bluuebunny/crossref_metadata_2025 huggingface dataset:
Analgesic efficacy of sustained release paracetamol in patients with osteoarthritis of the knee
Authors: Bacon T. H. , Hole J. G. , North M. , Burnett I.
Published: March 2003

Aims  Paracetamol is widely recommended as the initial treatment for pain associated with osteoarthritis (OA). A sustained release (SR) paracetamol formulation (Panadol Extend™) was compared with standard immediate release (IR) paracetamol (Panadol™) in patients with knee pain secondary to OA. The primary parameter for assessment of efficacy was patient‐assessed global pain relief as determined on day 8 of the treatment period.

Methods  A double‐blind, double‐dummy, randomized study was conducted. Patients (n=403) were treated for 7 days with paracetamol 4 g day−1 (SR paracetamol, two 665 mg tablets taken three times daily; IR paracetamol, two 500 mg tablets taken four times daily). Patients completed daily pain measurements and assessed global pain relief at the end of the study. Therapeutic noninferiority was defined on the basis of achieving statistical noninferiority for global pain relief.

Results  Analysis of the primary parameter for the intention to treat population showed that the difference in proportion of patients (SR−IR paracetamol) achieving a successful response on day 8 was −0.7%; 90% CI (−8.82%, 7.45%), P=0.890. For the per protocol population the difference in proportion was −3.0%; 90% CI (−11.61%, 5.66%), P=0.571. As the lower bound of the 90% CI for the treatment difference in each case was greater than the prespecified value (−15%), SR paracetamol was considered to be statistically noninferior to IR paracetamol in terms of pain relief. The treatments were not significantly different for any of the secondary parameters in either populations.

Conclusions  SR paracetamol taken three times daily was statistically and therapeutically noninferior to IR paracetamol taken four times daily in patients with knee pain due to OA. SR paracetamol may be more convenient for patients with chronic pain and has the potential to enhance compliance and therefore pain relief.
Pharmacokinetics of tibolone in early and late postmenopausal women
Authors: Timmer C. J. , Verheul H. A. M. , Doorstam D. P.
Published: March 2003

Aims  Tibolone is a tissue‐specific compound with favourable effects on bone, vagina, climacteric symptoms, mood and sexual well being in postmenopausal women, without stimulating the endometrium or breast. Since tibolone is used for the treatment of both young and elderly postmenopausal women, its pharmacokinetics were studied to investigate potential differences with age. In addition, the bioequivalence of the 1.25 and 2.5 mg tablets was evaluated.

Methods  Single doses of 1.25 or 2.5 mg of tibolone were given in a double‐blind, randomized, two‐way cross‐over study to women aged between 45 and 55 years or between 65 and 75 years of age.

Results  Age did not have a significant effect on Cmax, tmax, and t½ of tibolone and its metabolites and on the body weight standardized oral clearance (CL/F kg−1) of the 3α‐ and 3β‐hydroxy tibolones. In early postmenopausal women, significantly lower values were found for the AUC(0,16 h), and AUC(0,∞) of 3α‐hydroxy tibolone 24.6±6.6 vs 29.2±4.9 and 27.1±6.9 vs 32.3±6.5 ng ml−1 h for the 1.25 mg tablet, respectively, and 45.4±13.9 vs 55.7±14.1 and 49.6±14.6 vs 62.6±17.3 ng ml−1 h for the 2.5 mg tablet, respectively. When these values were adjusted for the significantly higher body weight of the early postmenopausal women, the differences disappeared. No significant differences between early and late postmenopausal women were found for the AUC(0,8 h), and AUC(0,∞) of 3β‐hydroxy tibolone. The rate of absorption of tibolone and the rates of absorption or formation of the 3α‐ and 3β‐hydroxy tibolones were significantly higher after the 1.25 mg dose than after the 2.5 mg tablet, resulting in increases of 32%, 27% and 17% for the dose normalized‐Cmax of tibolone and the 3α‐ and 3β‐hydroxy tibolones, respectively. tmax for tibolone and its metabolites was 12–27% less after 1.25 mg compared to 2.5 mg, which was statistically significant. The two formulations were bioequivalent with respect to the dose‐normalized AUC(0,∞) and the AUC(0,tfix) values for the 3α‐hydroxy tibolone (ratio point estimate [90%, confidence limits]: 1.08 [1.04, 1.14] and 1.08 [1.03, 1.13], respectively) and for the 3β‐hydroxy tibolone (1.07 [1.01, 1.14] and 1.04 [0.96, 1.12], respectively). Both formulations were also bioequivalent with respect to CL/F kg−1 and t½.

Conclusions  The pharmacokinetics of tibolone are similar in early (age 45–55 years) and late (65–75 years) postmenopausal women. The 2.5 and 1.25 mg tablets are bioequivalent with respect to the extent of absorption. The rate of absorption or formation of the metabolites of tibolone were not bioequivalent, but these differences are considered to have no clinical relevance in view of the chronic administration of tibolone.
Pharmacogenetics and the future of medical practice
Authors: Lindpaintner Klaus
Published: March 2003

Pharmacogenetic approaches are widely expected to bring about a “revolution” in medicine. While the application of molecular genetic approaches to disease research will provide us with new opportunities for progressively more targeted and, hopefully, more effective treatments, these developments will be evolutionary in nature and will, for their realization, still require the painstaking process that discovering and developing a new drug entails. It is also quintessential for the realization of these promises that we support a more rational understanding and more realistic expectations in the public at large through dialogue and information.
The value of inpatient pharmaceutical counselling to elderly patients prior to discharge
Authors: Al‐Rashed S. A. , Wright D. J. , Roebuck N. , Sunter W. , Chrystyn H.
Published: March 2003

Aims  The use of medication and information discharge summaries (MIDS) has become a standard procedure in many hospitals. We have evaluated if these summaries, together with in‐patient pharmaceutical counselling backed up with a simple medicine reminder card, may help with the delivery of seamless pharmaceutical care.

