bluuebunny/crossref_metadata_2025 huggingface dataset:
Lack of Functional Pit-1 and Pit-2 Expression on Hematopoietic Stem Cell Lines
Authors: Thomsen Silke , Vogt Birgit , von Laer Dorothee , Heberlein Christoph , Rein Alan , Ostertag Wolfram , Stocking Carol
Published: April 2003

Hematopoietic stem cells (HSC) are an important target for retroviral gene transfer. However, transduction efficiency in these HSC is extremely low compared to fibroblasts or more mature hematopoietic cells. This infection block was analyzed in the HSC line FDC-Pmix. The infection frequency with the amphotropic murine leukemia virus (MLV-A) is more than 100-fold lower in FDC-Pmix cells as compared to fibroblasts. Pseudotyping with the env of the 10A1 strain (MLV-10A1), which uses both the amphotropic receptor (Pit-2) and the receptor for gibbon ape leukemia virus (Pit-1), did not improve the infection efficiency. Vectors pseudotyped with VSV G protein were found to overcome the infection block in FDC-Pmix, confirming that the block is at the level of virus binding and possibly penetration. Accordingly, we could not detect virus binding of MLV-A or MLV-10A1 to FDC-Pmix cell lines. Northern blot analysis was performed to detect whether the defect is at the level of transcription. Surprisingly, similar levels of Pit-2 receptor transcripts were detected in all cell types. The overexpression of rat Pit-2 DNA in CHO but not in FDC-Pmix cells improved amphotropic infection frequency after introducing rat Pit-2 DNA into the cells. Taken together these results show that the inefficient infection of FDC-Pmix is due to a lack of functional receptors. Either the receptor protein is incorrectly processed in these cells or a cofactor is missing in FDC-Pmix cells that is necessary for efficient binding and/or penetration.
Cis-Active Elements of Friend Spleen Focus-Forming Virus: From Disease Induction to Disease Prevention
Authors: Baum Christopher , Hunt Nicholas , Hildinger Markus , Eckert Hans-Georg , Zaehres Holm , Richters Anke , John Judith , Löhler Jürgen , Ostertag Wolfram
Published: April 2003

The polycythemic strain of the Friend spleen focus-forming virus (SFFVp) is a replication-defective, acutely transforming retrovirus inducing a bistage erythroleukemia in susceptible mice. The first stage of the disease is an acute polyclonal erythroblastosis induced by the proliferation-promoting effect of gp55. gp55 is expressed from a spliced subgenomic message of SFFVp and stimulates the cellular receptor for erythropoietin. Using a selectable SFFVp that otherwise mimics the specificity of the disease, we demonstrate that the kinetics of the polyclonal expansion depends on the transcriptional strength of the retroviral cis-active elements. By exchanging gp55 for apathogenic genes, we show that SFFVp enhancer and splice signals can be successfully utilized for the development of retroviral vectors mediating very efficient transgene expression in hematopoietic cells. Apathogenic selectable SFFVp-based vectors carrying distinct enhancer alterations are a valuable tool to analyze transcriptional control of leukemia viruses in the absence of oncogenic proteins. Moreover, they might have therapeutic potential.
Coexistence of Chronic Myelogenous Leukemia and Multiple Myeloma
Authors: Tanaka Masahisa , Kimura Ryoko , Matsutani Akira , Zaitsu Ken , Oka Yoshitomo , Oizumi Kotaro
Published: April 2003

A case report of simultaneous presentation of chronic myelogenous leukemia (CML) and multiple myeloma (MM) in a 72-year-old female is described. Our case was typical of Ph1-positive and chimeric bcr-abl messenger RNA-positive CML. Furthermore, a marked IgG (κ-type) paraproteinemia was present. Fluorescence in situ hybridization showed that 97% of marrow nucleated cells were positive for bcr-abl fusion signal, when myeloma cells in the bone marrow amounted to 19.0%. In the literature survey, 4 similar cases with coexistence of CML and MM have been identified.
Remission Induction by Granulocyte Colony-Stimulating Factor in Hypoplastic Acute Myelogenous Leukemia Complicated by Infection
Authors: Takamatsu Yasushi , Miyamoto Toshihiro , Iwasaki Hiromi , Makino Shigeyoshi , Tamura Kazuo
Published: April 2003

A patient with hypoplastic acute myelogenous leukemia (AML) who achieved remission with granulocyte colony-stimulating factor (G-CSF) alone is reported. The 59-year-old male patient received antibiotics and G-CSF but not any antileukemic drugs because of ongoing pneumonia. After 2-week administration of G-CSF, he achieved complete remission and his pneumonia improved. Since leukemia relapsed after 3 months, he received G-CSF again for 5 weeks, but failed to be in remission this time. He underwent antileukemic chemotherapy and achieved second remission. When he suffered from a second relapse after 7 months, intensive chemotherapy was commenced but was stopped on the 2nd day since his general condition became very poor due to septicemia. He began to receive G-CSF again and achieved third complete remission after 3 weeks. In vitro studies showed that G-CSF did not stimulate proliferation of the patient’s blast cells although they expressed G-CSF receptor on their surface. Moreover, G-CSF induced differentiation of the blast cells into segmented neutrophils in vitro. According to the literature, in all of the 12 patients with AML who were reported to achieve remission by G-CSF the course was complicated by infection, and 7 of the patients were diagnosed as hypoplastic acute leukemia. It is suggested that not G-CSF alone but G-CSF with infection could induce remission, which might be related to a differentiation effect of G-CSF in this case. G-CSF is not only safe but also useful for remission induction therapy in hypoplastic acute leukemia.
Comparison of the Relative Quantities of γ-mRNAs and Fetal Hemoglobin in SS Patients with Different Haplotypes
Authors: Smetanina N.S. , Gu L.-H. , Huisman T.H.J.
Published: April 2003

