bluuebunny/crossref_metadata_2025 huggingface dataset:
The der(21)t(12;21) chromosome is always formed in a 12;21 translocation associated with childhood acute lymphoblastic leukaemia
Authors: KOBAYASHI HIROFUMI , SATAKE NORIKO , MASEKI NOBUO , SAKASHITA AKIKO , KANEKO YASUHIKO
Published: October 2003

We studied 116 patients (93 children and 23 adults) with acute lymphoblastic leukaemia (ALL) using fluorescence in situ hybridization (FISH) with the yeast artificial chromosome (YAC) clone, 964c10, which includes the recently described ETS‐like gene, TEL, on 12p13. FISH revealed that nine of the patients had a t(12;21), which had not been previously detected. The nine patients were all children, seven boys and two girls, aged 1–10 years (median 3 years), had an early B immunophenotype, and achieved complete remission, although two of them experienced haematological relapse. In addition to the t(12;21), FISH also revealed that three of the nine had a del(12p) in the other homolog of chromosome 12 or in the der(12) chromosome itself, and that two others had 12p translocations in the other chromosome 12 homolog. Although chromosomal rearrangements associated with the t(12;21) were heterogenous and complex, fusion of the sequences from chromosomes 12 and 21 on the der(21)t(12;21) chromosomes was consistent, suggesting that the TEL‐AML1 gene fusion on the der(21) chromosome may be critical in leukaemogenesis and that FISH or reverse transcriptase‐polymerase chain reaction (RT‐PCR) targeted to the chimaeric sequences on the der(21) will be most useful in detecting the t(12:21) or following a patient with the t(12;21), which is one of the most frequent chromosomal rearrangements in both Caucasian and Asian childhood ALL.
Graft failure after donor leucocyte infusion in relapsed chronic myeloid leukaemia: successful treatment with cyclophosphamide and antithymocyte globulin followed by peripheral blood stem cell infusion
Authors: K eil Felix , Kalhs Peter , Haas Oskar A. , Fritsch Gerhard , Mitterbauer Gerlinde , Brugger Stefan , Lechner Klaus , Schwarzinger Ilse , Mannhalter Christine , Linkesch Werner , Kurz Martin , Greinix Hildegard T.
Published: October 2003

We report a patient with chronic myeloid leukaemia who underwent allogeneic marrow transplantation (BMT) but had a molecular relapse 5 months and haematological relapse 15 months after BMT. Since therapy with α‐interferon had been ineffective he received leucocyte infusions from his sibling donor. He developed acute graft‐versus‐host disease and became aplastic 6 weeks later. Despite donor marrow infusion and cytokine stimulation marrow aplasia persisted for 13 weeks. Then, donors' peripheral blood stem cells were given after conditioning with cyclophosphamide and antithymocyte globulin resulting in trilineage engraftment of donor haemopoiesis. Since then, the patient has been in continuous molecular remission for 11 months.
A novel mutation in the ferrochelatase gene associated with erythropoietic protoporphyria
Authors: Imoto S. , Tanizawa Y. , Sato Y. , Kaku K. , Oka Y.
Published: October 2003

Erythropoietic protoporphyria (EPP) is a hereditary disorder caused by mutations of the ferrochelatase gene. We investigated a Japanese patient with a dominant form of erythropoietic protoporphyria for a ferrochelatase mutation. Sequence analysis of the proband's ferrochelatase cDNA revealed a T to C point mutation at nucleotide 557. This mutation resulted in the replacement of Ile by Thr at amino acid position 186, a novel mutation in erythropoietic protoporphyria. An increase in ferrochelatase activity was not observed in the crude extract of E. coli over‐expressing the mutant protein compared with the control, whereas a marked increase in activity was observed in that over‐expressing the wild type. Prediction of the secondary structure of ferrochelatase suggested that the Ile186 → Thr mutation changed the original β‐sheet structure to an α helix in the region including amino acid residue of mutation. We conclude that, in this patient, the Ile186 → Thr mutation had abolished enzyme activity, possibly by disrupting the secondary structure, thereby causing erythropoietic protoporphyria.
COLLECTION AND TRANSPLANTATION OF PERIPHERAL BLOOD PROGENITOR CELLS MOBILIzED BY G‐CSF ALONE IN CHILDREN WITH MALIGNANCIES
Authors: Diaz M. A. , Villa M. , Alegre A. , Lamana M. L. , De La Vega A. , Granda A. , Madero L.
Published: October 2003

Results of collection and transplantation of peripheral blood progenitor cells (PBPC) mobilized by G‐CSF in 31 children with different malignancies were analysed. A total of 43 aphereses were performed, following administration of granulocyte colony‐stimulating factor (G‐CSF), using a continuous flow blood cell separator (Cobe Spectra) through a central venous catheter. For patients weighing ≤25 kg the extracorporeal line was primed with red blood cells. The mean blood flow rate was 33.2ml/min (range 12–56ml/min). The mean number of mononuclear cells (MNC), granulocyte‐macrophage colony‐forming units (CFU‐GM) and CD34+ cells collected were 7.22×108/kg body weight (b.w.), 15.9×104/kg and 5.44×106/kg respectively.

The morbidity related to PBPC collection was low and the mean apheresis time was 302.7min (range 115–492). The volume of processed blood for apheresis (per kilogram body weight) ranged from 117 to 539.6 (mean 309.13ml/kg). 20 patients required only one apheresis to collect the minimum requirement of 5–7×108/kg of MNC.

