bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of IL-10 in the Crossregulation of Prostaglandins and Cytokines in Monocytes
Authors: Niho Yoshiyuki , Niiro Hiroaki , Tanaka Yosuke , Nakashima Hitoshi , Otsuka Takeshi
Published: April 2003

In the present study we have focused mainly on the role of IL (interleukin)-10 in the crossregulation of prostaglandins and cytokines in human monocytes. We first determined the effects of tumor necrosis factor-α (TNF-α) and IL-10 on monocyte prostaglandin E2 (PGE2) production. Unstimulated monocytes constitutively produced a small but significant amount of PGE2 in the culture supernatants. Both TNF-α and lipopolysaccharide (LPS) caused a remarkable increase in monocyte PGE2 production. On the other hand, IL-10 alone was without effect on constitutive PGE2 production but drastically inhibited LPS-induced PGE2 production in monocytes. Moreover, this inhibitory effect of IL-10 was not simply attributable to its inhibition of TNF-α production in LPS-stimulated monocytes. Next, we determined the effect of PGE2 on TNF-α mRNA expression in monocytes. Treatment of monocytes with or without PGE2 showed no detectable TNF-α mRNA. Activation of monocytes by LPS resulted in a remarkable accumulation of TNF-α mRNA and PGE2 efficiently inhibited this expression. Finally, we determined the effect of PGE2 on IL-10 mRNA expression in monocytes. Similar to TNF-α mRNA, unstimulated monocytes showed no detectable IL-10 mRNA. Interestingly, PGE2 alone drastically induced IL-10 mRNA. Besides, activation of monocytes by LPS resulted in a remarkable accumulation of IL-10 mRNA, and PGE2 further enhanced this expression. These results indicate that TNF-α and PGE2 are key molecules for the induction of IL-10 in monocytes, and that IL-10, in turn, plays a crucial role in terminating the inflammatory cascade via downregulation of production of proinflammatory molecules including TNF-α and PGE2.
Pentoxifylline Synergizes with All-Trans Retinoic Acid to Induce Differentiation of HL-60 Myelocytic Cells, but Suppresses tRA-Augmented Clonal Growth of Normal CFU-GM
Authors: Yang K.D. , Chao C.Y. , Shaio M.F.
Published: April 2003

All-trans retinoic acid (tRA) has been shown to promote terminal differentiation of promyelocytic leukemia cells, but frequently induce hyperleukocytosis and pulmonary leakage syndrome. Employing pentoxifylline (PTX), a phosphodiesterase inhibitor which could raise intracellular cAMP and modulate leukocyte activation, we sought to investigate if PTX could enhance tRA-induced promyelocytic leukemic cell differentiation but suppress tRA-augmented growth and activation of human granulocytes. tRA could significantly suppress clonal growth of U937 and HL-60 leukemic cells but enhanced the CFU-GM formation of normal bone marrow cells (22 ± 6 vs. 90 ± 16 CFU/well). PTX significantly augmented tRA suppression of clonal growth of U937 and HL-60 leukemic cells but suppressed tRA-augmented CFU-GM formation of normal bone marrow cells (90 ± 16 vs. 25 ± 9 CFU/well). In addition, PTX enhanced tRA-induced growth inhibition and differentiation of promyelocytic HL-60 leukemic cells, but suppressed respiratory burst activation by the immature granulocytic HL-60 cells and suppressed CD11b adhesion molecule expression by mature granulocytes. PTX similar to dibutyric cAMP promoted HL-60 myelocytic leukemic cell differentiation and growth inhibition, whereas PTX, in contrast to dibutyric cAMP which could augment phorbol myristate acetate (PMA)-elicited respiratory burst activity by immature granulocytes, suppressed the PMA-elicited respiratory burst activity by immature and mature granulocytes. PTX did not raise the intracellular cAMP level of HL-60 cells, but partly suppressed the dibutyric cAMP-elicited elevation of intracellular cAMP level. Results from these studies suggest that PTX might act through different signaling pathways to enhance tRA-induced myelocytic leukemic cell differentiation but prevent from hyperreactive normal granulopoiesis and granulocyte activation.
Effect of Three Days Starvation on the Phagocytic Activity of Rat Peritoneal Macrophages
Authors: Salman Hertzel , Bergman Michael , Bessler Hanna , Alexandrova Svetlana , Straussberg Rachel , Zahavi Izhar , Djaldetti Meir
Published: April 2003

To examine the effect of starvation on the phagocytic activity of rat peritoneal macrophages, 8 animals were kept for 3 days on water supply only. The cells showed an increased capacity for superoxide anion production and a marked decrease in their ability to engulf latex particles. The corticosterone level of the fasting animals was markedly increased, an observation suggesting that the alteration of macrophage function could be hormonally mediated. The findings in the present study indicate that the decreased phagocytic capacity of peritoneal macrophages following starvation may be one of the factors responsible for the increased susceptibility to infections in malnourished individuals.
Continuous Interleukin-2 Infusion Combined with Cyclophosphamide- Based Combination Chemotherapy in the Treatment of Hemato-Oncological Malignancies
Authors: Nagler A. , Ackerstein A. , Ben-Shahar M. , Or R. , Naparstek E. , Ben-Yosef R. , Slavin S.
Published: April 2003

