bluuebunny/crossref_metadata_2025 huggingface dataset:
Diabetes mellitus increases the in vivo activity of cytochrome P450 2E1 in humans
Authors: Wang Zaiqi , Hall Stephen D. , Maya Juan F. , Li Lang , Asghar Ali , Gorski J. C.
Published: March 2003

Aims  Cytochrome P450 2E1 (CYP2E1) is thought to activate a number of protoxins, and has been implicated in the development of liver disease. Increased hepatic expression of CYP2E1 occurs in rat models of diabetes but it is unclear whether human diabetics display a similar up‐regulation. This study was designed to test the hypothesis that human diabetics experience enhanced CYP2E1 expression.

Methods  The pharmacokinetics of a single dose of chlorzoxazone (500 mg), used as an index of hepatic CYP2E1 activity, was determined in healthy subjects (n = 10), volunteers with Type I (n = 13), and Type II (n = 8) diabetes mellitus. Chlorzoxazone and 6‐hydroxychlorzoxazone in serum and urine were analysed by high‐performance liquid chromatography. The expression of CYP2E1 mRNA in peripheral blood mononuclear cells was quantified by reverse transcriptase‐polymerase chain reaction.

Results  The mean ± s.d. (90% confidence interval of the difference) chlorzoxazone area under the plasma concentration‐time curve was significantly (P ≤ 0.05) reduced in obese Type II diabetics (15.7 ± 11.3 µg h ml−1; 9, 22) compared with healthy subjects (43.5 ± 16.9 µg h ml−1; 16, 40) and Type I diabetics (32.8 ± 9.2 µg h ml−1; 9, 25). There was a significant two‐fold increase in the oral clearance of chlorzoxazone in obese Type II diabetics compared with healthy volunteers and Type I diabetics. The protein binding of chlorzoxazone was not significantly different between the three groups. In contrast, Type 1 diabetics and healthy volunteers demonstrated no difference in the oral clearance of chlorzoxazone. The urinary recovery of 6‐hydroxychlorzoxazone as a percentage of the administered dose was not different between healthy, Type I and obese Type II diabetics. The elimination half‐life of chlorzoxazone did not differ between the three groups. CYP2E1 mRNA was significantly elevated in Type I and obese Type II diabetics compared with healthy volunteers. The oral clearance of chlorzoxazone, elimination half‐life, Tmax, and Cmax were not significantly influenced by weight, body mass index, serum glucose, serum cholesterol, or glycosylated haemoglobin.

Conclusions  There was a marked increase in hepatic CYP2E1 activity in obese Type II diabetics as assessed by chlorzoxazone disposition. Increased expression of CYP2E1 mRNA in peripheral blood mononuclear cells was found in both types of diabetes mellitus. Adverse hepatic events associated with Type II diabetes may be in part a result of enhanced CYP2E1 expression and activity.
Population pharmacokinetics of theophylline during paediatric extracorporeal membrane oxygenation
Authors: Mulla Hussain , Nabi Fazal , Nichani Sanjiv , Lawson Graham , Firmin R. K. , Upton David R.
Published: March 2003

Aims  To determine the population pharmacokinetics of theophylline during extracorporeal membrane oxygenation (ECMO) from routine monitoring data.

Methods  Retrospective data were collected from 75 term neonates and children (age range 2 days to 17 years) receiving continuous infusions of aminophylline (mean rate 9.2 ± 2.6 µg kg−1 min−1) during ECMO. A total of 160 plasma concentrations (range 1–8 per patient), sampled at time intervals ranging from 10 h to 432 h, were included. Population PK analysis and model building were carried out using WinNonMix Professional (Version 2.0.1). Cross‐validation was used to evaluate the validity and predictive accuracy of the model.

Results  A one‐compartment model with first order elimination combined with an additive error model was found to best describe the data. Of the covariables tested, bodyweight significantly influenced clearance and volume of distribution, whereas age was an important determinant of clearance, as adjudged by the differences in the −2 × log likelihood (P < 0.005) and the residual error value. The final model parameters were estimated as: clearance (l h−1) = 0.023 × bodyweight (kg) + 0.000057 × age (days) and volume of distribution (l) = 0.57 × bodyweight (kg). The interindividual variability in clearance and volume of distribution was 38% and 40%, respectively. The residual error corresponded to a standard deviation of 3.6 mg l−1. Cross‐validation revealed a median (95% confidence interval) model bias of 9.4% (2.9, 16.5%) and precision of 29.5% (24.8, 36.0%).

Conclusions  The estimated clearance is significantly lower, and volume of distribution higher, than previously reported in non‐ECMO patients of similar age. These differences are probably a result of the expanded circulating volume during ECMO and altered renal and hepatic physiology in this critically ill group. Large interindividual variability reflects the heterogeneous nature of patients treated on ECMO.
Changes in the pattern of sensitization to common contact allergens in Denmark between 1985–86 and 1997–98, with a special view to the effect of preventive strategies
Authors: Johansen J.duus , Menné T. , Christophersen J. , Kaaber K. , Veien N.
Published: March 2003

The objective of the present study is to describe any changes in the prevalence of sensitization to common contact allergens in patch-tested patients over a 12-year period. Attention is given to possible effects of preventive strategies introduced in Denmark regarding nickel and chromate sensitization during that period, and particular areas of concern are identified. Members of the Danish Contact Dermatitis Group collected patch-test results from consecutive eczema patients as well as information about exposures and demographic variables over a 6-month period in 1985–86. The investigation was repeated in 1997–98 in the same clinics, at the same time of year, using identical methods and patch-test substances, including nickel sulphate 5%, potassium dichromate 0·5% and fragrance mix 8%. Nickel was the most common contact allergen in both study periods, followed by the fragrance mix. In children 0–18 years of age, the frequency of nickel allergy decreased from 24·8% in the first study period to 9·2% in the second study period (P < 0·0008). Fragrance mix allergy doubled in frequency from 4·1% in 1985–86 to 9·9% in 1997–98, an increase that affected all age groups. Contact allergy to potassium dichromate decreased significantly from 3·0% in the first period to 1·2% in the second period (P = 0·001). The decrease was seen in both sexes and was most pronounced among those of working age. No other significant changes were found in the frequency of sensitization to common allergens over the 12-year observation period.
PUVA-induced lichen planus pemphigoides
Authors: Kuramoto N. , Kishimoto S. , Shibagaki R. , Yasuno H.
Published: March 2003