Methods  Elderly patients prescribed more than four items discharged to their own home received the standard discharge policy including a recently introduced MIDS and medicine reminder card. Each patient's GP was sent a copy on discharge. Pre‐discharge a pharmacist counselled study patients about their medicines and compliance. A research pharmacist visited patients in their home approximately 2–3 weeks and at 3 months post‐discharge to determine their drug knowledge, compliance, home medicine stocks and any healthcare related events.

Results  Forty‐three study and 40 control patients completed both visits. Their mean (s.d.) ages were 80.2 (5,7) and 81.1 (5,8) years and they were prescribed 7.1 (1.8) and 7.1 (2.3) items, respectively. At visit 1 knowledge (P < 0.01) and compliance (P < 0.001) was better in the study group. At visit 2 compliance had improved in the study group (P < 0.001). Unplanned visits to the GP and readmission to hospital amongst the study group were 19 and 5, respectively, which were both significantly less (P < 0.05) than 27 and 13 in the control group. At visit 2 for the study group the 24 unplanned GP visits and three re‐admissions were significantly (P < 0.05) less than the respective 32 and 15 in the control group. At visit 1, two study group patients had altered their own medication compared with 10 control patients. At visit 2 these reduced to 0 and 4, respectively.

Conclusions  In‐patient pharmaceutical counselling, linked to a medication and information discharge summary and a medicine reminder card, contributed to better drug knowledge and compliance together with reduced unplanned visits to the doctor and re‐admissions. A pharmaceutical domiciliary visit consolidated the improved healthcare outcomes.
Interaction between sotalol and albuterol after CABG: influence on postoperative arrhythmias and length of stay at an intensive care unit
Authors: Vader C. , Grouls R. , Van Gerven A. , Ackerman E. , Korsten H. , Schönberger J. , Ter Woorst F. , Botman C. , Leufkens H.
Published: March 2003

Atrial fibrillation (AF) represents the most common arrhythmic complication of coronary artery bypass grafting (CABG). Incidence rates vary between 17% and 33% [1]. Peak incidence occurs 2 to 3 days postoperatively [2]. Postoperative AF has been associated with increases in total length of stay (LOS), costs, morbidity and mortality.

The objective was to investigate if the pharmacodynamic interaction between albuterol and sotalol is associated with an increase in the incidence of AF and LOS in CABG‐patients at an Intensive Care Unit (ICU).

The study was a retrospective nested, matched case‐control study carried out in the ICU of the Catharina Hospital, Eindhoven, The Netherlands, in the period 1998–2000.

The subjects were patients hospitalised at an ICU after CABG, treated with sotalol with or without albuterol or albuterol/ipratropium bromide. Patients were matched by date of CABG, pre‐operative pulmonary function (Tiffeneau‐index >0.7) and pre‐operative CRASH score, a hospital score used as a prognostic value for post‐operative morbidity and mortality.

The results were analyzed using the Intensive Care Information System (ICIS; INAD Computers & Software BV, Eindhoven) and the Cardio Reporting System of the Catharina Hospital, Eindhoven.

Demographic characteristics of cases and controls (no statistically significant differences between both groups) are presented in Table 1.

Sotalol/albuterol

 (n=217)
Sotalol  (n=350)

Age (years, mean±s.d.)
64.8 ± 9.7
62.9 ± 9.4

Sex (Male)
167 (77.0%)
258 (73.7%)

Aortaocclusion  (min, mean±s.d.)
46.8 ± 29.1
40.1 ± 23.8

Extra corporal circulation  (min, mean±s.d.)
66.1 ± 38.5
56.9 ± 32.4

CRASH score (median %  mortality, range)
5 (0–19)
4 (0–20)

The concurrent use of sotalol and albuterol led to an increased risk for postoperative atrial fibrillation (OR 6.2; 95% confidence interval: 2.3, 17.7). The Mantel‐Haenszel odds ratio, corrected for age, was 1.67 (Table 2). LOS was significantly increased.

AF  present
AF  absent
LOS ICU  (days (median), range)

Sotalol/albuterol
18
199
1 (0–47)

Sotalol
5
345
1 (0–5)

The concurrent use of sotalol and albuterol leads to an increased risk for postoperative AF and is associated with an increased LOS at the ICU of our hospital.
Basal cell carcinoma: are early appointments justifiable?
Authors: Gordon P.M. , Cox N.H. , Paterson W.D. , Lawrence C.M.
Published: March 2003

In the U.K., patients with suspected skin tumours are usually referred from a general practitioner to a hospital-based dermatologist for treatment. The urgency of such referrals is currently a topic of political importance. We have studied case record data from 162 patients with basal cell carcinoma (BCC) to establish the relevance of referrals routinely being considered urgent. At presentation, mean tumour size (maximum dimension) was 7·4 mm (median 10) and mean duration of BCC was 20·5 months (median 12). There was no correlation (all P > 0·05) between tumour size and patient age (r = 0·1325), tumour size and duration (r = 0·4433), or tumour size and interval between referral and hospital consultation (r = 0·0695). If the slow growth rate of the average BCC is assumed to be linear, a reduction in referral interval from the mean value of 10·7 weeks in our patients down to the U.K. government target of 2 weeks would equate to a size difference in BCC of 0·7 mm, which is not therapeutically significant. Individual BCCs with rapid growth or other features of concern may require urgent referral, but evidence from our patients and from other studies is that this is not routinely necessary.
Measuring the activity of the disease in patients with cutaneous lupus erythematosus
Authors: Parodi A. , Massone C. , Cacciapuoti M. , Aragone M.G. , Bondavalli P. , Cattarini G. , Rebora A.
Published: March 2003