The combination of a cyclophosphamide (CTX)-based chemotherapy regimen and interleukin-2 (IL-2) has been shown to provide synergistic effects against malignancy in animal models. We therefore conducted a phase I–II trial combining CTX-based combination chemotherapy or CTX alone with high-dose IL-2 in patients with advanced and refractory malignant disease. Fifteen patients with hemato-oncological malignancies (malignant lymphoma 8, multiple myeloma 3, solid tumor 2, leukemia 2) were enrolled in the study. Continuous high-dose IL-2 infusion was shown to be safely administered, starting as soon as recovery of white blood cell count. All patients developed rebound lymphocytosis 24–48 h after termination of IL-2 infusion. Although grade IV toxicity was observed in 5 patients (7 episodes), all side effects completely subsided. Triple chemotherapy (CTX, etoposide and Ara-C) seemed rather toxic (in this group of heavily treated patients) while CTX alone was well tolerated. Four out of 13 (31%) evaluable patients had partial response and another patient (7%) had stabilization of disease progression lasting 2–8 months. Our conclusion is that the combination of CTX and continuous infusion of IL-2 is feasible and should be investigated in patients with various malignant neoplasms.
Interferon-Induced Vasospasm in Chronic Myeloid Leukaemia
Authors: Zeidman Aliza , Dicker Dror , Mittelman Moshe
Published: April 2003

The role of interferon (IFN) in the treatment of chronic myeloid leukaemia (CML) has been established. Many adverse effects have been reported, but vasospasm has been extremely rare. We report 2 CML patients who developed such complications. A 56-year-old female had been on IFN for 3 years with haematological and cytogenetic remission, when she developed an anginal syndrome followed by acute ischaemia. Coronary catheterization revealed normal arteries. After discontinuation of IFN her cardiac complaints disappeared and she needed no medication. A 61-year-old patient had been on IFN for 1 year when he presented with Raynaud’s phenomenon. No evidence of collagen vascular disease could be documented. IFN discontinuation and intravenous administration of iloprost (a prostacyclin analogue) resulted in the disappearance of the vascular complications. IFN appears to have a beneficial effect on the course and prognosis of CML. However, we have to be aware of the potential complications and adverse effects which can be related to IFN. Neither our experience nor the literature provides convincing recommendations regarding the management of such patients. We suggest proceeding with IFN at lower doses, especially in those who have achieved a cytogenetic response, as our first patient.
Familial Aggregation of Nonhematological Malignancies in Relatives of Patients with Hematological Neoplasms
Authors: Shpilberg Ofer , Modan Michaela , Modan Baruch , Chetrit Angela , Ramot Bracha
Published: April 2003

Familial aggregation of nonhematological malignant disorders (NHMD) was compared in 189 families of patients with hematological neoplasms (HN) with a control group of 36 families of patients with benign hematological disorders and a second group of 33 families of patients with diabetes mellitus. A self-administered questionnaire was used requesting from each family a full list of first- and second-degree relatives, their vital status, current age or age at death, and a list of their chronic diseases, including all malignant disorders. There was no evidence of a significantly increased tendency for developing NHMD among relatives of patients with HN as compared to controls (adjusted odds ratio of 0.88; 95% confidence interval 0.61–1.27). Moreover, in the HN group, no significant difference in the frequency of NHMD was found between the families with and without familial aggregation of HN. Based on the present analysis and our previous observations on familial aggregation of HN, we conclude that the increased aggregation of malignant disorders among relatives of patients with HN is unique to the hematopoietic system and might result from a genetic predisposition to HN in these families.
Clinical Outcome in Three Patients with Myelodysplastic Syndrome Showing Polyclonal Hematopoiesis
Authors: Karasawa Masamitsu , Tsukamoto Norifumi , Sakai Hirotaka , Okamoto Kiyoshi , Maehara Tadashi , Naruse Takuji , Morita Kimio , Sato Sadao
Published: April 2003

The clinical outcome of 3 myelodysplastic syndrome (MDS) patients with polyclonal hematopoiesis is reported. All patients were heterozygous for the phosphoglycerate kinase (PGK) gene. The presence of polyclonal hematopoiesis was determined by the X-chromosome-linked restriction fragment length polymorphism-methylation method using the PGK gene as a marker. The patients were initially diagnosed as having refractory anemia (RA), RA with ring sideroblasts (RARS), and RA with an excess of blasts (RAEB), respectively. Their pancytopenia persisted during the follow-up period of 11.4 years for the RA patient, 19.5 years for the RARS patient and 0.8 years for the RAEB patient. Although the RARS patient continues to be in good health, leukemic transformation occurred in the other 2 patients. A karyotype change from 46,XX to 45,XX,t(3;21),-7 was observed at the time of disease progression in the RA patient. The coexistence of a monoclonal MDS clone and normal bone marrow cells is thought to be the most probable reason for the polyclonal hematopoiesis of these patients.
Oral and Intravenous Itraconazole for Systemic Fungal Infections in Neutropenic Haematological Patients: Meeting Report
Authors: Prentice H.G. , Caillot D. , Dupont B. , Menichetti F. , Schuler U.
Published: April 2003

Effective prevention, or treatment, of invasive fungal infection in the neutropenic patient has hitherto been unsatisfactory because of either an inadequate anti-fungal spectrum of the agent or important toxicity. Itraconazole is effective against a broad spectrum of the opportunistic pathogens seen in Europe and North America. Prior problems with absorption, e.g. in the marrow transplant recipient, have been overcome with the introduction of an oral solution and an i.v. preparation. The deliberations of an expert meeting held in June, 1998 include recommendations on which patient requires one of these new preparations based on clinical trials, the dose and route. Important drug interactions are also detailed.
Adenovirus p53 Purging for Human Breast Cancer Stem Cell Products
Authors: Hirai Manabu , Kelsey LindaS. , Maneval DanielC. , Vaillancourt Mei , Talmadge JamesE.
Published: April 2003