A 72-year-old woman had suffered from parapsoriasis en plaque (large plaque type) controlled by topically applied psoralen ultraviolet A (PUVA) therapy. The parapsoriasis lesions gradually disappeared, but numerous tiny red papules with pruritus appeared over the forearms and lower legs 120 days after starting PUVA therapy. These papules developed to form violaceous plaques. Histological findings demonstrated the characteristics of lichen planus. Two months later, tense bullae developed on the plaques and on uninvolved skin of the limbs. These were subepidermal, with linear deposits of IgG and C3 along the basement membrane zone (BMZ) in immunofluorescence of peribullous skin, and immunodeposits of type IV collagen along the floor of the bullae. We therefore, diagnosed lichen planus pemphigoides (LPP). Using systemic and topical steroid therapy, the lesions rapidly resolved and there has been no recurrence. This case suggests that the combination of basal cell injuries caused by chronic inflammation and PUVA therapy could expose BMZ components to autoreactive lymphocytes and induce LPP.
Coexistence of psoriasis and linear IgA bullous dermatosis
Authors: Takagi Y. , Sawada S. , Yamauchi M. , Amagai M. , Niimura M.
Published: March 2003

Linear IgA bullous dermatosis (LABD) is characterized by IgA autoantibodies against components of the basement membrane zone (BMZ). A 97-kDa protein is one of the major autoantigens associated with this disease. We report a 68-year-old man who developed LABD after a 3-year history of psoriasis and in the context of active hepatitis C virus infection. He had been treated with cyclosporin for psoriasis for about 9 months. Histologically, there was a subepidermal blister containing neutrophils and eosinophils with lymphocytes infiltrating predominantly in the dermis. Direct immunofluorescent staining showed linear IgA deposition at the BMZ. The patient’s IgA autoantibodies bound exclusively to the epidermal side of 1 mol/L salt-split normal human skin. Immunoblot analysis identified a 97-kDa autoantigen in epidermal extracts. This appears to be the first case of LABD with IgA autoantibodies against a 97-kDa autoantigen, associated with psoriasis and hepatitis C virus infection.
Squamous cell carcinoma arising in Hailey–Hailey disease of the vulva
Authors: Cockayne S.E. , Rassl D.M. , Thomas S.E.
Published: March 2003

A 61–year-old woman, who was known to have Hailey–Hailey disease, presented with increasing vulval soreness. Biopsy showed vulval intraepithelial neoplasia (VIN) 3 and subsequent histology from a vulvectomy specimen showed extensive VIN with early invasive squamous cell carcinoma. This may be another example of chronic inflammation of the vulval area leading to the development of squamous cell carcinoma. However, in this case, chronic human papillomavirus may also have played a part, leading to VIN and reactivation of the Hailey–Hailey disease. We can find no previous reports of squamous cell carcinoma developing in the setting of Hailey–Hailey disease.
A randomized trial of amorolfine 5% solution nail lacquer combined with oral terbinafine compared with terbinafine alone in the treatment of dermatophytic toenail onychomycoses affecting the matrix region
Authors: Baran R. , Feuilhade M. , Datry A. , Goettmann S. , Pietrini P. , Viguie C. , Badillet G. , Larnier C. , Czernielewski J.
Published: March 2003

In view of recent advances in the development of antifungal agents, this study examined the possible synergy of two new antifungal agents, terbinafine and amorolfine. The study compared two different courses of terbinafine treatment combined with amorolfine 5% solution nail lacquer. Terbinafine was given orally for 6 (AT6 group) or 12 weeks (AT12 group) and amorolfine nail lacquer applied weekly for 15 months. A control group received terbinafine alone for 12 weeks. This was a randomized, prospective, open study of severe dermatophyte toenail onychomycosis with matrix region involvement. Nail samples were taken before the start of the study, at inclusion and at the visits at 6 weeks, 3, 9, 15 and 18 months. To assess the value of such combined therapy we chose an early parameter as the principal outcome variable, which was the result of mycological examination, including direct microscopy and culture, at 3 months (allowing a margin of 15 days). The secondary parameters of success were the mycological results at the later visits, clinical evaluation and a combined mycological–clinical response. Safety and tolerance were also assessed. Adverse events were recorded and liver function tests were performed monthly during the terbinafine treatment. Of the 147 patients included in the trial, 121 attended the 3-month visit, within a time limit of 15 days of 3 months after the beginning of treatment: 40 in the AT6 group, 40 in the AT12 group and 41 in the control group. In all, 32 of 121 patients (26·4%) had negative mycological results on direct microscopy and culture: 14 of 40 (35·0%) in the AT6 group, 11 of 40 (27·5%) in the AT12 group and seven of 41 (17·1%) in the control group. The cure rate for the global (mycological and clinical cure) response measured at 18 months in 145 patients was 44·0% (22 patients) in the AT6 group, 72·3% (34 patients) in the AT12 group and 37·5% (18 patients) in the terbinafine group. These results suggest that the combination of amorolfine and terbinafine may be of value in the treatment of severe onychomycosis. At the same time a pilot pharmacoeconomic analysis was performed demonstrating a better cost per cure ratio for the patients receiving combination treatment.
Treatment of verrucous carcinoma of vulva with acitretin
Authors: Mehta R.K. , Rytina E. , Sterling J.C.
Published: March 2003