The Systemic Lupus Activity Measure (SLAM) is a system proposed by rheumatologists to measure disease activity in their patients with systemic lupus erythematosus (LE). It involves scoring a group of clinical symptoms and laboratory findings, the maximum possible score being 84. In systemic LE, the mid-point is between 9 and 12. We applied SLAM to 176 patients with cutaneous LE. Ninety-seven had localized discoid LE (L-DLE), 59 had disseminated discoid LE (D-DLE) and 20 had subacute cutaneous LE (SCLE). Eighty-five patients had low activity disease (0–4 points), 72 mildly active disease (5–9 points), 15 moderately active disease (10–14 points) and only four had very active disease (≥ 15 points). The most frequent lesions in patients who scored more than 10 points were photosensitivity, cicatricial alopecia, Raynaud’s phenomenon and oral ulcers. Fifty patients were followed up for more than 5 years (mean follow-up 9 years). Nine of these had an increased SLAM score. Seven had L-DLE, one D-DLE and one SCLE. Seven of the 50 patients had photosensitivity, five cicatricial alopecia, five non-cicatricial alopecia, two Raynaud’s phenomenon and two oral ulcers. Three patients who started with L-DLE evolved to D-DLE. The SLAM system is useful in the monitoring of disease activity in patients with cutaneous LE. Over time, even L-DLE patients may develop active disease. Photosensitivity, alopecia, oral ulcers and Raynaud’s phenomenon seem to herald a worse prognosis.
Macroscopic and microscopic mucinosis in chronicsclerodermoid graft-versus-host disease
Authors: Ameen M. , Russell-Jones R.
Published: March 2003

Secondary cutaneous mucinosis is a well-recognized feature of connective tissue diseases such as lupus erythematosus and dermatomyositis. We report the first three cases of dermal mucinosis in association with severe chronic cutaneous graft-versus-host disease of the sclerodermoid variety. One patient had clinical changes due to abundant mucin accumulation within the papillary dermis (mucinoma). In the other two patients histological examination revealed extensive deposits of mucin predominantly within the reticular dermis. The microscopic appearances were striking, with numerous vacuolated spaces interspersed between collagen bundles. We speculate that this appearance is the result of ground substance becoming trapped within grossly sclerodermoid connective tissue.
Cerebriform nodular amelanotic metastases of malignant melanoma: a challenge in differential diagnosis of a rare variant
Authors: Assmann A. , Farthmann B. , Burkhardt O. , Dippel E. , Orfanos C.E. , Blume-Peytavi U.
Published: March 2003

High variability of the clinical appearance of malignant melanoma (MM) and its metastases render the differential diagnosis of solid amelanotic tumours difficult. We report a 71-year-old woman with several unusual cutaneous tumours of cerebriform morphology, suggesting skin metastases from occult internal cancer. Histopathological findings and thorough investigations, however, revealed a late-stage metastatic MM. We discuss the differential diagnosis of skin metastases of various origin and underline the difficulties for early detection of MM.
Mucosal human papillomavirus types in squamous cell carcinomas of the uterine cervix and subsequently on fingers
Authors: Forslund O. , Nordin P. , Hansson B.G.
Published: March 2003

Human papillomavirus (HPV), especially type 16, is causally involved in the pathogenesis of anogenital cancer. There is an increasing number of reports of HPV infections in squamous cell carcinoma (SCC) of the fingers. A search of the Swedish cancer register covering the period 1958–94 inclusive for women with a history of genital and upper extremity SCC revealed 63 cases. Archival material from both cervical and cutaneous lesions was traced and analysed for the presence of HPV DNA in 32 of these patients. A newly developed ‘neighbour primer’ polymerase chain reaction (PCR) for HPV 16 DNA, aimed at overcoming the obstacle of cross-linked target DNA, was shown to be superior to conventional general and type-specific HPV PCR tests. HPV DNA was significantly more frequently found in digital tumours than in tumours at other cutaneous sites of the upper extremities [67% (10 of 15) vs. 7% (three of 43); P < 0·001]. Among 13 patients with a history of both cervical and finger SCC, HPV 16 was found in cervical samples from seven patients. From five of these seven patients, HPV 16 was also present in the corresponding finger lesions. The results support the hypothesis of a possible transmission of patients’ genital HPV infections to fingers.
Widespread cutaneous necrosis occurring in association with the antiphospholipid syndrome: a report of two cases
Authors: Creamer D. , Hunt B.J. , Black M.M.
Published: March 2003

The antiphospholipid syndrome is an acquired prothrombotic state where thrombosis and/or pregnancy loss is related to the presence of antiphospholipid antibodies. Cutaneous necrosis secondary to intravascular thrombosis of small dermal vessels is a recognized but rare association with antiphospholipid syndrome. We report two patients with high circulating levels of anticardiolipin antibodies who developed widespread cutaneous necrosis as the first clinical manifestation of the antiphospholipid syndrome. The exact mechanism by which antiphospholipid antibodies mediate thrombosis is uncertain; however, proposed mechanisms of activity include endothelial cell activation, altered endothelial production of prostacyclin, activation of platelets, and modulation of the protein C and S pathways.
Erythrocytosis due to a mutation in the erythropoietin receptor gene
Authors: Percy Melanie J. , McMullin Mary Frances , Roques Anthony W. W. , Westwood Nigel B. , Acharya Jay , Hughes Anne E. , Lappin Terence R. J. , Pearson Thomas C.
Published: March 2003

