bluuebunny/crossref_metadata_2025 huggingface dataset:
Severe aplastic anaemia in association with a unique constitutional translocation 46,XY,t(6;10)(q13;q22)c
Authors: Hudson J. , Chown S. , Lawler M. , Duggan C. , Temperley I. J. , Secker‐Walker L.
Published: November 2003

Severe aplastic anaemia (SAA) is an uncommon disorder which may be associated with several congenital syndromes. However, it has rarely been described in association with a constitutional karyotypic abnormality. The breakpoint of the balanced t(6;10)(q13;q22) translocation described here does not disrupt any currently recognized gene of haemopoietic or stromal importance. This report also highlights the problems inherent in the use of bone marrow transplantation (BMT) for treating multiply transfused aplastic anaemia patients.
Granulocyte colony‐stimulating factor given in addition to interferon‐α to mobilize peripheral blood stem cells for autologous transplantation in chronic myeloid leukaemia
Authors: Archimbaud E. , Michallet M. , Philip I. , Charrin C. , Clapisson G. , Belhabri A. , Guilhot F. , Stryckmans P. , Adeleine P. , Fière D.
Published: November 2003

In order to potentially mobilize and harvest the Ph− cells observed in most patients with chronic myeloid leukaemia (CML) during interferon‐α (IF‐α) therapy, G‐CSF (filgrastim), 5 μg/kg/d, was administered subcutaneously together with IF‐α to 30 CML patients in haematological remission but with various degrees of cytogenetic remission, after IF‐α therapy. Peripheral blood stem cells (PBSC ) were harvested using standard aphereses from day 5 of G‐CSF. Patients underwent one to four (median three) aphereses. Median total yields/kg were 7.6 (range 3.8–25) × 108 MNC, 3.4 (0–140) × 106 CD34+ cells, and 17 (1.1–107) × 104 CFU‐GM. No patient had a significant increase in the percentage of Ph+ cells in the bone marrow under G‐CSF therapy. The percentage of Ph+ cells in apheresis products tended to decrease between the first and the last apheresis (P = 0.05). 14 patients who were not responsive to IF‐α were transplanted after conditioning with busulphan 16 mg/kg and melphalan 140 mg/m2. Median time to neutrophils > 0.5 × 109/l was 20 d (16–114 d) and to platelets > 50 × 109/l 18 d (12–149 d). Nine patients had a major cytogenetic response post graft, which correlated with the amount of Ph+ cells reinfused with the graft (P = 0.02). We conclude that this procedure is feasible, allowing the harvest of enough PBSC, some of them Ph− in patients who responded to IF‐α, to allow autologous transplantation.
Multiple myeloma: reduced plasma cell contamination in peripheral blood progenitor cell collections performed after repeated high‐dose chemotherapy courses
Authors: Omedè Paola , Tarella Corrado , Palumbo Antonio , Argentino Chiara , Caracciolo Daniele , Corradini Paolo , Dominietto Alida , Giaretta Fulvia , Ravaglia Roberta , Triolo Roberta , Triolo Sabrina , Pileri Alessandro , Boccadoro Mario
Published: November 2003

The possibility of reducing tumour cell contamination by cytotoxic drug courses prior to peripheral blood progenitor cell (PBPC) collection was evaluated in two consecutives groups of multiple myeloma (MM) patient candidates for autograft. All patients were at disease onset and received two VAD (vincristine, doxorubicin and dexamethasone) courses as initial debulking. In the first group (44 patients), mobilization and harvest were performed ‘upfront’, after a single cyclophosphamide (CY) administration of 4 g/m2; in the second group (17 patients), PBPC were collected at the end of a high‐dose sequential chemotherapy programme, including: CY 5 g/m2, etoposide (VP16) 2 g/m2, a chemotherapy‐free interval with three courses of high‐dose dexamethasone, a final mobilizing CY at 7 g/m2. G‐CSF was given following each high‐dose cytotoxic drug. Cytofluorimetric analysis was performed to quantify progenitors (CD34+ cells) and plasma cells, identified by the high CD38 expression and/or CD38 and CD138 coexpression. Large amounts of PBPC were collected in either group (median harvested CD34+/kg: 15.8 × 106 and 13.4 × 106, respectively; P = 0.9). Circulating plasma cells were significantly higher in patients mobilized ‘upfront’ compared to those who received the high‐dose sequence (median peak values of CD38bright/μl: 39 and 10, respectively; P = 0.02); a similar difference was observed in the amount of con‐taminating plasma cells in the harvest products (median CD38bright/kg: 7.4 × 106 and 1.3 × 106, respectively; P = 0.02). The results demonstrate that an in vivo purging approach is feasible in myeloma patients through repeated high‐dose chemotherapy courses; this may provide less‐contaminated material suitable for further in vitro purging procedures.
Hepatic GVHD after HLA‐haploidentical bone marrow transplantation in children with severe combined immunodeficiency: the effect of ursodeoxycholic acid
Authors: WULFFRAAT NICO M. , HADDAD ELIE , BENKERROU MALIKA , SPLIET WIM G. M. , PATEY NATACHA , FISCHER ALAIN , DE GRAEFF‐MEEDER BABETTER.
Published: November 2003

