A new myeloblastic leukemia cell line with double minute chromosomes. Induction of methotrexate resistance and dihydrofolate reductase gene amplification.

To test the relationship between DMs and drug resistance in newly established AML cell lines, KY821, and its clone KY821A3, the latter had lost DMs during cloning, were cultured in increasing concentrations of MTX. KY821 became resistant against 2 x 10(-4) M MTX, whereas KY821A3 did against 2 x 10(-5) M MTX in a same period. Enhanced enzyme activities of DHFR were correspondent to the increased DMs numbers and DHFR gene amplification in both resistant clones. The amplified DHFR gene was located on DMs by in situ hybridization. These data indicated that the presence of DMs in KY821 would facilitate the acquisition of drug resistance.     

Hematopoietic recovery in a patient with acute lymphoblastic leukemia after an autologous marrow graft purged by combined hyperthermia and interferon in vitro.

We describe a patient with acute lymphoblastic leukemia (ALL) in whom hemopoiesis recovered after an autologous marrow graft purged by in vitro hyperthermia. A 17-year-old woman was diagnosed as having ALL in April 1985. After clinical remission was induced, marrow cells were harvested. The marrow cells were treated with hyperthermia at 42.0 degrees C for 1 h in the presence of alpha-interferon to eliminate residual leukemic cells, and then cryopreserved. In January 1990, during her fourth remission she was treated with busulfan and cyclophosphamide, and then received the thawed autologous marrow. Her hematopoietic recovery was prompt with normal trilineage regeneration without any life-threatening complications. She is in good health without evidence of a leukemic relapse at 6 months after autologous bone marrow transplantation. This case suggests that human multilineage progenitor cells retain self-renewal capacity in vivo even after treatment with heat and alpha-interferon in vitro followed by the freezing and thawing procedures.     

Brain MR imaging in patients with hepatic cirrhosis: relationship between high intensity signal in basal ganglia on T1-weighted images and elemental concentrations in brain

In patients with hepatic cirrhosis, the globus pallidus and putamen show high intensity on T1-weighted MRI. While the causes of this high signal have been thought to include paramagnetic substances, especially manganese, no evidence for this has been presented. Autopsy in four cases of hepatic cirrhosis permitted measurement of metal concentrations in brain and histopathological examination. In three cases the globus pallidus showed high intensity on T1-weighted images. Mean manganese concentrations in globus pallidus, putamen and frontal white matter were 3.03 ± 0.38, 2.12 ± 0.37, and 1.38 ± 0.24 (μg/g wet weight), respectively, being approximately four- to almost ten-fold the normal values. Copper concentrations in globus pallidus and putamen were also high, 50 % more than normal. Calcium, iron, zinc and magnesium concentrations were all normal. The fourth case showed no abnormal intensity in the basal ganglia and brain metal concentrations were all normal. Histopathologically, cases with showing high signal remarkable atrophy, necrosis, and deciduation of nerve cells and proliferation of glial cells and microglia in globus pallidus. These findings were similar to those in chronic manganese poisoning. On T1-weighted images, copper deposition shows no abnormal intensity. It is therefore inferred that deposition of highly concentrations of manganese may caused high signal on T1-weighted images and nerve cell death in the globus pallidus. Received: 12 August 1996 Accepted: 17 December 1996     

Analysis of angiographical findings in patients with severe brain disorders progressing to brain death

In the present study, the authors analysed the serial angiographical findings progressing to brain death and their relation to the intracranial pressure (ICP) and the cerebral perfusion pressure (CPP). Seventy two patients, from four to eighty four years old (fourty six males and twenty six females) admitted in the Department of Emergency Medicine, University of Tokyo Hospital during the period from January, 1981 to April, 1986, were studied. Their underlying diseases were supratentorial primary brain lesions except two cases with asphyxias which progressed to brain death. ICP was continuously measured and CPP was calculated as the pressure gradient between the mean arterial blood pressure (MAP) and ICP. The direct carotid angiography was performed to follow the cerebral circulation. Fourty five patients were subjected to barbiturate (pentobarbital sodium) therapy. The degree of the intracranial filling staged as "Non-filling", "Siphon-filling", "Partial-filling", "Delayed-filling", "All-filling" correlated significantly with ICP and CPP. These relationships, however, disappeared once ICP exceeded MAP. When "Non-filling" angiogram was obtained, clinical signs had already showed brain death. On the other hand, minimal flow ("Siphon-filling", "Partial-filling", "Delayed-filling") were still demonstrated in six brain death cases while ICP was approaching its "peak" value. This study showed that clinical diagnosis of brain death preceded the Non-filling phenomenon, suggesting that, for the demonstration of the cerebral circulatory arrest, the angiograms should be performed after the clinical diagnosis of brain death was established and CPP became zero. The evaluation of the extremely slow and minimal filling is still a matter of discussion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of infrasound on the Rota-Rod Treadmill performance of rats

