Defining the load bearing axis of the lower extremity obtained from anterior-posterior digital radiographs of the whole limb in stance.

OBJECTIVE: The purpose of this study was to evaluate the reliability of inter-and intra-observer assessments of the mechanical alignment of the lower extremities with digitally computed radiographs of an anterior-posterior view of the whole leg using a new computer-assisted method. METHOD: Load bearing axis deviation of the lower extremities was quantitatively measured by three examiners in 105 knees of 73 subjects who had osteoarthritis of the knee with a Kellgren-Lawrence grade of 1 or more. A line representing the load bearing axis was drawn from the center of the femoral head to the center of the ankle and the alignment of the leg was assessed by measuring the width of the proximal tibia and the perpendicular distance from the middle of the proximal tibial condyle to the load bearing axis (Fujifilm OP-A). A ratio of the values was calculated and expressed as a percentage. RESULTS: The inter-observer mean difference was 2.9 % (SD, 2.7), and the intra-observer mean difference was 2.1% (SD, 2.2). The mean intraclass correlation coefficient (ICC) for inter-observer trials was 0.96; that for intra-observer trials was 0.99. CONCLUSION: Our computer-assisted method was reproducible, and should be considered an alternative method for the measurement of the alignment of the whole leg.

Rapid microglial activation induced by traumatic brain injury is independent of blood brain barrier disruption.

Following CNS injury, microglia respond and transform into reactive species exhibiting characteristic morphological changes that have been termed "activated" or "ameboid" microglia. In an attempt to establish that microglial reactions induced immediately after injury are caused by intrinsic mechanisms rather than infiltration of blood and its constituents, oxygenized Ringer's solution was perfused into the cerebral circulation of rats so that the circulating blood could be eliminated prior to injury induction. Under artificial respiration, a catheter was inserted from the cardiac apex into the ascending aorta, and oxygenized Ringer's solution was immediately perfused with a pulsatile blood pump, resulting in wash out of the circulating blood from the brain within 1 min. Subsequently, a cortical contusion was induced in the unilateral parietal cortex using a controlled cortical impact (CCI) device. At 5 min following the injury, the brain was fixed by perfusion of fixative through the catheter and removed. Coronal vibratome sections were then processed for CR3 immunohistochemistry to examine the microglial activation. It appeared that microglial activation with both morphological transformation and an increase in CR3 immunoreactivity was induced throughout the hemisphere ipsilateral to the injury side exclusively, even in rats with elimination of circulating blood. The microglial reactions did not differ substantially from those observed in the control rats with extensive BBB disruption. The present results thus provide direct evidence that the microglial activation induced immediately after injury is independent of infiltration of circulating blood induced by concurrent BBB disruption.

Role of the membrane form of human colony-stimulating factor-1 (CSF-1) in proliferation of multipotent hematopoietic FDCP-mix cells expressing human CSF-1 receptor.

Because IL-3-dependent multipotential FDCP-Mix cells expressing human colony-stimulating factor-1 (CSF-1) receptor did not proliferate in response to soluble CSF-1, we investigated whether their proliferation would be induced in co-culture with adherent cells expressing the membrane form of CSF-1 (MemCSF-1). FDCP-Mix cells with high CSF-1R expression (NAF21 cells) were placed on stromal MS-5 cells or STO fibroblasts expressing MemCSF-1 (2M-1 cells and STO-M2 cells, respectively), in absence of IL-3. NAF21 cells bound significantly to 2M-1 cells as compared to control FDCP-Mix cells. Adhesion of NAF21 cells was inhibited by anti-huCSF-1 antibodies, as well as anti-huCSF-1R antibodies. Interestingly, NAF21 cells proliferated on both 2M-1 and STO-M2 cells but with very different kinetics. Moreover, NAF21 cell proliferation was also supported by glutaraldehyde-fixed 2M-1 cells or highly concentrated MS-5 cell culture supernatant, but not by CSF-1 coated on culture dishes. These results strongly suggest that MemCSF-1/CSF-1R interaction mediates a specific adhesion of NAF21 cells to stromal cells and allows stimulation of hematopoietic cells by stromal cell-derived factors expressed in a membrane-bound form or concentrated within the extracellular matrix. Thus, cytokine receptors deficient in mitogenic signalling may nevertheless have a regulatory role in hematopoietic progenitor cell proliferation by acting as adhesion molecules.