Tumor cell (TC) contamination of stem cell products can contribute to relapse after high dose chemotherapy and stem cell rescue. A new purging technology using replication-deficient recombinant adenovirus (Adv) containing the p53 tumor suppressor gene (Adv-p53) has been suggested to reduce tumor contamination of autologous stem cell product. We demonstrate herein a safe and effective Adv-p53 purging procedure using four human breast cancer TC lines. Multiple parameters need to be achieved to successfully purge stem cell products, including a high cell:virus ratio, a small incubation volume, a long incubation time and 37°C rather than room temperature. These parameters are all interrelated and equally important for the inhibition of TC clonogenic growth. In our studies, we also observed that Adv could nonspecifically inhibit TC clonogenic growth, although Adv-p53 treatment led to a significantly greater inhibition of clonogenic growth by cells expressing mutated p53. The presence of peripheral stem cell (PSC) products was found to decrease the effect of Adv-p53 on TC clonogenic growth, suggesting that PSC products could compete with TC for infection by recombinant Adv. However, X-Gal staining after incubation with Adv containing-galactosidase demonstrated that PSC products were 2,000-fold more resistant to Adv infection than TC. We conclude that a 4-hour incubation of stem cell products (2 × 108/ml) with 4 × 1011 Adv-p53 particles is sufficient to completely purge TC with no effect on hematopoietic cell function.
Efficacy and Side Effects of Deferiprone (L1) in Thalassemia Patients Not Compliant with Desferrioxamine
Authors: Taher A. , Chamoun F.M. , Koussa S. , Saad M. Abi , Khoriaty A.I. , Neeman R. , Mourad F.H.
Published: April 2003

We report our experience with deferiprone (L1) (DFP) in 17 thalassemic patients, followed up in one center in Lebanon, who were initially on desferrioxamine and then shifted to DFP at a dose of 50–75 mg/kg/day as the sole chelator during 1-year follow-up. All 17 patients were compliant with therapy and there was no change in physical examination over the study period. Eight patients (47.1%) were positive for hepatitis C virus (HCV) antibodies. Urinary iron excretion was 21.8 ± 14 mg/24 h (mean ± SD) 1 week after starting DFP and dropped 12 months later to 13 ± 7.4 mg/24 h (p = 0.009, paired t test). The initial serum ferritin level was 3,863 ± 2,344 μg/l which dropped to 3,179 ± 2,075 at 12 months after starting therapy (p = 0.07). HCV-negative patients as a group exhibited a significant decrease in serum ferritin after 6 and 12 months of DFP therapy (3,942 ± 2,739 vs. 2,341 ± 1,179 and 2,681 ± 1,519 μg/l; p < 0.03 and p < 0.05, respectively). The most frequent side effects were joint pain, stiffness or swelling in 6 patients (35.3%), and nausea in 7 patients (41.2%), but these were well tolerated and did not require stopping treatment.
Long-Term Survival in Primary Plasma Cell Leukemia after Therapy with VAD, Autologous Blood Stem Cell Transplantation and Interferon-Alpha
Authors: Panizo Carlos , Rifón José , Rodríguez-Wilhelmi Pablo , Cuesta Braulia , Rocha Eduardo
Published: April 2003

Primary plasma cell leukemia (PCL) is a rare form of plasma cell neoplasm with a poor prognosis. Conventional melphalan-based treatments have been most disappointing. We report the case of a 62-year-old man with a primary form of PCL treated with VAD combination achieving an objective response, and who received high-dose melphalan and autologous peripheral blood stem cell (PBSC) transplantation followed by interferon-α. During the remission time, lasting for 3 years, an infiltration by large granular lymphocytes (LGL) was noted in peripheral blood. However, when the number of LGL declined, a bone marrow relapse was observed. The treatment for PCL and the possible role of these LGL on tumor cell control after autologous PBSC transplantation are discussed.
Monomorphic Agranular Natural Killer Cell Lymphoma/Leukemia with No Epstein-Barr Virus Association
Authors: Matano Sadaya , Nakamura Shinobu , Nakamura Shigeo , Annen Yusei , Hattori Noritaka , Kobayashi Kazumi , Kyoda Katsunori , Sugimoto Tatsuho
Published: April 2003

The conceptual view of natural killer (NK) cell malignancies has recently undergone a significant evolution. The majority of such diseases are associated with Epstein-Barr virus (EBV), while only a limited number of EBV-negative cases has been reported. We report an unusual case of NK cell lymphoma/leukemia showing a monomorphic histology, absence of intracytoplasmic azurophilic granules, and no EBV association. The patient was a 57-year-old woman who died 26 months after the diagnosis. Autopsy revealed tumor infiltration in the liver, spleen, lymph node, blood, and bone marrow. There was no involvement of the skin or nasal cavity throughout the clinical course. The tumor showed the monotonous proliferation of medium-sized cells without intracytoplasmic azurophilic granules. Phenotypic analysis showed CD2+, CD3/Leu4–, cytoplasmic CD3ε+, CD4–, CD5–, CD7+, CD8–, CD16–, CD38+, CD56+, CD57–, TdT–, granzyme B–, and TIA1+ phenotype. There were no detectable rearrangements of T cell receptor genes or immunogloublin heavy chain genes. Furthermore, there were no EBV-encoded small RNAs. These findings provide information to improve the understanding of poorly defined entities, i.e. aggressive NK cell lymphoma/leukemia and blastic NK cell lymphoma/leukemia.
Sequential Use of Deferiprone and Desferrioxamine in Primary School Children with Thalassaemia major in Turkey
Authors: Aydinok Y. , Nisli G. , Kavakli K. , Coker C. , Kantar M. , Çetingül N.
Published: April 2003