We have studied the relative levels of γ-mRNA [%γ/(γ + β)], Gγ- and Aγ-mRNAs [%Gγ/(Gγ + Aγ)], hemoglobin (Hb) F, and the Gγ and Aγ chains in some 50 patients with sickle cell anemia (SS) and with different haplotypes. As expected, the Hb F levels varied greatly and were high in patients with the Saudi Arabian-Indian haplotype. Similarly, the Gγ values varied greatly (from 19.5 to 76.5%) and depended on the haplotypes. A rare haplotype, named Mor, was found in 3 SS patients, 1 of whom was a homozygote Mor/Mor; this haplotype is associated with the lowest Gγ value (19.5% in the homozygote) and with a C→T mutation at position –202 of the Aγ promoter. The levels of γ-mRNA roughly parallel those of Hb F, but older patients have increased levels of mRNA, which appears not to be efficiently translated into Hb F. Similar observation have been reported for other hemoglobinopathies such as δβ-thalassemia heterozygotes and Hb Lepore heterozygotes. The relative quantity of Gγ-mRNA was closely related to that of the Gγ chain in the 15 patients who were studied; the Gγ- to Aγ-mRNA ratio did not change with age.
Determination of Concentration of Cytosolic NADH-Cytochrome b5 Reductase in Erythrocytes from Normal Chinese Adults, Neonates and Patients with Hereditary Methemoglobinemia by Double-Antibody Sandwich ELISA
Authors: Lan Fenghua , Tang Yuchai , Huang Changhui , Zhu Zhongyong
Published: April 2003

NADH-cytochrome b5 reductase (b5R), present in various tissues of the body, is a redox enzyme of multiple functions. The deficiency of the enzyme leads to hereditary methemoglobinemia. With rabbit anti-b5R antibody for plate coating and enzyme-labeled anti-b5R monoclonal antibody as reporter, we have developed a sandwich ELISA procedure for the determination of b5R concentration. This procedure is sensitive to a wide range of linearity, and convenient in coping with large numbers of samples. Using this novel method, cytosolic b5R concentration in the erythrocytes of 30 normal Chinese adults was estimated to be 25.63±8.54 ng/mg Hb. It was found that the concentration of red cell soluble b5R of five newborns was significantly lower than that of normal adults and soluble b5R was undetectable in the erythrocytes of 4 patients with type I hereditary methemoglobinemia. Our results demonstrated that the reduced b5R activity in red cell cytosol of both neonates and type I hereditary methemoglobinemic patients results largely or mainly from the lowered b5R concentration. Our novel method might be further exploited for in-depth investigation of the relationship between the qualitative and quantitative changes of b5R with regard to physiological and pathological conditions.
Epoetin Alfa in Low Hematocrit Patients to Facilitate Autologous Blood Donation in Total Hip Replacement: A Randomized, Double-Blind, Placebo- Controlled, Dose-Ranging Study
Authors: Mercuriali F. , Inghilleri G. , Biffi E. , Colotti M.T. , Vinci A. , Oriani G.
Published: April 2003

We investigated the safety and efficacy of preoperative epoetin alfa used in conjunction with preoperative autologous blood donation (PAD) in 40 anemic orthopedic surgical patients undergoing hip replacement surgery [hematocrit (Hct) ≤39%]. Patients were randomized 25–35 days before surgery to twice weekly intravenous (i.v.) epoetin alfa (300, 150, or 75 IU/kg) or placebo treatments supplemented with 200 mg i.v. iron. Epoetin alfa facilitated dose-dependent increases in PAD (4.3 units, 300 IU/kg; 3.4 units, 150 IU/kg; 3.0 units, 75 IU/kg; 2.1 units, placebo), and minimized reductions in Hct associated with repeated phlebotomy. All epoetin alfa doses were well tolerated and safe. Our results indicate that preoperative epoetin alfa dose-dependently stimulates erythropoiesis and facilitates PAD in anemic orthopedic patients, which may benefit patients by reducing their exposure to the risks of allogeneic transfusion, hastening recovery and discharge from hospital.
Gelatinous Bone Marrow Transformation Complicated by Self-Induced Malnutrition
Authors: Nonaka Daisuke , Tanaka Minoru , Takaki Kouichi , Umeno Morio , Okamura Takashi , Taketa Hirohisa
Published: April 2003

A 48-year-old woman with a history of anorexia nervosa was admitted to our hospital because of malaise, anorexia and edema in the face and legs. She was diagnosed with gelatinous bone marrow and iron deficiency anemia due to severe malnutrition. She was intravenously treated by saccharated ferric oxide and her anemia was improved, but her bone marrow still showed much gelatinous material.
Influence of α-Thalassemia on Cholelithiasis in SS Patients with Elevated Hb F
Authors: Haider M.Z. , Ashebu S. , Aduh P. , Adekile A.D.
Published: April 2003

