Takayoshi Asai

Herpes simplex lymphadenitis. Report of two cases with review of the literature.

We report two cases of herpes simplex lymphadenitis without widespread organ involvement in a 60-year-old man and a 67-year-old woman. Their complaints were high fever and generalized erythema followed after few days by generalized lymphadenopathy. This report describes the findings obtained by light and electron microscopy, immunohistochemistry, and in situ hybridization.In both instances, Cowdrys type A intranuclear inclusion bodies were found in T-immunoblasts in the background of T-zone hyperplasia with focal necrosis. Electron microscopic investigation revealed intranuclear and cytoplasmic virus particles with characteristics of the herpes group. Immunohistochemical staining utilizing anti-herpes simplex virus (HSV) antibody was positive and in situ hybridization with HSV-DNA probe revealed positive signals in the nucleus and in the cytoplasm of T-immunoblasts. Although rare, HSV lymphadenitis in the absence of generalized infection can occur.

Modulation of Polymorphonuclear Leukocyte Apoptosis in the Critically Ill by Removal of Cytokines with Continuous Hemodiafiltration

Delay of polymorphonuclear leukocyte (PMN) apoptosis caused by hypercytokinemia is considered to be a potential cause of tissue damage and resultant organ failure. We evaluated whether continuous hemodiafiltration using a polymethylmethacrylate membrane hemofilter (PMMA-CHDF), which can remove cytokines in the circulating blood, can modulate apoptosis in peripheral blood neutrophils and thereby reduce tissue damage and organ dysfunction in 25 critically ill patients. Following the completion of a 3-day PMMA-CHDF session, serum cytokine levels were significantly decreased and the percentage of apoptotic PMNs was significantly increased. A significant correlation was observed between the PMMA-CHDF-induced increase in the percentage of apoptotic PMNs and the degree of decrease in the serum interleukin-6 level. A significant correlation was also found between the increase in the percentage of apoptotic PMNs and improvement in sequential organ failure assessment score following PMMA-CHDF. These results suggest that PMMA-CHDF in critically ill patients with hypercytokinemia and concomitant delay in apoptosis of PMNs can alleviate the delay of PMN apoptosis through the removal of serum cytokines and thus may result in avoidance of organ dysfunction.

Expression of multidrug resistant gene (mdr-1/P-glycoprotein) in a megakaryoblastic cell line, CMK, and its enhancement during megakaryocytic differentiation.

Multidrug resistance is a severe clinical problem in the chemotherapy of malignant disease. Acute megakaryoblastic leukemia (AMKL) is a rare form of childhood leukemia, and is often resistant to many anti-cancer chemotherapeutic drugs. Here we report the expression of the mdr-1/P-glycoprotein in a cell line, CMK, established from a patient with AMKL. Expression of mdr-1 mRNA in CMK11-5 cells, a well differentiated subline, was higher than in CMK6 cells, a poorly differentiated subline. The level of P-glycoprotein was also higher in CMK11-5 cells. The cytokines interferon-gamma (IFN-gamma), GM-CSF and IL-3, which were shown to induce megakaryocytic differentiation of CMK cells, enhanced the expression of the mdr-1 mRNA and levels of P-glycoprotein. These results imply that differentiated megakaryocytic cells may have higher levels of the P-glycoprotein expression, suggesting a possible normal physiological function of P-glycoprotein in mature megakaryocytes.

Signalling defect in FMLP‐induced neutrophil respiratory burst in myelodysplastic syndromes