We describe a 60-year-old patient with verrucous carcinoma of the vulva, which recurred 6 years after simple vulvectomy and radiotherapy. Treatment with acitretin led to significant improvement and ongoing disease control has been achieved with low-dose maintenance therapy of 10 mg acitretin daily.
Neonatal thrombocytopenia: incidence and characterization of maternal antiplatelet antibodies by MAIPA assay
Authors: DE Moerloose  , Boehlen  , Extermann  , Hohlfeld
Published: March 2003

Neonatal thrombocytopenia (NNT) which is frequent in distressed newborns was uncommon in a non‐selected population of neonates. The aim of this prospective study was to determine the frequency of NNT and, in confirmed NNT, to search for maternal antiplatelet antibodies with a monoclonal antibody‐specific immobilization of platelet antigens (MAIPA) assay. Among the 8388 newborns studied, 40 (0.5%, 95% CI 0.3–0.6) had confirmed NNT, which was severe (platelet count < 50 × 109/l) in 10 cases (0.12%, 95% CI 0.05–0.20). Antiplatelet antibodies were detected in 10/31 studied mothers of thrombocytopenic newborns (32.3%): they were alloantibodies in five cases and autoantibodies in five other cases. Among these 10 newborns, seven had severe thrombocytopenia and four had bleeding complications. As controls, antiplatelet antibodies were also searched for in mothers of non‐thrombocytopenic newborns: antiplatelet antibodies were present in 8.5% (95% CI 5.9–11.7) of thrombocytopenic mothers (n = 400) and 3.2% (95% CI 0.7–9.0) of non‐thrombocytopenic mothers (n = 95). The difference was significant between the control groups and the group of mothers of thrombocytopenic newborns. In conclusion, our data indicate that an immune origin is frequent in NNT and should be looked for, particularly when the platelet count is < 50 × 109/l.
Acute leukaemia in Jehovah's Witnesses
Authors: Cullis  , Duncombe  , Dudley  , Lumley  , Apperley  , Smith
Published: March 2003

The refusal of Jehovah's Witnesses with leukaemia to accept transfusion provides a major clinical challenge because of the myelosuppressive effects of chemotherapy. Experience in treating five such patients is described. Two patients with acute lymphoblastic leukaemia (ALL) achieved remission following chemotherapy, the first without transfusion support, the second, a minor, receiving transfusion under a court order; the first patient remains in remission 5 years later, whereas the second subsequently relapsed and died. Of three patients with acute myeloid leukaemia (AML), two received chemotherapy: one died of anaemia during induction chemotherapy whereas the second eventually consented to transfusion but died of refractory leukaemia. The third patient died of anaemia despite erythropoietin. We feel Jehovah's Witnesses should not be denied antileukaemic therapy if they fully understand the risks involved. Minimizing phlebotomy, use of antifibrinolytic agents and growth factors may make chemotherapy feasible, especially in ALL where remission may be induced with less myelosuppressive agents. The outlook for those with AML treated with conventional chemotherapy appears poor; alternative approaches to treatment should be considered in these patients.
Lack of sequence variations in the C4b‐BP β‐chain in patients with type III protein S deficiency bearing the Ser 460 to Pro mutation: description of two new intragenic isomorphisms in the C4b‐BP β‐chain gene (C4BPB)
Authors: MorbŒuf Olivier , Aiach Martine , Gandrille Sophie
Published: March 2003

Type III protein S (PS) deficiency, characterized by low levels of free PS and normal total PS levels, is often associated with the Ser 460 to Pro substitution. However, some patients bearing this mutation have normal PS levels, suggesting that another gene defect may account for this phenotype. We postulated that this defect was located in the C4b‐BP β‐chain gene (C4BPB) and searched for a mutation in the coding regions of this gene in 35 propositi with type III PS deficiency and bearing the Ser 460 to Pro mutation. No mutations explaining the phenotype of type III PS deficiency were identified. We did, however, find two frequent nucleotide changes, one being located in the donor splice site of intron d and the second in the codon corresponding to Asn 137. We used these two polymorphisms to establish C4BPB gene haplotype in five informative type III PS‐deficient families and exclude a role of the C4BPB gene in this phenotype of three of them. Finally, increased C4b‐BP β‐chain levels were not responsible for the phenotype of type III PS deficiency as the C4BPB haplotype did not correlate with C4b‐BP β‐chain levels.
Cone and platelet analyser (CPA): a new test for the prediction of bleeding among thrombocytopenic patients
Authors: Kenet  , Lubetsky  , Shenkman  , Tamarin  , Dardik  , Rechavi  , Barzilai  , Martinowitz  , Savion  , Varon
Published: March 2003

The risk of bleeding among thrombocytopenic patients was evaluated using our new cone and platelet analyser (CPA) test. Using this test, adherence of platelets was quantitated on extracellular matrix and expressed as percent of surface coverage (SC) and the average size (AS) of aggregates. 42 thrombocytopenic patients with ITP (n=23), post chemotherapy (n=12) and others (n=7) were tested over a total of 82 visits. On each visit, complete blood count and CPA tests were performed and patients were evaluated for evidence of bleeding (found in 40 visits).

Bleeding patients had significantly lower platelet counts (27.4 ± 22.0 v 47.1 ± 21.0 × 109/l), lower haematocrit values (30.2 ± 8.1 v 35.2 ± 6.6%), lower MPV (6.83 ± 1.89 v 8.98 ± 1.13 fl), and lower SC (4.87 ± 3.95 v 10.33 ± 5.48%) and AS (33.99 ± 14.94 v 52.9 ± 24.34 μm2). Univariate analysis yielded platelet count 20.0 × 109/l, MPV  8fl, haematocrit <35%, SC < 5%, AS  40 μm2 as significantly associated with bleeding, whereas only MPV and SC were associated with bleeding (OR 6.95, CI 2.25–21.46 and OR 4.27, CI 1.29–14.16, respectively) by multivariate analysis. When taken together, 21/22 of patients (95%) with both low SC (<5%) and low MPV (<8.0 fl) had bleeding symptoms, whereas only 9/43 (21%) patients with both these parameters above these values experienced bleeding symptoms.