The effectiveness of the sequential use of deferiprone and desferrioxamine (DFO) in children with thalassaemia major was examined. Seven thalassaemic children in whom urinary iron induced by deferiprone was sufficient to maintain a negative iron balance were enrolled in the long-term trial. Deferiprone at a dose of 75 mg/kd/day in 3 divided doses was given for 4 school days a week. The group was given DFO at a dose of 40–50 mg/kg/day s.c. over 8–12 h with a battery-operated pump for 2 days at the weekend. In addition to the safety variables, they were monitored for serum ferritin levels at 2-month intervals and hepatic iron concentrations in liver tissues were determined at the beginning and the 6th month of therapy. The severity of hepatic damage was graded according to the Knodell hepatic activity index and the fibrosis was quantified. None of the patients suffered adverse effects of the therapy but a transient increase in serum ALT levels was noted. A nonsignificant decline in serum ferritin was observed (p = 0.08), a significant reduction in hepatic iron concentration was also determined (p = 0.03). The hepatic activity index in liver tissues of the patients at the 6th month of the sequential therapy significantly decreased (p = 0.03) whereas fibrosis scores did not differ significantly (p = 0.25).
Establishment and Characterization of a Fas-Resistant T Cell Line
Authors: Higuchi Masanori , Horiuchi Takahiko , Sawabe Takuya , Yoshizawa Shigeru , Nagasawa Kohei , Niho Yoshiyuki
Published: April 2003

Fas is a cell surface receptor that controls a signal transduction pathway leading to apoptosis. We established an antihuman Fas monoclonal antibody (mAb)-resistant variant, kit-225-FR, from the human T cell line, kit-225. Flow cytometric analysis revealed that the expression of Fas molecules on kit-225-FR was preserved. The defect in Fas molecule was not detected either by reverse transcription polymerase chain reaction (PCR) analysis of the Fas transcript or by PCR-single strand conformation polymorphism analysis of the Fas gene in kit-225-FR. Although kit-225-FR was resistant to a high concentration of anti-Fas mAb, apoptosis could be induced, as with the wild type, by exogenous C2-ceramide exposure. MORT1/FADD was expressed at wild-type level in kit-225-FR, as determined by Western blot analysis. It therefore appears that the apoptotic signal transduction in kit-225-FR is defective between FADD and the sphingomyelin-ceramide pathway. By comparing the differences from the wild-type kit-225, kit-225-FR would serve as a useful cell line for analyzing Fas-specific signal transduction pathways in detail.
Evaluation of the Left Ventricular Systolic and Diastolic Functions by Echocardiography in Patients with Acute Leukemia
Authors: Tunçkale Aydın , İlerigelen Barış , Aktuğlu Gülten
Published: April 2003

Eighteen patients with acute leukemia were studied to evaluate their left ventricular systolic and diastolic function by M mode, two-dimensional and Doppler echocardiographic techniques. The study group was compared with another group of 18 patients of similar age with iron deficiency anemia. The latter group had a comparable heart rate and levels of Hb. There were no differences in systolic function, whereas the diastolic function was lower in patients with acute leukemia (E/A ratio = 1.1 ± 0.3 vs. 1.42 ± 0.5, p < 0.05). It is concluded that left ventricular diastolic dysfunction is more common in patients with acute leukemia and may be an indicator of restrictive cardiomyopathy.
Plasma Thrombopoietin Concentrations in Response to Platelet Transfusion Therapy
Authors: Möller Per , Andersson Per-Ola , Andersson Ann-Christine , Wadenvik Hans
Published: April 2003

Studies performed in rabbit and mouse models and in a limited number of human subjects, show that transfused platelets bind thrombopoietin (TPO) and decrease its concentration in the circulation. The aim of the present study was to further examine this relationship. The material comprised 12 patients receiving a total of 21 transfusions, as part of the routine clinical treatment. Blood samples were collected from the patients immediately before and 30 min after completion of the platelet transfusion, and the corrected platelet count increment (CCI) was calculated. A commercially available ELISA kit was used to determine plasma TPO concentrations. Statistically significant reductions in median TPO concentration were observed in response to the platelet transfusions. Patients who were refractory to platelet transfusions showed the slightest decrease in TPO concentration. As for the linear regression between change in TPO level and CCI, only borderline significance was observed. Thus, our findings support the concept that platelets can remove TPO from the circulation.
Effect of Ozone on Red Blood Cell Enzymes and Intermediates
Authors: Zimran Ari , Wasser Gerhard , Forman Linda , Gelbart Terri , Beutler Ernest
Published: April 2003

Ozone autohemotherapy has been considered a form of alternative medicine and has not yet been subjected to the rigors of well-designed clinical trials. Despite encouraging anecdotal reports regarding the use of ozone in various disorders, there has been a concern that ozone per se may adversely affect red cell membranes and metabolites. The purpose of this study was to ascertain the effect of ozone administration at a concentration commonly used in autohemotherapy on a panel of red cell enzymes and intermediates, as well as its effect on red cell integrity. Since these parameters were unaffected by ozone, we suggest that clinical trials for the use of ozone autohemotherapy should be encouraged.
Current Results of Bone Marrow Transplantation in Patients with Acquired Severe Aplastic Anemia
Authors: Bacigalupo A. , Oneto R. , Bruno B. , Socié G. , Passweg J. , Locasciulli A. , van Lint M.T. , Tichelli A. , McCann S. , Marsh J. , Ljungman P. , Hows J. , Marin P. , Schrezenmeier H.
Published: April 2003