We describe two children with a severe combined immune deficiency (SCID) with B cells. Following a T‐cell‐depleted haploidentical bone marrow transplantation (BMT), they both developed a chronic graft‐versus‐host disease (GVHD) of the skin and a severe persisting hyperbilirubinaemia and elevated liver enzymes. The diagnosis of a vanishing bile duct syndrome was confirmed by liver biopsies. Because corticosteroids and cyclosporin A induced only a partial response, ursodeoxycholic acid (UDCA) was added to their treatment schedule. Serum bilirubin and liver enzymes returned to normal within months. A control liver biopsy showed normal and proliferating bile ducts without cholestatic damage. We conclude that UDCA was well tolerated and may be of value as an additional treatment for hepatic GVHD in SCID.
Action of human interleukin‐4 and stem cell factor on erythroid and mixed colony formation by peripheral blood‐derived CD34+c‐kithigh or CD34+c‐kitlow cells
Authors: SONODA YOSHIAKI , KIMURA TAKAFUMI , OHMIZONO YOSHIKAZU , SAKABE HIDEAKI , TANIMUKAI SHIGEATSU , YOKOTA SHOUHEI , CLARK STEVEN C. , ABE TATSUO
Published: November 2003

We studied the interaction of interleukin (IL)‐4 and other burst‐promoting activity (BPA) factors, such as IL‐3, granulocyte/macrophage colony‐stimulating factor (GM‐CSF), IL‐9 and stem cell factor (SCF), on erythroid burst‐forming unit (BFU‐E) and erythrocyte‐containing mixed (CFU‐Mix) colony formation in serum‐free culture. IL‐4 alone did not support mixed colony formation in the presence of erythropoietin (Epo). However, IL‐4 showed weak but significant BPA when peripheral blood (PB)‐derived CD34+c‐kitlow cells were used as the target population. The BPA of IL‐4 was much weaker than that of IL‐3, which exerted the most potent activity, as previously reported. When CD34+c‐kithigh cells were used as the target, four factors known to have BPA, IL‐3, GM‐CSF, IL‐9 and SCF, could express BPA. In contrast, IL‐4 alone failed to support erythroid burst formation. Interestingly, IL‐4 showed a remarkable enhancing effect with SCF in promoting the development of erythroid burst and erythrocyte‐containing mixed colonies from CD34+c‐kitlow and CD34+c‐kithigh cells. Delayed addition of SCF + Epo or IL‐4+Epo to the cultures initiated with either IL‐4 or SCF alone clearly demonstrated that SCF was a survival factor for both BFU‐E and CFU‐Mix progenitors. In contrast, the survival effect of IL‐4 was much weaker than that of SCF, and appeared to be more important for progenitors derived from CD34+c‐kitlow cells than for those derived from CD34+c‐kithigh cells. It was recently reported that CD34+c‐kitlow cells represent a more primitive population than CD34+c‐kithigh cells. Taken together, these results suggest that IL‐4 helps to recruit primitive progenitor cells in the presence of SCF.
Quantitation and characterization of human megakaryocyte colony‐forming cells using a standardized serum‐free agarose assay
Authors: HOGGE DONNA , FANNING STEPHEN , BOCKHOLD KATHRYN , PETZER ANDREAS , LAMBIE KAREN , LANSDORP PETER , EAVES ALLEN , EAVES CONNIE
Published: November 2003

Human progenitors of the megakaryocyte (Mk) lineage were detected by their ability to generate colonies containing from 3 to >100 Mk, detectable as glycoprotein IIb/IIIa+ cells in APAAP‐stained whole mount agarose cultures. Optimal growth conditions were achieved through the use of a defined serum substitute and a suitable cocktail of recombinant cytokines. Under these culture conditions, the smallest Mk‐containing colonies (CFC‐Mk) were detectable within a week followed by colonies containing larger numbers of Mk over the ensuing 2 weeks. The total number of CFC‐Mk at 18–21 d was linearly related to the number of cells plated. Variation in the cytokines added showed that thrombopoietin (TPO) or IL‐3 alone would support the formation of large numbers of CFC‐Mk. However, optimal yields of colonies containing cells of both Mk and non‐Mk lineages required the addition of other growth factors, of which a combination of IL‐3, IL‐6, GM‐CSF and Steel factor (SF) ± TPO was the best of those tested. The further addition of erythropoietin to this combination reduced the number of large ‘pure’ Mk colonies seen and in their place a corresponding number of mixed erythroid‐Mk colonies became detectable. Flt3‐ligand alone was unable to support the growth of CFC‐Mk nor did it enhance their growth when combined with other factors. Plating of FACS‐sorted subpopulations of CD34+ marrow cells in both serum‐free agarose and methylcellulose assays demonstrated that most CFC‐Mk are generated from CD34+ cells that are CD45RA− and CD71+, approximately half of which are CD41+. Thus, CFC‐Mk are more similar to primitive clonogenic erythroid progenitors than to their granulopoietic counterparts in their expression of CD34, CD45RA and CD71. Taken together, these findings support the concept that some erythroid and Mk progenitors may share a common developmental pathway. The availability of sensitive and reproducible procedures for isolating and detecting human Mk progenitors should facilitate future investigations of their biology and role in a variety of haematological conditions.
Clinical, haematological and molecular studies in patients with chromosome translocation t(7;11): a study of four Chinese patients in Taiwan
Authors: HUANG SHANG‐YI , TANG JIH‐LUH , LIANG YUH‐JIN , WANG CHIU‐HWA , CHEN YAO‐CHANG , TIEN HWEI‐FANG
Published: November 2003

Translocation t(7;11)(p15;p15) is an uncommon but recurrent chromosome aberration in acute myeloid leukaemia (AML), which occurs mostly in oriental patients and in AML M2 or, occasionally, M4 subtype. Recently, a consistent chimaeric fusion transcript NUP98‐HOXA9 was found in several cases of t(7;11). Four AML cases with the chromosome abnormality in Taiwan are described. They were all adults with ages ranging from 30 to 41 years (median 36 years). Three of them were diagnosed as having AML M2; the remaining one as M4. Marked dyserythropoiesis was demonstrated in two patients. All four patients showed pan‐myeloid antigen CD13 on the leukaemic cells, but none coexpressed lymphocyte‐associated antigens and neither of the two patients studied for CD34 expression had positive staining. NUP98‐HOXA9 fusion transcript was detected in both patients who had molecular analysis and the breakpoints on chromosome 11 and 7 respectively were similar to those previously reported. They all received conventional induction chemotherapy, but only one achieved a complete remission (CR) with short duration. This study and others reported in the literature suggest a racial or geographical predisposition among oriental patients to AML with t(7;11) and that this is associated with a poor prognosis. The molecular detection of NUP98‐HOXA9 fusion transcript would be a useful method for the diagnosis of t(7;11) and also for monitoring disease status after treatment.
Allogeneic MHC‐mismatched activated natural killer cells administered after bone marrow transplantation provide a strong graft‐versus‐leukaemia effect in mice
Authors: Zeis M. , Uharek L. , Glass B. , Steinmann J. , Dreger P. , Gassmann W. , Schmitz N.
Published: November 2003

