Successful T-cell-replete peripheral blood stem cell transplantation from HLA-haploidentical microchimeric mother to daughter with refractory acute lymphoblastic leukemia using reduced-intensity conditioning.

A 16-year-old girl with refractory acute lymphoblastic leukemia underwent reduced-intensity hematopoietic stem cell transplantation from her two-locus-mismatched haploidentical mother, who was microchimeric for the patient's hematopoietic cells. The conditioning regimen comprised melphalan, fludarabine, and low-dose total body irradiation. Non-T-cell-depleted peripheral blood stem cells were infused with graft-versus-host disease (GVHD) prophylaxis consisting of tacrolimus, prednisolone, and short-course methotrexate. Complete donor-type engraftment without evidence of residual leukemia was confirmed on day 22. Severe GVHD was not observed despite rapid cessation of immunosuppression. The patient remains well in continuous remission 15 months after transplant. This successful experience suggests that maternal hematopoietic stem cell transplants for children, in the presence of microchimerism, may be associated with hyporesponsiveness to the inherited paternal HLA antigens (IPA); preventing severe GVHD.

[Childhood t(8;21) acute myelocytic leukemia: a comparison of clinical features and risk factors with adult cases].

Despite the abundance of reports describing adult cases of t(8;21) acute myelocytic leukemia (AML), childhood cases have received little attention. We retrospectively investigated 14 childhood cases of t(8;21) AML, and compared their clinical characteristics with those of adult cases, focusing on the risk factors for poor prognosis. Seventy-one percent of the patients had fever. Their mean leukocyte count was 12,700/microliter, and they showed decreased NAP activity. The cell surface showed positivity for CD13, 33, 19, 34, and HLA-DR. The complete remission rate was 100%, and relapse was observed in three of the patients. Bone marrow eosinophilia was present in a smaller proportion of the childhood cases than in the adult cases. Although an increased leukocyte count, tumor formation, and other risk factors have been reported in adults, there was no correlation between these factors and prognosis in our childhood cases. As children who showed AML relapse had TdT-positive blasts, detectable blast TdT activity may be a risk factor for relapse in childhood cases of t(8;21) AML. However, to confirm this, a study with a larger subject base should be conducted.

Soluble stem cell factor receptor (CD117) and IL-2 receptor alpha chain (CD25) levels in the plasma of patients with mastocytosis: relationships to disease severity and bone marrow pathology.

Systemic mastocytosis is a disease of mast cell proliferation that may be associated with hematologic disorders. There are no features on examination that allow the diagnosis of systemic disease, and mast cell-derived mediators, which may be elevated in urine or blood, may also be elevated in individuals with severe allergic disorders. Thus, the diagnosis usually depends on results of bone marrow biopsy. To facilitate evaluation, surrogate markers of the extent and severity of the disease are needed. Because of the association of mastocytosis with hematologic disease, plasma levels were measured for soluble KIT (sKIT) and soluble interleukin-2 receptor alpha chain (sCD25), which are known to be cleaved in part from the mast cell surface and are elevated in some hematologic malignancies. Results revealed that levels of both soluble receptors are increased in systemic mastocytosis. Median plasma sKIT concentrations as expressed by AU/mL (1 AU = 1.4 ng/mL) were as follows: controls, 176 (n = 60); urticaria pigmentosa without systemic involvement, 194 (n = 8); systemic indolent mastocytosis, 511 (n = 30); systemic mastocytosis with an associated hematologic disorder, 1320 (n = 7); aggressive mastocytosis, 3390 (n = 3). Plasma sCD25 levels were elevated in systemic mastocytosis; the highest levels were associated with extensive bone marrow involvement. Levels of sKIT correlated with total tryptase levels, sCD25 levels, and bone marrow pathology. These results demonstrate that sKIT and sCD25 are useful surrogate markers of disease severity in patients with mastocytosis and should aid in diagnosis, in the selection of those needing a bone marrow biopsy, and in the documentation of disease progression. (Blood. 2000;96:1267-1273)

Combined measurement of the c-erbB-2 protein in breast carcinoma tissues and sera is useful as a sensitive tumor marker for monitoring tumor relapse.