We have analyzed 2,002 patients grafted in Europe between 1976 and 1998 from an identical twin (n = 34), from an HLA-identical sibling (n = 1,699) or from an alternative donor (n = 269), which included unrelated and family mismatched donors. The proportions of patients surviving in these three groups are, respectively, 91, 66 and 37%: major causes of failure were acute graft-versus host disease (GvHD) (11%), infection (12%), pneumonitis (4%), rejection (4%). In multivariate Cox analysis, factors predicting outcome were patient’s age (p < 0.0001), donor type (p < 0.0001), interval between diagnosis and bone marrow transplantation (BMT) (p < 0.0005), year of BMT (p = 0.0005) and female donor for a male recipient (p = 0.02). Patients were then divided in two groups according to the year of BMT: up to or after 1990. The overall death rate dropped from 43 to 24% (p < 0.00001). Improvements were seen mostly for grafts from identical siblings (from 54 to 75%, p < 0.0001), and less so for alternative-donor grafts (from 28 to 35%; p = 0.07). Major changes have occurred in the BMT protocol: decreasing use of radiotherapy in the conditioning regimen (from 35 to 24%; p < 0.0001) and increasing use of cyclosporin (with or without methotrexate) for GvHD prophylaxis (from 70 to 98%; p < 0.0001). In conclusion, the outcome of allogeneic BMT for patients with severe aplastic anemia has considerably improved over the past two decades: young patients, grafted early after diagnosis from an identical sibling, have currently an over 80% chance of long-term survival. Transplants from twins are very successful as well. The risk of complications with alternative donor transplants is still high.
Results of Immunosuppression in Aplastic Anaemia
Authors: Marsh Judith C.W.
Published: April 2003

The availability of immunosuppressive agents, such as antilymphocyte globulin and cyclosporin, has had a major impact on the outcome of patients with aplastic anaemia. The majority of patients will show a response to immunosuppressive therapy. For those who fail to respond to an initial course, a second or further courses of immunosuppression can be given. Improvement in survival may also be due to the improved quality of supportive care available today for patients with aplastic anaemia. The additional use of haemopoietic growth factors such as granulocyte colony stimulating factor with immunosuppressive therapy requires further prospective randomised studies in order to evaluate whether they are of benefit for patients with aplastic anaemia. Because of the rarity of this condition, the use of multicentre prospective randomised studies is crucial for further evaluation of treatment modalities in aplastic anaemia.
Possible Association between Cytomegalovirus Infection and Gastrointestinal Bleeding in Hemophiliac Patients
Authors: Nogueira Eliana , Arruda Valder R. , Annicchino Bizzacchi Joyce M. , Costa Fernando F. , Castro Ozelo Margareth , Lúcio Rossi Claudio , Costa Sandra C.B.
Published: April 2003

Cytomegalovirus (CMV) infection is of major concern in immunocompromised and immunosuppressed patients. Prior to the introduction of HIV-1 antibody screening and efficient virucidal processes to inactivate viruses, individuals with a factor VIII or factor IX deficiency had a high risk of contracting HIV-1 infection through the infusion of contaminated blood products. In addition, blood products were also frequently associated with alterations in immune function. This study investigated the frequency of active CMV infection and its clinical relevance in Brazilian hemophiliacs. One hundred hemophiliacs were screened for the presence of CMV-DNA in their blood using nested PCR. Twenty-five out of 100 patients (25%) were positive for CMV-DNA and 24 of these 100 patients (24%) were HIV-1 positive; 6 of these 24 (25%) were positive for CMV-DNA. A similar frequency was observed among HIV-1-negative patients. In 60 hemophiliacs, the clinical relevance of the CMV infection was assessed. Twenty-one patients were positive for CMV-DNA. Of these, 10 had gastrointestinal bleeding compared to only 9 of 39 patients who were CMV-DNA negative (p = 0.05; χ2 test). These data indicate a high prevalence of active CMV infection in Brazilian hemophiliac patients, irrespective of whether the patients were or were not infected by HIV-1. There was a possible association between the presence of CMV and the occurrence of gastrointestinal bleeding.
Type I and Type II T Cell Profiles in Chronic Myelogenous Leukemia
Authors: Tsuda Hiroyuki , Yamasaki Hiroshi
Published: April 2003

T cell immunity is considered to play an important role in the control of cell growth in chronic myelogenous leukemia (CML) although information regarding the characteristics of T lymphocytes in CML patients is limited. Using flow cytometric analysis of intracellular cytokine expression at the single-cell level, we analyzed helper T and cytotoxic T subsets in 8 CML patients. The percentage of interferon-γ (IFN-γ) single-positive CD4 cells (Th1) and that of interleukin-4 (IL-4) single-positive CD4 cells (Th2) was markedly decreased in pretreated CML patients compared to normal controls. In addition, the percentage of IFN-γ/IL-4 double-positive CD4 cells (Th0) was also reduced. Consequently, the percentage of IFN-γ/IL-4 double-negative CD4 cells was markedly increased. Similarly, a reduction in IFN-γ single-positive CD8 cells (Tc1) and IFN-γ/IL-4 double-positive CD8 cells (Tc0) and an increase in IFN-γ/IL-4 double-negative CD8 cells were observed in pretreated CML patients. Imbalance of these parameters was markedly improved following cytoreduction therapy. Our findings directly indicate anergic states in CML patients. Determination of the factors that affect Th and Tc profiles may lead to further understanding of immunological states and the development of effective immunotherapy.
Down-Regulation of CD98 in Melphalan-Resistant Myeloma Cells with Reduced Drug Uptake
Authors: Harada Naoko , Nagasaki Akitoshi , Hata Hiroyuki , Matsuzaki Hiromitsu , Matsuno Fumihiko , Mitsuya Hiroaki
Published: April 2003