Chronic hemolysis, with consequent hyperbilirubinemia, predisposes SS patients to pigment gallstones. The other factors which influence the development of stones in these patients have not been identified. We have carried out a combined prospective and retrospective study of SS patients in Kuwait and specifically investigated the influence of coexistent α-thal trait on the prevalence of gallstones. A total of 45 patients (30 males, 15 females) with ages ranging from 1 to 16 years (mean 7.2 ± 3.1) were studied. Most were either homozygotes for the Saudi Arabia/India haplotype (86.7%) or compound heterozygotes for this and the Benin haplotype (11.1%). They were screened for gallstones with ultrasonography. α-Globin genotypes were determined using a combination of PCR and allele-specific oligonucleotide hybridization techniques to identify the common α-thalassemia alleles in this population. Gallstones were detected in 7 (15.6%) patients (4 males, 3 females), whose mean age (10.5 ± 5.5 years) was significantly higher than that (6.8 ± 3.2 years) of those without stones (p < 0.01). The mean total Hb of the former (8.4 ± 0.8 g/dl) was also significantly (p < 0.05) lower than in the latter (9.5 ± 1.3 g/dl), while the difference in mean Hb F levels was not significant. None of the 4 α-thal homozygotes had gallstones while 2 of 13 heterozygotes and 5 of the 23 patients without coexistent α-thal had. The differences in these proportions are statistically significant (χ2 = 20.4, p < 0.001). It therefore appears that coexistent α-thal decreases the chance of developing gallstones in Arab SS patients. This may be related to less hemolysis in such patients as shown by their higher mean Hb level.
Complete Blood Counts from Umbilical Cords of Healthy Term Newborns by Two Automated Cytometers
Authors: Walka M.M. , Sonntag J. , Kage A. , Dudenhausen J.W. , Obladen M.
Published: April 2003

Complete blood counts (CBC) of umbilical cord blood from 123 healthy term newborns were simultaneously performed with two different cytometers using laser as a light source. Medians (95% range) were: WBC 14.2 (7.8–24.3) × 109/l, platelets 265 (174–363) × 109/l, Hb 15.7 (12.5–18.2) g/dl, RBC 4.6 (3.9–5.5) × 1012/l, MCV 106 (95–113) fl, PCV 0.49 (0.40–0.56), and MCH 33.8 (30.3–36.4) pg. Reticulocytes were 149 (95–212) × 109/l or 3.3 (2.0–4.7)%; erythroblasts 5 (0–24) per 100 WBC or 0.53 (0.00–3.20) × 109/l. The counters agreed well except for MCHC. WBC counts showed the smallest difference irrespective of erythroblast number; platelets showed the largest difference. The lower limit for normal Hb should be fixed at 12.5 g/dl for the adequate diagnosis of anaemia from cord blood of term newborns.
A Human Myeloid Cell Line Producing Stem Cell Growth Factor, KPB-M15, Secretes Another Growth Factor Active on Murine Hematopoietic Progenitor Cells
Authors: Hiraoka Atsunobu , Nagasawa Tetsuji , Ohta Naomi , Sugimura Atsushi , Shimizu Norihide , Ooi Kiyomoto , Shimizu Shin
Published: April 2003

Human stem cell growth factor (SCGF) produced by a myeloid cell line, KPB-M15, exhibits species-specific hematopoietic activities. However, KPB-M15-conditioned medium induced colony formation of mouse bone marrow cells. KPB-M15-derived colony-stimulating activity (CSA) was purified through Butyl-Toyopearl 650c and Cu2+ chelating-Sepharose 6B chromatography. TSK-G3000SW gel filtration of the purified preparation presented 3 distinct peaks around Vo, 150 kD and 85 kD. Gel fractions extracted from SDS-PAGE had macrophage colony-stimulating factor (M-CSF)-specific amino acid sequences. PCR, Northern hybridization and ELISA demonstrated that KPB-M15 cells secreted a significant amount of M-CSF and IL-6. Anti-M-CSF but not anti-IL-6 antibody abrogated CSA in KPB-M15-CM. IL-6 hardly synergized with M-CSF to enhance colony formation. Collectively, M-CSF is a sole CSA for murine hematopoietic progenitor cells in KPB-M15-CM. This is the first report of a human myeloid cell line, KPB-M15, constitutively producing M-CSF in addition to SCGF and IL-6. It can be useful in investigating the mechanism of production of M-CSF.
In vivo Administration of Granulocyte Colony-Stimulating Factor Increases the Surface Expression of Sialyl-Lewisx on Neutrophils in Healthy Volunteers
Authors: Ohsaka Akimichi , Saionji Katsu
Published: April 2003

We examined the in vivo effect of granulocyte colony-stimulating factor (G-CSF) on the surface expression of putative counterligands for endothelial selectins on neutrophils in healthy volunteers. G-CSF (50 µg/m2/day) was administered subcutaneously to 5 healthy volunteers for 4 days. The expression of surface antigens on neutrophils was determined by flow cytometry and monoclonal antibodies. G-CSF administration increased the number of leukocytes, mainly of neutrophils, which was associated with an increase in the expression of the high-affinity Fc receptor for IgG (FcRI, CD64) and CD14 on neutrophils. G-CSF administration decreased the surface expression of L-selectin on neutrophils, whereas it increased the expression of sialyl-Lewisx but not Lewisx on neutrophils. These findings suggest that G-CSF participates in the neutrophil-endothelial cell interactions in vivo by modulating the expression of adhesion molecules and ligands for endothelial selectins on neutrophils.
New Strategies for Allogeneic Bone Marrow Transplantation and Organ Allografts
Authors: Ikehara Susumu
Published: April 2003