All patients subsequently underwent autografting with PBPC after myeloablative therapy. Days to achieve an absolute count of neutrophils (ANC) <0.5×109/l and a platelet count of 20×1092/l without platelet support were 9.5 and 18, respectively. The number of CD34+ cells infused correlated highly with engraftment kinetics. The extramedullary toxicity was low and manageable.
Cyclic change of cytokines in a patient with cyclic thrombocytopenia
Authors: Kimura Fumihiko , Nakamura Yukistugu , Sato Ken , Wakimoto Naoki , Kato Takashi , Tahara Tomoyuki , Yamada Muneo , Nagata Naokazu , Motoyoshi Kazuo
Published: October 2003

The serial change of various cytokines in the serum from a patient with cyclic thrombocytopenia is described. Interleukin 7, stem cell factor, and transforming growth factor β1 synchronized with the platelet count, and there was a significant positive correlation between the three cytokines and the platelet count. Levels of macrophage colony‐stimulating factor, thrombopoietin, platelet‐associated IgG and erythropoietin changed reciprocally with the platelet count, and there was a significant negative correlation between the platelet count and these cytokines except erythropoietin. No cyclic change was observed in IL‐3, IL‐6, IL‐11, granulocyte‐macrophage colony‐stimulating factor, or leukaemia inhibitory factor. These observations suggest that this disease involves two cyclic changes: megakaryocytopoiesis and platelet destruction, in both of which the cytokines play an important role.
The Labelling Index of Primitive Plasma Cells Determines the Clinical Behaviour of Patients with Myelomatosis
Authors: Joshua Doug , Petersen Amarette , Brown Ross , Pope Belinda , Snowdon Leonie , Gibson John
Published: October 2003

For patients with multiple myeloma the most important laboratory correlate of prognosis and disease activity is the bromodeoxyuridine (BrdUrd) plasma cell labelling index (LI). However, the traditional immunofluorescent microscope LI technique, like other manual enumeration assays, can suffer from poor precision and accuracy. In this study the LI of different subpopulations of plasma cells (CD38++) as determined by flow cytometry was correlated with disease state. The mean LI of the total CD38++ population was significantly higher (2.7±0.4%) than the LI determined by the traditional slide technique (0.6±0.1%) for 65 samples tested. Primitive plasma cells (CD38++, CD45++) had a higher labelling index than mature plasma cells (CD38++, CD45−) (7.0±1.3% v 1.8%±0.3%) and in one patient the LI of the primitive plasma cells was 46%. In addition, the LI of the mature plasma cells was lower than the total plasma cell population. As expected, there was a significant difference between the LI of patients in plateau phase and progressive disease but this difference was greatest when the LI of the primitive plasma cells was studied (9.2±2.9% v 2.2±0.7%; z=19.9, P<0.001). This study has raised some concerns about the sensitivity and accuracy of the traditional labelling index and has shown that the increased LI associated with progressive disease is almost entirely attributable to an increase in the LI of the primitive plasma cell subpopulation and that the LI of primitive plasma cells provides a more clinically significant correlation with disease status than the traditional assay.
Parenteral iron increases serum erythropoietin concentration during the ‘early anaemia’ of 10–20‐day‐old mice
Authors: Bechensteen Anne Grete , Halvors en Sverre
Published: October 2003

Parenteral iron abolishes the ‘early anaemia’ in all mammals tested and increases packed cell volume (PCV) and haemoglobin concentration (Hb) significantly above adult levels. In the present study we examined the effect of parenteral iron upon serum immunoreactive erythropoietin (siEpo) concentration in young mice (age 6–24 d) and in adult iron‐deficient mice.

Young BALB/CJ mice, from 5 to 23 d of age, and adult (60–100 d old) mice, some of which were iron deficient due to a low iron diet from the time of weaning, were given iron subcutaneously (iron sorbitol, Fe+++, 1.2 mg iron/100 g body weight/injection). Iron was given as one single dose and the animals killed 20–24 h later. Control mice were similarly treated with saline.  In young mice, from the age 10 d to weaning at 20 d, the siEpo level, measured 20–24 h after a single subcutaneous dose of iron, was 5 times greater than that in control mice (P < 0.0001). In contrast, there was no change in siEpo 20–24 h after a single subcutaneous dose of iron in adult mice, whether iron‐deficient anaemic or normal.

Thus, during the ‘early anaemia’ in young mice over 10 d old, iron increased siEpo levels, whereas it had no such effect in iron‐deficient and normal adult mice, This suggests that between ages 10 and 20 d, iron has additional effects in the regulation of erythropoiesis, which may not occur in adult or in 6‐d‐old mice.
In vivo effect of platelet factor 4 (PF4) and tetrapeptide AcSDKP on haemopoiesis of mice treated with 5‐fluorouracil
Authors: Aidoudi Sallouha , Guigon Martine , Lebeurier Isabelle , Caen Jacques P. , Han Zhong Chao
Published: October 2003