We conclude that the CPA test and the parameter SC (<5%) together with MPV (8 fl) might be used as independent predictors of bleeding in the management of thrombocytopenic patients.
Distinction of eosinophilic leukaemia from idiopathic hypereosinophilic syndrome by analysis of Wilms' tumour gene expression
Authors: Menssen Hans D. , Renkl Hans‐J. , Rieder Harald , Bartelt Simone , Schmidt Alexander , Notter Michael , Thiel Eckhard
Published: March 2003

In patients presenting with immature eosinophilic precursors it is notoriously difficult to distinguish acute eosinophilic leukaemia (EoL) from the benign idiopathic hypereosinophilic syndrome (HES), based on morphological, cytochemical and immunophenotyping criteria, alone. Cytogenetic analysis or fluorescence in situ hybridization (FISH) can help in discriminating between these rare haematological disorders, but often treatment decisions cannot wait for the results of these time‐consuming techniques. Recently, we and others found Wilms' tumour (WT1) gene expression to be increased in virtually all patients with acute leukaemias, whereas normal haemopoietic progenitors express the WT1 gene at much lower levels or not at all. To determine whether detection of WT1 gene expression is useful to distinguish EoL from HES patients, we analysed, by RT‐PCR, bone marrow or blood mononuclear cells from EoL (n=3), HES (n=3) and reactive eosinophilia patients (n=4) for WT1 gene expression. Using our WT1‐RT‐PCR protocol, we found WT1 gene expression to be restricted to EoL patients. By detecting WT1 mRNA transcripts in the cerebrospinal fluid using RT‐PCR, we were also able to diagnose isolated CNS‐relapsed leukaemia, initially confused with bacterial meningitis, in an EoL patient. In conclusion, we show that WT1‐RT‐PCR is a powerful complementary diagnostic tool to distinguish acute eosinophilic leukaemia from the hypereosinophilic syndromes. This observation needs confirmation in a larger series of EoL and HES patients.
Clonal T‐cell proliferation causing pure red cell aplasia in chronic B‐cell lymphocytic leukaemia: successful treatment with cyclosporine following in vitro abrogation of erythroid colony‐suppressing activity
Authors: Yamada Osamu , Yun‐Hua Wang , Motoji Toshiko , Mizoguchi Hideaki
Published: March 2003

Clonal T cells that cause pure red cell aplasia were demonstrated in a patient with chronic B‐cell lymphocytic leukaemia. The T‐cell fraction was enriched by the immunomagnetic method and shown to have T‐cell receptor‐β gene rearrangement. Cryopreserved T cells, obtained during the period of anaemia, suppressed autologous erythroid progenitor cell growth from remission marrow. The colony‐suppressing activity was markedly improved by the addition of cyclosporine to the culture medium. The patient achieved haematological remission by cyclosporine monotherapy. Cyclosporine treatment may improve erythropoiesis in both a T‐cell‐dependent and a T‐cell‐independent manner.
Evidence for a critical role of DNA topoisomerase IIα in drug sensitivity revealed by inducible antisense RNA in a human leukaemia cell line
Authors: Towatari Masayuki , Adachi Koichi , Marunouchi Tohru , Saito Hidehiko
Published: March 2003

To examine the role of human DNA topoisomerase IIα (topo IIα) in drug resistance, we selectively inhibited topo IIα gene expression in U937 human monocytic leukaemia cells stably transfected with a plasmid that allowed for Zn‐mediated conditional expression of a human α‐topo IIα antisense sequence. Expression of topo IIα mRNA was reduced to <30%, whereas no significant alteration of topo IIβ mRNA expression was observed. Under these conditions, drug sensitivity to the topo‐II‐directed agents, etoposide and daunorubicin, was reduced to approximately 50%, whereas sensitivity to 4‐hydroperoxy‐cyclophosphamide (4‐HC) was not altered. This suggests that a reduced amount of topo IIα mRNA may be sufficient for the resistance to topo II inhibitors in leukaemia cells.
Aprotinin complements heparin bonding in an in vitro model of cardiopulmonary bypass
Authors: Bannan Sally , Martin Paul G.
Published: March 2003

The relative contribution of full‐dose aprotinin, used with heparin‐bonded surfaces, to contact activation during cardiopulmonary bypass was examined. In vitro Carmeda‐bonded cardiopulmonary bypass circuits were perfused with whole blood anticoagulated with heparin (3.3 U/ml). Aprotinin (300 kIU/ml) was added to the circuits of one set of experiments. Samples were taken prior to perfusion and at 30, 60, 120 and 360 min. The activated coagulation time was extended in the aprotinin experiments, significantly at 30 min (P = 0.003) and 120 min (P = 0.001). Thrombin–antithrombin complexes and prothrombin fragment F1+2 were both higher in the non‐aprotinin experiments at 120 min (P = 0.02 each) and 360 min (P = 0.005 and 0.001, respectively). Plasma leucocyte elastase was raised in the non‐aprotinin experiments in comparison to the aprotinin experiments at each timepoint (30 min, P = 0.04; 60 min, P = 0.006; 120 min, P = 0.001; 360 min, P = 0.0001), as was interleukin‐8 at 120 min (P = 0.05) and 360 min (P = 0.0001). No differences were found for the platelet activation marker P‐selectin. Platelet and white blood cell counts fell significantly in the non‐aprotinin experiments compared with the aprotinin experiments at 360 min (P = 0.05 and 0.03, respectively). It would appear that the use of aprotinin has additional haemostatic beneficial effects to those found with heparin‐bonded circuits in terms of effects on contact activation and inflammation.
Liver iron and fibrosis during long‐term treatment with deferiprone in Swiss thalassaemic patients
Authors: TÖndury Petrign , Zimmermann Arthur , Nielsen Peter , Hirt Andreas
Published: March 2003

Serum ferritin levels, hepatic histology and iron concentration were studied in a ‘veteran’ group of seven Swiss β‐thalassaemic patients after 93–99 months of treatment with the oral iron chelator deferiprone (L1), and another four patients who had received 54–82 months of L1 therapy. Despite continuous compliance, unexplained resurgence of serum ferritin levels occurred in 4/7 patients of the ‘veteran’ group after 4–5 years on L1. In three of these a concomitant increase of liver iron was also observed.

