations of higher venous thromboembolic risk among users of combined oral contraceptives (COCs) containing desogestrel (DSG) than levonorgestrel (LNG). Sixty‐three current users of monophasic 30 μg oestrogen COCs containing either LNG or DSG omitted one pill‐free interval (PFI), switching immediately either to the opposite formulation for one cycle or continuing with the same pill. Venesection followed the initial PFI after one cycle (21 tablets) and two cycles (42 tablets) of continuous pill taking, and after the following PFI. Protein S was lower in users of DSG than LNG formulations after the first PFI (mean ± SD, 0·67 ± 0·09 vs 0·76 ± 0·10, P < 0·001) and after one cycle (0·61 ± 0·09 vs 0·76 ± 0·09, P < 0·0001). Protein S decreased when switching from LNG to DSG pills (0·77 ± 0·07–0·65 ± 0·06, P < 0·0001), mirrored by an increase at switching from DSG to LNG formulations (0·61 ± 0·08–0·73 ± 0·10, P < 0·005). Mean protein S levels remained within the normal range. Three different markers of thrombin generation remained unaltered. Potential explanations for COC‐related thrombotic events are ‘acquired resistance to activated protein C’ or inhibition of fibrinolysis. A potential role has been described for protein S deficiency in both. A further triggering factor is a probable prerequisite for actual thrombosis, but pill‐takers whose levels of protein S were in the lowest percentiles may be at greatest risk.
A Val193Met mutation in GPIIIa results in a GPIIb/IIIa receptor with a constitutively high affinity for a small ligand
Authors: Fullard John , Murphy Ronan , O'Neill Sarah , Moran Niamh , Ottridge Brian , Fitzgerald Desmond J. 
Published: March 2003

We have identified a patient designated as (GTa) with Glanzmann's Thrombasthenia (GT) diagnosed on the basis of a prolonged bleeding time and failure of the patient's platelets to aggregate. The number of glycoprotein (GP)IIb/IIIa receptors on the platelet surface was 37% of normal and those receptors displayed a defect in soluble fibrinogen binding. Nevertheless, GTa platelets showed increased adhesion to solid‐phase fibrinogen and binding affinity for the RGD‐mimetic 3H‐SC52012, a non‐peptide GPIIb/IIIa antagonist. Dithiothreitol (DTT) and ADP enhanced the affinity for [3H]‐SC52012 in normal platelets, but had little effect in GTa platelets. These findings suggested that GTa platelets were locked in an altered affinity state. Genetic analysis showed that GTa was a compound heterozygote for the GPIIIa gene. One allele showed a deletion at the 3′ end of exon 3 resulting in a premature stop codon. The second GPIIIa allele had a G to A transition at nucleotide 577, resulting in a Val193Met substitution. HEK 293T cells transfected with mutant GPIIb/IIIaV193M bound [3H]‐SC52012 with a higher affinity than wild‐type GPIIb/IIIa, and this was not increased by DTT. The mutant receptor distinguishes between platelet adhesion and aggregation, and demonstrates the phenotype that may be expected when platelet aggregation alone is inhibited.
Burkitt's acute lymphoblastic leukaemia transformatation after myelodysplastic syndrome
Authors: Ikeda Takashi , Sato Ken , Yamashita Takuya , Kanai Yoshiaki , Kuwada Naruo , Matsumura Takuya , Nakamura Yukitsugu , Kimura Fumihiko , Motoyoshi Kazuo 
Published: March 2003

We describe a patient with myelodysplastic syndrome (MDS) that transformed to Burkitt's acute lymphoblastic leukaemia (ALL). The leukaemic blasts were negative for peroxidase staining, and expressed CD10, CD19, CD22, CD38, human leucocyte antigen (HLA)‐DR and surface immunoglobulin (sIg) M, but neither sIgD nor sIgG were expressed. Chromosomal study during the ALL phase showed t(8;22)(q24;q11) in addition to the karyotypes determined during the MDS phase. Furthermore, overexpression of c‐myc mRNA was confirmed in ALL blasts. These findings indicate that MDS transformed to Burkitt's ALL through multiple cytogenetic evolutions, the final event of which seems to be overexpression of the c‐myc gene.