Purification of peripheral blood hematopoietic cells using Campath-1M monoclonal antibody.

The purpose of this study was to develop a method for exhaustive depletion of T cells in preparation for allogeneic peripheral blood (PB) hematopoietic stem cell (HSC) transplantation. Our goal was to achieve 4 logs of T-cell reduction while minimizing the loss of colony-forming progenitor cells. Campath-1M, a rat IgM cytolytic to T cells, B cells, and monocytes in the presence of human complement, was chosen for the investigation. To determine the best target population for Campath treatment, three different PB mononuclear cell (MNC) populations were examined: 1) total MNC isolated with Ficoll-sodium diatrizoate; 2) MNC after T-cell depletion by sheep red blood cell rosetting and high capacity Percoll density gradient fractionation (E-MNC); and 3) B/Null cells, a population of E-MNC obtained by lysing monocytes with L-phenylalanine methyl ester (PME). Of the three target MNC populations, Campath-treated B/Null cells had the highest cloning efficiencies (CE) for granulocyte-macrophage colony-forming units (CFU-GM) and erythroid burst-forming units (BFUe) (56.7 and 38.5 colonies per 5 x 10(3) cells, respectively) and the highest enrichments for CFU-GM (108.2-fold, range 18- to 440-fold) and BFUe (178.5-fold, range 78- to 314-fold). Recoveries of progenitor cells were comparable for Campath-treated MNC (49.7% and 84.5%, respectively, for CFU-GM and BFUe) and Campath-treated B/Null (44.8% and 76.1%), and significantly higher than those from Campath-treated E-MNC (19.5% and 34.3%). The T-cell content in the Campath-treated B/Null cells were reduced by 5.0 logs from the starting MNC, as determined by limiting dilution analysis (LDA). We conclude that Campath-1M can be used for T-cell depletion of PB HSC. The best results were obtained when T cells and monocytes were removed prior to Campath treatment. This cell population is highly enriched for hematopoietic cells and may prove useful for in vitro studies as well allogeneic transplantation.

Diagnostic and prognostic values of plasma levels of fibrinolytic markers in gastric cancer.

In the present study, we determined the plasma and tissue concentrations of tissue-type plasminogen activator, urokinase-type plasminogen activator, plasminogen activator inhibitor-1, plasminogen activator inhibitor-2 and urokinase-type plasminogen activator receptor in 32 patients with pathology-proved gastric cancer. The plasma levels of the same markers were compared in 37 patients with benign gastric ulcer in order to find out if these plasma levels could be used to evaluate the prognostic value in patients with gastric cancer. Plasma plasminogen activator inhibitor-1 was significantly higher in gastric cancer than in benign gastric disease (p < 0.0005), whereas plasma urokinase-type plasminogen activator was significantly lower in patients with gastric cancer than in those with benign ulcer (p = 0.003). There was no significant correlation between tissue and plasma concentrations of the same parameters. The plasma and tissue levels of fibrinolytic parameters were not affected by tumor size or distant metastasis, whereas tumor tissue concentration of urokinase-type plasminogen activator receptor and plasminogen activator inhibitor-2 were significantly higher in N0 than in N1 and N2, and tissue plasminogen activator inhibitor-1 was significantly higher in N0 than in N1. Plasma levels of the five fibrinolytic parameters could not take the place of the corresponding tissue concentrations on the diagnosis and prediction of prognosis in patients with gastric cancer. Tissue concentrations of urokinase-type plasminogen activator receptor and plasminogen activator inhibitor-2, especially the latter, can be used to predict lymph node involvement in patients with gastric cancer.

Improved sonographic visualization by fluid challenge method of renal lithiasis in the nondilated collecting system. Experience in seven cases.

Ultrasound is an effective means of detecting renal stones in patients with hydronephrosis. The filled urinary bladder in a normal person under hydration frequently results in the sonographic appearance of minimal or moderate hydronephrosis. For optimal visualization of stones and associated acoustic shadows, six patients with radiographically opaque renal stones were evaluated with fluid-loaded renal ultrasonography, which was compared to conventional renal sonogram in basal state. Although ultrasound in basal state can be used for detection of intrarenal lithiasis, fluid-challenged renal sonogram appears more accurate for the determination of contour, location, size, and number of small stones. In one case, this fluid-loaded ultrasound well demonstrated the tramline appearance of an isolated renal artery, which mimics stone before fluid challenge.