Familial erythrocytosis, associated with high haemoglobin levels and low serum erythropoietin (Epo), has been shown to co‐segregate with a sequence repeat polymorphism at the 5′ region of the erythropoietin receptor (EpoR) in a large Finnish family.  We have investigated the cause of erythrocytosis in an English boy. Sequencing of the cytoplasmic region of the EpoR detected a de novo transition mutation of G to A at nucleotide 6002. This mutation resulted in the formation of a stop codon at amino acid 439 with the loss of 70 amino acids from the carboxy terminus. The mutation (G6002A) has arisen independently in a Finnish family and de novo in this English boy. Patients with unexplained erythrocytosis and low serum Epo levels should be investigated for EpoR mutations.
Cyclosporin A therapy in hypoplastic MDS patients and certain refractory anaemias without hypoplastic bone marrow
Authors: JonÁšovÁ Anna , NeuwirtovÁ Radana , ČermÁk Jaroslav , VozobulovÁ Věra , MocikovÁ Katarina , ŠiškovÁ Magda , HochovÁ Ivana
Published: March 2003

We report 17 cytopenic patients with myelodysplastic syndrome (MDS) of refractory anaemia (RA) subtype with hyper‐, normo‐ or hypo‐cellular bone marrow (BM), who were treated with cyclosporin A (CyA). Substantial haematological response was observed in 14 patients (82%): their anaemia improved and all transfusion‐dependent patients achieved transfusion independence. Complete trilineage recovery was observed in four patients (23%). The CyA therapy has not yet failed in any of the 14 successfully treated patients during follow‐up times ranging from 5 to 30 months. CyA was well tolerated in 14 patients; serious side‐effects required termination of the therapy in three patients in whom the blood count rapidly deteriorated to former levels upon cessation of therapy. Two patients benefited from a combination therapy of CyA and erythropoietin. Six patients experienced various autoimmune phenomena. CyA could thus offer an alternative treatment for certain MDS patients with RA regardless of hyper‐, normo‐ or hypo‐cellularity of bone marrow (BM). The mechanism of the beneficial effect of CyA is discussed and remains the subject of an ongoing study.
Quantitative expression of thrombopoietin receptor on leukaemia cells from patients with acute myeloid leukaemia and acute lymphoblastic leukaemia
Authors: Takeshita Akihiro , Shinjo Kaori , Izumi Masakazu , Ling Pan , Nakamura Satoki , Naito Kensuke , Ohnishi Kazunori , Ohno Ryuzo
Published: March 2003

Using a non‐isotopic ligand binding assay, we quantitatively examined the amount of human thrombopoietin (TPO) receptor (TPO‐R) on leukaemia cells from 128 patients with acute myeloid leukaemia (AML) or acute lymphoblastic leukaemia (ALL). The TPO‐R was expressed in 53 (47%) of 114 AML cases, and an in vitro treatment with TPO led to proliferation (stimulation index >1.5) of leukaemia cells in 13 (20%) of 66 AML cases examined. The TPO levels had no relation to the FAB classification except for FAB‐M7 AML. All five FAB‐M7 cases expressed TPO‐R, and one of three FAB‐M7 examined showed in vitro proliferative response to TPO. Although there was no significant correlation (r = 0.3125) between the amount of TPO‐R and the proliferative response, all of the AML cases which showed the in vitro response had TPO‐R expression. There was no relationship between TPO‐R amount and CD phenotypes, or the amount of granulocyte‐colony stimulating factor (G‐CSF) receptor. TPO‐R was also expressed in two (14%) of 14 cases of ALL, and only these two cases had in vitro proliferative response to TPO. One had only lymphoid antigens, and the other had both lymphoid and myeloid antigens. Our results suggest that some leukaemia cells express functionally active TPO‐R.
Acute promyelocytic leukaemia with t(11;17)(q23;q12‐21) and a good initial response to prolonged ATRA and combination chemotherapy
Authors: Culligan Dominic J. , Stevenson David , Chee Yen‐Lin , Grimwade David
Published: March 2003

Acute myeloid leukaemia (AML) of FAB subtype M3 is associated with t(15;17)(q22;q21) and a relatively good prognosis when treated with all‐trans retinoic acid (ATRA) and combination chemotherapy. Rarely, alternative balanced translocations have been described in this subtype of AML. The translocation t(11;17)(q23;q21) leading to a PLZF/RARα rearrangement has been described in a very small number of cases and has been associated with a poor response to ATRA and an adverse prognosis. We describe a case of AML FAB type M3 with this translocation who entered morphological and cytogenetic complete remission after concurrent prolonged ATRA and one course of induction chemotherapy and remains in morphological and molecular remission at 10 months after presentation. This diagnosis therefore may not always be associated with a poor initial response to treatment.
In vivo induction of functional FcγRI (CD64) on neutrophils and modulation of blood cytokine mRNA levels in cancer patients treated with G‐CSF (rMetHuG‐CSF)
Authors: Michon Jean M. , Gey Alain , Moutel Sandrine , Tartour Eric , Meresse Valerie , Fridman Wolf , Teillaud Jean‐Luc
Published: March 2003

Neutrophils from 13 children who received G‐CSF for the collection of peripheral blood progenitors while they were in haematological steady state were studied at various times after G‐CSF injection for FcγR expression (FcγRI or CD 64, FcγRII or CD32, and FcγRIII or CD16) and for their ability to exert antibody‐dependent cell cytotoxicity (ADCC) through FcγRI. Changes in IFNγ, IL8, IL10, MCP1 and TNFα mRNA levels in peripheral blood cells were also studied 4 h and 24 h after the first G‐CSF injection. FcγRI expression increased strongly after 24 h and then remained at the same level throughout treatment. In contrast, FcγRIII expression sharply decreased at day 1 and diminished even further thereafter. No change in FcγRII was observed. ADCC exerted by neutrophils through FcγRI started to increase after 24 h with the peak level at day 5. Cytokine mRNA analyses indicated a reproducible and strong increase of IL8 mRNA (11/13 children) after 24 h, whereas the changes in the mRNA levels of the other cytokines tested were more heterogenous (IFNγ: three; IL10: six; MCP1: five; TNFα: four, of the 13 children). Therefore this study opens the way to an optimized therapeutic schedule for the combined use of G‐CSF and monoclonal antibodies in adjuvant immuno‐intervention.
A de novo translocation 46,X,t(X;15) causing haemophilia B in a girl: a case report
Authors: Schröder  , Poetsch  , Gazda  , Werner  , Reichelt  , Knoll  , Rokicka‐Milewska  , Zieleniewska  , Herrmann
Published: March 2003