In vivo effects of platelet factor 4 (PF4) and tetrapeptide N‐acetyl‐Ser‐Asp‐Lys‐Pro (AcSDKP) on haemopoietic progenitors were studied in mice treated with 5‐fluorouracil (5‐FU). The mice were injected with PF4 (40 μg/kg) or AcSDKP (4 μg/kg) twice at 6 h intervals, and 20 h after the second injection they were given one injection of 5‐FU (150 mg/kg). 6, 8 and 13 d later the high proliferative potential‐colony forming cell (HPP‐CFC), burst‐forming unit erythroid (BFU‐E), colony forming unit granulocyte‐macrophage (CFU‐GM) colony forming unit megakaryocyte (CFU‐MK), and megakaryocytes (MK) were examined. The results showed that the administration of PF4 or AcSDKP resulted in a significant increase in the number of HPP‐CFC on days 6–8 and BFU‐E and CFU‐GM on day 8 when compared to 5‐FU alone. Furthermore, PF4 was found to increase significantly the number of CFU‐MK and MK on day 8, which was not observed with AcSDKP. However, both molecules had no obvious effect on peripheral blood cells. These data indicate that PF4 or AcSDKP accelerate the recovery in vivo of HPP‐CFC, CFU‐GM and BFU‐E after 5‐FU treatment but their effect may be different on megakaryocytic progenitors and suggests that both molecules may have a haemoprotective effect against chemotherapeutic agents.
Spontaneous complete remission and recovery of donor haemopoiesis without GVHD after relapse and apparent marrow graft rejection in poor‐prognosis myelodysplastic syndrome
Authors: Beguin Yves , Collignon Joelle , Laurent Christian , Fillet Georges
Published: October 2003

We report a patient with poor‐prognosis myelodysplastic syndrome (MDS) after successful treatment of lymphoma, who was given an allogeneic BMT, engrafted and achieved complete remission, but later had a relapse of his MDS with complete disappearance of donor haemopoiesis. After two episodes of CMV pneumonia and continued prophylactic use of ganciclovir thereafter, he experienced a spontaneous complete disappearance of all signs of MDS, including myelofibrosis, and a complete return to donor haemopoiesis. This case is the first one to suggest a graft‐versus‐leukaemia effect (GVL) in MDS patients. It depicts the complex relationship between GVL, graft‐versus‐host disease (GVHD) and graft rejection. It could also constitute a clinical illustration of the possible antileukaemic effect of CMV infection and its treatment with ganciclovir.
Cytogenetic findings in adult de novo acute myeloid leukaemia. A population‐based study of 303/337 patients
Authors: Preiss Birgitte S. , Kerndrup Gitte B. , Schmidt Kai G. , Sørensen Anne G. , Clausen Niels‐Aage T. , Gadeberg Ole V. , Mourits‐Andersen Torben , Pedersen Niels T. ,   on behalf of the AML Study Group Region of Southern Denmark
Published: October 2003

Summary. During a 10‐year period (1992–2001) in the region of Southern Denmark, 337 patients aged 15 years or older (range 16–93 years, median 67 years) were diagnosed with acute myeloid leukaemia (AML). Cytogenetic analysis was carried out in 90%, of whom 53% had clonal chromosome aberrations. Some 24% and 31% had only numerical or structural abnormalities respectively. The remaining patients showed both types of abnormalities. Ploidy levels in decreasing order were: pseudodiploidy, 41%; hyperdiploidy, 32%; and hypodiploidy, 27%. Pseudodiploidy characterizes type M3 (70%) and hypodiploidy M6 (56%). Recurrent cytogenetic abnormalities – t(8;21), t(15;17) and inv(16) – were found in 3·3%, 3·3% and 2·0% of all patients respectively. Prognostically intermediate and adverse aberrations were found in 39% and 44%, respectively, of those with an abnormal karyotype. Rare recurrent aberrations were found in two patients in this material. A previously described non‐recurrent abnormality was found to be recurrent in one patient [der(20)t(11;20)(q13.2;p13)]. New, previously undescribed abnormalities were found in 41 patients. Statistically significant correlations were found between t(15;17) and young age (P < 0·001), inv(16) and young age (P < 0·006), −17 and M6 (P = 0·007), and M6 and complex karyotype with five or more unrelated aberrations (P = 0·004). We conclude that this truely population‐based cytogenetic study of adult AML showed distributions of chromosome abnormalities that differ from those described so far.
Chlorambucil in combination with induction and maintenance rituximab is feasible and active in indolent non‐Hodgkin's lymphoma
Authors: Martinelli Giovanni , Laszlo Daniele , Bertolini Francesco , Pastano Rocco , Mancuso Patrizia , Calleri Angelica , Vanazzi Anna , Santoro Paola , Cavalli Franco , Zucca Emanuele
Published: October 2003

Summary. We investigated the toxicity and efficacy of the chimaeric anti‐CD20 antibody rituximab in combination with standard‐dose chlorambucil in newly diagnosed and relapsed/refractory indolent B‐cell lymphoma patients. A total of 29 patients (15 newly diagnosed and 14 relapsed/refractory) with low‐grade or follicular B‐cell non‐Hodgkin's lymphoma (NHL) were included in this phase II study. Therapy consisted of chlorambucil 6 mg/m2/d for 6 consecutive weeks in combination with a standard 4‐weekly rituximab administration schedule in the induction phase. Patients responding to the induction therapy received four additional cycles with chlorambucil (6 mg/m2/d for 2 weeks/month) plus rituximab (once a month). Twenty‐six patients (89%) completed the treatment; only one patient discontinued treatment because of haematological toxicity. At the end of the study, the dose of chlorambucil had to be reduced in seven patients (27%) and six patients (23%) required a delay in further treatment, as a result of toxicity during consolidation therapy. Only one patient was withdrawn from the study because of progressive disease; the 27 patients evaluable for response at the end of consolidation achieved a clinical response (63% complete response and 26% partial response). A significant CD4+ and CD56+ depletion was observed after induction and during consolidation therapy; two herpes zoster virus infections and one perianal abscess represented major infectious morbidities registered during the study. Based on our preliminary data, the combination of chlorambucil with rituximab seemed to be well tolerated and active. Its definitive role in the treatment of low‐grade NHL should be further evaluated in randomized trials.
Familial polycythemia vera with Budd–Chiari syndrome in childhood
Authors: Cario Holger , Pahl Heike L. , Schwarz Klaus , Galm Christoph , Hoffmann Martin , Burdelski Martin , Kohne Elisabeth , Debatin Klaus‐Michael
Published: October 2003