Allogeneic lymphocytes administered with an unmanipulated bone marrow transplant provide a strong antileukaemic effect, the so‐called graft‐versus‐leukaemia (GVL) effect. On the other hand, T‐cell‐mediated graft‐versus‐host‐disease (GVHD) observed after transplantation of unmanipulated BM graft causes substantial morbidity and mortality. The aim of the present study was to determine the antileukaemic potential of enriched IL‐2 activated NK cells administered 2 h after BMT. Balb/c (H‐2d) mice were given a dose of A20 (H‐2d, B‐cell leukaemia) cells 2 d prior to lethal total body irradiation (TBI) and transplantation of either syngeneic or allogeneic anti‐Thy1.2 (CD90) depleted bone marrow cells. Either syngeneic (Balb/c, H‐2d) or allogeneic (C57BL/6, H‐2b) enriched and IL‐2 (200 U/ml for 24 h) activated NK cells were given 2 h after BMT. Injection of A20 leukaemia into normal Balb/c recipients led to death after a median of 14 d. A lethal dose of TBI followed by either syngeneic or allogeneic Thy1.2‐depleted BMT resulted in a modest antileukaemic effect. The adoptive transfer of syngeneic enriched and IL‐2 preincubated NK cells given at time of BMT exerted a significantly better GVL effect. However, the infusion of allogeneic enriched NK cells resulted in a stronger GVL effect. These results clearly demonstrate that allogeneic NK cells are superior to syngeneic NK cells in their potential to eradicate residual leukaemia cells after BMT without mediating clinical overt GVHD. This experimental setting may offer a strategy for treatment of haematological malignancies in a phase of minimal residual disease.
Flow cytometry of leucocyte alkaline phosphatase in normal and pathologic leucocytes
Authors: RAMBALDI ALESSANDRO , MASUHARA KENSAKU , BORLERI GIAN‐MARIA , AMARU RICARDO , GIANNÌ MAURIZIO , TERAO MINEKO , BARBUI TIZIANO , GARATTINI ENRICO
Published: November 2003

Leucocyte alkaline phosphatase (LAP) is an enzyme expressed on the external aspect of the neutrophilic granulocyte plasma membrane, and represents a specific marker for the fully differentiated granulocyte. In this report we characterize 1B12.1, a monoclonal antibody raised against human bone alkaline phosphatase, by its ability to recognize the LAP protein. As assessed by Western blot analysis, following electrophoresis under non‐reducing conditions, the antibody specifically reacts with LAP upon forced expression of the protein in simian COS‐7 fibroblasts. In addition, the 1B12.1 antibody recognizes partially purified LAP isolated from peripheral blood granulocytes. With this antibody we developed a quantitative flow‐cytometry‐based method for the determination of LAP. Double fluorescence flow cytometry demonstrated that the LAP protein was present in relatively high amounts in neutrophilic granulocytes, but not in monocytes, natural killer cells, or B and T lymphocytes of normal individuals. The protein was completely absent in granulocytes obtained from chronic myeloid leukaemia and paroxysmal nocturnal haemoglobinuria patients. Higher than normal levels of LAP protein were evident in neutrophilic granulocytes of patients suffering from polycythaemia vera, essential thrombocythaemia and severe aplastic anaemia. However, the highest amounts of LAP protein were present in the granulocytes of normal individuals treated with G‐CSF for the isolation of peripheral blood stem cells.
Congenital leukaemia with a mixed phenotype of megakaryoblasts and erythroblasts: a case report and characterization of the blasts
Authors: MORI TETSUYA , KANEKO HIDEO , KUMAGAI MASA‐AKI , MIYAUCHI JUN , KANEKO YASUHIKO , FUJIMOTO JUNICHIRO , TSUNEMATSU YUKIKO
Published: November 2003

We present a congenital leukaemia with a mixed phenotype of megakaryoblasts and erythroblasts. A newborn male with exopthalmus and multiple skin nodules, had bone marrow blasts which expressed CD41b, CD42b, glycophorin‐A and haemoglobin, but monocyte or lymphoid markers were negative. The patient achieved a complete remission with chemotherapy. Blasts cultured for a few months expressed erythroid markers but lost the megakaryocytic phenotype, although addition of phorbol ester induced the latter phenotype. Spontaneous colony formation was observed in semi‐solid culture and the number of colonies was increased by erythropoietin. Detailed studies further indicated the heterogeneity of congenital leukaemia.
Cell birth and death in childhood acute lymphoblastic leukaemia: how fast does the neoplastic cell clone expand?
Authors: Hirt Andreas , Leibundgut Kurt , Ridolfi Lüthy Annette , Von der Weid Nicolas , Wagner Hans‐Peter
Published: November 2003