[Plasma cell leukemia presenting with circulating villous lymphocytes and an indolent clinical course].

A 73-year-old man was admitted to our hospital in July 1996 because of lymphoctyosis and lumbago. Physical examination revealed hepatomegaly and anemia. Hematologic examination showed a hemoglobin concentration of 9.6 g/dl and a leukocyte count of 32,700/microliter with 74% abnormal mononuclear cells. In Wright-Giemsa stained blood films, these cells had short villi arising from 1 or 2 poles. Immunophenotyping of peripheral mononuclear cells showed moderate to strong expression of CD10, CD24, CD38, and sIg lambda, but not of CD19, CD20, or CD25. Southern blot analysis of the peripheral mononuclear cells demonstrated rearranged monoclonal bands in the C lambda. Urine immunoelectrophoresis detected a monoclonal band identifiable as lambda-type Bence Jones protein. In addition, bone X-ray studies disclosed multiple osteolytic lesions. A diagnosis of plasma cell leukemia was made, and the patient was placed on chemotherapy consisting of cyclophosphamide and prednisolone. No notable improvement in laboratory findings was seen but the patient experienced an indolent clinical course. He died of pneumonia in January 1998. The morphological and clinical findings were unusual for a case of plasma cell leukemia. This case study suggested that signs of lymphocytosis require immunophenotypic and electron microscopic studies for the differential diagnosis of plasma cell leukemia.

Age-related changes in myelopoietic response to lipopolysaccharide in senescence-accelerated (SAM) mice.

The effects of in vivo lipopolysaccharide (LPS) administration on myelopoiesis were examined in senescence-accelerated (SAM) mice. Young mice injected with LPS exhibited: (a) increased femoral proliferative pool size; (b) transient reduction in femoral non-proliferative pool size and number of femoral colony forming unit-granulocyte macrophages (CFU-GMs); (c) marked increase in splenic CFU-GMs; and (d) transient increase in S-phase of femoral CFU-GMs. The responses of old mice after LPS administration differed from those of young mice in the following points: (a) no recovery of the femoral non-proliferative pool or femoral CFU-GMs, (b) less significant augmentation of the femoral proliferative pool and splenic CFU-GMs, and (c) prolonged reduction in S-phase of femoral CFU-GM. Injection of LPS into mice resulted in a hyperproduction of colony-stimulating activity (CSA) in bone followed by production of colony-inhibitory activity (CIA) in young mice and in contrast, an excessive CIA secretion from bone without an increase in CSA levels in old mice. These imbalances in the regulatory factors derived from non-hemopoietic cells in the bones may lead to an inappropriate response of myelopoiesis in aged SAM mice after LPS administration, which may play a key role in infections.

Therapy-related megakaryoblastic leukemia with pituitary involvement following treatment for non-Hodgkin's lymphoma.

A case of a 66-year-old Japanese man developed therapy-related megakaryoblastic leukemia with pituitary involvement after chemotherapy for non-Hodgkin's lymphoma. Alkylating agents had been administered for the treatment of non-Hodgkin's lymphoma and 6 years later, megakaryoblastic leukemia with myelofibrosis and myelodysplasia developed. The blast cells expressed CD41, and immature antigens also. These findings were compatible with therapy-related megakaryoblastic leukemia. An autopsy revealed blast-cell infiltration into multiple organs including the posterior pituitary lobe. Therapy-related megakaryoblastic leukemia is very rare, and pituitary involvement may be associated with immaturity of blast cells.

Comparison of erythropoietic response to androgen in young and old senescence accelerated mice.

In this study, to clarify whether the functional capacity of hemopoietic progenitor cells and the micro-environment of aged mice are identical with those of the young, we investigated the changes in the number of hemopoietic progenitor cells and the production of regulatory cytokines from splenic cells as well as changes in the serum levels of cytokine in senescence-accelerated mice (SAM) after administration of 19-nandrolone decanoate (19-ND), a synthetic androgenic anabolic steroid. 19-ND induced an increase in erythroid colony-forming units (CFU-E), erythroid burst-forming units (BFU-E), and granulocytic-macrophage committed progenitor cells (CFU-GM) in bone marrow and spleen; especially remarkable increases were observed in the splenic CFU-E in both young and old mice. Antigen expression analysis of hemopoietic organs revealed that total TER-119+ cells per spleen of young and old mice with androgen treatment rose 2.6- and 3.2-fold over their respective control values. The responsiveness of hemopoietic progenitor cells to androgen did not change with age. Injection of 19-ND into young and old mice markedly enhanced the erythropoietin levels but not IL3 and GM-CSF levels in the serum of both groups. Cytokine production assessed by pokeweed mitogen-stimulated spleen condition medium showed an age-related decline. Androgen treatment could not influence IL-3 and GM-CSF production of spleen. These findings suggest that the spleen of both old and young mice served as the major site of regenerative repopulation of hemopoietic progenitors, especially the late erythroid progenitors in 19-ND-treated mice. The proliferative reserve of erythropoiesis with androgen treatment in aged mice was not reduced more than that in treated-young mice.