Summary. Polycythemia vera is a myeloproliferative disorder that, in most cases, occurs sporadically with a median age at presentation of 60 years. Familial cases are very rare and usually manifest in elderly family members. The Budd–Chiari syndrome, characterized by the obstruction and occlusion of the suprahepatic veins, is a rare typical complication in polycythemia vera patients. To date, only two children or adolescents with polycythemia vera and Budd–Chiari syndrome have been described. Here, we report an 11‐year‐old girl with Budd–Chiari syndrome as the initial symptom of familial polycythemia vera, which was also found in the girl's grandmother. Details of the diagnostic procedures used and the clinical course are reported. The patient underwent orthotopic liver transplantation and is being treated with hydroxyurea. The available literature on familial polycythemia vera and polycythemia vera in childhood with and without Budd–Chiari syndrome is reviewed.
Predictive value of early 18F‐fluoro‐deoxyglucose positron emission tomography in chemosensitive relapsed lymphoma
Authors: Schot Bart , Van Imhoff Gustaaf , Pruim Jan , Sluiter Wim , Vaalburg Willem , Vellenga Edo
Published: October 2003

Summary. 18F‐fluoro‐deoxyglucose (FDG) positron emission tomography (PET) might be a better tool than computerized tomography (CT) in predicting long‐term treatment outcome in patients with relapsed chemosensitive lymphoma who are candidates for autologous stem cell transplantation (ASCT). We studied patients with recurrent or persistent aggressive non‐Hodgkin's lymphoma (NHL) and Hodgkin's disease (HD), who were treated with three courses of second‐line induction chemotherapy [DHAP–VIM (dexamethasone, cytarabine, cisplatin followed by etoposide, iphosphamide and methotrexate)‐DHAP], followed by myeloablative therapy and ASCT if chemosensitive. FDG‐PET was performed in parallel to conventional diagnostic methods before starting, and after two courses of, second‐line therapy. Of 68 relapsed lymphoma patients, 46 chemosensitive patients (33 NHL and 13 HD) were included, of whom 39 were transplanted. After DHAP‐VIM, the second PET scan was normalized in 15/46 patients; progression‐free survival at 2 years was 62% for PET‐negative patients versus 32% for PET‐positive patients (P = 0·048). The relative risk for progressive disease in patients with < 90% intensity reduction was 2·85 (95% confidence interval 1·15–7·05, P = 0·018). Early FDG‐PET may help to predict the long‐term treatment outcome of ASCT in chemosensitive patients with relapsed lymphoma and identify those patients who need extra or alternative treatment. Disappearance or > 90% reduction of intensity of abnormal FDG uptake after two courses of reinduction therapy was correlated with a favourable outcome.
Causes of death in sickle cell disease: an autopsy study
Authors: Manci Elizabeth A. , Culberson Donald E. , Yang Yih‐Ming , Gardner Todd M. , Powell Randall , Haynes Johnson , Shah Arvind K. , Mankad Vipul N. ,   Investigators of the Cooperative Study of Sickle Cell Disease
Published: October 2003

Summary. More precise analysis of causes of death is needed to focus research efforts and improve morbidity and mortality in sickle cell disease. In this study, the morphological evidence of the cause of death was studied in 306 autopsies of sickle cell disease, which were accrued between 1929 and 1996. The most common cause of death for all sickle variants and for all age groups was infection (33–48%). The terminal infection was heralded by upper respiratory tract syndromes in 72·6% and by gastroenteritis in 13·7%. The most frequent portal of entry in children was the respiratory tract but, in adults, a site of severe chronic organ injury. Other causes of death included stroke 9·8%, therapy complications 7·0%, splenic sequestration 6·6%, pulmonary emboli/thrombi 4·9%, renal failure 4·1%, pulmonary hypertension 2·9%, hepatic failure 0·8%, massive haemolysis/red cell aplasia 0·4% and left ventricular failure 0·4%. Death was frequently sudden and unexpected (40·8%) or occurred within 24 h after presentation (28·4%), and was usually associated with acute events (63·3%). This study shows that the first 24 h after presentation for medical care is an especially perilous time for patients with sickle cell disease and an acute event. Close monitoring and prompt aggressive treatment are warranted.
DNA microarray analysis of stage progression mechanism in myelodysplastic syndrome
Authors: Ueda Masuzu , Ota Jun , Yamashita Yoshihiro , Choi Young Lim , Ohki Ruri , Wada Tomoaki , Koinuma Koji , Kano Yasuhiko , Ozawa Keiya , Mano Hiroyuki
Published: October 2003

Summary. Myelodysplastic syndrome (MDS) is a clonal disorder of haematopoietic stem cells. Despite the high incidence of MDS in the elderly, effective treatment of individuals in its advanced stages is problematic. DNA microarray analysis is a potentially informative approach to the development of new treatments for MDS. However, a simple comparison of ‘transcriptomes’ of bone marrow mononuclear cells among individuals at distinct stages of MDS would result in the identification of genes whose expression differences only reflect differences in the proportion of MDS blasts within bone marrow. Such a ‘population shift’ effect has now been avoided by purification of haematopoietic stem‐like cells that are positive for the cell surface marker AC133 from the bone marrow of healthy volunteers and 30 patients at various stages of MDS. Microarray analysis with the AC133+ cells from these individuals resulted in the identification of sets of genes with expression that was specific to either indolent or advanced stages of MDS. The former group of genes included that for PIASy, which catalyses protein modification with the ubiquitin‐like molecule SUMO. Induction of PIASy expression in a mouse myeloid cell line induced apoptosis. A loss of PIASy expression may therefore contribute directly to the growth of MDS blasts and stage progression.
Successful treatment of refractory pure red cell aplasia associated with lymphoproliferative disorders with the anti‐CD52 monoclonal antibody alemtuzumab (Campath‐1H)
Authors: Ru Xiuqing , Liebman Howard A.
Published: October 2003