Hepatic histology revealed significantly higher degrees of fibrosis in 6/11 hepatitis C (HC)‐positive patients (fibrosis scores 1–5, mean 3.0) than in the HC‐negative group (fibrosis score 0–2, mean 0.8). Two HC‐negative patients had no detectable fibrosis after 98 and 93 months on deferiprone. Therefore the hepatic pathology in these patients cannot definitely be attributed as a side‐effect of deferiprone. Chronic active hepatitis C and the accumulation of iron are the major causative factors to be considered.
Red cell morphology and malaria anaemia in children with Southeast‐Asian ovalocytosis band 3 in Papua New Guinea
Authors: O'donnell  , Allen  , Mgone  , Martinson  , Clegg  , Weatherall
Published: March 2003

Southeast‐Asian ovalocytosis (SAO) was diagnosed in children from Madang, Papua New Guinea, by detection of the SAO band 3 gene variant using the polymerase chain reaction. SAO band 3 was present in 16/241 (6.6%) children living in the community and 32/389 (8.2%) children with acute Plasmodium falciparum malaria (P = 0.42). SAO band 3 was detected in 8.2% (23/281) of α+‐thalassaemia homozygotes, 9.4% (20/214) of heterozygotes and 2.4% (2/85) of children with a normal α‐globin genotype (P = 0.12). The most consistent feature of SAO band 3 on microscopy of thin blood films was red cells with two or more linear or irregularly‐shaped pale regions. In children living in the community, these were present in 15 with SAO band 3 (sensitivity 93.8%) and only two normals (specificity 99.1%). The presence of 20% ovalocytosis was a poorer indicator of SAO band 3 (sensitivity 68.8% and specificity 100%). Haematological data were similar in SAO band 3 and normal children. However, in children with acute malaria, haemoglobin levels and red cell counts were significantly lower in SAO band 3 than normal children. The degree of ovalocytosis was lower in children with SAO band 3 during acute malaria, suggesting that a selective loss of ovalocytes may contribute to malaria anaemia in Southeast‐Asian ovalocytosis.
Human myeloma cells shed the interleukin‐6 receptor: inhibition by tissue inhibitor of metalloproteinase‐3 and a hydroxamate‐based metalloproteinase inhibitor
Authors: Hargreaves Philip G. , Wang Fengfei , Antcliff Jennifer , Murphy Gillian , Lawry John , Russell R. Graham G. , Croucher Peter I.
Published: March 2003

Interleukin‐6 (IL‐6) is the major growth factor for human myeloma cells, exerting its effect through the IL‐6 receptor (IL‐6R). A soluble form of IL‐6R (sIL‐6R) has been identified, which increases the sensitivity of myeloma cells to IL‐6. In patients with multiple myeloma (MM), serum concentrations of sIL‐6R are elevated and associated with poor prognosis. The present study was undertaken to determine whether proteolytic cleavage of IL‐6R could contribute to sIL‐6R release from human myeloma cells, and also to identify the class of proteinase responsible for this event. Human myeloma cell lines were shown to express IL‐6R upon their surface and also to release sIL‐6R into culture supernatants. In addition, phorbol 12‐myristate 13‐acetate (PMA) stimulated a loss of IL‐6R from the cell surface, with a corresponding increase in the concentration of sIL‐6R in the supernatant. Inhibitors of serine and cysteine proteinases, and tissue inhibitor of metalloproteinase (TIMP) ‐1 and TIMP‐2, were shown to have no effect on the magnitude of sIL‐6R release. In contrast, TIMP‐3 and a hydroxamate‐based metalloproteinase inhibitor (BB‐94), inhibited both constitutive and PMA‐induced release of sIL‐6R. Myeloma cells freshly isolated from the bone marrow of a patient with MM were also shown to express IL‐6R upon their surface, and to shed this receptor in response to PMA. These data demonstrate that increased proteolytic cleavage of IL‐6R, mediated by a non‐matrix‐type metalloproteinase, is likely to contribute to the elevated concentrations of sIL‐6R found in the serum of patients with MM. Inhibition of sIL‐6R release by hydroxamate‐based metalloproteinase inhibitors may represent a novel therapeutic approach to the treatment of MM.
Clinical and haematological consequences of recurrent G6PD mutations and a single new mutation causing chronic nonspherocytic haemolytic anaemia
Authors: Vulliamy Tom J. , Kaeda Jaspal S. , Ait‐Chafa Dahlila , Mangerini Rosa , Roper David , Barbot Jose , Mehta Athul B. , Athanassiou‐Metaxa  , Luzzatto Lucio , Mason Philip J.
Published: March 2003

We have determined the causative mutation in 12 cases of glucose‐6‐phosphate dehydrogenase deficiency associated with chronic non‐spherocytic haemolytic anaemia. In 11 of them the mutation we found had been previously reported in unrelated individuals. These mutations comprise seven different missense mutations and a 24 base pair deletion, G6PD Nara, previously found in a Japanese boy. Repeated findings of the same mutations suggests that a limited number of amino acid changes can produce the CNSHA phenotype and be compatible with normal development. The one new mutation we have found, G6PD Serres, is 1082 C → T causing a 361 Ala → Val substitution in the dimer interface where most other severe G6PD mutations are found. Now that several patients with the same mutation have been reported we can compare the resulting clinical phenotypes. For each mutation we find a reasonably consistent clinical picture, ranging from mild (G6PD Clinic) through moderate (G6PD Nashville) to severe (G6PD Beverly Hills and G6PD Nara).
Thioguanine versus mercaptopurine for therapy of childhood lymphoblastic leukaemia: a comparison of haematological toxicity and drug metabolite concentrations
Authors: Lancaster  , Lennard  , Rowland  , Vora  , Lilleyman
Published: March 2003

As a prelude to a nationwide randomized trial of thioguanine (TG) versus mercaptopurine (MP) for childhood lymphoblastic leukaemia we compared a pilot group of 23 children taking TG with a matched group taking MP. We assessed drug tolerance based on haematological toxicity and measured erythrocyte (RBC) concentrations of thioguanine nucleotides (TGN).