[A case of chronic myelogenous leukemia presenting multiple extramedullary tumors localized in cranial dura].

A 64-year-old woman had been given a diagnosis of Ph-positive chronic myelogenous leukemia (Ph+ CML) in October 1992 and accordingly treated with interferon-alpha busulfan, and hydroxyurea. She was admitted to our hospital with a one-day history of consciousness disturbance on May 30, 1993. Two weeks before admission, she had received chemotherapy consisting of vincristine and predonisolone because of progressive thrombocytopenia, basophilia, and leukocytosis accompanied by a heightened degree of cell immaturity in peripheral blood and bone marrow. Cranial computerized tomography on admission disclosed tumoral masses in the left frontal lobe and the right temporal lobe. Moreover, lumbar puncture ezinkns disclosed blastoid cells in cerebrospinal fluid. Based on these laboratory findings, the diagnosis was blastic crisis CML, 46XX t(9; 22; 17) (q34; q11; q23), cytogenetic aberration and extramedulary brain disease Although the patient underwent the same combined chemotherapy again, her unconsciousness did not resolve. She died of cerebellar herniation on the 7th hospital day. Post mortem examination revealed three extramedullary tumors localized in cranial dura. This was a rare case of CML presenting multiple extramedullary tumors localized in cranial dura.

A novel growth-factor-dependent myeloid cell line derived from mouse bone marrow cells contains progenitors endowed with high proliferative potential.

Constitutive expression of human colony-stimulating factor-1 receptor (CSF-1R) confers long-lasting CSF-1-dependent proliferation to mouse myeloid cell lines. We developed mice transgenic for human CSF-1R because mouse CSF-1 cannot activate human CSF-1R. Then bone marrow cells from transgenic mice were plated onto MS-5 stromal cells expressing the membrane form of human CSF-1 (2M-1 cells) in order to combine the hematopoietic supporting properties of stromal cells and the proliferative effects of CSF-1. Thus, we were able to derive a hematopoietic cell line, called 47.10, that grew indefinitely under these conditions, whereas no cell line could be developed from nontransgenic mice. Proliferation of 47.10 cells is severely affected by neutralizing anti-CSF-1R monoclonal antibodies. Morphologic and cytofluorometry analysis established that most 47.10 cells are immature myelomonocytic cells. Consistent with this phenotype, the myeloid transcription factor PU.1, but not the erythroid transcription factor GATA-1, is expressed in 47.10 cells. A few 47.10 cells (3-5%) do not express lineage specific markers; they differentiate spontaneously to lineage-positive cells after replating on 2M-1 cells. In agar cultures, 47.10 cells form 7- and 14-day colonies in response to a cocktail of granulocyte/macrophage colony-stimulating factor (2.5 ng/mL), interleukin-3 (1 ng/mL), and mouse CSF-1 (10 ng/mL). Under these conditions, about 0.5% of 47.10 cells formed large 14-day colonies (>1 mm) composed of mature monocytes and granulocytes, reflecting the presence of progenitors endowed with high proliferative potential (HPP-47.10 cells). In conclusion, we have characterized a novel continuous myeloid cell line presenting a hierarchical structure similar to that of the bone marrow progenitor cell compartment.

The effects of nonsteroidal anti-inflammatory drugs on immune functions of human peripheral blood mononuclear cells.