Summary.Acquired pure red cell aplasia (PRCA) is a rare, but significant, complication of lymphoproliferative disorders. It is characterized by anaemia, absence of red cell precursors in the bone marrow and normal granulopoiesis and megakaryopoiesis. We describe two patients with refractory pure red cell aplasia associated with chronic lymphocytic leukaemia (CLL) and a large granular CD8 T‐lymphocytic leukaemia (LGL) respectively. Both patients had failed multiple treatment regimens for PRCA and were transfusion dependent. Both patients were subsequently treated with the anti‐CD52 humanized monoclonal antibody, alemtuzumab, receiving total doses in excess of 300 mg. Response to treatment, as documented by a rapid increase in the reticulocyte count, occurred as early as the third infusion. At the time of this report, both patients remain in complete remission with normal haemoglobin levels. Alemtuzumab appears to be an effective and well‐tolerated therapy for pure red blood cell aplasia associated with lymphoproliferative disorders.
Two novel factor V null mutations associated with activated protein C resistance phenotype/genotype discrepancy
Authors: Dargaud Yesmin , Trzeciak Marie C. , Meunier Sandrine , Angei Christele , Pellechia Dorothee , Négrier Claude , Vinciguerra Christine
Published: October 2003

Summary.  Activated protein C (APC) resistance phenotype/genotype discrepancy is a very rare event. The objective of this study was to characterize the molecular mechanisms in two cases of APC phenotype/genotype discrepancy. An approach using direct sequencing of each exon and splicing junctions of the factor V gene showed that two novel factor V null mutations combined with heterozygous factor V Leiden mutation were responsible for this discrepancy. Our results suggest the necessity to use both phenotypic and genotypic analyses in some cases to determine an accurate diagnosis.
A multicentre assessment of the endogenous thrombin potential using a continuous monitoring amidolytic technique
Authors: Lawrie Anorew S. , Gray Elaine , Leeming Dave , Davidson Simon J. , Purdy Gordon , Iampietro Romina , Craig Steve , Rigsby Peter , Mackie Ian J.
Published: October 2003

Summary. This study assessed the inter‐laboratory imprecision associated with the measurement of the endogenous thrombin potential (ETP). The initial studies used techniques that had evolved in each of the participating laboratories. Samples from normal healthy subjects (n = 10), two patients receiving coumarin therapy [International Normalized Ratio ∼2·0 and ∼4·0] and a further two subjects receiving treatment with unfractionated heparin (anti‐Xa 0·07 iu/ml and 0·31 iu/ml) were assayed relative to a lyophilized normal plasma that had arbitrarily been assigned a potency of 100%. Considerable variation in potency estimates was observed between the centres, although individual laboratories using fully automated techniques achieved acceptable levels of imprecision as assessed by the coefficient of variation (CV) (intra‐assay CV < 9·5%, inter‐assay CV < 12·5%). A second study to assess a similar range of samples, using a standardized assay protocol and incorporating appraisal of two chromogenic substrates, CBS.0068 or Pefachrom® TG, demonstrated markedly improved agreement in potency estimates between centres and good correlation (r > 0·96) between the chromogenic substrates. Our data demonstrates that an automated ETP method can be standardized between laboratories and suitable levels of imprecision achieved, using different analysers (COBAS Mira at two centres and an ACL‐300R) and two thrombin substrates. This indicates that more widespread use of ETP measurements in clinical laboratories is feasible.
Issues facing brothers of sexually abused children: implications for professional practice
Authors: Hill Andrew
Published: October 2003

This paper draws on the author's experience of undertaking play therapy with the brothers of sexually abused children. It illustrates the range of difficulties that such boys may face and identifies implications for professional practice. Difficulties are understood as arising from the struggle to develop a coherent narrative about the abuse, often in the face of incomplete or contested information. The outcome is influenced by the nature of the child's attachment relationships with his non‐abusing parents or carers, his abused sibling and with the abuser himself. This struggle for a coherent narrative about the sexual abuse can have a significant impact on the development of a sense of masculine identity, particularly when the abuser is the boy's own father and/or the abused child is male. In the context of increasing professional awareness of the need to support siblings of sexually abused children it is argued that there should be a specific focus on gender and identity issues for boys.
Worries and problems of young carers: issues for mental health
Authors: Cree Viviene E.
Published: October 2003

This paper reports on a research study which explored the worries and problems of young carers in Edinburgh. Sixty‐one young carers took part in the study, conducted between April and June 2002. Findings indicate that young carers identify significant worries and problems in relation to their well‐being, and that these come over and above any ‘normal’ adolescent difficulties. It is suggested that these findings may have important implications for young carers’ mental health, now and in the future, and contain important lessons for child and family social work in general.
Parenting fostered adolescents: skills and strategies
Authors: Lipscombe Jo , Farmer Elaine , Moyers Sue
Published: October 2003