Summary. Myelodysplastic syndromes (MDS) are clonal haematological disorders and MDS neutrophils have various abnormal functions which cause an increased risk of infective mortality. We examined luminol‐dependent chemiluminescence and cytoplasmic Ca2+ increase in order to characterize the mechanisms of a signalling defect in MDS neutrophil respiratory burst. In MDS patients, chemiluminescence stimulated with n‐formyl‐l‐methionyl‐l‐leucil‐ l‐phenylalanine (FMLP) and calcium ionophore A23187 was defective (17·2 ± 13·7 v 44·3 ± 16·6, P= 0·001; 42·2 ± 21·3 v 82·0 ± 23·6, P < 0·05, respectively), but phorbol 12‐myristate 13‐acetate (PMA) chemiluminescence was normal (73·4 ± 26·9 v 79·5 ± 23·8, P= 0·52). There were no statistical significances in cytoplasmic Ca2+ increase stimulated with FMLP and recombinant human interleukin‐8 (rhIL‐8) compared with controls (251·1 ± 104·3 v 272·7 ± 41·2, P= 0·295; 238·6 ± 65·0 v 253·9 ± 38·3, P= 0·567, respectively). Flow cytometric analysis of MDS neutrophils disclosed that most MDS patients showed normal neutrophil cytoplasmic Ca2+ response to FMLP and rhIL‐8. However, two patients with refractory anaemia with excess of blasts displayed a significant decrease of both chemiluminescence and cytoplasmic Ca2+ response to FMLP, and they also displayed low expression of FMLP receptor. These data suggest that most MDS patients have low FMLP chemiluminescence which is not due to a defect in the FMLP receptor. It is proposed that defective FMLP chemiluminescence in MDS results from a putative defect in protein kinase C‐ and Ca2+‐independent cell‐signalling mechanisms. Only a small group of patients have numerical or structural defects in the FMLP receptor, causing significant decrease of neutrophil respiratory burst.

Platelet calmodulin correlates with platelet turnover

Abstract: We measured the calmodulin content in platelets in 13 normal persons and in 62 patients with hematological diseases. The level of platelet calmodulin was higher in patients with idiopathic thrombocytopenic purpura (ITP), systemic lupus erythematosus, myeloproliferative disorders, acute leukemia in a recovery phase, aplastic anemia, thrombosis and hyper‐splenism as compared to the controls. Among the patients with ITP, calmodulin was lower in responders than in nonresponders and those at the initial diagnosis. We also measured the volume, life‐span and aggregation of the platelets and demonstrated a significant relationship between the calmodulin level and the platelet volume, and a negative relationship between the calmodulin level and platelet life‐span, there was no correlation between the calmodulin level and platelet aggregation. We thus conclude that platelet calmodulin is inversely correlated with platelet turnover.

Analysis for the Optimal Blood Draw Speed to Collect Sufficient Peripheral Blood Mononuclear Cells by COBE Spectra

Abstract:  In recent years the procedures for peripheral blood mononuclear cell (PBMNC) harvests have gradually been increasing. These PBMNCs are collected for several treatments, for example, donor lymphocyte infusion (DLI), immunotherapy for solid carcinoma, and regeneration therapy for ischemic limbs. In order to analyze the optimal procedure for collecting PBMNCs safely and efficiently, we evaluated 129 PBMNC apheresis procedures from April 1996 to May 2003, without hemopoietic stem cell mobilization by G‐CSF. In every case, PBMNC collections were performed with a COBE Spectra cell separator (Gambro BCT). The median apheresis volume was 5550 mL. The median of blood draw speed was 48.1 mL/min. The median TNC (total nuclear cell) number in products was 50.4 ×  103/µL. In the regression analysis, no significant correlation was seen between the blood draw speed and the concentrations of TNC in products (Y = aX + b, a = 0.842497, b = 17.11352, r = 0.222032, P = 0.012464). A positive correlation was seen between WBC on apheresis day and the concentrations of TNC (Y = aX + b, a = 0.009822, b = 3.224679, r = 0.550431, P = 2.93 × 10−11). A significantly higher correlation was seen between the MNC (mononuclear cells) on apheresis day and the concentrations of TNC (Y = aX + b, a = 0.028278, b = 13.09266, r = 0.696988, P = 9.486 × 10−9). This study has shown evidence that a higher increment of blood draw speed does not provide a higher concentration of products. An adequate apheresis speed is about 40 mL/min. If we want to obtain sufficient cell counts, it is very important to obtain sufficient volume with a moderate blood draw speed, therefore protecting against side‐effects.