The median tolerated dose of TG was 30 mg/m2 compared to 55 mg/m2 for MP. There was no difference in the pattern of anaemia or neutropenia between the two groups, but dose‐limiting thrombocytopenia was more evident in the TG children (P < 0.001), four of whom had a decrease in platelet count to <20 × 109/l compared to only one on MP. The median RBC TGN concentration for those on 40 mg/m2 TG was 1726 pmol/8 × 108 RBCs compared with 308 pmol/8 × 108 RBCs for those on 75 mg/m2 MP (P < 0.0001). There was an inverse correlation between RBC TGNs and neutrophil count in the MP group but not in those on TG. No correlation between metabolite concentration and thrombocytopenia was found in either group.

These results provide further evidence that TG has a selective effect on platelets. They also showed that RBC TGN were, on average, 5‐fold higher in those taking TG but did not obviously relate to myelotoxicity as found in children on MP. The higher concentrations seen may partly reflect the erythrocyte's ability to metabolize TG directly to TGN by pathways not open to MP.
Mobilizing peripheral blood stem cells with high‐dose G‐CSF alone is as effective as with Dexa‐BEAM plus G‐CSF in lymphoma patients
Authors: KRÖger  , Zeller  , Fehse  , Hassan  , KRÜger  , Gutensohn  , LÖlliger  , Zander
Published: March 2003

We compared retrospectively the efficacy of granulocyte colony stimulating factor (G‐CSF) alone with chemotherapy plus G‐CSF in mobilizing CD34‐positive cells in patients with malignant lymphoma. 35 patients underwent peripheral blood stem cell (PBSC) collection following mobilization either with 24 μg/kg G‐CSF for 4 consecutive days (n = 18) or Dexa‐BEAM chemotherapy plus 5 μg/kg G‐CSF (n = 17). High‐dose G‐CSF was well tolerated with only slight bone pain and/or myalgia. The Dexa‐BEAM therapy required hospitalization with a median duration of 21 d. The median number of apheresis procedures in both groups was two (range two to four), resulting in a median of 5.3 and 5.1 × 106 CD34+ cells/kg. No patients in the G‐CSF group, but one in the Dexa‐BEAM group, failed to reach the target of collecting >2.0 × 106 CD34+ cells/kg. The number of CFU‐GM (10.4 v 6.0 × 105/kg) and of BFU‐E (10.6 v 4.5 × 105/kg; P = 0.04) was higher in the G‐CSF group than in the Dexa‐BEAM group. A subset analysis of CD34+ cells was performed in 16 patients showing a higher mean of Thy‐1 (CD90w) coexpression in the G‐CSF than in the Dexa‐BEAM group (4.8 v 1.8%, P = 0.12). Additionally the percentage of CD34+/CD38− cells was higher in the G‐CSF group (10.6% v 8.8%). However, these differences were not stastistically significant. The median time to leucocyte and platelet engraftment after high‐dose chemotherapy was slightly shorter in the G‐CSF than in the Dexa‐BEAM group (9 v 10 and 12 v 13.5 d, respectively). These results demonstrate that high‐dose G‐CSF is as effective as Dexa‐BEAM plus G‐CSF in mobilizing peripheral blood stem cells and produces prompt engraftment. The major advantages of G‐CSF mobilization were the safe outpatient self‐application and the fixed‐day apheresis.
Homozygous missense mutation (band 3 Fukuoka: G130R): a mild form of hereditary spherocytosis with near‐normal band 3 content and minimal changes of membrane ultrastructure despite moderate protein 4.2 deficiency
Authors: Inoue Takafumi , Kanzaki Akio , Kaku Mayumi , Yawata Ayumi , Takezono Masami , Okamoto Naoto , Wada Hideho , Sugihara Takashi , Yamada Osamu , Katayama Yasuyuki , Nagata Naokazu , Yawata Yoshihito
Published: March 2003

The characteristics of phenotypic expression were studied in a Japanese family with hereditary spherocytosis and an extremely rare homozygous missense mutation of the band 3 gene (band 3 Fukuoka: G130R). The homozygous unsplenectomized proband was a 29‐year‐old male with compensated haemolytic anaemia (red cell count 4.21 × 1012/l, reticulocytes 278 × 109/l, and indirect bilirubin 44 μmol/l). His red cell band 3 (B3) protein demonstrated a 9.3% reduction and his protein 4.2 (P4.2) level was substantially reduced (45.0%), compared to normal subjects. P4.2 protein was composed mostly of a wild type (72 kD) with a trace of 68 kD peptide. The binding properties of the mutated B3 to normal P4.2 were significantly impaired, which probably resulted in the substantial reduction of P4.2 in this proband, since no abnormalities were detected on the P4.2 gene. Electron microscopy (EM) using the freeze‐fracture method demonstrated a mild decrease in intramembrane particles (IMPs) of near‐normal size (8 nm in diameter) with no substantial increases in their oligomerization. Their distribution on the membrane P face was almost normal, although most of the IMPs could represent the homozygously mutated B3 protein. EM (quick‐freeze deep‐etching method) disclosed a skeletal network of near‐normal size and size distribution of the skeletal units, suggesting that the mutated B3 protein itself did not have much effect on the skeletal network in situ. Therefore the reduced P4.2 content (45% of that of normal subjects), which remained on the red cell membrane of this proband, appeared to be nearly sufficient for maintaining the normal structure of the skeletal network and IMPs in situ, contrary to the marked abnormalities in both IMPs and the skeletal network in complete P4.2 deficiencies.
Arsenic induces apoptosis in B‐cell leukaemic cell lines in vitro: activation of caspases and down‐regulation of Bcl‐2 protein
Authors: Akao Yukihiro , Mizoguchi Hiromi , Kojima Seiji , Naoe Tomoki , Ohishi Nobuko , Yagi Kunio
Published: March 2003