Intensive postremission chemotherapy in Taiwanese adults with acute myelogenous leukemia.

Intensive postremission chemotherapy has produced disease-free survival comparable to that of bone marrow transplantation in patients with acute myelogenous leukemia (AML), but its efficacy was unknown in Taiwan. We assessed the efficacy of intensive postremission chemotherapy, consisting of high-dose arabinoside-C (HiDAC) with or without transplantation of peripheral blood stem cells, in 33 AML patients from a single institute in Taiwan. Toxic reactions, treatment outcome, prognostic factors, and the size of the peripheral blood stem-cell harvest after HiDAC were analyzed. After a median follow-up of 21 months, 18 patients remained in continuous complete remission. The actuarial leukemia-free survival at 4 years was 51%. Relapse occurred in 12 patients, at a median of 12 months after initial diagnosis. All 6 patients with acute promyelocytic leukemia remained disease free after HiDAC therapy. Age, sex, and number of remission-induction or intensive consolidation chemotherapy courses had no effect on the risk of relapse. Intensive postremission chemotherapy can effectively prolong the duration of remission in young (< 60 years of age) adults with AML.

Definitive diagnosis of choledochal cyst by 99mTc-pyridoxylideneglutamate sequential scintiphotography.

Sequential supine scintiphotography of the abdomen was performed after intravenous administration of 99mTc-pyridoxylideneglutamate (99mTc-PG) to verify the presence of choledochal cyst in an 18-month-old boy who presented with a mass in the right upper quadrant of the abdomen. The total body radioactivity effect, which is similar to the total body opacification during intravenous cholangiography or urography, was clearly demonstrated as a cold area corresponding to the cyst and assisted in the diagnosis. The specific finding was sequential filling of the gallbladder and cyst. In view of the relatively low radiation dose and good physical characteristics of 99mTc for imaging, 99mTc-PG sequential scintiphotography is useful for the definitive preoperative diagnosis of choledochal cyst in infants and children.

Real time ultrasound diagnosis of non-vaterian duodenal adenoma--a brief case report.

Non-vaterian duodenal adenomas are rare lesions, usually present with bleeding and abdominal pain. Sonography is often the first imaging procedure in patients with vague upper abdominal complaints, unsuspected duodenal pathology may be encountered. It is now widely accepted as the first choice for the investigation of abdominal masses. Oral water load is recognised as a good procedure for the detection of a bowel-related mass during sonographic examination. In our case, we used the real-time ultrasound with water ingestion to detect the location of the duodenal lesion and its relationship to the neighboring structure. This report demonstrates that conventional real-time ultrasound is still a simple method to diagnose duodenal lesions.

c-Kit mediates chemoresistance and tumor-initiating capacity of ovarian cancer cells through activation of Wnt/ß-catenin-ATP-binding cassette G2 signaling.

Cisplatin and paclitaxel are standard chemotherapy for metastatic ovarian cancer, but with limited efficacy. Cancer stem/progenitor cells (or tumor-initiating cells, TICs) are hypothesized to be chemoresistant, and the existence of TICs in ovarian cancer has been previously demonstrated. However, the key signals and molecular events regulating the formation and expansion of ovarian tumor-initiating cells (OTICs) remain elusive. Here, we show that c-Kit is not just a marker of OTICs, but also a critical mediator of the phenotype that can be a viable target for the treatment of ovarian cancer. In contrast to non-OICs, c-Kit was overexpressed in OTICs. Moreover, the use of small interfering RNA to inhibit c-Kit expression markedly attenuated the number and size of OTIC subpopulations, inhibited the expression of stem cell markers and decreased the tumorigenic capabilities of OTICs. Imatinib (Gleevec), a clinical drug that blocks c-Kit kinase activity, also demonstrated its inhibition potency on OTICs. In addition, cisplatin/paclitaxel, which killed non-OTICs, with c-Kit knockdown or imatinib revealed that this was critically required for intervening ovarian cancer progression and recurrence in vitro and in xenograft tumors in vivo. Similar results were obtained with OTICs derived from ovarian carcinoma patients. Studies into the mechanisms suggest an important role for the activation of Wnt/ß-catenin and ATP-binding cassette G2 downstream of c-Kit. The tumor-promoting microenvironment, such as hypoxia, could promote OTICs via upregulation of c-Kit expression. These results unravel an integral role for c-Kit in ovarian neoplastic processes and shed light on its mechanisms of action.