Although melphalan has been used as a therapeutic reagent for multiple myeloma, many patients become refractory. To elucidate the mechanism of resistance to melphalan, we generated a melphalan-resistant myeloma cell line, KHM-11EMS, by treating a parental line, KHM-11, with a mutagen, ethylmethanesulfonate. KHM-11EMS is 55 times more resistant to melphalan. γ-Glutamylcysteine synthetase, P-glycoprotein, multidrug-resistance-associated protein, lung-resistance-related protein and the Bcl-2 family of proteins were not responsible for the drug resistance in KHM-11EMS. Intracellular incorporation of melphalan to myeloma cells was determined by using [14C]-labeled melphalan. Accumulation of melphalan in KHM-11EMS was 43% of KHM-11, while the efflux rates were comparable in both cell lines. The uptake of melphalan was inhibited by the addition of L-phenylalanine, indicating that melphalan is incorporated through the L-phenylalanine transporter as reported previously. Expression of CD98, which was recently cloned as an L-phenylalanine transporter, was 6-fold decreased in KHM-11EMS, suggesting that CD98 may be correlated with the incorporation of melphalan. CD98 expression and incorporation of melphalan were analyzed in fresh purified myeloma cells from 5 patients. All myeloma cells from 4 cases expressed CD98 at a high level and incorporated melphalan. However, tumor cells from 1 case expressed CD98 at low levels and did not incorporate melphalan. Taken together, reduced melphalan uptake could be responsible for the drug resistance in KHM-11EMS, and down-regulation of CD98 may be related to this phenomenon. Further investigation of the correlation between impaired drug uptake and down-regulation of CD98 in myeloma cells should be important to understand the mechanism of resistance to melphalan.
Expression of Wild-Type p53 and Bcl-2 Family Genes Oscillates with Recurrent Remission and Relapse in an Unusual Case of Low-Grade Lymphoma
Authors: Aizman Irina , Many Amira , Peller Shoshana , Ramot Bracha , Kaufmann Yael
Published: April 2003

Downregulation of apoptosis has been proposed as a mechanism of clonal expansion in low-grade B cell neoplasms. We have previously described an unusual case of CD5+ B cell lymphoma characterized by cycles of leukemic phase alternating with spontaneous remission. In the present study, we examined the involvement of apoptosis-related proteins in the progression of this cyclic lymphoma ex vivo. During the leukemic phases, the clonal cells were activated blasts expressing elevated levels of wild-type (wt) p53, Bcl-2, Bcl-xL, and Bax, while Bak expression increased during the decline of lymphocytosis. Bax heterodimerized with Bcl-2 but not with Bcl-xL. The anti-apoptotic Bcl-2/Bax heterodimers peaked during early leukemic phases and declined during regression. The elevation in Bcl-2, Bcl-xL and Bax expression during early leukemic phases seems to result from cell activation since a similar increase was induced by activating the remission phase leukemic cells in culture. The data suggest that wt p53, Bcl-xL, and Bcl-2/Bax heterodimers support the accumulation of activated leukemic cells during the leukemic phases, while Bax and Bak may be involved in their decline during regression.
Neutropenia Accompanying Parvovirus B19 Infection after Gynecologic Surgery
Authors: Honda Ken-ichi , Ishiko Osamu , Tsujimura Akemi , Hino Masayuki , Hirai Kouzo , Itoh Fumihiro , Tanaka Tetsuji , Ogita Sachio
Published: April 2003

The hematologic data and symptoms of 7 patients seropositive for parvovirus B19 IgM antibody after gynecologic surgery were analysed. Parvovirus may have been transmitted by fibrin glue prepared from heat-treated human plasma and used for hemostasis during surgery. The peripheral blood neutrophil count decreased to below 1 × 109/l between postoperative day (POD) 10 and 18, but recovered spontaneously to within the normal range. G-CSF injection was effective in preventing neutropenia or obtaining a prompt recovery. The reticulocyte count fell below 10 × 109/l between POD 13 and 19, and also recovered spontaneously. A slapped-cheek rash was not observed in any of the 7 patients.
Trisomy 14 with Thrombocytosis and Monocytosis
Authors: Kaya Hiroyasu , Nakamura Shinobu , Okafuji Kazuhiro , Terasaki Yasushi , Maeno Koji , Tanaka Nobuyoshi , Ohtake Shigeki , Matsuda Tamotsu
Published: April 2003

It has been reported that trisomy 14 is associated with myeloid malignancies, but a case with increased platelet count has also been reported. However, the clinical significance of trisomy 14 is still uncertain. We report a patient with trisomy 14 with thrombocytosis and a gradual increase in monocytosis. He was treated with hydroxyurea, cytarabine and aclarubicin in low doses and his quality of life was maintained for a period of about 1 year from blastic crisis. Hydroxyurea, cytarabine or aclarubicin in low doses may be the treatment of choice for trisomy 14 patients with respect to the patients’ quality of life.
Consequences of Total and Subtotal Myeloperoxidase Deficiency: Risk or Benefit ?
Authors: Kutter D. , Devaquet P. , Vanderstocken G. , Paulus J.M. , Marchal V. , Gothot A.
Published: April 2003

A group of 100 totally or subtotally myeloperoxidase (MPO)-deficient individuals was compared to a reference population of 118 probands selected at random. Data for a protective effect of the deficiency against cardiovascular damage are presented. On the other hand, a significantly higher occurrence of severe infections and chronic inflammatory processes was noted among the deficient patients. An increased incidence of cancer among the MPO-deficient individuals was not demonstrated.
Oral Contraceptives Can Cause Falsely Low Vitamin B12 Levels
Authors: Gardyn J. , Mittelman M. , Zlotnik J. , Sela B.A. , Cohen A.M.
Published: April 2003

Serum vitamin B12 radioimmunoassays may give falsely low results in patients with folate deficiency, multiple myeloma, megadose of vitamin C and following radioisotope organ scan. We evaluated 10 consecutive healthy women on oral contraceptives (OC) who had falsely low vitamin B12 levels, as reflected by normal urine methylmalonic acid and plasma homocysteine. After 1-month cessation of OCs, vitamin B12 returned to the normal range in all women. Transcobalamin I (TCI) blood level was decreased in 60% of patients. OCs may cause temporary low vitamin B12 blood levels of no clinical significance that can be associated with low TCI levels
Hydroxyurea as a Cause of Drug Fever
Authors: Braester Andrei , Quitt Miriam
Published: April 2003