We showed that arsenic inhibited the cell growth of four B‐cell leukaemia cell lines of 11 various cell lines in vitro. In two of these four lines, KOCL44 and LyH7, apoptosis was identified by morphological and nucleosomal DNA fragmentation studies. Three of the four B‐cell lines that were growth inhibited were acute infantile leukaemia with t(11;19)(q23;p13) translocations involving the MLL gene that encodes the transcriptional factor Drosophila trithorax. The arsenic‐induced apoptosis in KOCL44 and LyH7 cells was found to be linked to caspases by Western blot and enzymological analyses. The amount of Bcl‐2 was reduced during apoptosis in LyH7 as judged by Western blot analysis. We concluded that combined activation of the caspases and down‐regulation of Bcl‐2 could determine the fate of B‐cell leukaemic cells in response to arsenic.
Elevated TFPI in malignant disease: relation to cancer type and hypercoagulation
Authors: Iversen Nina , Lindahl Anne‐Karin , Abildgaard Ulrich
Published: March 2003

We have previously reported high levels of the coagulation inhibitor TFPI in the blood of patients with gastrointestinal cancer. TFPI is not an acute‐phase reactant, but high levels have also been reported in patients with septicaemia and disseminated intravascular coagulation (DIC). To study its relationship with other types of malignancy, TFPI activity was first determined in plasma samples from 214 patients with various malignancies. In a second cohort of 83 patients, total and free TFPI antigen, protein C, antithrombin, fibrin monomer and D‐dimer were also measured. Elevated TFPI activity and antigens were found in about half of the patients with solid tumours. In contrast, elevated TFPI was rare in haematological malignancies (12%). In the 18 patients with acute nonlymphocytic leukaemia (ANLL), elevated free TFPI was found only in patients who also had DIC. No correlation was found between TFPI levels and fibrin monomer or D‐dimer levels. Only four out of 20 patients with solid tumours had normal levels of fibrin monomer and D‐dimer, yet three out of these four had elevated TFPI. In conclusion, elevated TFPI in ANLL is related to the coexistence of DIC. In solid tumour disease increased TFPI may reduce protective fibrin formation, but the pathogenic mechanism is as yet unknown.
Antibodies to prothrombin in antiphospholipid syndrome and inflammatory disorders
Authors: Guerin Jan , Smith Owen , White Barry , Sweetman Gertrude , Feighery Conleth , Jackson John
Published: March 2003

Antiphospholipid antibodies associated with the antiphospholipid syndrome (APS) have been shown to bind plasma proteins, particularly beta 2‐glycoprotein I (β2‐GPI). In this study the incidence of antibodies to solid‐phase prothrombin was examined in patients with antiphospholipid syndrome and a variety of other inflammatory disorders.

Significantly elevated levels of IgG anti‐prothrombin (anti‐PT) antibodies were detected in 63% of patients with APS (n = 27, median 22 arbitrary units: AU), 33% with SLE (n = 92, median 14 AU), 45% with rheumatoid factor (n = 22, median 16 AU), 21% with carotid artery stenosis (n = 21, median 15 AU), 32% with stroke (n = 38, median 13 AU), 67% of patients with a false positive serology for syphilis (n = 21, median 24 AU), 37% with HIV (n = 30, median 14 AU), 29% with syphilis (n = 14, median 19 AU) and 3% with infectious mononucleosis (n = 30, median 9 AU). In addition, a group of lupus anticoagulant (LA) positive patients (n = 48) was examined for antibodies to prothrombin, β2‐GPI and cardiolipin. 10 (21%) patients had raised levels of IgG anti‐PT antibodies, 30 (62%) had significantly elevated levels of anti‐β2‐GPI antibodies and 15 (31%) had elevated levels of anticardiolipin antibodies (ACA). Of the LA‐positive patients, 15 (43%) were identified with definite APS, eight (23%) with probable APS, two (6%) with possible APS and 10 (28%) patients had no clinical evidence of APS.

In conclusion, antibodies to prothrombin were found in a variety of inflammatory disorders and were therefore not specific for the APS. However, identification of the plasma proteins recognized by antibodies from patients with APS may provide insight into the pathogenic mechanisms involved in the heterogenous clinical manifestations of the APS.
Telomerase activity and the expression of telomerase components in acute myelogenous leukaemia
Authors: XU Dawei , Gruber Astrid , Peterson Curt , Pisa Pavel
Published: March 2003

In 95 leukaemic cell samples from 66 patients with acute myelogenous leukaemia (AML) (47 de novo and 19 secondary AML) telomerase activity was determined and the expression of the telomerase components: telomerase reverse transcriptase (hTERT), telomerase RNA template (hTR) and telomerase‐associated protein (TP1) evaluated by RT‐PCR. Compared to peripheral blood mononuclear cells (PBMC) from normal adult 87% (82/95) of patient samples exhibited elevated telomerase activity. hTERT, but not hTR and TP1 expression strongly correlated with the levels of telomerase activity (r = 0.47, P < 0.0001). The levels of telomerase activity were significantly higher at time of relapse or progression than at time of diagnosis (P = 0.003), and correlated to CD34 expression and chromosomal abnormalities of leukaemic cells (P = 0.01 and P = 0.001 respectively). The rate and duration of complete remission (CR) did not correlate with the levels of telomerase activity at diagnosis. Among eight patients in first relapse, however, two of three with low levels of telomerase activity re‐entered CR, whereas none of five patients with high telomerase activity achieved a second CR. Taken together, telomerase activation/up‐regulation in AML is a disease progression‐associated event. Undifferentiated status and chromosomal aberration also lead to the up‐regulation of telomerase activity in AML.
Th1/Th2 balance alteration in the clinical course of a patient with pure red cell aplasia and thymoma
Authors: Fujisao Shoji , Tsuda Hiroyuki
Published: March 2003