It is generally accepted that the nonsteroidal anti-inflammatory drugs (NSAIDs) exhibit anti-inflammatory effects primarily through inhibition of prostaglandin (PG) synthesis. However, effects of NSAIDs on immune responses are not fully understood. This study investigated effects of indomethacin and a new NSAID (d-2-[4-(3-methyl-2-thienyl)phenyl]propionic acid, termed as M-5011 in this study) on cytokine production, lymphocyte proliferation, activities of natural killer (NK) and lymphokine activated killer (LAK) cells and secretion of immunoglobulin (Ig). Both indomethacin and M-5011 augmented interleukin (IL)-2 production, whereas they suppressed IL-6 production both at the protein and mRNA levels. These two NSAIDs augmented proliferation of phytohemagglutinin (PHA)-stimulated PBMC and enhanced NK and LAK cell activities. In contrast, indomethacin was more potent than M-5011 in inhibition of both PG synthesis and Ig secretions by pokeweed mitogen (PWM)-stimulated PBMC. These results suggest that these two NSAIDs equally augment cell-mediated immunity, whereas indomethacin was more potent than M-5011 in the inhibition of humoral immunity and PG synthesis.

[Severe hemolytic anemia with tear drop red cells as initial manifestation of Wilson's disease].

A 16-year-old girl was admitted for a detailed examination of hemolytic anemia in November 1995. Initial laboratory findings included a total bilirubin concentration of 1.46 mg/dl, hemoglobin of 9.1 g/dl, and a reticulocyte count of 89/1000 percent. The plasma haptoglobin concentration was below 10 mg/dl. A blood smear showed many dacryocytes and a few echinocytes and codocytes. GOT was 71 IU/l; GPT, 44 IU/l; and LDH, 812 IU/l; the results of a hepaplastin test were 45% of normal. On further investigation, the level of serum ceruloplasmin was found to be 4 mg/dl, and of serum copper, 43 micrograms/dl. Urinary copper excretion was markedly increased, at 345 micrograms per day. Slit-lamp examination of both corneas revealed obvious Kayser-Fleischer rings. A liver biopsy sample showed fibrosis histologically and an elevated copper concentration of 535 micrograms/g dry weight and 183 micrograms/g wet weight. In family studies, the patient's asymptomatic 5-year-old sister was observed to have metabolic abnormalities consistent with Wilson's disease. These findings suggested that the patient's hemolytic anemia with red cell deformities was due to abnormal copper metabolism associated with Wilson's disease.

[Plasma cell leukemia (IgG kappa) presenting bilateral neurosensory hearing loss and left sixth cranial nerve plasy].

A 30-year-old man who had been given a diagnosis of IgG-kappa multiple myeloma by another hospital and treated with melphalan, prednisone, and cyclophosphamide 6 months earlier, was admitted to our hospitaly in July 1994 because of progressively impaired hearing in both ears, vertigo, and worsening fatigue. Peripheral blood examination showed a white blood cell count 25,000/microliter, with 77.5% atypical plasma cells. Examination at the time of hospitalization also revealed retinal hemorrhages and serum hyperviscosity. The diagnosis was plasma cell leukemia with hyperviscosity syndrome. Subsequent treatment consisted of vincristine, doxorubicine, and prednisone and repeated plasmapheresis. This resulted in a partial response and a reduction of serum viscosity but no reversal of hearing loss. One month after admission, left sixth cranial nerve plasy was demonstrated. Cranial computed tomography studies disclosed a tumoral mass in the sphenoid sinus. The patient received local radiotherapy and intensive chemotherapy, but exhibited no notable alleviation of his cranial nerve palsy. He died of septicemia and progressive disease in August 1994. This case was rare in that it involved plasma cell leukemia and bilateral neurosensory hearing loss associated with serum hyperviscosity and sixth cranial nerve plasy due to plasmacytoma within the sphenoid sinus.

Antibiotic susceptibility of enterohemorrhagic Escherichia coli O157:H7 isolated from an outbreak in Japan in 1996.

The antibiotic susceptibilities of 43 strains of Escherichia coli O157:H7 identified in the summer of 1996 in Japan were investigated. Growth of 90% of O157 strains was inhibited at a concentration of < or = 0.5 micro/ml by several agents including fosfomycin with glucose-6-phosphate.

Appearance of fosfomycin resistant Rahnella aquatilis clinically isolated in Japan.