In humans, the success rate of bone marrow transplantation (BMT) across major histocompatibility complex (MHC) barriers is not high due to: (1) graft-versus-host reaction (GvHR); (2) graft rejection, and (3) incomplete T cell recovery. In mice, GvHR can be prevented if T cell- depleted bone marrow cells (BMCs; <2% T cells) are used. Graft rejection can be prevented by either bone grafts (to recruit donor-derived stromal cells) or the injection of donor BMCs via the portal vein (p.v; to induce donor-specific tolerance). T cell functions are recovered by BMT plus bone grafts if the thymic functions of recipients are not completely lost. After the complete loss of thymic functions (due to aging), BMT plus embryonal thymus grafts should be carried out.Recently, we have found that persistent donor-specific tolerance can be induced if allogeneic hemopoietic stem cells are injected via the p.v. Based on these findings, we have established new strategies for organ allografts. Without irradiation, donor BMCs should be injected from the p.v. injection on day 0 plus i.v. injection on day 5, and an immunosuppressant (CsA or FK506) should be used on days 2 and 5. Without using immunosuppressants, sublethal irradiation (7 Gy) followed by skin allografts plus allogeneic BMC injection via the p.v. should be carried out. This leads to a 100% acceptance of skin and pancreas allografts for more than 300 days. The recipient mice show mixed allogeneic chimerism, and spleen cells from the recipients show tolerance to both donor-type and host-type MHC determinants in the assays for mixed lymphocyte reaction and generation of cytotoxic T lymphocytes. We have confirmed that these strategies are applicable to other animals such as pigs and rats. We therefore believe that they will become viable and valuable strategies for human organ allografts.
Antigenic Phenotyping of Lymphoid Cells and B Cell Gene Rearrangement in Type B Gastritis and in Gastritis Not Associated with Helicobacter pylori Colonization
Authors: Fritscher-Ravens A. , Petrasch S. , Tiemann M. , Wacker H. , Dörr T. , Kemmeries G. , Hilgers W. , Reinacher-Schick A. , Kunstmann E. , Schmiegel W.
Published: April 2003

Marginal-zone B cells of the mucosa-associated lymphoid tissue (MALT) are the normal counterpart of the neoplastic cells in MALT lymphoma. In both cases these lymphocytes express surface immunoglobulins, but are negative when stained for B cell associated antigens like CD10 and CD23. Furthermore, the B cell gene rearrangement has been found in Helicobacter pylori associated chronic gastritis and in extranodal type of marginal-zone lymphoma. The aim of this study was to quantify the number of IgM-, CD10-, and CD23-positive lymphocytes in patients with type B gastritis and to compare the results with the antigen profile of mononuclear cells in patients with gastritis not associated with H. pylori. Additionally, the immunoglobulin heavy-chain (IgH) gene rearrangement in H. pylori positive and H. pylori negative gastritis was studied. From 23 patients with a positive urease test and/or histologically proven H. pylori infection and chronic gastritis and from 22 patients with H. pylori negative chronic gastritis mucosa biopsy specimens were taken. Single-cell suspensions were obtained following enzymatic digestion. For immunocytochemistry, an alkaline phosphatase-antialkaline phosphatase method was applied. IgH gene rearrangement in formalin-fixed, paraffin-embedded specimens was determined by polymerase chain reaction in 11 patients with chronic gastritis. An increase in mu-positive plasma cells and B lymphocytes was detected in patients with H. pylori positive gastritis as compared with patients with H. pylori negative gastritis (10.0 vs. 3.9%, p < 0.001, and 4.3 vs. 1.6%, p < 0.01, respectively). In both groups, the proportion of CD10- and CD23-positive lymphocytes was <1%. IgH gene rearrangement was not restricted to type B gastritis; single bands were also present in 3 of 7 patients with H. pylori negative chronic gastritis. Our finding of IgH gene rearrangement in some of the patients with H. pylori negative chronic gastritis indicates that additional factors may be critical for these genotypical changes and for the pathogenesis of gastric MALT lymphoma.
Function and X Chromosome Inactivation Analysis of B Lymphocytes in Myelodysplastic Syndromes with Immunological Abnormalities
Authors: Okada Masaya , Okamoto Takahiro , Takemoto Yoshinobu , Kanamaru Akihisa , Kakishita Eizo
Published: April 2003

To investigate the pathogenesis of immunological abnormalities (IA) in myelodysplastic syndrome (MDS), we examined B cells for their ability to produce cytokine and their X chromosome inactivation pattern (XCIP). An IA was defined as being positive for at least one autoimmune laboratory test (e.g. antinuclear antibody, rheumatoid factor). Seventy-three MDS patients [65 with refractory anemia (RA), 3 with RA with excess blasts (RAEB), and 5 with RAEB in transformation] were examined; 47 had IA and 26 had no IA. To examine the function of B cells in MDS, the production of interleukin (IL)-6 and IL-10 was measured in cultures of purified B cells with or without stimulators. Both IL-6 and IL-10 production rates in patients with IA were significantly higher than in patients without IA and normal controls. The skewing of XCIP of B cells was analyzed by using the polymerase chain reaction, and the skewing rate of B cell XCIP was quantitatively assayed by compared to control T lymphocytes. The skewing rate of B cells was higher in patients with IA than in those without IA and normal controls. Therefore, a small population of B cells in patients with IA might be derived from MDS clones, and be associated with the induction of IA.
The Synergistic Effect of Thrombopoietin in Erythropoiesis and Myelopoiesis from Human Bone Marrow Cells
Authors: Liang D.-C. , Shih L.-Y. , Kuo M.-C. , Chen S.-H. , Liu H.-C. , Shimosaka A.
Published: April 2003