c-erbB-2 protein levels in tissue extracts and sera were determined in a retrospective analysis of 158 patients who underwent surgical resection of breast carcinoma by means of a sandwich enzyme immunometric assay (EIA) using monoclonal antibodies (MAbs) directed to the extracellular domain of the c-erbB-2 oncogene protein (ErbB-2). In the analysis of tissue extracts, 48 samples (30.3%) showed ErbB-2 levels exceeding 18.0 ng/mg protein (group A), while in 110 samples these levels were below 18.0 ng/mg protein (group B). Immunohistochemical examination of resected tissues using anti-c-erbB-2 antibody revealed positive staining in 93.8% (45/48) in group A and 13.6% (15/110) in group B (p < 0.00001). The proportion of patients who preoperatively showed a serum ErbB-2 value above 5.4 ng/ml was 52.1% (25/48) in group A and 10.0% (11/110) in group B (p < 0.00001). Thus, the level of ErbB-2 in tissue extracts was significantly associated with immunohistochemistry and ErbB-2 levels in preoperative sera. During follow-up, 48 patients (30.3%) developed recurrent disease: 17 in group A (35.4%) and 31 in group B (28.2%). From an ROC analysis based on the postoperative serum ErbB-2 levels in patients either with or without relapse, the cutoff value of serum ErbB-2 for tumor relapse was determined to be 6.5 ng/ml. The sensitivity of serum ErbB-2 in patients with relapsed breast cancer was 58.3% (21/36) overall, 84.6% (11/13) in group A and 43.5% (10/23) in group B. In the analysis of serum samples taken before relapse, 90.9% (10/11) of the subjects in group A and 26.7% (4/15) of those in group B were shown to be positive for serum ErbB-2. Serum ErbB-2 in group A was a more sensitive marker than other tumor markers such as CEA, CA15-3, and NCC-ST-439. Thus, the determination of ErbB-2 in tissue extracts of breast carcinoma may be useful for assessing c-erbB-2 protein expression in the primary tissue and indicates that serum ErbB-2 may be a sensitive marker for monitoring tumor relapse.

A phase III randomized study comparing doxifluridine and 5-fluorouracil as supportive chemotherapy in advanced and recurrent gastric cancer.

We conducted a phase III randomized study to investigate effects of supportive chemotherapy with oral doxifluridine (group A, 75 patients) or 5-fluorouracil (group B, 75 patients) in advanced gastric cancer when intensive chemotherapy was not an option. Although there were no significant differences between the groups with regard to survival, hospital-free survival and time to progression, median values of 3 endpoints were superior in group A. Secondary analysis showed that group A patients with prior chemotherapy tended to have longer survival and hospital-free survival and significantly longer time to progression.

Measurement of KIT ligand/stem cell factor: clinical and biochemical significance.

Stem cell factor (SCF) and its receptor KIT play an important role in various biologic phases, such as hematopoiesis, reproduction, and regeneration. It has been possible to measure both soluble SCF and soluble KIT using enzyme-linked immunosorbent assay since 1993 and 1995, respectively. Although the significance of interaction of soluble SCF with soluble KIT has not yet been elucidated, in certain diseases proteins fluctuate in human sera. We found that serum SCF levels were fivefold higher in patients with chronic renal failure than levels in healthy controls. We review the results of the analysis of SCF. In addition, possible pathologic mechanisms in various clinical abnormalities and the clinical potential for recombinant human SCF are discussed.

Serum c-erB-2 levels in monitoring of operable breast cancer patients.