Cost-effectiveness of postremission intensive therapy in patients with acute leukemia.

BACKGROUND: We assessed the cost-effectiveness of high-dose arabinoside (HiDAC)-based and allogeneic stem-cell transplantation (alloSCT)-based therapy in patients with acute leukemia. PATIENTS AND METHODS: We analyzed the outcome, cost and cost-effectiveness of 106 patients treated from January 1994 to January 2002 [94 acute myelogenous leukemia (AML)/12 acute lymphoblastic leukemia (ALL)]. Forty-two young patients at either intermediate or unknown cytogenetic risk received postremission intensive therapy (24 HiDAC-based/18 alloSCT-based therapy). RESULTS: After a median follow-up of 50 months, the estimated 7-year overall survival for the HiDAC-based group showed a tendency to be higher than the alloSCT-based group (48% versus 28%, P = 0.1452). The HiDAC-based group spent a significantly lower total cost ($US51,857 versus 75,474, P = 0.004) than the alloSCT-based group. Cost-effectiveness analysis showed that the mean cost per year of life saved for the HiDAC-based group is considerably less expensive than the alloSCT-based group ($US11,224 versus 21,564). The reduced total cost for the HiDAC-based group originated from lower cost in room fees, medication, laboratory and procedure, but not in blood transfusion and professional manpower fees. CONCLUSION: For the postremission therapy in young AML patients at either intermediate or unknown cytogenetic risk, cost-effectiveness of HiDAC-based therapy compares favorably with that of alloSCT-based therapy, which deserves further clinical trials.

Pretreatment prognostic factors and treatment outcome in elderly patients with de novo acute myeloid leukemia.

BACKGROUND: Elderly patients with acute myeloid leukemia (AML) generally have an unfavorable clinical course and are under-represented in clinical trials. The aim of this study was to analyze the prognosis and treatment outcome of elderly AML patients. PATIENTS AND METHODS: We studied 205 AML patients aged 65 years or older at our hospital. Prior to study initiation, we designated 13 variables to be analyzed for their impact on complete remission (CR) rate and overall survival (OS). RESULTS: Induction regimen (standard chemotherapy) and good performance status (PS) (Eastern Cooperative Oncology Group PS 0-1) independently influenced the achievement of CR. Multivariate analysis also determined five poor prognostic factors for OS: poor PS (score 2-4), presence of comorbidities, elevated serum lactate dehydrogenase level (> or =2x upper normal limit), extreme leukocytosis (> or =100 x 10(9)/l) and marked thrombocytopenia (< or =20 x 10(9)/l). Age was not an independent contributing factor in terms of either CR attainment or OS duration. Low-risk patients, who possessed one or less non-leukocytosis poor prognostic factor, had significantly longer disease-free survival and OS than their high-risk counterparts. CONCLUSIONS: Elderly AML patients should be risk-stratified at diagnosis. Anthracycline-based induction chemotherapy would be the best therapeutic option for such patients.

Effect of L-phenylalanine methyl ester on the colony formation of hematopoietic progenitor cells from human bone marrow.

We have previously shown that L-phenylalanine methyl ester (PME) is capable of removing monocytes and enhancing the growth of hematopoietic colonies from human peripheral blood (PB) mononuclear cells (MNC). In the present study, we further compared the effect of PME on the colony formation of bone marrow (BM) and PB. Low density (less than or equal to 1.077 g/ml) MNC were obtained by Ficoll-diatrizoate density gradient centrifugation. Granulocyte/macrophage colony-forming units (CFU-gm) and erythroid burst-forming units (BFU-e) were cultured in agarose with conditioned media (CM) and/or interleukin 3 (IL-3), granulocyte colony-stimulating factor (G-CSF) and granulocyte/macrophage-CSF (GM-CSF). Treatment of BM MNC with 5 mM PME for 15 min at room temperature yielded a nucleated cell recovery of 44.8 +/- 5.0% (mean +/- SE; N = 8). CFU-gm were enriched 2.7-fold (range 2.0 to 4.8). Using CM or CM supplemented with G-CSF or GM-CSF has minimal effect on the enrichment. Leukocyte differentials revealed that 94.3 +/- 3.05% of the monocytes, as well as 91.2 +/- 1.60% of the cells in the neutrophilic maturation series were removed by PME. Incubation for 40 min in PME abolished CFU-gm formation. BFU-e were not enriched by the PME treatment. In contrast, 40 min incubation of PB MNC produced higher enrichment of CFU-gm than that obtained from 15 min of treatment, although lower cell recovery was obtained with the longer treatment time. In conclusion, we have demonstrated that phagocytic cells can be removed from BM or PB MNC by PME treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Sonographic diagnosis of a small fistulous communication between a subphrenic abscess and a perforated duodenal ulcer.