Summary The present study was designed to evaluate the effects of infrasound on behavioral performance in rats. The rats were divided into two groups, one selected for good performance (six rats: superior group) and the other for poor performance (six rats: inferior group) on the Rota-Rod Treadmill.Exposure conditions were as follows:Exp. 1 Control (150 min), Exp. 2 Exposure to infrasound with a main frequency of 16 Hz and a sound pressure level of 105 dB (S.P.L.) (70 min), Exp. 3 Exposure to infrasound with a main frequency of 16 Hz and a sound pressure level of 95 dB (S.P.L.) (70 min), Exp. 4 Exposure to infrasound with a main frequency of 16 Hz and a sound pressure level of 85 dB (S.P.L.) (150 min), Exp. 5 Exposure to infrasound with a main frequency of 16 Hz and a sound pressure level of 75 dB (S.P.L.) (150 min), Exp. 6 Exposure to Pink Noise of 72 dB (A) (70 min).Comparison of the pre-exposure endurance time (Maximum: 2 min) on the Rota-Rod Treadmill with endurance after exposure to infrasound showed that the endurance time of the superior group after exposure to 16 Hz, 105 dB was not reduced. The endurance of the inferior group was reduced by exposure to 16 Hz, 105 dB after 10 min, to 16 Hz, 95 dB after 70 min, and to 16 Hz, 85 dB after 150 min.     

Urine levels of CD46 (membrane cofactor protein) are increased in patients with glomerular diseases

Soluble membrane cofactor protein (MCP, CD46) has not been detected by conventional ELISA in human urine. Here, we established a highly sensitive assay method for determination of urinary MCP (uMCP) using monoclonal antibody-coated paramagnetic beads. This method enabled us to detect less than 0.05 ng/ml of purified membrane and recombinant soluble MCP, a sensitivity 10-fold higher than that of conventional ELISA. In normal subjects, the levels of uMCP were <0. 05 ng/ml. The levels of uMCP were elevated in patients with IgA nephropathy and more prominently in patients with rapidly progressive glomerulonephritis. The levels of uMCP were correlated significantly with those of serum MCP (sMCP) and N-acetyl-beta-glucosaminidase and nonsignificantly with those of beta(2)-microglobulin, total urine protein, or serum creatinine. The properties of uMCP were inconsistent with those of the reported sMCP, since uMCP showed three bands on SDS-PAGE/immunoblotting with molecular mass profiles different from those of sMCP. uMCP exhibited factor I cofactor activity for cleavage of C3b comparable to that of sMCP. The origin of uMCP, however, remains to be determined. These results, taken together with the parameter correlation profiles, suggested that uMCP is secreted or produced secondary to tubular or glomerular damage. The physiological role and clinical significance of uMCP are now within the scope of our investigation by establishment of this assay.     

Reduced-intensity unrelated cord blood transplantation for patients with advanced malignant lymphoma.

We report the results of reduced-intensity unrelated cord blood transplantation (RI-UCBT) in patients with advanced malignant lymphoma. Twenty patients (median age, 46.5 years; range, 27-66 years) underwent RI-UCBT with a preparative regimen consisting of fludarabine 125 mg/m2 , melphalan 80 mg/m 2 , and 4 Gy of total body irradiation. The median infused total cell dose was 2.75 x 10(7)/kg (range, 2.3-3.4 x 10(7)/kg). Graft-versus-host disease (GVHD) prophylaxis was composed of cyclosporine or tacrolimus alone. Fifteen patients achieved primary neutrophil engraftment after a median of 20 days. Eight patients developed grade II to IV acute GVHD, and 2 developed chronic GVHD. Of the 16 patients with evaluable disease, 10 achieved a complete response. Primary disease recurred in 1 patient, and transplant-related mortality within 100 days occurred in 8 of 20 patients. The estimated 1-year probability of progression-free survival was 50%. These data suggest that RI-UCBT is a feasible option for patients with refractory lymphoma who lack an HLA-matched donor.     

Contractions in normal and atherosclerotic rabbit aortas

Length-tension relationships and contractile responses to agonists associated with cyclic nucleotide levels in aortas from atherosclerotic rabbits were studied. The same number of tissues from healthy rabbits was also studied. Diffuse lesions were produced by denudation of the vascular endothelium of rabbit aorta using a balloon catheter and then feeding these rabbits a high cholesterol diet for two months. Stretch-passive tension curves and length-active tension curves which developed in the presence of 60 mM KCl significantly shifted to the left. The altered length-tension relationships indicate a decreased distensibility in these strips and this is attributed to an enhanced accumulation of collagen. Dose-response curve analysis revealed a marked hypersensitivity to serotonin as indicated by greater pD2 values (negative logarithm of half maximum dose) than seen in the controls. In addition, the mean absolute values of the maximum contraction induced by serotonin and KCl were significantly higher than in the controls. Although cyclic AMP levels in the strips with a pathology were lower and cyclic GMP/cyclic AMP ratios were higher than the controls in the basal state, serotonin induced no marked alterations in the cyclic nucleotides levels in the strips from either the diseased or control rabbits. These mechanical and pharmacological alterations of atherosclerotic vessels may relate to clinical events such as coronary spasm.     

Infective endocarditis caused by Granulicatella elegans originating in the oral cavity

We studied the pheno- and genotypes of an oral Granulicatella elegans strain in comparison with those of a blood-derived isolate which caused infective endocarditis. The two isolates exhibited identical biochemical characteristics and had the same drug MICs. Their genotypes were indistinguishable, indicating that these were from the same clone. The transmission of G. elegans from the oral cavity thus should be noted as a possible cause of infective endocarditis.