This paper discusses findings from a recently completed study of adolescent foster care, which included a detailed assessment of the parenting approaches and strategies used by the foster carers of adolescents in long‐term placements. Sixty‐eight foster carers were interviewed at two points in time. The first interview was conducted two months after the start of a new adolescent placement and the second after the placement had been continuing for a year, or at the point of disruption if this occurred earlier. The interview schedules were designed specifically for this study and were based upon well‐established techniques developed in other studies of parenting. They enabled the researchers to make summary ratings for each carer on established dimensions of parenting such as control and discipline, responsiveness and the level of engagement with the child. The researchers assessed how these strategies changed and developed in relation to the young person's behaviour and whether these approaches influenced either the likelihood of placement disruption or the quality of the placement for the child. This paper describes the parenting strategies used by the foster carers, highlighting the areas of parenting that significantly affected the placement outcomes and the corresponding implications for policy and practice.
Between a rock and a hard place: the role of relatives in protecting children from the effects of parental drug problems
Authors: Barnard Marina
Published: October 2003

Using qualitative data gathered from 62 problem drug using parents in Glasgow, Scotland, this paper explores the role played by the extended family in protecting children from some of the negative effects of parental drug use. However, the data also indicated that the intervention of the extended family, whilst often critical, was not without its tensions and difficulties for parents, for the extended family and for the children too. What seemed to obtain in most cases was a complex and volatile mix of practical and emotional concerns over children's appropriate care and issues of responsibility and obligation to the child. These in turn were overlaid by expressions of anxiety, worry, anger and disappointment over both the parent's drug problem and its profound effect on the family. Strained family dynamics and a lack of a supportive welfare infrastructure can all compromise the ability of kin to provide a stable, nurturing environment for children over time. In a worst case scenario it can lead to a breakdown of these care arrangements, leaving these already vulnerable children exposed to further rupture, emotional damage and instability. If we are to avoid such outcomes it is crucial that we first identify and then work to rectify those factors that compromise the abilities of extended family carers to look after these children both in the short and the longer term.
Broadening constructions of family violence: mothers’ perspectives of aggression from their children
Authors: Jackson Debra
Published: October 2003

This qualitative study used feminist insights to explore experiential aspects of the phenomenon of child‐to‐mother violence from the perspectives of six women survivors. Participants took part in conversational style audiotaped interviews lasting between two and four hours. Feminist analysis revealed that fear and violence had become a feature of mothering for these women. Three themes were identified: It was only a matter of time: feeling intimidated and under threat; He just punched me: physical violence from child to mother; and Other men in the house: violence directed to the mothers by friends and associates of their children. Currently the literature overwhelmingly constructs family violence as elder abuse, spousal/partner violence, or child abuse, and generally fails to acknowledge, or address, violence from children to parents. However, findings from this study suggest that the discourse about family violence needs to be broadened to include child‐to‐mother violence. Implications of this study for practice and research are discussed.
Differential activities, subcellular distribution and tissue expression patterns of three members of Slingshot family phosphatases that dephosphorylate cofilin
Authors: Ohta Yusaku , Kousaka Kazuyoshi , Nagata‐Ohashi Kyoko , Ohashi Kazumasa , Muramoto Aya , Shima Yasuyuki , Niwa Ryusuke , Uemura Tadashi , Mizuno Kensaku
Published: October 2003

Background:  Cofilin, a key regulator of actin filament dynamics, is inactivated by phosphorylation at Ser‐3 by LIM‐kinases and is reactivated by dephosphorylation by a family of protein phosphatases, termed Slingshot (SSH).

Results:  We have identified two novel isoforms of SSHs, termed SSH‐2L and SSH‐3L and characterized them in comparison with SSH‐1L that was previously reported. SSH‐1L and SSH‐2L, but not SSH‐3L, tightly bound to and co‐localized with actin filaments. When expressed in cultured cells, SSH‐1L, SSH‐2L and SSH‐3L decreased the level of Ser‐3‐phosphorylated cofilin (P‐cofilin) in cells and suppressed LIM‐kinase‐induced actin reorganization, although SSH‐3L was less effective than SSH‐1L and SSH‐2L. In cell‐free assays, SSH‐1L and SSH‐2L efficiently dephosphorylated P‐cofilin, whereas SSH‐3L did do so only weakly. Using deleted mutants of SSH‐1L and SSH‐2L, we found that the N‐terminal and C‐terminal extracatalytic regions are critical for cofilin‐phosphatase and F‐actin‐binding activities, respectively. In situ hybridization analyses revealed characteristic patterns of expression of each of the mouse Ssh genes in both neuronal and non‐neuronal tissues; in particular, expression of Ssh‐3 in epithelial tissues is evident.

Conclusion:  SSH‐1L, SSH‐2L and SSH‐3L have the potential to dephosphorylate P‐cofilin, but subcellular distribution, F‐actin‐binding activity, specific phosphatase activity and expression patterns significantly differ, which suggests that they have related but distinct functions in various cellular and developmental events.
Negative regulation of Lyn protein‐tyrosine kinase by c‐Cbl ubiquitin‐protein ligase in FcɛRI‐mediated mast cell activation
Authors: Kyo Shinkou , Sada Kiyonao , Qu Xiujuan , Maeno Koichiro , Miah S. M. Shahjahan , Kawauchi‐Kamata Keiko , Yamamura Hirohei
Published: October 2003

Background: Recent studies have demonstrated that c‐Cbl functions as a ubiquitin‐protein ligase toward immune receptors and non‐receptor protein‐tyrosine kinase Syk by facilitating their ubiquitination and subsequent targeting to proteasomes. However, it was not clear whether Src family kinase Lyn is regulated by the Cbl family of ubiquitin‐protein ligases.