In 23 children with untreated precursor B‐cell acute lymphoblastic leukaemia (ALL), the daily growth rate of the malignant cell clone was calculated. Cell birth expanded the leukaemic cell clone an average 10–11% per day, programmed cell death or apoptosis reduced the leukaemic cell mass by some 4% per day. From these two variables a net increase in the size of the leukaemic cell population of 6.9 ± 7.3% (range −1.2–27.3%) per day could be calculated. The daily growth rate correlated negatively with the logarithm of the duration of clinical symptoms before the diagnosis of ALL was established (r=−0.680; P = 0.0004). A long history, especially in children with undefined bone pain and arthralgias, was associated with a very slow expansion of the neoplastic cell clone.
Escape of HIV-1 is associated with lack of V3 domain-specific antibodies in vivo
Authors: Schreiber M , Muüller H , Wachsmuth C , Laue T , Hufert F T , Van Laer M D , Schmitz H
Published: November 2003

This study was performed to analyse correlates of viral escape in AIDS patients. Peripheral blood mononuclear cells (PBMC) from HIV− donors were inoculated with AIDS patients' serum to detect neutralization-resistant cell-free virus. Infectious virus was detected by polymerase chain reaction (PCR) and analysed by sequencing the V3 region. The escaped virus species was compared with all V3 virus variants found in the patients' PBMC and plasma. In one patient escaped virus was also compared with variants found in CD4+ T cells isolated by FACS from blood, spleen and lymph node. The frequency of the virus variants was determined by cloning and sequence analysis of 20 V3 clones for each PCR amplification. To monitor anti-V3 antibodies by ELISA, each V3 sequence was expressed as fusion with glutathione S-transferase (GST-V3). In our AIDS patients, a V3-directed antibody response against the infectious virus V3 loop was not detectable. In contrast, virus variants unable to infect the donor PBMC in vitro were well recognized by homologous V3-directed antibody. After an interval of 1 year the frequency of these variants clearly decreased, while at the same time the escaped variants grew out and finally represented the predominant viral species both in plasma and PBMC. The infectious variants lacking V3 antibody response were also predominant in CD4+ T cells in spleen and lymph node. Our data indicate that the escape of virus variants is closely related to the lack of V3-directed antibody.
Protection against anti-glomerular basement membrane (GBM)-mediated nephritis in C3- and C4-deficient mice
Authors: SHEERIN N S , SPRINGALL T , CARROLL M C , HARTLEY B , SACKS S H
Published: November 2003

Mice rendered completely deficient of the complement components C3 or C4 were used to determine the influence of complement activation in the heterologous phase of the anti-GBM disease model. In wild-type animals the disease is characterized by a neutrophil infiltrate, capillary thrombosis, proteinuria and C3 and C4 deposited within the glomerulus. The early infiltration of neutrophils into the glomeruli is greater in wild-type mice (2·8 ± 0·3) compared with C3-deficient (1·4 ± 0·2) and C4-deficient (1·2 ± 0·003) mice. Deficiency also protects against the subsequent development of proteinuria (2·99 ± 1·11 mg/24 h, 0·059 mg/24 h and 0·327 ± 0·14 mg/24 h in wild-type, C3-deficient and C4-deficient mice, respectively) and decreases glomerular capillary thrombosis in both C3- and C4-deficient mice. The degree of protection is greater in the C3-deficient than the C4-deficient animals, suggesting both classical and alternative pathway involvement. These studies support a critical role for complement in the development of anti-GBM disease. However, the protective effect of complement deficiency can be broken if the dose of nephritogenic antibody is increased.
B cells and monocytes are not developmentally affected in a case of reticular dysgenesis
Authors: DE LA CALLE-MARTÍN O , BADELL I , GARCÍA A , GELPÍ C , CASAMITJANA N , ESTELLA J , RODRÍGUEZ-SÁNCHEZ J L
Published: November 2003

We report a patient with reticular dysgenesis (RD) who received an HLA-identical marrow graft and remained free of infection in spite of incomplete haematological recovery. Mixed chimerism was achieved and resulted from the presence of autologous B cells and monocytes and grafting of donor T cells. Granulocyte recovery was impaired. The B cells were CD5+ (B1 cells) and appeared to be functional, since serum immunoglobulin levels became normal after the graft. The findings described here suggest that in some cases the defect selectively affects different cell types, including the more abundant leucocyte populations, granulocytes and T lymphocytes. However, B cells and monocytes appear to be relatively spared in this case of RD. Furthermore, the present case may provide insight into the mechanism involved in the expansion of distinct B cell subpopulations (B1 and B2 cells).
Intracellular cytokine expression in whole blood preparations from normals and patients with atopic dermatitis
Authors: FERRY B , ANTROBUS P , HUZICKA I , FARRELL A , LANE A , CHAPEL H
Published: November 2003

In recent years, the importance of characterizing the role of cytokines in a wide range of clinical conditions has resulted in development of new methods to assess cytokine expression in clinical samples. The use of anti-cytokine MoAbs and flow cytometry to detect cytokines intracellularly at the single-cell level has the potential to quantify cytokine production in different diseases. For this technique to be useful in a clinical setting, rapid throughput of clinical samples and a cheap, reliable assay would be required, therefore the development of the above technique using unseparated whole blood samples would be advantageous. Using this technique, only one study to date (Maino et al., 1996) has used unseparated whole blood as the source of cells for detecting intracellular cytokines. In clinical practice, whole blood may be optimal, since this most closely approximates conditions in vivo: as no purification of blood mononuclear cells is required, very little blood is needed to detect a number of cytokines simultaneously in various lymphocyte subpopulations, and the assay can be applied to samples from infants and children. In this study we describe an intracellular cytokine assay using unseparated whole blood from normals. In activated CD8− T cells, IL-2 and interferon-gamma (IFN-γ) were optimally induced after 10 h stimulation with phorbol 12-myristate acetate (PMA)/ionomycin, and in CD8+ T cells IL-2 was optimally induced after 10 h and IFN-γ after 6 h. The levels of IL-2 and IFN-γ in CD8+ and CD8− T cells in four healthy individuals were consistent on four occasions over a 3-month period. In a large group of 34 normal subjects, there was considerable heterogeneity in CD3/IL-2+ (range 9·7–41·3) and CD3/IFN-γ+ cells (10·1–44), expressed as a percentage of total lymphocytes. In patients with atopic dermatitis (n = 5) there was a significantly decreased percentage of CD3+/CD8+ peripheral blood T cells expressing IFN-γ and an increased percentage of CD3+/CD8− T cells expressing IL-4 compared with non-atopic dermatitis controls (n = 5). Possible applications of this technique are discussed.
The in vivo induction of lymphocyte apoptosis in MRL-lpr/lpr mice treated with FTY720
Authors: Suzuki S , Li X-K , Shinomiya T , Enosawa S , Amemiya H , Amari M , Naoe S
Published: November 2003