We report a case of a fistula between a subphrenic abscess and a perforated duodenal ulcer diagnosed by sonography and confirmed by CT. The sonographic findings included a subphrenic fluid collection connected to the anterior aspect of the superior duodenum by a nonpulsatile, anechoic tubular lesion. Manual compression of the upper epigastrium resulted in movement of echogenic debris from the antrum and superior duodenum through the fistulous tract into the abscess.

Sonographic findings of intraabdominal foreign bodies due to retained gauze.

The ultrasonic images obtained in 2 patients with retained gauze in the abdomen are presented. Identification of a cystic mass with highly irregular internal echoes in patients with previous laparotomy should alert the radiologist to the possibility of foreign body retention.

Spatiotemporal analysis of event-related fMRI data using partial least squares.

Partial least squares (PLS) has proven to be a important multivariate analytic tool for positron emission tomographic and, more recently, event-related potential (ERP) data. The application to ERP incorporates the ability to analyze space and time together, a feature that has obvious appeal for event-related functional magnetic resonance imaging (fMRI) data. This paper presents the extension of spatiotemporal PLS (ST-PLS) to fMRI, explaining the theoretical foundation and application to an fMRI study of auditory and visual perceptual memory. Analysis of activation effects with ST-PLS was compared with conventional univariate random effects analysis, showing general consensus for both methods, but several unique observations by ST-PLS, including enhanced statistical power. The application of ST-PLS for assessment of task-dependent brain-behavior relationships is also presented. Singular features of ST-PLS include (1) no assumptions about the shape of the hemodynamic response functions (HRFs); (2) robust statistical assessment at the image level through permutation tests; (3) protection against outlier influences at the voxel level through bootstrap resampling; (4) flexible analytic configurations that allow assessment of activation difference, brain-behavior relations, and functional connectivity. These features enable ST-PLS to act as an important complement to other multivariate and univariate approaches used in neuroimaging research.

A case of 'superwarfarin' poisoning.

A young adult ingested 10 mg brodifacoum (a potent rodenticide) in an attempt to commit suicide. Prolonged bleeding was noted for over 6 months. It required large doses and prolonged use of vitamin K1 to combat this 'superwarfarin' poisoning.

A novel splicing acceptor mutation of the factor VIII gene producing skipping of exon 25.

A gross deletion in the factor VIII (FVIII) mRNA was determined by reverse transcriptase polymerase chain reaction (RT-PCR) for a patient with moderately severe hemophilia A. Sequencing of the RT-PCR product depicted a 177-bp deletion ranging from nucleotide (nt) 6724 to nt 6900 of FVIII cDNA, exactly corresponding to the whole exon 25. Further study of the genomic DNA revealed the presence of a single base pair substitution (G >A) at position -1 of intron 24. The absolute consensus AG doublet of the intron 24 splicing acceptor changed to AA. In the novel splice site mutation, exon 24 was erroneously spliced to exon 26, skipping exon 25. The FVIII antigen level was normal despite the markedly reduced functional activity. Since exon 25 corresponds to part of the C2 domain, we speculate that for this patient the aberrant C2 domain markedly reduces binding affinity of FVIII protein to the phospholipid membrane, thus severely impairing the protein function.

Diagnostic value of glycosylated hemoglobin in intravascular hemolysis after cardiac surgery.