Results:  Aggregation of the high affinity IgE receptor (FcɛRI) induces the rapid ubiquitination of Lyn in rat basophilic leukaemia RBL‐2H3 cells. Treatment of cells with a proteasome inhibitor enhances the ubiquitination of Lyn. Stimulation of FcɛRI results in the association of Lyn with c‐Cbl and Cbl‐b, both of which then become tyrosine phosphorylated. Co‐transfection study shows that both c‐Cbl and Cbl‐b could induce the ubiquitination of activated Lyn in COS cells. Furthermore, over‐expression of membrane‐anchored form of c‐Cbl inhibits the FcɛRI‐mediated degranulation and cytokine gene production in RBL‐2H3 cells by the down‐regulation of the kinase activity of Lyn through the enhanced ubiquitination.

Conclusions:  These results demonstrate that Lyn is down‐regulated by c‐Cbl‐mediated ubiquitination and subsequent degradation in proteasome after FcɛRI stimulation in mast cells. Targeting of c‐Cbl in the lipid raft results in the inhibition of FcɛRI‐mediated mast cell activation.
On multiple solutions of the atmosphere–vegetation system in present‐day climate
Authors: Claussen Martin
Published: October 2003

An asynchronously coupled global atmosphere–biome model is used to assess the stability of the atmosphere–vegetation system under present‐day conditions of solar irradiation and sea‐surface temperatures. When initialized with different land‐surface conditions (1, the continents, except for regions of inland ice, completely covered with forest; 2, with grassland; 3, with (dark) desert; and 4, with (bright) sand desert), the atmosphere–biome model finds two equilibrium solutions: the first solution yields the present‐day distribution of subtropical deserts, the second reveals a moister climate in North Africa and Central East Asia and thereby a northward shift of vegetation particularly in the south‐western Sahara. The first solution is obtained with initial condition 4, and the second with 1, 2, 3. When comparing these results with an earlier study of biogeophysical feedback in the African and Asian monsoon area, it can be concluded that North Africa is probably the region on Earth which is most sensitive considering bifurcations of the atmosphere–vegetation system at the global scale.
Influences of local feedbacks on land–air exchanges of energy and carbon
Authors: Raupach M. R.
Published: October 2003

Land–air exchanges of energy and matter are modulated by several feedback processes at both small and large space and time scales, with implications for the linked carbon, water and energy cycles. This paper studies the influences of four local feedbacks, occurring at single‐patch spatial scales and subdiurnal temporal scales, on the surface energy balance (SEB) and land–air carbon fluxes. The feedbacks are: (i) radiative feedback, the modulation of available energy through the effect of surface temperature, Ts, on outgoing longwave radiation; (ii) physiological feedback, the interaction between vegetation physiology and the SEB through Ts; (iii) aerodynamic feedback, the modulation of turbulent heat and moisture transfer by atmospheric stability; and (iv) Convective Boundary Layer (CBL) feedback, the coupling between the daytime evolution of the SEB and CBL through saturation deficit.

It is found that radiative feedback is significant only over very smooth surfaces. Physiological feedback is positive with respect to Ts at moderate to high temperatures, pushing stomata towards complete closure and the SEB towards very low evaporation rates. The SEB is quite sensitive to whether or not such closure occurs. Aerodynamic feedback, on the other hand, is negative with respect to Ts at these temperatures, reducing Ts and attenuating the tendency for heat‐induced stomatal closure. CBL feedback alone does not dampen the sensitivity of the SEB to physiological feedback and stomatal closure. However, when aerodynamic feedback is included, this sensitivity is greatly reduced.
Interactions between the atmosphere and terrestrial ecosystems: influence on weather and climate
Authors: Pielke Roger A. , Sr . , Avissar RonI. , Raupach Michael , Dolman A. Johannes , Zeng Xubin , Denning A. Scott
Published: October 2003

This paper overviews the short‐term (biophysical) and long‐term (out to around 100 year timescales; biogeochemical and biogeographical) influences of the land surface on weather and climate. From our review of the literature, the evidence is convincing that terrestrial ecosystem dynamics on these timescales significantly influence atmospheric processes. In studies of past and possible future climate change, terrestrial ecosystem dynamics are as important as changes in atmospheric dynamics and composition, ocean circulation, ice sheet extent, and orbit perturbations.
Variations of ecosystem gas exchange in the rain forest mesocosm at Biosphere 2 in response to elevated CO2
Authors: Rosenthal YaiR.
Published: October 2003

The effects of elevated CO2 on tropical ecosystems were studied in the artificial rain forest mesocosm at Biosphere 2, a large‐scale and ecologically diverse experimental facility located in Oracle, Arizona. The ecosystem responses were assessed by comparing the whole‐system net gas exchange (NEE) upon changing CO2 levels from 900 to 450 ppmV. The day‐NEE was significantly higher in the elevated CO2 treatment. In both experiments, the NEE rates were similar to values observed in natural analogue systems. Variations in night‐NEE, reflecting both soil CO2 efflux and plants respiration, covaried with temperature but showed no clear correlation with atmospheric CO2 levels. After correcting for changes in CO2 efflux we show that the rain forest net photosynthesis increased in response to increasing atmospheric CO2. The photosynthetic enhancement was expressed in higher quantum yields, maximum assimilation rates and radiation use efficiency. The results suggest that photosynthesis in large tropical trees is CO2 sensitive, at least following short exposures of days to weeks. Taken at face value, the data suggest that as a result of anthropogenic emissions of CO2, tropical rain forests may shift out of steady state, and become a carbon sink at least for short periods. However, a better understanding of the unique conditions and phenomena in Biosphere 2 is necessary before these results are broadly useful.
Evidence that local land use practices influence regional climate, vegetation, and stream flow patterns in adjacent natural areas
Authors: Stohlgren Thomas J. , Chase Thomas N. , Pielke Roger A. , Sr . , Kittel Timothy G. F. , Baron JilL. S.
Published: October 2003