We report on a patient with essential thrombocythemia treated with hydroxyurea who became febrile 3 weeks after the treatment was started. After drug withdrawal, the fever resolved but after rechallenge there was recurrence of the fever. Although hydroxyurea-induced fever is rare, this drug must be added to the list of drugs that produce fever and the physicians should be aware of this possibility.
Consider ‘Hyperviscosity Syndrome’ in Unexplained Breathlessness
Authors: Coppell Jason
Published: April 2003

A 58-year-old man presenting with increasing shortness of breath over several months is reported here. All investigations were repeatedly normal until he suddenly and rapidly deteriorated, warranting admission, and a diagnosis was made of early multiple myeloma associated with the hyperviscosity syndrome. The plasma viscosity was extremely high compared to the low concentration of paraprotein present, and the possible mechanisms are discussed. Although uncommon, the hyperviscosity syndrome is an important cause of dyspnoea which is readily reversible, and should be considered when investigations of cardiac and pulmonary function are normal.
Secretion of Adrenomedullin by Renal Tubular Cell Lines
Authors: Sato Kyoko , Imai Taihei , Iwashina Masatora , Marumo Fumiaki , Hirata Yukio
Published: April 2003

Adrenomedullin (AM) is a novel and potent vasodilator peptide originally isolated from human pheochromocytoma. The present study was designed to study whether AM is produced by and secreted from renal tubular cell lines and whether arginine vasopressin (AVP) affects AM secretion from these cell lines. Three renal tubular cell lines derived from different species (LLCPK1, MDCK, and MDBK) secrete AM-like immunoreactivity (AM-LI) into culture medium, the immunological and physicochemical properties of which are similar to that of synthetic human AM as evaluated by reverse-phase high-performance liquid chromatography. Among the three cell lines, AVP in combination with a phosphodiesterase inhibitor (isobutylmethylxanthine) stimulated AM-LI secretion most potently from MDCK cells in a time- and dose-dependent manner. In MDCK cells, a V2 receptor agonist (deamino-D-Arg8-vasopressin) dose-dependently stimulated AM-LI secretion in the same manner as AVP. Furthermore, the AVP-induced AM-LI secretion was blocked by a V2 receptor antagonist (OPC31260), but not by a V1 receptor antagonist (OPC21268). These data indicate that AM is secreted from renal tubular cell lines and that AVP stimulates AM secretion via V2 receptors, suggesting its autocrine/paracrine role in renal function.
Compartmental Localization of Complement Component Transcripts in the Normal Human Kidney
Authors: Song D. , Zhou W. , Sheerin S.H. , Sacks S.H.
Published: April 2003

Local synthesis of complement components may play a crucial role in the pathogenesis of renal disease. Previous reports have shown that a number of complement components are produced by renal tissue both in vitro and in disease states. In the present study, we focused on the topographical distribution of components of the alternative and classical activation pathways in normal human kidney. As a whole, the normal renal cortex has the capacity to express the genes corresponding to most components of both complement pathways. There appears to be relatively high expression of transcripts for factor D and properdin in glomeruli, whilst factor B expression is greater within the medulla. Components C2, C3, and C4 and factor H are expressed predominantly in cortical tubule-rich fractions, and C1q is similarly expressed in all fractions. These results suggest that there may be differing emphasis on the alternative and classical pathways of complement activation in different regions within normal kidney.
Stress Response in a Leporine Renal Cell Model
Authors: Henle Kurt J. , Jethmalani Sunita M. , Nolen Greg T. , Wang Sho-Ya , Nowak Grazyna , Schnellmann Rick G.
Published: April 2003

It is well established that renal proximal tubule (RPT) cells grown under standard in vitro conditions attenuate many of their in vivo properties and functions. Thus, the study of renal stress response mechanisms requires an appropriate cell culture model. In the present study, we compared the heat stress (10 min, 45°C) response of freshly isolated RPT cells with that of RPT cells grown in vitro for 6 days under two different culture conditions: (1) SHAKE conditions, where oxygen levels and physiological functions are maintained via continuous media motion [Nowak G, Schnellmann RG: Am J Physiol 1996;271:C2072–2080] and (2) STILL conditions, involving standard cell culture which leads to partial hypoxia and a marked reduction in physiological functions. The freshly isolated RPT cells progressively synthesized heat shock proteins (HSPs) and stress glycoproteins (SGs) during a 3-hour culture period in vitro. Under these conditions, heat stress did not further increase HSP and SG synthesis. In RPT cells grown under SHAKE conditions, HSP70 synthesis was detected 1 h after heat stress and decreased below detection by 3 h. In contrast, the uptake of radiolabeled mannose into (glycoprotein) GP62 (Mr 62,000), GP50, and GP38 was observed in control SHAKE cultures and was not further increased after heat stress. These results are consistent with immunohistochemistry studies, where similar changes in HSP70 and GP50 expression were noted. RPT cells grown under STILL conditions showed both increased synthesis of HSP70 and increased glycosylation of GP62, GP50, and GP38 as early as 1 h after heat stress, but in contrast to SHAKE conditions, this heat-induced stress response further intensified at 3 h after heat stress. By 7 h after heating, HSP synthesis returned to control levels, while glycosylation of GP62 and GP50 remained elevated. Based on our results, we conclude that freshly isolated RPT cells exhibit a stress response that may be caused by acute cell isolation/culture stress. While this stress response unfolds, freshly isolated RPT cells appear unable to respond to additional heat stress. RPT cells grown under SHAKE and STILL conditions exhibit high rates of SG glycosylation, especially that of GP62, possibly reflecting a ‘stress’ condition associated with growth on plastic substrate. Concurrently, RPT cells from STILL cultures show a higher capacity for responding to acute heat stress than SHAKE cultures, evidenced by the transiently increased HSP synthetic rates. The interpretation of the renal stress response capacity, therefore, must be linked to a specific culture condition.
Potassium Depletion Stimulates mRNA Expression of Proximal Tubule AT1 Angiotensin II Receptors
Authors: Burns Kevin D. , Smith Iain B.
Published: April 2003