The in vitro treatment of lpr thymocytes with FTY720 resulted in a dose-dependent reduction in cell viability due to apoptosis. In order to study the effect of FTY720 in vivo, lpr mice received an oral daily dose of 1 mg/kg FTY720 for 14 days, beginning at 16 weeks of age which was when the animals were developing massive lymphoadenopathy. Compared with untreated lpr mice, FTY720-treated lpr mice had significantly prolonged lives. At 24 weeks of age, treated mice demonstrated markedly reduced weights of the spleen and lymph nodes, and the proportion of CD3+B220+ and CD4−CD8− cells in the thymus, spleen and lymph nodes decreased markedly. In addition, in these mice the percentage of CD4+CD8+ and CD3−B220− cells in the thymus and the percentage of CD4+CD8−, CD4−CD8+, CD3+B220− and CD3−B220+ cells in the spleen returned to almost the normal values observed in wild-type mice. Histological observation 1 day after the final administration of FTY720 revealed a remarkable infiltration of neutrophils in the lymphoid organs. Apoptotic cells were detected in all the lymphoid organs using in situ DNA nick-end labelling. Electron microscopy showed that the apoptotic cells were ingested by phagocytes. FTY720 therapy is thus highly effective in Fas-mutant animals with abnormally expanding lymphocytes.
Bone mass and metabolism in women aged 45–55
Authors: Garton Mark , Martin James , New Susan , Lee Sue , Loveridge Nigel , Milne John , Reid David , Reid Ivan , Robins Simon
Published: November 2003

OBJECTIVE Changes in calcium homeostasis and bone mass around the climacteric are poorly understood. We examined relations between endocrine factors and indices of bone mass and metabolism in healthy women approaching the menopause.

DESIGN Cross‐section study.

PATIENTS Sixty‐eight spontaneously menstruating women aged 45–55.

MEASUREMENTS Bone density measured at lumbar spine (LS) and femoral neck (FN) using dual energy X‐ray absorptiometry and distal non‐dominant forearm using peripheral quantitative computed tomography. We recorded menstrual history, physical activity and dietary calcium, and measured serum calcium, phosphate, alkaline phosphatase, osteocalcin, vitamin D, fT3, T4, TSH, PTH, FSH and oestradiol (E2), and urinary pyridinoline (PYD) and deoxypyridinoline (DPD) excretion.

RESULTS Using serum FSH level as a marker of ovarian function, 63 subjects could be classified into one of three groups: group A (serum FSH <10 U/l, n = 29), group B (10–35 U/l, n = 27) and group C (>35 U/l, n = 7). Bone density fell with declining ovarian function at the LS, FN and forearm trabecular (but not cortical) sites. Serum PTH was lower in group A vs B (mean (SD) 2.68 (0.97) vs 3.52 (1.17) pmol/l, P < 0.05), but similar to group C (2.90 (1.09) pmol/l, P = NS). Serum phosphate was elevated in group C compared to groups A and B (1.17 (0.15) vs 1.04 (0.11) and 1.05 (0.13) mmol/l, P < 0.05), and urinary PYD (61.1 (8.0) vs 50.4 (11.6) and 43.9 (8.1) μmol/mol creatinine) and DPD (15.9 (3.9) vs 12.0 (3.6) and 11.4 (3.6) μmol/mol creatinine) excretion were also increased. There were no significant differences in vitamin D metabolites or osteocalcin. Multivariate analysis suggested serum osteocalcin was positively related to physical activity and serum 1,25‐dihydroxycholecalciferol levels. Serum free T3 was positively correlated with urinary DPD excretion, and inversely related to serum PTH. In all subjects, serum PTH was related to body weight (r = 0.38, P = 0.002).

CONCLUSIONS Declining ovarian function before menopause is accompanied by reductions in bone mass and altered calcium metabolism. Free T3 may regulate bone resorption and indirectly modulate PTH release.
Regular audit of colposcopic biopsies from women with a mildly dyskaryotic or borderline cervical smear results in fewer cases of CINIII
Authors: JONES M. H. , JENKINS D. , SINGER A.
Published: November 2003

Two years after introducing mandatory review of cases in which the cervical smear was discrepant with subsequent colposcopic or histological finding, the predictive accuracy of a first abnormal smear and the need for treatment were analysed. The results were compared with performance figures prior to this form of audit policy. Over 12 months 415 women referred for colposcopy were studied. Three per cent of patients with a single borderline smear and 6% with mild dyskaryosis had cervical intraepithelial neoplasia grade III (CINIII) revealed in histopathological examinations after colposcopy. Only 25% with a borderline smear and 33% with mild dyskaryosis required treatment. Of women with moderate dyskaryosis, 18% had a biopsy showing CINIII and 46% were treated. Of women with severe dyskaryosis in their cervical smear, 61% were shown to have CINIII or invasive cancer on biopsy and 90% were treated. Regular audit improved cytological prediction of grade of epithelial abnormality found on biopsy, allowing accurate, safe surveillance for minor smear abnormalities.
DNA ploidy and pS2 protein expression in breast cancer
Authors: IOAKIM‐LIOSSI A. , KARAKITSOS P. , ARONI K. , MARKOPOULOS C. , DELIVELIOTI K. , GOGAS J. , KYRKOU K.
Published: November 2003