VINBLASTINE-LOADED PLATELET TRANSFUSION IN AN ALLOIMMUNIZED PATIENT

Department of Haematology, JEAN-PIERRE N c Stobhill General Hospital, Clasgow G21 3 U W R. L. C. CUMMINC E. H. HORN R. B. HOGC Lefrere. J.J.. Courouche, A.M.. Girot. R. . Bertrand. Y. & Soulier. J.P. (1986) Six cases of hereditary spherocytosis revealed by human parvovirus infection. British journal ojHaernatolog!l. 62 ,65 3-658. Mortimer. P.P. (1983) Hypothesis: The aplastic crisis of hereditary spherocytosis is due to a single transmissible agent. journal oJ Clinical Pathology. 36, 4 4 5 4 4 8 . Mortimer. P.P.. Humphries. R.K.. Moore, J.G.. Purcell. R.H. & Young. N.S. ( 1 983) A human parvovirus-like virus inhibits haematopoietic colony formation in vitro. Nature. 302, 4 2 6 4 2 9 . REFERENCES Cossart. Y.E.. Field, A.M.. Cant, B. & Widdons, D. (1975) Parvoviruslike particles in human sera. Lancet. i, 72-73.

DECREASED SERUM 1α,25-DIHYDROXYVITAMIN D3 LEVELS IN LEUKAEMIA

istics of our case deserve additional comments. The myelodysplastic changes seen in apparently non-leukaemic bone marrow cells and the hypoplastic presentation coupled with pancytopenia suggest that AML might have followed a preleukaemic state. Cases of AML following myelodysplastic syndromes (MDS) have been recently recognized (Hehlmann et al, 1981; Eridani et d, 1985; Neame et al, 1985). This, like other evidences (Editorial, 1983), may indicate that some cases of MDS are the result of a defect of the pluripotential stem cell. A hypoplastic presentation has been reported in both AML (Howe et al, 1982) and common ALL (Breatnach et al, 198 1). Hypocellular acute and leukaemia and monosomy 7 are regarded as bad prognostic characteristics in AML (Borgstrom et al, 1980; Howe et ul, 1982). The excellent response to therapy in this patient suggests that intensive post-remission chemotherapy schemes designed for AML might be of value in these cases.

Board certification of medical doctors and medical technologists in transfusion medicine in Japan.

The Japan Society of Blood Transfusion (JSBT) organized a Board for certification of medical doctors in Transfusion Medicine (Board certified medical doctors by the JSBT) and a Board for certification of medical technologists for transfusion medicine (qualified medical technologists in Transfusion Medicine). For certified medical doctors the JSBT co-ordinated with the Japanese Association of Medical Science and with the Japan Medical Association, and for qualified medical technologists the JSBT co-organized with the Japanese Society of Laboratory Medicine, the Japanese Association of Medical Technologist and the College of Clinical Pathology of Japan. By May 2001, 259 of the certified medical doctors and 875 of the qualified medical technologists have been certified and are participating actively to increase the level of transfusion practice at individual facilities.

Magnetic resonance imaging of reticulo‐endothelial system in patients with idiopathic thrombocytopenic purpura

Idiopathic thrombocytopenic purpura (ITP) is characterized by accelerated platelet destruction in the reticulo‐endothelial system (RES). We performed magnetic resonance imaging (MRI) to estimate the degree of activated RES. MRI was performed with a Gyroscan S‐15 (1.5 tesla) in 7 healthy volunteers and 22 patients with ITP. The 22 patients included 19 who were at initial diagnosis or were nonresponders to the therapy (non‐DX group), and 3 who were responders. For the non‐DX group, the T1 relaxation time of the spleen was initially significantly shorter than for healthy volunteers, but normalized after responding to the therapy. The initially shorter T1 values of the spleen for ITP patients correlated with a low platelet count (P < 0.05). This condition may indicate foam cells or fatty components due to platelet destruction. There was no significant relationship between the sequestration in 111In‐scan and T1 values of the liver or spleen. However, MRI is a noninvasive method, and it may be a clinically useful tool in the evaluation of RES in patients with ITP. Am. J. Hematol. 56:52–58, 1997.