Glycosylated hemoglobin (GH), hemoglobin (Hb), hematocrit, LDH and serum bilirubin were measured pre-operatively, and 1-10 days and 11-42 d post-operatively in 42 patients undergoing cardiac surgery. Their mean age was 39.90 years, ranging from 4 to 68 yr. In the early post-operation days, Hb and hematocrit, but not GH percentage, were significantly decreased. This demonstrates that, at this stage, acute blood loss rather than hemolysis is more prominent. In the later post-operative days, GH, but not Hb or hematocrit, decreased significantly. This indicates the presence of chronic hemolysis with bone marrow compensation. In our study, the incidence of chronic mild hemolysis after cardiac surgery was very high (68.8%). We conclude that GH determination is a simple, easy and sensitive method to detect chronic hemolysis and we suggest measuring it in every case with suspicion of hemolysis.

Acute febrile neutrophilic dermatosis (Sweet's syndrome) in hairy cell leukemia: a case report.

Sweet's syndrome is a cutaneous reactive process that is usually associated with fever, but rarely occurs in patients with hairy cell leukemia (HCL). We report the case of a patient with HCL who developed typical Sweet's syndrome five months after the diagnosis of HCL. Skin eruptions and constitutional symptoms subsided rapidly after short-term systemic adrenocorticosteroid treatment without recurrence, until the patient died from disease exacerbation and uncontrolled infection. According to his manifestations, chemical allergens, microorganisms or HCL progression were unlikely to have been the precipitating factors for development of Sweet's syndrome. Because immunologic disorders and opportunistic infection are not uncommon in patients with HCL, a skin biopsy should be taken as early as possible when cutaneous lesions and fever develop in order to establish a correct diagnosis. Hence, unnecessary and prolonged use of antibiotics is avoided and prompt relief of the symptoms by appropriate management can be achieved.

[X-linked dyskeratosis congenita with aplastic anemia--genetic and hematologic studies].

Two brothers in a Chinese family were affected with dyskeratosis congenita (DC). The proband's stepbrother died of pancytopenia and rectal adenocarcinoma. The proband with progressive pancytopenia was studied in genetics and hematology. Family investigations suggest that the disorder was inherited as an X-linked recessive transmission. No specific chromosome aberrations were detected in both peripheral blood and bone marrow cells. Bone marrow biopsy revealed markedly hypocellular marrow without prominent reticulin fibers. Hematopoiesis was assessed by assay of granulomonocytic (CFU-GM) colonies and clusters in vitro. Numbers of CFU-GM were profoundly reduced in marrow cultures from our patient. Although a defect in the stromal microenvironment and lymphocyte mediated suppression could not be excluded. It is possible that the defect is intrinsic to the hematopoietic stem cells. The familial disease described by this paper favors that the dyskeratosis congenita is X-linked with constitutional aplastic anemia. It is believed that the pancytopenia with hypocellular marrow in DC is an inherited rather than an acquired condition.

Combined cold- and warm-antibody autoimmune hemolytic anemia--review of the literature and a case report.

This report described the second case in the literature of Waldenstrom's macroglobulinemia with combined cold- and warm- antibody autoimmune hemolytic anemia (CCW-AIHA). Both IgG and IgM red cell auto-antibodies (auto-Abs) were present in the patient's serum. The IgM auto-Abs had a high thermal amplitude and anti-I blood group specificity. The IgG auto-Abs did not reveal blood group specificity. This patient depended on steroids to control hemolysis, but responded well to chemotherapy and plasmapheresis. CCW-AIHA has rarely been reported. In this study, the literature is reviewed and the characteristic serological and clinical features of this rare category of autoimmune hemolytic anemia (AIHA) are discussed.

Immunophenotyping of myeloma cells with immunoalkaline phosphatase technique.

An immunoalkaline phosphatase (IAP) technique was used to determine the class of cytoplasmic immunoglobulin presenting in the malignant plasma cells of patients with multiple myeloma. Using monoclonal antibodies against different human immunoglobulins (Igs) as the primary antibodies, and calf intestinal phosphatase as the enzymatic indicator. The presence of monoclonal immunoglobulin was demonstrated within the cytoplasm of malignant plasma cells. These results correlate well with the electrophoretic patterns of the Igs present in the serum samples of these patients. This IAP technique is suggested as a practical method for evaluating the immunophenotype of multiple myeloma.