The DNA content of ductal breast carcinomas of varying histological grade was measured using static image cytometry and correlated with pS2 expression in the tumour cells. Our study was performed on imprint of surgical biopsies of 60 women with ductal breast cancer. A statistically significant difference was observed between pS2+ expression and grade of malignancy (P<0.001). The percentage of euploid tumours significantly decreased from grade I to grade II to grade III (P=0.01). The percentage of aneuploid tumours increased from pS2+ to pS2− breast tumours (P<0.001). These findings may be indicative of pS2 and DNA ploidy alterations and tumour aggressiveness.
A pilot study of cervical screening in an inner city area—lessons for a national programme
Authors: HERITY B. , McDONALD P. , JOHNSON Z. , CARROLL B. , CODY M. , DUIGNAN N. , McGEE D. , O'KELLY F. , HURLEY M.
Published: November 2003

The objectives of this study were to examine aspects of organization of a proposed national screening programme based in general practice. The target population of women aged 25–59 years and their general practitioners (GPs), in a defined inner city area, was identified from a population register of persons eligible for free medical services; a computerized system was developed for invitations and record linkage of cytology results. Smears were examined in one laboratory and follow up of women with abnormal smears was undertaken by one gynaecologist. A random sample of non‐responders was surveyed by questionnaire. Response following two invitations was only 20%. Practices with male doctors only had significantly lower reponse rates (P<0.001) than those with a female doctor/nurse. A survey of non‐responders showed that over 20% of addresses were incorrect and 16% of those interviewed were ineligible for smear tests. A preference for a female to undertake smears was expressed by 67%, and 77% believed that the purpose of the cervical smear was to detect cancer. An accurate population register, health promotion, support for GP practices, provision of alternative venues for smear tests, development of computer systems, accurate data entry and fail‐safe follow up are aspects of a cervical screening service which must be addressed prior to setting up a national service.
LDL‐apheresis significantly reduces urinary apo(a) excretion
Authors: KOSTNER K. M. , JANSEN M. , MAURER G. , DERFLER K.
Published: November 2003

Abstract. Increased plasma Lp(a) is an established risk factor for atherosclerosis. We recently described the presence of apo(a) fragments in urine and the significant correlation between urinary apo(a) concentrations and plasma Lp(a). Here we investigated urinary apo(a) in patients suffering from familial hypercholesterolaemia (FH), treated with LDL apheresis. Before treatment, plasma Lp(a) levels and urinary apo(a) normalized to creatinine were >2‐fold increased in FH patients (P <0.0001) as compared to controls. LDL‐apheresis led to a reduction of plasma Lp(a) by 75% and a concommittant immediate reduction of urinary apo(a) by 45%. We conclude that a steady state condition for urinary apo(a) is rapidly achieved via LDL‐apheresis.
Relationship between low‐density lipoprotein size and apolipoprotein A‐I‐containing particles: the ECTIM study
Authors: Luc G. , Bard J.‐M. , Poulain P. , Arveiler D. , Evans A. E. , Cambien F. , Fruchart J.‐C. , Ducimetière P.
Published: November 2003

It is now established that small dense low‐density lipoproteins (LDLs) are more common among patients with coronary heart disease than among control individuals. Small LDL size is also associated with a high‐risk profile, including increased levels of triglycerides and decreased high‐density lipoprotein (HDL)‐cholesterol. Furthermore, some human HDL particles contain both apolipoprotein (apo) A‐I and apoA‐II (LpA‐I:A‐II) while others contain apoA‐I but are devoid of apoA‐II (LpA‐I). We have investigated the relationship between LDL size, measured by non‐denaturing gradient‐gel electrophoresis, and HDL parameters, particularly LpA‐I and LpA‐I:A‐II levels, in healthy control subjects (n = 408). LDL size was positively and significantly correlated with HDL‐cholesterol (r = 0.43), apoA‐I (r = 0.32) and LpA‐I (r = 0.29), whereas no correlation was observed with apoA‐II and LpA‐I:A‐II. The determination of the mean apoA‐I and LpA‐I in the quintiles of LDL size distribution revealed a progressive increase in apoA‐I and LpA‐I from the first quintile (small LDLs), 137 and 42 mg dL−1 respectively, to the fifth quintile (large LDLs), 161 and 54 mg dL−1 respectively. Conversely, no evolution of apoA‐II and LpA‐I:A‐II was observed. Multivariate analysis showed that not only triglycerides, but also HDL‐cholesterol, apoA‐I and LpA‐I, are determinants of LDL size, depending on the model used. Thus, part of the variation in size of LDLs is associated with the metabolism of HDL independently of that of triglycerides. Thus, a low concentration of LpA‐I combined with the presence of small LDLs could contribute to the high‐risk profile observed in subjects with small LDLs.
Effect of colchiceine and ursodeoxycholic acid on hepatocyte and erythrocyte membranes and liver histology in experimentally induced carbon tetrachloride cirrhosis in rats
Authors: NAVA‐OCAMPO A. A. , SUSTER S. , MURIEL P.
Published: November 2003