BACKGROUND: Various methods and criteria are used to determine protein overexpression of c-erbB-2 and the clinical utility of c-erbB-2 is under investigation. We have reported previously that the levels of cytosol c-erbB-2 in breast cancer were significantly different between the clinical stages. METHODS: The levels of c-erbB-2 protein were determined in sera from 210 breast cancer patients using a sandwich enzyme immunoassay between November 1996 and March 1998. The cut-off level was set at 5.4 ng/ml for healthy female blood donors. RESULTS: First, serum c-erbB-2 levels were analyzed in 73 preoperative breast cancer patients with stage I-IIIB disease. The range and median values were 2.3-32.3 and 4.8 ng/ml, respectively. The positive rate was 38%. Overexpression of serum c-erbB-2 was significantly associated with tumor size, clinical stage, histological grade, lymphatic invasion, nodal status and overexpression of cytosol c-erbB-2, but not with hormonal receptor status and other clinico-pathological factors. Second, c-erbB-2, CEA and CA15-3 in sera were examined in 157 postoperative breast cancer patients. In the 137 disease-free patients, specificities of c-erbB-2, CEA and CA15-3 were 72, 93 and 99%, respectively, but in the 20 first recurrent patients, these sensitivities were 80, 25 and 25%, respectively. CONCLUSIONS: These results suggest that serum c-erbB-2 protein is a useful marker for predicting aggressive behavior and first recurrence of breast cancer.

Significance of stem cell factor and soluble KIT in patients with systemic lupus erythematosus.

To determine the significance of stem cell factor (SCF) and soluble KIT (sKIT) in the serum of patients with systemic lupus erythematosus (SLE), levels of SCF and sKIT in patients with SLE were estimated, and their correlations with clinical parameters were examined. The sKIT levels in SLE patients (n = 106) were significantly lower than those in healthy controls (n = 40). A significant negative correlation was found between the SCF and sKIT levels of SLE patients. Although the SCF levels correlated with the titre of anti-RNP antibody, no significant relationship was found between SCF levels and blood cell counts, such as white blood cell, red blood cell and platelet counts. sKIT levels were significantly correlated with the platelet count, and were negatively correlated with the white blood cell count, titre of anti-DNA antibody, and SLE activity index (SLEDAI). sKIT levels were also negatively affected by high doses of corticosteroid. These results indicate that serum sKIT levels may be more closely related than SCF to the haematological abnormalities in SLE patients, and may reflect the clinical status of SLE patients and the effectiveness of high-dose corticosteroid treatment.

Elevated SCF levels in the serum of patients with chronic renal failure.

Serum stem cell factor (SCF) and soluble KIT (sKIT) levels were estimated in patients with chronic renal failure (CRF) and anaemia, and compared with clinical parameters of blood cells and renal function. Serum SCF levels in CRF patients were 5-fold higher than those in healthy controls. However, serum sKIT levels in haemodialysis (HD)-CRF patients were only slightly higher than those of healthy controls. In untreated CRF patients and healthy controls, serum SCF levels were significantly correlated with blood urea nitrogen (BUN), creatinine. haemoglobin, red blood cell (RBC) count and sKIT. In untreated CRF patients, serum SCF levels were significantly correlated with BUN, creatinine, and sKIT. These results suggest that serum SCF levels increased with the deterioration of renal function and might be related to erythropoiesis.

[Postoperative analgesic effects of epidural bolus administration of bupivacaine and morphine in patients after vaginal hysterectomy and vaginoplasty].

This study was conducted to evaluate the postoperative analgesic effects of epidural bolus administration of bupivacaine and morphine in patients after vaginal hysterectomy and vaginoplasty. A total of 100 patients who had received the elective vaginal hysterectomy and vaginoplasty were investigated retrospectively by separating them into two groups. Fifty patients who had received epidural analgesics (E group) showed significantly lower pain scores for 12 hrs after the surgery compared with 50 patients who had received intramuscular buprenorphine hydrochloride (C group). The patients in E group needed significantly less supplemental analgesics than those in C group for 2 hrs after the operation. The incidence of side effects was not significantly different between the two groups. But the incidence of postoperative complications was significantly lower in the patients in E group than those in C group. These results show that epidural bolus administration of bupivacaine and morphine is effective for postoperative pain relief and decreases the incidence of postoperative complications after vaginal hysterectomy and vaginoplasty.

The incidence of colorectal cancer during pregnancy in Japan: report of two cases and review of Japanese cases.