Haemophilia B is an X‐linked recessive bleeding disorder caused by mutations in the factor IX gene with an incidence of 1:25 000–30 000. Usually female carriers are clinically normal, and severe phenotypic expression of the disease in females is extremely rare. In this report we describe a girl with a clinically severe course of haemophilia B who had no signs of Turner syndrome or any other dysmorphic features. Cytogenetic and molecular studies in the patient and her parents showed a de novo translocation 46,X,t(X;15)(q27.1;p11.2) in the patient, indicating a possible break near the factor IX gene. The structurally normal X chromosome was late replicating and inactivated in all metaphases as shown by high‐resolution R‐banding. By fluorescence in situ hybridization (FISH) with YAC and cosmid probes we could further characterize the breakpoint region on the X chromosome and the involvement of the factor IX gene.
Fas‐mediated apoptosis with normal expression of bcl‐2 and p53 in lymphocytes from aplastic anaemia
Authors: Callera Fernando , Garcia Aglair B. , FalcÃO Roberto P.
Published: March 2003

In order to investigate the involvement of apoptosis in the pathogenesis of aplastic anaemia (AA) we measured the expression of the Fas receptor (membrane protein that triggers apoptosis), Fas ligand (FasL), bcl‐2 (cytoplasmatic protein that blocks apoptosis) and p53 (nuclear protein that induces apoptosis) in CD3 and CD19 lymphocytes from the peripheral blood or bone marrow of controls, patients with AA, aplastic anaemia in complete remission (AA‐CR) and multiply transfused patients without aplastic anaemia. The Fas receptor was overexpressed in both T and B lymphocytes from the peripheral blood and bone marrow from patients with AA. These abnormalities were not detected in AA‐CR or multiply transfused patients. CD3/FasL cells were not increased and no FasL expression was detected in B lymphocytes. Bcl‐2 was highly expressed in lymphocytes from controls, AA, AA‐CR and multiply transfused patients (> 99% of positive cells) whereas p53 was not detected in any group. To further characterize the functional activity of the Fas receptor we performed a Fas‐induced apoptosis assay in peripheral blood lymphocytes using an anti‐Fas monoclonal antibody. The crosslinking of the Fas receptor transduced an increased apoptotic signal in lymphocytes from AA patients, but not in lymphocytes from controls, AA‐CR patients or multiply transfused patients. Taken together, these data suggest that a Fas‐based mediated apoptosis without the apparent participation of bcl‐2 or p53 is a possible mechanism of lymphocyte depletion in patients with AA. In addition, these findings suggest that Fas expression is a continuous event occurring from progenitor bone marrow cells to mature cells.
Humic acid reduces protein‐C‐activating cofactor activity of thrombomodulin of human umbilical vein endothelial cells
Authors: Yang Hsin‐Ling , Hseu You‐Cheng , Lu Fung‐Jou , Tsai Horng‐Der
Published: March 2003

Humic acid in the drinking water of blackfoot disease endemic areas in Taiwan has been implicated as one of the aetiological factors of the disease. For this report we examined the effects of humic acid on the expression of thrombomodulin (TM) cofactor activity by cultured human umbilical vein endothelial cells (HUVEC).

Incubation of HUVEC with humic acid (HA) isolated from the drinking water, as a synthetic humic acid polymer (SHA) or with commercial HA, resulted in a dose‐dependent reduction of cell surface thrombomodulin activity. Characterization of the mechanism by which humic acid reduced the protein C activation indicated that inhibition was not caused by production or release of a protein C inhibitor. Kinetic analysis showed that binding affinities of TM to thrombin and of TM–thrombin complex to protein C was unchanged upon humic acid treatment. However, the cell surface TM activity was reduced by humic acid, which functions as an irreversible noncompetitive inhibitor of thrombin binding. Down‐regulation of TM was inhibited by non‐selective protein kinase C inhibitors and a selective inhibitor. These results suggest that protein kinase C is intricately involved in HA‐induced TM down‐regulation. Down‐regulation of TM was also inhibited by free radical scavengers. All these changes occurred in the absence of significant cytotoxic effect.

In conclusion, our results suggest that HA induces down‐regulation of TM by directly increasing permeability of the cell membrane, thus causing elevation in [Ca2+]i. This species functions as a second messenger to activate protein kinase C, and/or Ca‐dependent enzymes eventually inducing down‐regulation of TM. Attenuation of vascular endothelial cell TM cofactor activity by humic acid may play a role in the humic acid‐induced thrombotic vascular disorders of blackfoot disease.
Bone marrow and peripheral blood involvement in mantle cell lymphoma
Authors: Cohen  , Kurtin  , Donovan  , Hanson
Published: March 2003

The peripheral blood smears, bone marrow aspirates and biopsies of 46 patients with mantle cell lymphoma were reviewed. The diagnosis of mantle cell lymphoma was established in all cases on extramedullary tissue samples using standard morphologic, phenotypic and molecular genetic criteria. 27/35 patients (77%) had circulating lymphoma cells (median 20% of all circulating white blood cells; range 5–90%) identified by morphology at some point during the course of their disease. No statistical difference in survival was detected in patients with or without peripheral blood involvement. Lymphoma was identified in bone marrow aspirate specimens from 33/40 patients (83%) and in bone marrow biopsy specimens from 39/43 patients (91%). The pattern of marrow biopsy involvement was nodular (31 cases; 82%), interstitial (19 cases; 50%), paratrabecular (17 cases, 45%) and diffuse (12 cases; 32%). Although the median survival of patients with ≥ 50% bone marrow involvement was 13 months, and the median survival of patients with ≤ 50% was 49 months, no statistically significant differences between these small subgroups were observed. Mantle cell lymphoma frequently involves the peripheral blood and bone marrow. Its appearance is distinctive but variable, and immunophenotypic studies as well as morphologic confirmation by a biopsy of tissue other than bone marrow is still required for diagnosis.
Epitope mapping of HTLV envelope seroreactivity in LGL leukaemia
Authors: Loughran, JR Thomas P. , Hadlock Kenneth G. , Perzova Raisa , Gentile Teresa C. , Yang Qing , Foung Steven K. H. , Poiesz Bernard J.
Published: March 2003