We sought to determine whether recombinant human thrombopoietin (TPO) acts synergistically with recombinant human erythropoietin (EPO) and/or recombinant human interleukin-3 (IL-3) on erythroid burst formation and granulocyte-macrophage colony formation from human bone marrow (BM). BM cells were from 5 adults and 15 children who underwent bone marrow examination because of a clinical suspicion of malignancy; their bone marrows as well as the complete blood counts were normal and were cultured in a methylcellulose system. TPO has a synergistic effect with EPO or EPO + IL-3 on erythropoiesis of human BM, as the addition of TPO to EPO significantly gave rise to more erythroid bursts (p = 0.0001) and the addition of TPO to EPO + IL-3 might give rise to more erythroid bursts (p = 0.05). TPO also has a synergistic effect with recombinant human granulocyte colony-stimulating factor (G-CSF) on myelopoiesis of human BM, since the addition of TPO to G-CSF gave rise to significantly more granulocyte-macrophage colonies (p = 0.0001). Besides its well-known significant role in megakaryopoiesis, TPO also has synergistic effects on erythropoiesis and myelopoiesis.
Graft Failure of Autologous Peripheral Blood Stem Cell Transplantation due to Acute Metabolic Acidosis Associated with Total Parenteral Nutrition in a Patient with Relapsed Breast Cancer
Authors: Sawada Michio , Tsurumi Hisashi , Hara Takeshi , Goto Hideko , Yamada Toshiki , Oyama Masami , Moriwaki Hisataka
Published: April 2003

A 32-year-old female had been diagnosed as having relapsed breast cancer and liver metastasis. She underwent high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation (PBSCT) with 5.8 × 106/kg CD34+ cells. She was supported by total parenteral nutrition (TPN) without vitamins throughout these therapies. Hematopoietic recovery was not observed by day 28 after PBSCT, necessitating a second PBSCT on day 29 using the back-up material of 4.4 × 106/kg CD34+ cells. On the next day, she suddenly developed severe metabolic acidosis, heart failure and deep coma. After immediate infusion of thiamine, heart failure and coma rapidly improved. The neutrophil count reached 0.5 × 109/l on day 9 and the platelet count 50 × 109/l on day 15 after the second PBSCT. This is a rare graft failure due to acute metabolic acidosis or thiamine deficiency associated with TPN.
Aplastic Anemia before Bone Marrow Transplantation and Antilymphocyte Globulin
Authors: Heimpel Hermann
Published: April 2003

In severe aplastic anemia, disease-dependent mortality was high before allogeneic bone marrow transplantation (BMT) and immunosuppressive therapies (IST) including antilymphocyte globulin became available. However, under supportive therapy alone, spontaneous remissions were observed in up to 20% of severe cases, reflecting the natural course of the disease. Therefore, in evaluating new forms of treatment, one has to keep in mind that remission is not necessarily response, and that final proof of utility of any new therapy still requires a randomized study design. Transition to leukemia or myelodysplasia was rarely observed if the initial diagnosis was accurate. The much higher incidence of leukemias in patients treated by IST, but not by BMT is probably due to the better life expectancy of patients at risk, rather than to a leukemogenic potential of IST itself. ‘Outdated’ therapeutic modalities, such as androgens or splenectomy, may still be justified as an adjuvant therapy in selected cases.
Cell Cycling Stress in the Monocyte Line as a Risk Factor for Progression of the Aplastic Anaemia/Paroxysmal Nocturnal Haemoglobinuria Syndrome to Myelodysplastic Syndrome
Authors: Nissen Catherine , Genitsch Alexander , Sendelov Silvia , Dalle Carbonare Verena , Wodnar-Filipowicz Aleksandra
Published: April 2003

Severe aplastic anaemia (SAA) causes permanent stem cell damage from which patients do not recover after treatment with antilymphocyte globulin (ALG). To produce peripheral blood values compatible with life, the few remaining stem and precursor cells are put under stress. We defined a ‘stress factor’ (SF) for various haematopoietic lines as the ratio of the corresponding peripheral blood (PB) value to the total colony number in short-term bone marrow cultures from 86 patients with different outcomes. Both values are expressed as percentage of normal, hence SF averages 1 in normal steady-state haematopoiesis. SF was elevated in all patients, from 2-to 40-fold, with wide variations in different patient groups and striking differences between haematopoietic lineages. In long-term disease-free survivors after ALG (group 1) the mean total colony count was 19% of normal, with a significantly higher proportion of erythroid burst-forming units compared to normal. They had ineffective erythropoiesis with haemoglobin (Hb) values below, and reticulocyte counts above normal; platelet counts were 67% of normal. In contrast, monocyte counts were in the high normal range, resulting in a high SF (18.7 ± 1.9) for monocytes. In patients who developed paroxysmal nocturnal haemoglobinuria (PNH) after ALG (group 2), ineffective erythropoiesis, reflecting haemolysis, was more pronounced and they had striking relative monocytosis, resulting in a significantly higher SF for monocytes (33.7 ± 5.7) compared with group 1 (p < 0.0001). High monocyte counts most likely reflect the relative resistance of nucleated cells to complement, compared with red cells and platelets. Patients who developed myelodysplastic syndrome (MDS) or acute myeloid leukaemia (AML) after ALG, with or without PNH (group 3), had the highest SF for monocytes (39 ± 10). They also had neutrophil counts in the upper range, or above normal, resulting in a high SF for neutrophils: 32 ± 19. In patients with persisting or relapsing-remitting pancytopenia without a clinically detectable clonal disorder (group 4), all values were strikingly similar to those of the PNH group. In patients who achieved normal PB values after uncomplicated bone marrow transplantation (group 5), the SF averaged 3, but they also had ineffective erythropoiesis and mild relative monocytosis, a possible sign of occult PNH. We conclude that all patients after treatment of SAA have ineffective erythropoiesis and relative monocytosis, and that these abnormalities probably reflect PNH. We suggest that the resulting high SF for the leukocyte – particularly the monocyte line – predisposes to the development of MDS/AML. We discuss how these results may provide some of the missing pieces in the puzzle of SAA/PNH.
Flow Cytometric Quantification of Phagocytosis in Acute Myeloid Leukemia
Authors: Bassøe Carl-Fredrik
Published: April 2003