Colorectal cancer during pregnancy is very rare. We report two additional cases of rectal carcinoma. A 34-year-old woman with an obstructive adenocarcinoma of the rectal region was diagnosed at labor. A 35-year-old woman with an adenocarcinoma of the rectal region was diagnosed at 32 weeks of gestation and underwent a cesarean section and rectal resection at 35 weeks of gestation. A retrospective review of the Japanese literature was performed to identify patients who appeared to have primary colorectal cancer during pregnancy. Thirty-six patients with colon cancer (75.0%), 10 (20.8%) with rectal cancer, and two (4.2%) of unknown sites have been reported in Japanese series. The average age of the mother was 32.2 years. The calculated incidence of colorectal cancer among Japanese pregnant women was one case per 502,316 live births during the years between 1986 through 1995. Although the majority of colorectal cancers diagnosed during pregnancy are rectal carcinomas, the patients in Japan were predominantly complicated by colon cancer. The fetal risk seems small, because there were no cases of colorectal cancer metastatic to the products of conception.

NADH dehydrogenase deficiency in an apoptosis-resistant mutant isolated from a human HL-60 leukemia cell line.

An apoptosis-resistant mutant (VC-33) was selected from HL-60 by alternating exposure to camptothecin and etoposide. VC-33 cells demonstrated resistance to apoptosis as induced not only by camptothecin and etoposide but by a variety of other agents as well, including 1-beta-D-arabinofuranosylcytosine, hydroxyurea, calcium ionophore (A23187), cycloheximide, and UV irradiation. In an effort to identify the mechanism of such apoptosis resistance, a mRNA differential display analysis was used. Among a total of 12 bands with reduced expression in VC-33 cells, 1 cDNA clone was isolated that was hybridized to the wild-type transcript but not to the VC-33 transcript on Northern blotting. Partial sequence of this gene revealed 98% homology to mitochondrial NADH dehydrogenase subunit 5. When cell growth and intracellular ATP levels under glucose starvation were measured, VC-33 cells were found to be more sensitive than wild-type cells. Thus, NADH dehydrogenase deficiency may contribute, at least in part, to the mechanism of resistance to apoptosis in VC-33 cells.

Amino acid control of asparagine synthetase: relation to asparaginase resistance in human leukemia cells.

Complete amino acid deprivation in mammalian cells causes a significant enhancement in gene expression for a number of important cellular activities; among these is asparagine synthetase (AS). The data presented demonstrate that, in both nonleukemic (rat Fao hepatoma cells) and human leukemia cells (MOLT-4, NALL-1, and BALL-1), AS mRNA levels, protein content, and enzymatic activity are induced after incubation in an otherwise complete tissue culture medium that is deficient in a single amino acid or in medium that has been depleted of the amino acid asparagine by the addition of asparaginase. Complete amino acid deprivation results in a concerted increase in AS mRNA, protein, and enzymatic activity, which, in conjunction with previously published research, suggests that the mechanism of this cellular response involves transcriptional control of the AS gene. Asparaginase treatment is a standard component of acute lymphoblastic leukemia therapy for which the effectiveness is related to the inability of these cells to upregulate AS activity to a sufficient level. With regard to the asparaginase sensitivity of the three human leukemia cell lines, there was a trend toward an inverse relation to the degree of AS expression. Selection for asparaginase-resistant MOLT-4 sublines resulted in enhanced AS mRNA and protein content regardless of whether the cells had been selected by asparaginase treatment directly or asparagine was removed from the culture medium. Collectively, the data illustrate that further advances in asparaginase therapy will require additional knowledge of amino acid-dependent regulation of AS gene expression and, conversely, that asparaginase resistance represents a model system for investigating metabolite control in a clinically relevant setting.

Successful bone marrow transplantation from an HLA-identical unrelated donor in a patient with hemophagocytic lymphohistiocytosis.

We report a 3-year-old girl with hemophagocytic lymphohistiocytosis (HLH) who received BMT from an HLA-identical unrelated donor when the disease was active. She had entered remission in response to chemotherapy consisting of etoposide and methylprednisolone. After a relapse, her disease became refractory to chemotherapy, and splenectomy was performed with marginal improvement. She underwent BMT from an HLA-identical unrelated donor, conditioned with CY, TBI and anti-thymocyte globulin (ATG). She has been in complete remission for 18 months since the BMT. This result suggests that BMT from an HLA-identical unrelated donor should be considered for HLH even if the disease is active.