Sera from approximately 50% of patients with large granular lymphocyte (LGL) leukaemia react with a recombinant human T‐cell leukaemia/lymphoma virus (HTLV) transmembrane envelope protein, p21e. Two immunodominant epitopes within env p21e have been defined by reactivity against recombinant proteins GD21 and BA21. In this study sera from 41 patients with LGL leukaemia were examined for reactivity against these recombinant HTLV env proteins. Overall, 21/41 (51%) sera reacted to p21e. Only two sera reacted to GD21. The predominant immunoreactivity against p21e was directed against the BA21 epitope, with 19/41 (46%) sera being BA21 positive. Seroconversion to BA21 protein was also documented. PCR analyses confirmed the low incidence of protypical HTLV sequences (2/41, 5%). These data document an association between BA21 seroreactivity and LGL leukaemia. This finding raises the possibility that such BA21 seroreactivity could be due to cross‐reactivity to a cellular or retroviral antigen sharing some amino acid homology with the transmembrane glycoprotein of HTLV.
Failure of oral pamidronate to reduce skeletal morbidity in multiple myeloma: a double‐blind placebo‐controlled trial
Authors: Brincker Hans , Westin Jan , Abildgaard Niels , Gimsing Peter , Turesson Ingemar , Hedenus Michael , Ford John , Kandra Albert
Published: March 2003

In order to study whether oral bisphosphonate therapy might prevent or reduce skeletal‐related morbidity in patients with newly diagnosed multiple myeloma who required chemotherapy, 300 patients were included in a randomized multi‐centre trial. Patients were given oral pamidronate at a dose of 300 mg daily, or placebo, in addition to conventional intermittent melphalan/prednisolone (and in some cases alpha‐interferon) treatment. With a median treatment duration of about 550 d, no statisticallly significant reduction in skeletal‐related morbidity (defined as bone fracture, related surgery, vertebral collapse, or increase in number and/or size of bone lesions) could be demonstrated. Pamidronate treatment also did not have any influence on patient survival or on the frequency of hypercalcaemia. However, in patients treated with pamidronate there were fewer episodes of severe pain (P = 0.02) and a decreased reduction of body height of 1.5 cm (P = 0.02). The overall negative result of the study is attributed to the very low absorption of orally administered bisphosphonates in general.
Methylation of the p16INK4A gene in multiple myeloma
Authors: Tasaka Taizo , Asou Hiroya , Munker Reinhold , Said Jonathan W. , Berenson James , Vescio Robert A. , Nagai Masami , Takahara Jiro , Koeffler H. Phillip
Published: March 2003

The p16INK4A (p16) binds to both cyclin D‐CDK4 and cyclin D‐CDK6 and inhibits the progression of the cell cycle from G1 to S phase. Loss of expression of this protein can occur by several mechanisms including structural alterations. Recent studies have suggested that the loss of expression of p16 can occur by hypermethylation of the gene. The methylation status of the p16 gene in multiple myeloma was examined in three myeloma cell lines (U266, RPMI8226 and IM9) and 16 primary myeloma samples using methylation‐specific polymerase chain reaction (MSP). The U266 and RPMI8226 cell lines contained a completely methylated p16 gene and the IM9 line had a partially methylated p16 gene. Identical results were obtained by another polymerase chain reaction (PCR)‐based methylation assay system as well as Southern blotting after using a methylation‐sensitive restriction enzyme. The U266 cell line expressed no p16, and the IM9 had weak expression as determined by reverse transcript (RT‐)PCR. The U266 cells began to express, and IM9 increased the accumulation of, the p16 RNA after treatment with the demethylating agent 5′‐aza‐2‐deoxycytidine (10−6–10−5 M). This suggested that the levels of methylation of the p16 gene detected by the MSP technique correlated with the regulation of transcription of this gene. Examination of the primary myeloma samples showed that eight of 16 (50%) contained a methylated p16 gene. We have previously found that alterations of the p16 gene, such as deletions and point mutations, are rare in primary multiple myeloma; none of the 16 samples included in this study had p16 gene alterations. Our results suggest that methylation of the p16 gene may contribute to the development and/or progression of multiple myeloma.
Chemotherapeutic drug‐induced apoptosis in human leukaemic cells is independent of the Fas (APO‐1/CD95) receptor/ligand system
Authors: Mcgahon Anne J. , Costa Pereira Ana P. , Daly Lisa , Cotter Thomas G.
Published: March 2003