Among recent clinical isolates in Japan, strain CU264 was discovered which formed unusual colonies. This strain was identified as Rahnella aquatilis which is usually found in water. The antibiotic susceptibilities against tetracycline, carbenicillin, chloramphenicol, streptomycin, kanamycin, gentamicin, sulphonamide, neomycin, fosfomycin, rifampicin, norfloxacin and nalidixic acid, were investigated. The result demonstrated that the strain was highly resistant to fosfomycin only. It was further shown that this resistance was transmissible with low frequency to Serratia marcescens whereas it was not transmissible to Escherichia coli.

In vitro effects of immunosuppressive agents on cytokine production by HTLV-I-infected T cell clones derived from the ocular fluid of patients with HTLV-I uveitis.

The present study was designed to investigate the in vitro effects of potential therapeutic agents on cytokine production by five HTVL-I-infected T cell clones (TCC) established from the ocular fluid of patients with HTLV-I uveitis. Each of the five HTLV-I-infected TCC was cultured at 1 x 10(6) cells/ml with or without an immunosuppressive agent (hydrocortisone, FK506, rapamycin, indomethacin, or prostaglandin E2) for 22 hr in humidified 5% CO2 in air at 37 C. The production of various cytokines in the culture supernatant from each TCC was measured by ELISA. The HTLV-I-infected TCC produced high amounts of IL-1 alpha, IL-3, IL-6, IL-8, TNF-alpha, IFN-gamma, and GM-CSF, and low but significant levels of IL-2 and IL-10 without any stimuli. Hydrocortisone severely depressed the production by these TCC of all the cytokines except for IL-2, which was slightly increased. Prostaglandin E2 depressed the production of IL-1 alpha, while it up-regulated the production of IL-6, TNF-alpha, and IFN-gamma. Rapamycin depressed the production of IL-6 and TNF-alpha, and FK506 depressed the production of TNF-alpha. Hydrocortisone also severely depressed the cytokine production by PHA-stimulated peripheral blood mononuclear cells obtained from healthy volunteers. Of the immunosuppressive agents tested, hydrocortisone exhibited the strongest suppression of cytokine production by HTLV-I-infected TCC. This result was in agreement with the in vivo effects of hydrocortisone in patients with HTLV-I uveitis. These TCC will be useful in investigating the effects of potential therapeutic agents for HTLV-I uveitis in vitro.

Release of interleukin-1 beta by dermatan sulfate suppresses hepatocyte growth.

Among glycosaminoglycans, dermatan sulfate (DS) is the strongest inhibitor of DNA synthesis in adult rat hepatocytes in primary culture stimulated with insulin and epidermal growth factor. Hyaluronate also inhibited DNA synthesis, whereas chondroitin-6 sulfate, 4-sulfate or heparin had no effect on DNA synthesis in hepatocytes. Analysis of growth regulatory factors in hepatocyte culture medium treated with DS revealed that interleukin-1 (IL-1) was released into the medium. IL-1 beta mRNA was detected in DS-treated hepatocytes by reverse transcriptase-polymerase chain reaction, but not in untreated hepatocytes. For a marked inhibition of DNA synthesis, more than 10 hr of exposure to DS before cultured hepatocytes started DNA synthesis, was required. Similarly, more than 10 hr was required after the addition of DS before IL-1 beta mRNA was detected. These findings suggest that DS in the medium induced the production of IL-1 beta which, in turn, reduced DNA synthesis in hepatocytes.

[Daily oral low-dose etoposide therapy for aged patients with relapsed aggressive lymphoma].

Two patients with relapsed Non-Hodgkin's lymphoma (NHL) were treated with oral administration of etoposide. In these patients, long-term hematological remission was obtained. One patient was a 76-year-old man, who was successfully treated with CHOP for diffuse large cell NHL stage III B. One year after obtaining CR, he was admitted to our hospital for enlargement of lymph node. Rebiopsy of lymph node made a diagnosis of relapse from NHL. A new regimen of oral administration of etoposide treatment was employed. Hematological remission was obtained and continued for 3 years. Without interfering with his quality of life. The other patient was a 74-year-old man, who was treated with 6 cycles of CHOP for diffuse large cell NHL stage IV B. The patient attained complete remission following an additional 2 cycles of COMLA therapy. Eight years later, he was admitted for enlargement of lymph node. Rebiopsy of lymph node provided the basis for a diagnosis of relapse from NHL. Oral administration of etoposide treatment was started. Hematological remission was obtained and has been continued until now. These results show that oral administration of etoposide treatment is effective for some patients with recurrent NHL.