Soluble c-kit molecule in serum from healthy individuals and patients with haemopoietic disorders.

The proto-oncogene, c-kit, encodes a transmembrane tyrosine kinase receptor (KIT) and plays an important role in haemopoiesis. We have identified a 95 kD soluble form of KIT (S-KIT) in culture supernatant of human megakaryoblastic cell line, CMK. To study the physiological significance of S-KIT, we have established a sensitive sandwich ELISA system. Serum samples from healthy individuals contained detectable amounts of S-KIT. Next, we determined a total of 220 samples from 134 patients with haemopoietic disorders. A considerable number of patients with acute myeloid leukaemia (AML), especially those with more immature phenotypes (M0, M1 or M2) had elevated levels of serum S-KIT. Those levels decreased to the normal range after effective chemotherapy. In chronic myeloid leukaemia, patients with myeloid blastic crisis showed markedly elevated levels of serum S-KIT. In contrast, S-KIT levels decreased in cases with either acute or chronic lymphoid leukaemia. There was a tendency for patients with severe aplastic anaemia to show decreased levels, but it was not significant. In myelodysplastic syndrome, S-KIT levels appeared to vary by subsets, with higher concentration in more advanced forms of the disease. Although the functional role of S-KIT is not yet elucidated, these results suggest that the serum S-KIT levels may reflect the pathological states of various haematological disorders.

[The effects of postoperative continuous epidural analgesia on the early recovery from lower abdominal surgery].

This study was conducted to evaluate the effects of continuous epidural analgesia (CEA) on the incidence of postoperative complications and the early recovery after lower abdominal surgery. A total of 109 patients who had received elective lower abdominal surgery were investigated retrospectively by separating them into two groups. Compared to 35 patients who had received standard analgesic techniques without epidural analgesia, 74 patients who had been administered CEA with buprenorphine, mepivacaine and droperidol for 24 hrs after surgery could sit on the bed significantly earlier. But the patients with CEA could not stand on the floor and could not walk significantly earlier than the patients without CEA. The overall postoperative complication rate was not significantly different between the patients with and without CEA. These results show that postoperative CEA exerts a beneficial effects on the early recovery after the lower abdominal surgery, but the effect is not so strong as in upper abdominal surgery group. The results also suggest that CEA does not decrease the incidence of postoperative complications.

Life-style and subsite of gastric cancer--joint effect of smoking and drinking habits.

To clarify the effects of life-style on gastric cancer by subsite focusing on the proximal part (cardia and fundus) and the distal part (pyloric antrum), a case-control study was conducted at the Aichi Cancer Center in Nagoya, Japan from 1988-1991. This study compared 668 histologically confirmed gastric cancer cases [123 cardia, 218 middle (body), 256 antrum, and 71 unclassified] with 668 controls using a common questionnaire about life-styles as related to smoking, drinking, dietary habits and frequency of food intake. Controls were selected from among outpatients of the same hospital. Controls free of cancer and other specific diseases were matched with cases for sex, age (within 2 years), and time of hospital visit (within 2 months). A Western-style breakfast decreased the risk of antrum cancer, while consumption of greasy food increased the risk of cardia cancer. Fresh vegetables decreased the risk of cancer in both cardia and antrum. Habitual smoking is associated with increased risk of gastric cancer and it is more prominent in cardia cancer, especially in those who are drinkers. Results obtained from this study suggest that risk factors and relative risks of gastric cancer varied by subsite to a considerable degree. Furthermore, the joint effect of smoking and drinking may play an important role in the development of gastric cancer, especially of cardia cancer.

[The effects of postoperative continuous epidural analgesia on the early recovery from gastrectomy].

After gastrectomy, 103 patients were investigated concerning the effects of postoperative continuous epidural analgesia on the postoperative complications and early recovery. Sixty-seven patients who received epidural analgesia after the operation (group E) were able to sit on the bed, stand on the floor and walk themselves significantly earlier than 36 patients who were given analgesics intramuscularly or transrectally (group C). The incidence of postoperative complications was significantly less in the patients of group E than those of group C. These results show that postoperative continuous epidural analgesia decreases the incidence of postoperative complication and supports the early recovery from the operation.