The potential role of the Fas (CD95/APO‐1) receptor/ligand system in chemotherapeutic drug‐induced apoptosis was examined in a number of human leukaemic cell lines. Flow cytometric profiles of doxorubicin‐treated HL‐60, K562, U937 and Jurkat cells failed to show any significant increase in Fas or Fas ligand expression over 24 h, despite the induction of significant levels of apoptosis in these cells. Although preincubation of human leukaemic cells with a neutralizing anti‐Fas IgG antibody blocked anti‐Fas IgM‐induced apoptosis, this strategy failed to inhibit chemotherapeutic drug‐induced apoptosis. To determine whether recruitment of the Fas/Fas ligand complex during drug‐induced apoptosis was a cell‐specific event we utilized the CEM cell line. Doxorubicin treatment of CEM cells over 24 h failed to show any up‐regulation in Fas or Fas ligand protein levels as detected by flow cytometry. Furthermore, neutralizing anti‐Fas IgG Ab failed to inhibit chemotherapeutic drug‐induced apoptosis in CEM cells. The present studies do, however, demonstrate a role for anti‐Fas IgM Ab in producing a cytotoxic synergistic effect when used in combination with chemotherapeutic drugs. Low‐dose anti‐Fas IgM treatment in combination with doxorubicin, methotrexate, camptothecin and etoposide produced an augmented cytoxicity in CEM cells. Taken together these observations demonstrate that although recruitment of the Fas/APO‐1/CD95 receptor/ligand system is not a necessary requirement for chemotherapeutic drug‐induced apoptosis, combination of anti‐Fas IgM and drug treatment produces a synergistic cytotoxic effect which may prove useful in the treatment of human leukaemias.
Novel enzymatic abnormalities in AML and CML in blast crisis: elevated leucocyte leukotriene C4 synthase activity paralleled by deficient leukotriene biosynthesis from endogenous substrate
Authors: Stenke Leif , Sjölinder Mikael , Miale Thomas D. , Lindgren Jan Åke
Published: March 2003

Leukotrienes (LT) are inflammatory mediators which can also exert regulatory effects on human myelopoiesis. We have studied the LT‐producing capacity of freshly isolated leucocyte suspensions (containing blast cells in variable proportions) from 41 patients with acute myeloid leukaemia (AML) or chronic myeloid leukaemia (CML) in blast crisis (CMLbc) at diagnosis or relapse/resistant disease. Leucocyte suspensions from 19/29 AML patients (66%), and 2/12 CMLbc patients (17%; P = 0.012) demonstrated deficient capacity to synthesize LT from endogenous substrate after ionophore A23187 stimulation. Thus, these cells produced < 8 pmol LTB4 + LTC4/106 cells (< 20% of mean LT formation in leucocyte suspensions from 18 healthy subjects). Addition of exogenous arachidonic acid did not normalize the LT synthesis in poor‐producing cell suspensions. Purified, morphologically mature granulocytes from two AML patients also failed to produce normal amounts of LT. In leucocyte suspensions from the remaining 20 AML/CMLbc patients A23187 provoked LT biosynthesis, with markedly increased production of LTC4, but decreased LTB4 formation. Furthermore, elevated conversion of exogenous LTA4 to LTC4 was noted in the patient samples, independent of their capacity to produce LT after A23187 stimulation. The percentage of blast cells in patient white blood cell differential counts correlated inversely with ionophore‐induced LT synthesis, but positively with the conversion of exogenous LTA4 to LTC4. The results suggest elevated LTC4 synthase activity and suppressed 5‐lipoxygenase activity as novel enzymatic features of myeloid leukaemia patients with immature phenotype.
Prognostic factors for survival and response after high‐dose therapy and autologous stem cell transplantation in systemic AL amyloidosis: a report on 21 patients
Authors: Moreau Philippe , Leblond Véronique , Bourquelot Priscille , Facon Thierry , Huynh Anne , Caillot Denis , Hermine Olivier , Attal Michel , Hamidou Mohamed , Nedellec Gérard , Ferrant Augustin , Audhuy Bruno , Bataille Régis , Milpied Noël , Harousseau Jean‐Luc
Published: March 2003

We retrospectively investigated the feasibility and the toxicity of autologous stem cell transpantation (ASCT) in 21 cases of systemic amyloidosis (AL). The conditioning regimens consisted of high‐dose melphalan (HDM) alone (n = 18) or in combination with 12 Gy total body irradiation (n = 3). Toxic death rate was high: 9/21 patients (43%) died within the first month following ASCT, and 10/12 surviving patients achieved a response. With a median follow‐up of 14 months, the OS and the EFS rates at 4 years were 57.1% (±10.8) and 29.9% (±14.5) respectively for the whole group. The major prognostic factor for both response and survival was the number of clinical manifestations at the time of ASCT, of the following five criteria, i.e. creatinine clearance <30 ml/min, nephrotic syndrome with urinary protein excretion > 3000 mg/24 h, congestive heart failure, neuropathy, or hepatomegaly associated with alkaline phosphatase level > 200 IU/l. For patients presenting with two or more clinical manifestations the 4‐year OS and EFS were both 11.1% compared with 91.7% and 46.3% respectively in patients with fewer than two clinical manifestations at the time of ACST. We conclude that ASCT is feasible in AL in a subset of patients with fewer than two clinical manifestations at the time of ASCT. Given the severe extra‐haematological toxicity, ASCT should not be considered in other cases.
Ontogeny‐associated changes in the cytokine responses of primitive human haemopoietic cells
Authors: Zandstra  , Conneally  , Piret  , Eaves
Published: March 2003

Time course studies revealed that the combination of Flt‐3 ligand (FL), Steel factor (SF) and interleukin‐3 (IL‐3) did not elicit as large an amplification of the long‐term culture‐initiating cell (LTC‐IC) population in serum‐free cultures of CD34+CD38− cord blood (CB) cells as was obtained in similar cultures of adult human CD34+CD38− bone marrow (BM) cells (4‐ v 90‐fold maximum increases), even though both total and colony‐forming cell (CFC) numbers initially increased more rapidly in CB cultures. Multifactorial analysis of the short‐term (10 d) effects of different cytokines identified FL and IL‐6 in combination with the soluble IL‐6 receptor (sIL‐6R) as most important for expanding the CB LTC‐IC population. In contrast, their counterparts in adult BM were most effectively stimulated by FL, SF and IL‐3. For rapid generation of increased numbers of CFC, SF with either FL or IL‐6/sIL‐6R were found to be the most important contributors in cultures of CD34+CD38− CB cells, whereas, in analogous BM cultures, IL‐6/sIL‐6R and TPO (in addition to FL, SF and IL‐3) were required. These findings reinforce the principle of altered cytokine responsiveness as a hallmark of early haemopoietic cell differentiation and demonstrate how cytokine requirements may change during human ontogeny. Identification of conditions for optimizing the expansion of different subsets of primitive CB cells has additional important implications for clinical transplantation and gene transfer.
Lupus anticoagulant, anticardiolipin antibodies and hepatitis C virus infection in thalassaemia
Authors: Giordano Paola , Galli Monica , Del Vecchio Giovanni C. , Altomare Maria , Norbis Francesca , Ruggeri Luisa , Petronelli Maria , DE Mattia Domenico
Published: March 2003