Dietary potassium (K+) deficiency is associated with blood pressure elevation and impaired urinary sodium excretion. Since angiotensin II is a potent stimulator of tubular sodium transport, we studied the effect of low [K+] on expression of kidney AT1 angiotensin receptors. In rabbits fed a K+-deficient diet for 14 days, plasma [K+] was significantly reduced compared to rabbits fed a standard diet (control: 4.06 ± 0.12 vs. K+-deficient: 2.66 ± 0.19 mmol/l; p < 0.001; n = 6–9). By Northern hybridization or RNase protection assays, dietary K+ deficiency caused an increase in mRNA expression for AT1 receptors in kidney cortex (43.5 ± 12.9% increase vs. control; p < 0.04; n = 8), and in proximal tubule segments in suspension (76.4 ± 28.8% increase vs. control; p < 0.005; n = 6). K+ deficiency had no effect on AT1 receptor mRNA expression in liver, or on mRNA expression of β-actin in kidney cortex, proximal tubule suspensions, or liver. To determine if low extracellular [K+] might directly modulate AT1 receptor mRNA expression, primary cultures of rabbit proximal tubule cells were incubated for 1, 3, 6 or 24 h in media with or without 5 mmol/l K+. Incubation of cells in 0 mmol/l K+ caused a 99.2 ± 32.9% increase in AT1 receptor mRNA expression at 3 h (p < 0.001; n = 14), returning to control levels by 24 h. Incubation of proximal tubule cells in 0 mmol/l K+ also caused a significant increase in basolateral membrane specific binding of [125I]-angiotensin II (p < 0.05; n = 4). These results indicate that dietary K+ deficiency and low extracellular [K+] stimulate expression of kidney AT1 angiotensin II receptors. Increased AT1 receptor mRNA and protein expression in proximal tubule may promote enhanced sodium reabsorption in K+ deficiency.
Biosynthesis of Methylguanidine in the Hepatic Peroxisomes and the Effect of the Induction of Peroxisomal Enzymes by Clofibrate
Authors: Takemura Katsumi , Aoyagi Kazumasa , Nagase Sohji , Gotoh Michihiro , Hirayama Aki , Ueda Atsushi , Tomida Chie , Koyama Akio
Published: April 2003

A state of peroxidation is one of the factors contributing to uremia. For example, we have reported that certain species of reactive oxygen, particularly the hydroxyl radical, play an important role in the biosynthesis of methylguanidine which contributes to toxicity in patients with uremia. However, it is uncertain which enzymes are involved in the synthesis of methylguanidine from creatinine. In this study, we attempt to show methylguanidine synthesis in the presence of peroxisomal enzymes that catalyze the β-oxidation of fatty acids. In addition, we investigate the effect of clofibrate, which induces peroxisomal enzymes or glutathione peroxidase activity, on methylguanidine synthesis in the peroxisomal fraction. Male Wistar rats were fed with the chow containing 0.5% clofibrate to induce peroxisomal enzymes and control rats were fed with ordinary laboratory chow. Peroxisomal fractions were obtained from liver homogenates by centrifugation, and incubated with creatinine in 0.1 M potassium phosphate buffer pH 7.4 at 37°C. Results show that methylguanidine is synthesized from creatinine concomitant with the synthesis of hydrogen peroxide from endogenous substrates in the peroxisomal fraction. This methylguanidine synthesis is inhibited by the addition of dimethylsulfoxide, glutathione, or sodium azide (p < 0.01). The rate of methylguanidine synthesis in clofibrate-treated rats was significantly less than that in control rats (p < 0.02). These results suggest that methylguanidine is synthesized in the peroxisomal fraction, and reactive oxygen species which are generated through this enzymatic reaction, participate in methylguanidine synthesis. Moreover, the induction of a scavenger system, especially glutathione peroxidase takes precedent over the generation of reactive oxygen species in peroxisomes treated with clofibrate.
The Expression of Water Channels AQP1 and AQP2 in a Large Series of ADPKD Kidneys
Authors: Devuyst Olivier
Published: April 2003

We read with interest the paper by Hayashi et al. [1] reporting the expression of AQP2 and AQP3 in cyst-lining epithelial cells from ADPKD kidneys. The authors suggest that about 30% of the cysts in ADPKD are derived from collecting-duct cells and that these cells remain well differentiated in terms of AQP expression. However, these conclusions are limited by the small number of ADPKD kidneys (n = 2) studied and by the fact that both AQP2 and AQP3 are located in the same cells – the principal cells of the collecting duct.
Intradialytic Cardiovascular Stability with Low Calcium Dialysate Levels
Authors: Navarro Juan F. , Teruel José L.
Published: April 2003

Over the last few years, the use of calcium salts has become an alternative method to phosphate binders containing aluminum in the control of hyperphosphatemia in dialysis patients, hypercalcemia being the main limitation to this therapeutical approach [1]. On this basis, a reduction in the calcium dialysate (CD) concentration has been proposed in order to prevent the appearance of hypercalcemia [2, 3]. However, the use of low CD concentrations may have negative effects on cardiovascular stability during hemodialysis (HD) [4, 5].