Phagocytosis was studied by flow cytometry (FCM) in 15 patients with acute myeloid leukemia (AML). The pattern of phagocytosis differed markedly between AML and controls. The percentage of phagocytosing AML leukocytes was below that of the controls (p < 0.01). The phagocytic capacity of a subpopulation of leukemic cells was diminished, but compensated by the phagocytosis of a few prey by each of many immature leukocytes. Phagocytosis by immature and mature AML leukocytes was receptor dependent, and both carried functional complement receptors. Attachment to the cell surface was not rate-limiting in phagocytosing AML leukocytes.
CD8 Expression in a Case of Chronic Lymphocytic Leukemia with Trisomy 12
Authors: Bayo Hanza María Carolina , Palacios María Fernanda , Dourisboure Ricardo , Slavutsky Irma , Chena Christian , Galmarini Carlos , Sarmiento Marcela , de Bracco María M.E. , Scolnik Mariano
Published: April 2003

We report a case of B-cell chronic lymphocytic leukemia (B-CLL) with aberrant expression of the T-cell-associated antigen CD8, as revealed by two-color flow-cytometric analysis. DNA studies showed immunoglobulin heavy-chain gene rearrangement, but not of γ-chain T-cell receptor, confirming the B-cell origin of the neoplastic cells. Ploidy analysis showed a tetraploid population and high S-phase fraction. B-CLL cells also carried trisomy 12, detected by fluorescence in situ hybridization. The identification of more cases with the same features would be necessary to establish the prognosis of this subtype of B-CLL.
Gene Rearrangements in Bone Marrow Cells of Patients with Acute Myelogenous Leukemia
Authors: Schmetzer H.M. , Braun S. , Wiesner D. , Duell T. , Gerhartz H.H. , Mittermueller J.
Published: April 2003

At diagnosis, clonal gene rearrangement probes [retinoic acid receptor (RAR)-α, major breakpoint cluster region (M-bcr), immunoglobulin (Ig)-JH, T cell receptor (TcR)-β, myeloid lymphoid leukemia (MLL) or cytokine genes (GM-CSF, G-CSF, IL-3)] were detected in bone marrow samples from 71 of 153 patients with acute myelogenous leukemia (AML) (46%): in 41 patients with primary AML (pAML) (58%) and in 30 patients with secondary AML (42%). In all cases with promyelocytic leukemia (AML-M3) RAR-α gene rearrangements were detected (n = 9). Gene rearrangements in the Ig-JH or the TcR-β or GM-CSF or IL-3 or MLL gene were detected in 12, 10, 16 and 12% of the cases, respectively, whereas only few cases showed gene rearrangements in the M-bcr (6%) or G-CSF gene (3%). Survival of pAML patients with TcR-β gene rearrangements was longer and survival of pAML patients with IL-3 or GM-CSF gene rearrangement was shorter than in patients without those rearrangements. No worse survival outcome was seen in patients with rearrangements in the MLL, Ig-JH or M-bcr gene. In remission of AML (CR), clonal gene rearrangements were detected in 23 of 48 cases (48%) if samples were taken once in CR, in 23 of 26 cases (88%) if samples were taken twice in CR and in 23 of 23 cases (100%) if samples were studied three times in CR. All cases with gene rearrangements at diagnosis showed the same kind of rearrangement at relapse of the disease (n = 12). Our data show that (1) populations with clonal gene rearrangements can be regularly detected at diagnosis, in CR and at relapse of AML. (2) Certain gene rearrangements that are detectable at diagnosis have a prognostic significance for the patients’ outcome. Our results point out the significance of gene rearrangement analyses at diagnosis of AML in order to identify ‘poor risk’ patients – independently of the karyotype. Moreover, the persistence of clonal cells in the further course of AML can be studied by gene rearrangement analysis.
Thrombopoietic Cytokines in Patients with Iron Deficiency Anemia with or without Thrombocytosis
Authors: Akan Hamdi , Güven Nilüfer , Ayogdu İsmet , Arat Mutlu , Beksaç Meral , Dalva Klara
Published: April 2003

Iron deficiency anemia is a cause of reactive thrombocytosis. A moderate increase in platelet numbers is common but sometimes counts may exceed 1,000 × 109/l. The mechanisms causing reactive thrombocytosis are unclear. In this study, we evaluated 15 women with iron deficiency anemia and thrombocytosis (platelets >450 × 109/l) and 16 women with iron deficiency anemia with normal platelet counts. Serum samples were taken before oral iron replacement therapy, after 1 and 3 months and at the end of replacement therapy. Thrombopoietin, erythropoietin (EPO), leukemia inhibitory factor, interleukin-6 and interleukin-11 levels were assayed. There was no change in the levels of thrombopoietic cytokines except for EPO. The correlation between high EPO levels and high platelet counts may suggest that EPO increases platelet counts, but the same EPO level changes can also be demonstrated in women with iron deficiency anemia but normal initial platelet counts. The fact that the levels of other cytokines remained unchanged during treatment suggests that either these cytokines have no effect on reactive thrombocytosis or the change in platelet counts in our patients is in a narrow range and is thus not affected by the cytokine levels.
The Inflammation Meter: Novel Technology to Detect the Presence of Infection/Inflammation in Patients without Leukocytosis but with Increased Leukocyte Adhesiveness/Aggregation
Authors: Maharshak Nitsan , Kassirer Michael , Zeltser David , Rotstein Riva , Rogowski Ori , Shapira Itzhak , Deutsch Varda , Arber Nadir , Eldor Amiram , Berliner Shlomo
Published: April 2003