We present evidence that land use practices in the plains of Colorado influence regional climate and vegetation in adjacent natural areas in the Rocky Mountains in predictable ways. Mesoscale climate model simulations using the Colorado State University Regional Atmospheric Modelling System (RAMS) projected that modifications to natural vegetation in the plains, primarily due to agriculture and urbanization, could produce lower summer temperatures in the mountains. We corroborate the RAMS simulations with three independent sets of data: (i) climate records from 16 weather stations, which showed significant trends of decreasing July temperatures in recent decades; (ii) the distribution of seedlings of five dominant conifer species in Rocky Mountain National Park, Colorado, which suggested that cooler, wetter conditions occurred over roughly the same time period; and (iii) increased stream flow, normalized for changes in precipitation, during the summer months in four river basins, which also indicates cooler summer temperatures and lower transpiration at landscape scales. Combined, the mesoscale atmospheric/land‐surface model, short‐term trends in regional temperatures, forest distribution changes, and hydrology data indicate that the effects of land use practices on regional climate may overshadow larger‐scale temperature changes commonly associated with observed increases in CO2 and other greenhouse gases.
Terrestrial models and global change: challenges for the future
Authors: Hurtt George C. , Moorcroft PauL. R. , And Stephen W. Pacala , Levin Simon A.
Published: October 2003

A wide variety of models have illustrated the potential importance of terrestrial biological feedbacks on climate and climate change; yet our ability to make precise predictions is severely limited, due to a high degree of uncertainty. In this paper, after briefly reviewing current models, we present challenges for new terrestrial models and introduce a simple mechanistic approach that may complement existing approaches.
Residues near the N‐terminus of protein B control autocatalytic proteolysis and the activity of soluble methane mono‐oxygenase
Authors: Callaghan Anastasia J. , Smith Thomas J. , Slade Susan E. , Dalton Howard
Published: October 2003

Soluble methane mono‐oxygenase (sMMO) of Methylococcus capsulatus (Bath) catalyses the O2‐dependent and NAD(P)H‐dependent oxygenation of methane and numerous other substrates. During purification, the sMMO enzyme complex, which comprises three components and has a molecular mass in excess of 300 kDa, becomes inactivated because of cleavage of just 12 amino acids from the N‐terminus of protein B, which is the smallest component of sMMO and the only one without prosthetic groups. Here we have shown that cleavage of protein B, to form the inactive truncated protein B′, continued to occur when intact protein B was repeatedly separated from protein B′ and all detectable contaminants, giving compelling evidence that the protein was cleaved autocatalytically. The rate of autocatalytic cleavage decreased when the residues flanking the cleavage site were mutated, but the position of cleavage was unaltered. Analysis of a series of incremental truncates showed that residue(s) essential for the activity of sMMO, and important in determining the stability of protein B, lay in the region Ser4–Tyr7. Protein B was shown to possess intrinsic nucleophilic activity, which we propose initiates the cleavage reaction via a novel mechanism. Proteins B and B′ were detected in approximately equal amounts in the cell, showing that truncation of protein B is biologically relevant. Increasing the growth‐medium copper concentration, which inactivates sMMO, did not alter the extent of in vivo cleavage, therefore the conditions under which cleavage of protein B may fulfil its proposed role as a regulator of sMMO remain to be identified.
Structural basis for poor uracil excision from hairpin DNA
Authors: Ghosh Mahua , Rumpal Nidhi , Varshney Umesh , Chary Kandala V. R.
Published: October 2003

Two‐dimensional NMR and molecular dynamics simulations have been used to determine the three‐dimensional structures of two hairpin DNA structures: d‐CTAGAG GATCCUTTTGGATCCT (abbreviated as U1‐hairpin) and d‐CTAGAGGATCCTTUTGGATCCT (abbreviated as U3‐hairpin). The 1H resonances of both of these hairpin structures have been assigned almost completely. NMR restrained molecular dynamics and energy minimization procedures have been used to describe the three‐dimensional structures of these hairpins. This study and concurrent NMR structural studies on two other d‐CTAGAGGA TCCTUTTGGATCCT (abbreviated as U2‐hairpin) and d‐CTAGAGGATCCTTTUGGATCCT (abbreviated as U4‐hairpin) have shed light upon various interactions reported between Echerichia coli uracil DNA glycosylase (UDG) and uracil‐containing DNA. The backbone torsion angles, which partially influence the local conformation of U12 and U14 in U1 and U3‐hairpins, respectively, are probably locked in the trans conformation as in the case of U13 in the U2‐hairpin. Such a stretched‐out backbone conformation in the vicinity of U12 and U14 is thought to be the reason why the Km value is poor for U1‐ and U3‐hairpins as it is for the U2‐hairpin. Furthermore, the bases U12 and U14 in both U1‐ and U3‐hairpins adopt an anti conformation, in contrast with the base conformation of U13 in the U2‐hairpin, which adopts a syn conformation. The clear discrepancy observed in the U‐base orientation with respect to the sugar moieties could explain why the Vmax value is 10‐ to 20‐fold higher for the U1‐ and U3‐hairpins compared with the U2‐hairpin. Taken together, these observations support our interpretation that the unfavourable backbone results in a poor Km value, whereas the unfavourable nucleotide conformation results in a poor Vmax value. These two parameters therefore make the U1‐ and U3‐hairpins better substrates for UDG compared with the U2‐hairpin, as reported earlier [Kumar, N. V. & Varshney, U. (1997) Nucleic Acids Res. 25, 2336–2343.].