Peripheral T helper subsets in a patient with acquired pure red cell aplasia (PRCA) and thymoma were analysed by flow cytometry. Thymectomy resulted in a transient but definite improvement in anaemia. A continuous remission was attained by cyclosporin A (CsA) therapy. The changes in Th1/Th2 ratios corresponded with the haematological changes. Specifically, Th2 type lymphocytes were dominant during a haemopoietic suppression and elevation of the Th1/Th2 ratio was concomitant with haematological improvement. These observations suggest that polarization of Th subsets to Th2, induced by clonally expanded (γδ‐type) T cells in thymoma, are involved in the pathogenesis of PRCA.
Prevalence of the C282Y and H63D mutations in the HFE gene in patients with hereditary haemochromatosis and in control subjects from Northern Germany
Authors: Nielsen  , Carpinteiro  , Fischer  , Cabeda  , Porto  , Gabbe
Published: March 2003

Mutation analysis was performed for two HFE mutations (C282Y, H63D) in unrelated patients with hereditary haemochromatosis (n = 92), family members of patients (n = 34), and unrelated controls (n = 157) from Northern Germany. 87/92 patients (94.6%) revealed the C282Y mutation in homozygous form, five were heterozygous. No H63D mutation was found in 174 chromosomes of patients homozygous for C282Y, whereas four of the heterozygote patients also carried the H63D mutation. Among the control group, 9.6% were heterozygotes for C282Y. 2/157 subjects were homozygous, 37/157 were heterozygous for the H63D mutation, but showed no signs of iron overload.
Reactivation of Behçet's disease in the course of multicentric HHV8‐positive Castleman's disease: long‐term complete remission by a combined chemo/radiation and interferon‐α therapy regimen
Authors: Strohal Robert , Tschachler Erwin , Breyer Stefan , Uthman Aumed , Simonitsch Ingrid , Tratting Sigfried , Scheithauer Werner , Stingl Georg , Kornek Gabriela V.
Published: March 2003

We used a new combined chemo‐ (COP/ABVD), radiation and interferon‐α (10 × 106 IU s.c. 3× per week/12 months) therapy regimen to treat severe multicentric Castleman's disease (CD) complicated by relapsing Behçet's disease (BD) manifestations. More than 16 years after diagnosis of CD the patient remains in very good clinical condition, with remission of all CD and BD manifestations 13 months after discontinuation of the interferon‐α treatment. In addition, our clinicopathological, immunohistological and virological data suggest a pathogenetic link between CD and BD via activation of pre‐existing BD‐specific plasma cells due to CD‐related HHV8‐induced overexpression of interleukin‐6.
Composition and function of peripheral blood stem and progenitor cell harvests from patients with severe active rheumatoid arthritis
Authors: Snowden  , Nink  , Cooley  , Zaunders  , Keir  , Wright  , Milliken  , Brooks  , Biggs
Published: March 2003

High‐dose chemotherapy with autologous stem cell rescue has been proposed as an intensive therapy for severe rheumatoid arthritis (RA). In view of previous observations of abnormal haemopoiesis in RA patients, the composition and function of peripheral blood stem cell harvests (PBSCH) was investigated. Compared with PBSCH from healthy allogeneic donors mobilized with the same dose of G‐CSF (filgrastim; 10 μg/kg/d, n = 14), RA PBSCH (n = 9) contained significantly fewer mononuclear cells (375 v 569  × 106/kg, P  = 0.03) and CD34+ cells (2.7 v 5.8  × 106/kg, P = 0.003). However, there were increased proportions of CD14+ cells (P = 0.006) and CD14+CD15+ cells (the phenotype of previously described ‘abnormal’ myeloid cells, P = 0.002) in the RA PBSCH which translated into 3.5‐ and 7‐fold increases respectively on a per CD34+ cell basis. There were no differences in T‐cell activation status as judged by proportions of CD4+ and CD8+ expressing CD45RA, CD45RO, HLA‐DR and CD28 (RA PBSCH, n = 7, donor PBSCH, n = 5, P = 0.2–0.7). Phytohaemagglutinin responses determined fluorocytometrically with induction of CD69 expression were reduced in CD4+ and CD8+ cells following filgrastim administration in 3/3 RA patients tested. Compared with bone marrow as a potential source of CD34+ cells, PBSCH contained 11‐fold more T cells (P < 0.0005), 8‐fold more B cells (P < 0.0005) and 4‐fold more monocytes (P = 0.02). In short‐term methylcellulose culture there were no differences in colony counts (CFU‐GM, CFU‐GEMM, BFU‐E) per CD34+ cell from PBSCH from RA patients (n = 11) and healthy donors (n = 10). Long‐term culture initiator cells were cultured successfully from cryopreserved PBSCH from RA patients (n = 9). In conclusion, PBSCH from RA patients differed significantly in composition from normal individuals, but in vitro studies support normal stem and progenitor cell function. Changes in T‐cell function occur during mobilization in RA patients. This work provides reassurance for the use of PBSCH as haematological rescue and baseline data for clinical trials of graft manipulation strategies in patients with RA.
Elevated serum soluble Fas ligand in natural killer cell proliferative disorders
Authors: Kato Kei , Ohshima Koichi , Ishihara Shigehiko , Anzai Keizo , Suzumiya Junji , Kikuchi Masahiro
Published: March 2003

We evaluated the serum level of soluble Fas ligand (sFasL) in patients with natural killer lymphocyte proliferative disorders (NK‐ LPD). The serum sFasL level was elevated in neoplastic groups of aggressive NK leukaemia, indolent NK leukaemia and NK lymphoma, all of which contained clonal EBV‐DNA. In NK leukaemia the serum sFasL level was significantly higher than that found in others. However, it was not elevated in the patients with reactive NK‐LPD and in one patient with NK leukaemia in remission. These findings indicate that the serum sFasL level is a useful indicator in evaluating disease activity.