Colchiceine and ursodeoxycholic acid (UDCA) are drugs currently in use as therapy for different types of liver damage. We evaluated their ability to reverse the damage induced by carbon tetrachloride (CCl4) in rats. Six groups were analysed: (1) CCl4 (0.4 g kg−1, i.p., three times a week) for 13 weeks; (2) CCl4 for 8 weeks followed by colchiceine (60 μg kg−1) + CCl4 for 5 weeks; (3) CCl4 for 8 weeks and thereafter UDCA (25 mg kg−1) + CCl4 for 5 weeks. Groups 4, 5 and 6 were appropriate controls of colchiceine, UDCA and vehicles respectively. Na+,K+‐ and Ca2+‐ATPase activities and the cholesterol–phospholipid (CH/PL) ratio from erythrocyte and hepatocyte membranes were quantified. Membrane enzymatic activities and CH/PL ratios were affected more in group 1 than groups 2 and 3. We concluded that colchiceine and UDCA were effective drugs in this model of liver damage.
Bovine ferritin iron bioavailability in man
Authors: Skikne B. , Fonzo D. , Lynch S. R. , Cook J. D.
Published: November 2003

The bioavailability of ferritin iron was evaluated in human subjects using radiolabelled [55Fe]ferritin isolated from bovine spleen and liver. Preliminary studies with bovine spleen ferritin labelled in vitro demonstrated an inappropriately high absorption compared with ferritin labelled in vivo, and the latter was therefore used in all subsequent absorption studies. In 10 subjects, geometric mean absorption from 5 mg of ferritin iron was 3.8% when taken without and 3.2% when taken with food (P >0.05). These values were significantly lower than absorption from the same dose of iron given as ferrous sulphate, which averaged 24.1% without and 8.2% with food. When the iron dose was increased 10‐fold, absorption of ferritin iron averaged only 0.6–0.7% with or without food as compared with 7.9% without and 2.6% with food when the iron was given as ferrous sulphate. In a further study, mean absorption from bovine spleen ferritin of 4.0% did not differ significantly from the mean of 2.7% observed with bovine liver ferritin. These findings confirm previous studies indicating that ferritin iron is poorly absorbed. Furthermore, its use as a pharmaceutical iron preparation cannot be advocated.
Bone mineral density of vertebrae, proximal femur and os calcis in normal Greek subjects as assessed by dual‐energy X‐ray absorptiometry: comparison with other populations
Authors: Hadjidakis D. , Kokkinakis E. , Giannopoulos G. , Merakos G. , Raptis S.A.
Published: November 2003

Dual energy X‐ray absorptiometry (DXA) is an established method for the detection of even small changes in bone mineral density (BMD). It thus allows the earliest possible diagnosis of osteopenia, with consequent prompt estimation of fracture risk. However, for proper evaluation of densitometry results it is essential that a comparison with reference BMD values of normal age‐ and sex‐matched persons from the same population be performed. For this purpose we determined bone density of the L2–L4 vertebrae, the L3 vertebra in the lateral projection, the proximal femur and the os calcis in a cross‐sectional study of 168 men and 244 women from the Greek population. The age range of the subjects was 20–80 years. Peak bone mass for both sexes was attained in the 30–35 year age group for the vertebrae and in the 25–30 year age group for the proximal femur and os calcis. Mean annual vertebral bone loss calculated on cross‐sectional data ranged from 0.1% to 0.22% for women <50 years and from 1.3% to 1.6% for those >50 years, whereas in men the range was from 0.36% to 0.64% for the whole age spectrum. Regarding femoral neck, the values were 0.3% (women <50 years), 1.2–1.5% (>50 years) and 0.6–0.8% for men. Total bone loss between ages 20 and 70 was 29.5% for the vertebrae and 32% for the femoral neck in women, whereas the values for men were 19.5% and 29% respectively. A positive correlation was observed between bone density, body weight and body height in both sexes. Body mass index correlated significantly with density only in postmenopausal women. Compared with North American, Finnish and German populations, Greek men presented with lower BMD values in the decades above 40 years. Greek women exhibited lower vertebral BMD values than those from the USA, Germany and Japan (50–60 age group), whereas they did not differ from those of Finnish women. However, femoral neck BMD in Greek women was higher than in Japanese women in all age groups.
Delayed growth response of Mountain Birch seedlings to a decrease in fertilization and temperature
Authors: Weih M.
Published: November 2003

1. The effects of previous‐year environment on current growth response were tested in seedlings of Mountain Birch (Betula pubescens Ehrh. ssp. czerepanovii[Orlova] Hämet‐Ahti), a subarctic tree species with indeterminate shoot growth.

2. Mountain Birch seedlings were pot‐grown outdoors in subarctic Sweden for 2 years and showed a clear delay in growth response when fertilization and temperature were reduced after the first year. The seedlings were grown under four experimental treatments (two temperatures and two nutrient availabilities) in 1994, and under low‐temperature/low‐nutrient conditions in 1995.

3. When nutrient supply and/or temperature was reduced in 1995 compared to 1994, the seedlings maintained the high relative growth rates (RGR) of the previous growing season, although the internal plant N accumulation rate was lower than in 1994. This resulted in decreasing plant N concentration (PNC), and a poor relationship between RGR and PNC during 1995. The high RGR in 1995 was achieved in response to phenotypic adjustments (e.g. number of foliar buds) to a more favourable environment in the past, and by dilution of the internal nutrient storage.