Anticardiolipin antibodies (ACA) and lupus anticoagulant (LA) have been detected in patients with hepatitis C virus (HCV) infection and have been associated in autoimmune diseases (i.e. systemic lupus erythematosus) with an increased risk of thromboembolic events. Because of the high prevalence of HCV infection and the thrombotic risk described in thalassaemia we decided to investigate the prevalence of ACA and LA in a cohort of 68 thalassaemia patients. We found a high prevalence (34%) of β2‐glycoprotein I independent ACA in our thalassaemia patients which was related to HCV infection. None of patients developed any complications related to antiphospholipid antibodies (APL); therefore the clinical significance of positivity for APL in patients with HCV infection is at present unclear. In conclusion, the results of our study indicate that ACA in the serum of HCV‐infected thalassaemic patients exhibit the characteristics of natural autoantibodies rather than those of the pathogenic autoantibodies that are found in patients with systemic lupus erythematosus.
Unbalanced X‐chromosome inactivation in haemopoietic cells from normal women
Authors: Tonon Laura , Bergamaschi Gaetano , Dellavecchia Claudia , Rosti Vittorio , Lucotti Claudia , Malabarba Lucia , Novella Annunziata , Vercesi Elena , Frassoni Francesco , Cazzola Mario
Published: March 2003

We studied X‐chromosome inactivation patterns in blood cells from normal females in three age groups: neonates (umbilical cord blood), 25–32 years old (young women group) and >75 years old (elderly women). Using PCR, the differential allele methylation status was evaluated on active and inactive X chromosomes at the human androgen receptor (HUMARA) and phosphoglycerate kinase (PGK) loci. A cleavage ratio (CR) ⩾ 3.0 was adopted as a cut‐off to discriminate between balanced and unbalanced X‐chromosome inactivation. In adult women this analysis was also performed on hair bulbs. The frequency of skewed X‐inactivation in polymorphonuclear (PMN) cells increased with age: CR ⩾ 3.0 was found in 3/36 cord blood samples, 5/30 young women and 14/31 elderly women. Mathematical analysis of patterns found in neonates indicated that X‐chromosome inactivation probably occurs when the total number of haemopoietic stem cell precursors is 14–16. The inactivation patterns found in T lymphocytes were significantly related to those observed in PMNs in both young (P < 0.001) and elderly women (P < 0.01). However, the use of T lymphocytes as a control tissue for distinguishing between skewed inactivation and clonal proliferation proved to be reliable in young females, but not in elderly women, where overestimation of the frequency of clonal myelopoiesis may appear.
Shift from Rh‐positive to Rh‐negative phenotype caused by a somatic mutation within the RHD gene in a patient with chronic myelocytic leukaemia
Authors: CHÉrif‐Zahar Baya , Bony Viviane , Steffensen Rudi , Gane Pierre , Raynal Virginie , Goosens Dominique , Skaaning Laursen Jens , Varming Kim , Jersild Casper , Cartron Jean‐Pierre
Published: March 2003

We report a female patient whose Rh phenotype shifted from RhD‐positive to RhD‐negative over a 3‐year period (1991–94), during which time she was treated with mastectomy (1992) and local irradiation for a low‐grade recurrent breast cancer. She was diagnosed with chronic myeloid leukaemia in 1994, and has since then received chemotherapy. The patient was repeatedly typed as O, RhD‐positive between 1965 and 1991 and was repeatedly found RhD‐negative after 1994. Bcr‐Abl transcripts typical of Ph1 chromosome were detected. Molecular analysis indicated that the patient was heterozygous at the RH locus, carrying one haplotype in which the RHD gene exhibited a single nucleotide deletion (G600) resulting in a frameshift and premature stop codon, and a normal RHCE gene (allele Ce). The second haplotype contained only the RHCE gene (allele ce) and was normal. Further analysis carried out on total leucocytes, purified neutrophils, EBV‐lymphoblastoid cell line and cultured erythroblasts indicated that the G600 deletion was restricted to the myeloid lineage. No modification of other blood group antigens could be detected. These findings suggest a somatic mutation which most probably occurred in a stem cell common to the myeloid lineage.
Establishment and characterization of a mantle cell lymphoma cell line
Authors: Jeon HO Jong , Kim Chul Woo , Yoshino Tadashi , Akagi Tadaatsu
Published: March 2003

A mantle cell lymphoma (MCL) cell line (JeKo‐1) was established from peripheral blood mononuclear cells of a patient with a large cell variant of MCL showing leukaemic conversion. JeKo‐1 cells were Epstein‐Barr virus negative and showed a B‐cell phenotype with IgM+, IgD+, CD3−, CD5+, CD10−, CD19+, CD20+ and CD23−; they overexpressed cyclin Dl, Bcl‐2, c‐Myc and Rb proteins. Bcl‐1/JH gene rearrangement was confirmed by polymerase chain reaction, although karyotypic analysis showed 40/41 chromosomes devoid of apparent t(11;14)(q13;q32) translocation. JeKo‐1 cells were highly tumourigenic in SCID mice.