Objectives: To reveal the presence of infection/inflammation in patients with relatively normal white blood cell count (WBCC) by using the leukocyte adhesiveness/aggregation test (LAAT). Methods: The LAAT was performed by using a simple slide test and image analysis (Inflamet™), the WBCC, by an electronic cell analyzer, C-reactive protein, by Laser nephelometry and CD11b/CD18 by whole blood flow cytometry. Results: Forty out of a cohort of 121 patients with nonviral acute febrile illness had a WBCC within normal limits. The intensity of the inflammatory response in these individuals as judged by either C-reactive protein, or fibrinogen concentrations, erythrocyte sedimentation or polymorphonuclear leukocyte CD11b/CD18 expression was similar to that observed in patients with a leukocytic response. Our present finding that 63% out of the group with documented infection/inflammation and no leukocytosis had a significantly increased LAAT suggest that the lack of leukocytosis is in part a pseudoleukopenia, or is associated with some degree of uncompensated tissue leukostasis. Conclusions: The lack of a leukocytic response in a patient with nonviral infection/inflammation is by no means a sign of a less inflammatory response. The increased state of leukocyte adhesiveness/aggregation might help to disclose the presence of inflammation in these individuals.
Significance of Glomerular Deposition of Protein S in Various Glomerulopathies
Authors: Suzuki Satoru , Gejyo Fumitake , Takahashi Hoyu , Ueno Mitsuhiro , Nishi Shin-ichi , Kimura Hideki , Arakawa Masaaki
Published: April 2003

To elucidate the relationship between glomerular deposition of protein S (PS) and renal lesions or dysfunction, 30 patients with various glomerulopathies were examined. Glomerular PS deposition was found in 20 patients (group A), and other 10 patients showed no deposition (group B). PS was found mainly along the capillary loops and segmentally in the mesangium. Group A showed significantly more severe proteinuria than group B (p < 0.05). Group A patients showed significant decreases in glomerular filtration rate (p < 0.01). Patients in group A had significantly lower plasma levels of plasmin-alpha2-plasmin inhibitor complexes (p < 0.05) and thrombin-antithrombin III complexes (p < 0.01) than those in group B. Group A showed significant decreases in the mean values of plasma total PS (p < 0.01) and protein C (PC) antigens (p < 0.01) and C4b-binding protein (C4bp; p < 0.05) as compared with group B patients. There was a positive correlation between plasma PS and C4bp (p < 0.02). Histologically, group A showed a significantly higher incidence of glomerular deposition of factor XIII (subunit a), alpha2-plasmin inhibitor, PC (p < 0.05), and C4bp (p < 0.01). The present study demonstrates that glomerular PS deposition indicates the existence of PC and C4bp in the glomeruli and suggests that the glomerular PS deposition may modify the activation of fibrinolytic and coagulation systems within the glomeruli in various glomerulopathies.
Edema, Oncotic Pressure, and Free Entropy: Novel Considerations for the Treatment of Edema through Attention to Thermodynamics
Authors: Diskin Charles J. , Gupta Ram B. , Ravis William , Stokes Thomas J. , Dansby Linda M. , Carter Thomas B. , Thomas Selby G.
Published: April 2003

Over 170 years after Richard Bright and a century after Ernest H. Starling, the development, location, and severity of edema in patients with renal impairment continue to baffle the predictions of most nephrologists. While much of the phenomenon can be explained by levels of serum proteins, or hydrostatic pressures, there are stunning exceptions well known to any practicing nephrologist. Some of the derangement is undoubtedly due to unmeasured but well-known variables, such as membrane permeability; however, other factors such as free entropy of plasma are also clearly involved. The study of other polyelectrolyte colloids, similar to plasma proteins, for industrial purposes has led to the identification of various phenomena such as counterion condensation that can result in loss of entropy and consequently osmotic pressure. Variables known to result in a loss of free entropy, such as pH, oxidation products and ligand binding, are discussed. Older equations developed by van’t Hoff and Donnan might require replacement by newer mathematical models such as the nonlinear Poisson-Boltzmann equation or the Monte Carlo simulator. Attempts to restore free entropy to plasma would be a more physiological treatment of edema than diuretic use. Implications are noted for future drug development to treat edema.
Biocompatibility Differences with Respect to the Dialyzer Sterilization Method
Authors: Müller Thomas F. , Seitz Michael , Eckle Ilsebill , Lange Harald , Kolb Gerald
Published: April 2003

The impact of the method of sterilization (steam vs. ethylene oxide, ETO) on indices of biocompatibility is investigated using polysulfone membranes. Eight patients were treated with a random choice of the high-flux membranes F60S (steam) and F60 (ETO) and the low-flux membrane F6 (ETO). Blood samples were taken prior to and 5, 15, 30, 60, and 180 min after the start of hemodialysis. White blood cell count, platelet count, and plasma concentrations of polymorphonuclear neutrophil elastase, complements C3a and C5a, and β2-microglobulin were determined. The dialysis procedure was associated with a significant decrease in white blood cell count and β2-microglobulin level and a significant increase in polymorphonuclear neutrophil elastase and complement C3a and C5a levels. However, the steam-sterilized F60S membrane had a significantly lower impact on the biocompatibility indices than the ETO-sterilized F60 and F6 membranes (p < 0.05 or p < 0.001 for the individual markers). We conclude that using steam instead of ETO for sterilization may improve the biocompatibility of membranes.
Fatal Sepsis from Vibrio vulnificus in a Hemodialyzed Patient
Authors: Stabellini N. , Camerani A. , Lambertini D. , Rossi M.R. , Bettoli V. , Virgili A. , Gilli P.
Published: April 2003

Vibrio vulnificus, a particularly virulent halophilic vibrio, has been isolated from the blood and skin necrotic lesion of a hemodialyzed patient with sepsis. The patient has had exposure of the skin to seawater. Various chronic conditions including renal failure have a great risk for developing septicemia due to V. vulnificus. It is necessary to inform persons with liver diseases or immunocompromising conditions of hazards associated with the consumption of undercooked seafood and seawater exposure.
Lupus Anticoagulant Cofactors and the Kidney
Authors: Cheng Hwee-Ming
Published: April 2003