4. The effects of delayed responses found in this study indicate problems for the interpretation of results from growth studies performed under any climate with great year‐to‐year variability, such as the subarctic, because a delay in growth response could distort the relationships between plant growth, resource availability and climate. Predictions of current growth are therefore meaningless if the previous history of the plants is not taken into account.
Protein splicing: its chemistry and biology
Authors: Anraku Yasuhiro
Published: November 2003

Protein splicing is a chemical reaction in which a spliced intervening polypeptide is excised from a precursor protein and the flanking N‐ and C‐terminal regions are ligated with the peptide bond to produce two mature proteins. This unique autocatalytic reaction was first discovered in the yeast VMA1 protein, a 120 kDa spliced polypeptide encoded by the VMA1 gene of Saccharomyces cerevisiae. The VMA1 protein catalyses a self protein splicing post‐translationally to yield the 70 kDa catalytic subunit of the vacuolar H+‐ATPase and the 50 kDa DNA endonuclease. Accumulating evidence has indicated that splicing precursors distribute widely in many organisms covering eukarya, bacteria and archaea. This article argues and summarizes current chemical and biological views on protein splicing.
Initiation mechanisms in replication of filamentous phage DNA
Authors: Horiuchi Kensuke
Published: November 2003

Filamentous phage DNA replication occurs in two steps. First, an RNA‐primed minus strand is made on the plus strand. Then, a new plus strand is made on the resulting double strand. The RNA primer which initiates synthesis of the minus strand is produced at a specific site on the plus strand template by the host RNA polymerase holoenzyme. Despite a lack of sequences similar to the promoter consensus, the DNA replication origin has a much higher affinity for the holoenzyme than the transcriptional promoters. The non‐template strand of the single‐stranded −10 region of the origin appears to be responsible for this high affinity. The recognition mechanisms seem to share common features with those in transcriptional promoters. Plus‐strand synthesis is initiated by a specific nick introduced into the negatively supercoiled replicative form by the phage‐encoded initiator protein. The nicking reaction is preceded by an ordered series of protein‐induced DNA conformational changes.
The role of DNA demethylation during development
Authors: Weiss Alona , Cedar Howard
Published: November 2003

The somatic genomic methylation pattern which plays a role in the suppression of basal gene activity is established anew in each generation through developmentally regulated de novo and demethylation steps. Demethylation appears to be carried out by a nucleotide exchange reaction which may involve RNA molecules, and is directed to specific loci in the genome through interactions between cis acting elements and trans acting factors.
The efficiency and timing of initiation of replication of multiple replicons of Saccharomyces cerevisiae chromosome VI
Authors: Yamashita Minoru , Hori Yuji , Shinomiya Tomoyuki , Obuse Chikashi , Tsurimoto Toshiki , Yoshikawa Hiroshi , Shirahige Katsuhiko
Published: November 2003

Background:

A complete set of nine ARSs was identified (the tenth ARS in this paper), mapped on chromosome VI of Saccharomyces cerevisiae, and characterized for functional elements.

Results:

The level of activity of all ARSs as chromosomal replication origins was determined by neutral/neutral 2D gel‐electrophoresis. These origins were classified into three groups: (i) three high frequency origins used once nearly every cell cycle, (ii) four intermediate frequency origins used once in two to three cycles and (iii) two low frequency origins used in fewer than 5% of cell cycles. These variations in initiation frequency among origins of chromosome VI are present in three common laboratory wild‐type strains. Each origin is initiated at a fixed time and passively replicated by incoming replication forks at a fixed time during a synchronous S phase. Replication of each arm of the chromosome starts from one major origin located one‐fifth (left arm) and one‐third (right arm) of the distance from the centromere, and expands sequentially in both directions. Two telomere vicinity origins are replicated last. Time of initiation and replication of the last replicating origin, Ori609, was remarkably variable from cell to cell.

Conclusions:

Chromosome VI of S. cerevisiae contains nine replication origins that comprise five active replicons under normal cell growth conditions. A clear correlation was found between the efficiency of initiation and the order of replication. The timing of initiation of most origins, except for the first and last, is coincident with the time of passive replication by incoming forks from neighbouring origins.
Pain after hysterectomy: a comparison between four currently available procedures
Authors: Roushdy M. , Farag O. , Mosaad M. , Zayed M.
Published: November 2003

Objective To compare the postoperative pain of four types of hysterectomy. Design Prospective study. Setting Cairo University Faculty of Medicine Hospital and the authors' private practice. Subjects A total of 427 patients underwent hysterectomies for different indications. Interventions Patients were assigned to either total abdominal hysterectomy, laparoscopic‐assisted vaginal hysterectomy, subtotal laparoscopic hysterectomy, or vaginal hysterectomy. Measures Linear pain score 12 and 24 h after surgery, dose of meperidine required in the first 24 h, multidimensional pain score 8 days after surgery, and dyspareunia assessed after 3 months of surgery. Results Both laparoscopic procedures were followed by the least postoperative pain and earliest return to work. Subtotal laparoscopic hysterectomy was followed by the least dyspareunia. Abdominal hysterectomy was followed by the highest pain score and latest return to normal physical activity. Conclusion Laparoscopic hysterectomy had the best patient acceptance, with the least postoperative pain.
Microendoscopy in the operating room
Authors: Van Der Wat J.
Published: November 2003

Objective

To test the capabilities of microlaparoscopes in the performance of gynaecological endoscopic surgery.

Design

A prospective study involving 118 cases. The surgical procedures performed were stratified according to surgical skills as defined by the American Society of Reproductive Surgeons.

Setting

A fully equipped endoscopy operating room.

Subjects

Patients presenting for gynaecological diagnostic and surgical procedures.

Interventions

Microlaparoscopy using a 2‐mm fibreoptic laparoscope, and performance of endoscopic surgery using microinstrumentation where possible.

Results

Microlaparoscopy can be used for level I procedures. A break‐point occurred with level II procedures, as a quarter of cases could not be completed using the microlaparoscope. This was mainly because of lens contamination with blood and peritoneal fluid.

Conclusion

Microendoscopy can be used to perform level I and some level II procedures with precision and confidence. It is atraumatic and well tolerated.
Rupture of a tubal pregnancy following successful treatment with methotrexate
Authors: Cane P. , Wijngaarden W. , Powell M. C.
Published: November 2003

We report a case of a woman not wanting surgery for an ectopic pregnancy, whose HCG titres successfully fell to an undetectable level whereupon the ectopic ruptured. This case illustrates the need for a careful prolonged follow‐up with serial HCG measurements when methotrexate is employed for the treatment of ectopic pregnancy.