We wish to supplement the useful comment by Stratta et al. [1] on the importance of including lupus anticoagulant (LA) tests when detecting for antiphospholipid autoantibodies (aPL), since monotesting solely with solid-phase aPL immunoassays may not be clinically relevant.
Mesangial Expression of a Nonmuscle Myosin Heavy Chain, SMemb, Is Associated with Glomerular Sclerosis and Renal Prognosis in IgA Nephropathy
Authors: Mise Naobumi , Kimura Kenjiro , Nagai Ryozo , Ohba Shigeyoshi , Suzuki Naoe , Miyashita Kazuhisa , Tojo Akihiro , Goto Atsuo , Omata Masao
Published: April 2003

To characterize the phenotypic alteration in mesangial cells in human glomerulonephritis, we investigated the expression of nonmuscle-type myosin heavy chain, SMemb, and α-smooth muscle actin (α-SM actin) in IgA nephropathy. The expression of SMemb and α-SM actin was examined by immunohistochemistry in biopsy specimens from 45 patients with IgA nephropathy. We examined a total of 489 glomeruli representing all patients enrolled, and found that mesangial expression of SMemb and α-SM actin was associated with mesangial proliferation. Only mesangial expression of SMemb showed a significant relationship with mesangial matrix accumulation. Semiquantitative evaluation using composite expression scores showed that the expression of SMemb was elevated in the patients with poor renal prognosis. The expression of α-SM actin showed no significant relationship with renal prognosis. These results suggest that mesangial expression of SMemb is an important factor in the progression of IgA nephropathy, and that SMemb and α-SM actin are associated with the activation of mesangial cells by different mechanisms.
Effects of Chitosan-Coated Dialdehyde Cellulose, a Newly Developed Oral Adsorbent, on Glomerulonephritis Induced by Anti-Thy-1 Antibody in Rats
Authors: Yagi Mikio , Kato Shinichiro , Nishitoba Tsuyoshi , Sato Hiromitsu , Kobayashi Nami , Iinuma Noriko , Nagano Nobuo
Published: April 2003

The effects of chitosan-coated dialdehyde cellulose (chitosan DAC), a newly developed oral adsorbent for urea and ammonia, were examined in a glomerulonephritis model in rats. Mesangial proliferative glomerulonephritis accompanied with proteinuria was induced by an intravenous injection of anti-rat Thy-1.1 monoclonal antibody (OX-7). The proliferation of mesangial cells and an accumulation of extracellular matrix components such as type I collagen and fibronectin were observed in the glomeruli 9 days after OX-7 injection; these were improved in rats fed a diet containing chitosan DAC (10% content) for 9 days compared with those in rats fed a normal diet. Chitosan DAC treatment decreased the elevated urinary protein and blood urea nitrogen at days 8–9 to the normal levels; the increased fecal excretion of nitrogen might participate in this phenomenon. In addition, chitosan DAC treatment showed an increase in fecal water content associated with a decrease in urinary volume. These therapeutic effects may be due to the reduction of proteinic factor expression and the compensational function of chitosan DAC for kidney. These results suggest that chitosan DAC treatment may be useful for ameliorating mesangial proliferative glomerulonephritis.
Apoptosis and Expression of Bax Protein and Fas Antigen in Glomeruli of a Remnant-Kidney Model
Authors: Hattori Tomohisa , Shindo Shoichiro , Kawamura Hideki
Published: April 2003

The role of apoptosis in glomerular cell depletion associated with a decrease in renal function is still controversial. To examine the involvement of apoptosis in renal disease, the occurrence of apoptosis during the progression of renal insufficiency as well as the expression of Bax protein and Fas antigen that are related to the apoptosis were investigated using five-sixths nephrectomized rats, one of the progressive renal disease models. Serum creatinine was significantly elevated to a level approximately five-fold higher than that in the sham-operated group on day 1 after the five-sixths nephrectomy and then maintained at a level approximately two- to three-fold higher until day 56 and then elevated further to a level eight-fold higher on day 96 after nephrectomy as compared with the sham-operated group. The total number of glomerular cells was significantly increased from day 7 to day 56 after nephrectomy and then returned to the level of the sham-operated group by day 96. The number of PCNA-positive cells (a marker of proliferating cells) in the glomeruli was significantly increased from day 7 to day 28 after nephrectomy; the highest level was observed on day 7, and the numbers then decreased gradually. Apoptotic cells, which were represented by TUNEL-positive cells, as well as apoptotic bodies were persistenly increased with time after nephrectomy in the glomeruli of nephrectomized rats; apoptotic cells could hardly be observed in the sham-operated group. Therefore, glomerular cell proliferation appeared to begin immediately after nephrectomy and to continue until day 28 at a level high enough to overcome the decrease in the number of glomerular cells due to apoptosis, since the total number of glomerular cells was apparently high until day 56. On day 96, the decrease in the number of glomerular cells probably becomes predominant over cell proliferation, since apoptosis continuously increased with time after nephrectomy. The events on day 96 may be associated with the severely decreased renal function which was represented by the explosive increase in the serum creatinine level on the same day. The number of Fas antigen positive glomerular cells was increased from day 1 after nephrectomy and reached a plateau on day 21. The number of Bax protein positive glomerular cells was generally increased with time after nephrectomy, but the number was slightly decreased on day 21. The theory that the expression of Bax protein is correlated with apoptosis appears to fit the case of progressive renal disease. These results suggest that apoptosis is involved in the cell depletion of progressive renal insufficiency.