A radio-frequency ion trap with string electrodes.

A radio-frequency (rf) ion trap with string electrodes is introduced. In this trap configuration, the rf electrodes are made of narrow metal strings, by which a negligibly small portion of light-induced fluorescence (LIF) is blocked. Then the LIF collection solid angle can be maximized. In the demonstration, barium ions are trapped and laser-cooled in the rf trap with string electrodes successfully, and the crystallization is confirmed by the LIF spectrum.

Risk factor analysis for thrombotic microangiopathy after reduced-intensity or myeloablative allogeneic hematopoietic stem cell transplantation.

Thrombotic microangiopathy (TMA) impairs long-term survival after allogeneic hematopoietic stem cell transplantation (HSCT). As the allogeneic HSCT procedure has developed, addressing risk factors for TMA has become more complicated. The aim of this study was to investigate the impact of transplant-associated factors on TMA incidence in patients who have undergone HSCT in various settings. One hundred twenty-three consecutive allogeneic HSCT patients with hematologic diseases receiving myeloablative and reduced-intensity conditioning were evaluated retrospectively. Of 123 patients, 22 (17.9%) developed TMA after HSCT. Multivariate analysis showed the significance of GVHD grade II-IV, and the use of FK506 and the use of high-dose busulfan (Bu) (16 mg/kg) persisted. The hazard ratios of the use of FK506, the use of high-dose Bu (16 mg/kg), and GVHD grade II-IV for TMA were 8.7 (95% CI 2.0-37), 5.7 (95% CI 1.5-21), and 3.4 (95% CI 1.3-9.1), respectively. In the present study, reduced-intensity conditioning did not have an advantage over myeloablative conditioning in decreasing the incidence of TMA after HSCT. Our results also showed that high-dose Bu (16 mg/kg) for the conditioning and FK506 for the prophylaxis of GVHD might contribute more significantly to TMA onset after HSCT than other agents.

T-lymphocyte subset analysis using the automated hematology analyser CELL-DYN 4000 for patients with hematological disorders.

We evaluated the performance of a fully-automated hematology analyser CELL-DYN 4000 (CD4000; Abbott Laboratories, CA, USA) for T-lymphocyte subset analysis using the samples from 58 patients with hematological disorders. A flow cytometer (CytoronAbsolute; Ortho Diagnostics, NJ, USA) was used for comparison. Good correlations were shown between CD4000 and CytoronAbsolute for the of CD3(+), CD4(+), and CD8(+) T-lymphocyte counts (for each of correlations, r = 0.992, 0.992, and 0.969, respectively). This good correlation was sustained even in 24 cases in which the CD4(+) T-lymphocyte counts were less than 200/microl. In this group the correlation coefficients (r-values) for the comparisons between CD3(+), CD4(+), and CD8(+) T-lymphocytes measured by CD4000 and CytoronAbsolute were 0.991, 0.976, and 0.990, respectively. The CD4000 T-lymphocyte assay is a completely automated method, and does not require opening of the sample tube, which reduces exposure to any biohazards such as HIV. Furthermore, the method is rapid and requires only 7 min for completion of the analysis.

[Acute myelomonocytic leukemia complicated with multiple lower intestinal ulcers induced by nonsteroidal anti-inflammatory drugs].

We report an 18-year-old woman with acute myelomonocytic leukemia, who developed massive lower intestinal bleeding following induction chemotherapy. Colonoscopy revealed multiple circular ulcers but no infectious colitis or infiltration of leukemia. The biopsy specimen showed mild non-specific inflammatory changes and scattered apoptosis bodies. She took nonsteroidal anti-inflammatory drugs (NASIDs) for pyrexia and pharyngalgia for a long time. We concluded these were signs of ulcers induced by NSAIDs. Despite discontinuance of NSAIDs, melena did not improve. Transarterial embolization therapy using microcoils was tried with unsatisfactory results. Finally, colonoscopic clipping therapy and continuous arterial injection of vasopressin were performed. Subsequently, her condition improved markedly. In conclusion, NSAID-induced intestinal bleeding is not limited to the upper GI tract but may occur in the lower GI tract after long-term NSAID use. The possibility of lower GI tract complications from NSAID should be kept in mind.

Cord blood transplantation for adult patients with hematological malignancies.

BACKGROUND: Recently, clinical studies of cord blood transplantation (CBT) in adults after myeloablative or nonmyeloablative conditioning regimens showed cord blood (CB) could effectively restore hematopoiesis and was associated with acceptable levels of graft versus host disease (GVHD). METHODS: This study reports the results of cord blood transplantation in 7 adults with hematological malignancies. RESULTS: Median age was 56 years (range, 43-69 years). HLA match was 4 of 6 in 4 cases and 5 of 6 in 3 cases. Median nucleated cell dose was 2.74 x 10(7) cells/kg (range, 2.13-3.80) and CD34+ cell dose was 1.15 x 10(5) cells/kg (range, 0.44-2.79). Three patients had primary graft failure. There was one early death at day 24 after CBT due to pneumonia. Three patients with engraftment are alive and free of disease at day 390, day 348 and day 164 after CBT. Acute GVHD grade II occurred in 2 cases with engraftment, and chronic GVHD occurred in 1 of 3 evaluable patients. Six patients with and without engraftment received more than 2.0 x 10(7) cells/kg nucleated cells. Three patients without engraftment received CD34+ cell dose less than that of 3 patients with engraftment. CONCLUSIONS: It is considered that graft CD34+ cell dose besides nucleated cell dose is important for engraftment. We believe that adult patients without suitable related or unrelated bone marrow donors should be considered as candidates for CBT by the choice of CB including both sufficient nucleated cell dose and CD34+ cell dose.

Selective activation of STAT3 in human monocytes stimulated by G-CSF: implication in inhibition of LPS-induced TNF-alpha production.

Lipopolysaccharide (LPS) induced tumor necrosis factor (TNF)-alpha production in human monocytes, which was dependent on activation of extracellular signal-regulated kinase (ERK), p38, c-Jun NH(2)-terminal kinase (JNK), and nuclear factor (NF)-kappa B. LPS-induced TNF-alpha production was inhibited by granulocyte colony-stimulating factor (G-CSF) and interleukin (IL)-10. G-CSF, like IL-10, exerted the inhibitory effect even when simultaneously added with LPS. Among the signaling pathways, signal transducer and activator of transcription 3 (STAT3) was selectively activated in monocytes stimulated by G-CSF or IL-10. G-CSF-mediated inhibition of LPS-induced TNF-alpha production as well as G-CSF-induced STAT3 phosphorylation and suppressor of cytokine signaling 3 mRNA expression were prevented by pretreatment of monocytes with AG-490, an inhibitor of Janus kinase 2. G-CSF did not affect LPS-induced activation of ERK, p38, JNK, and NF-kappa B, indicating that G-CSF affects the pathway downstream or independently of these signaling molecules. G-CSF-induced, but not IL-10-induced, STAT3 phosphorylation was attenuated in the presence of LPS. These findings suggest that G-CSF, like IL-10, inhibits LPS-induced TNF-alpha production in human monocytes through selective activation of STAT3, and the immunomodulation observed in vivo by G-CSF administration may be partly ascribed to the direct effect of G-CSF on monocyte functions.

[Primary malignant lymphoma arising from the nipple and areolar area of the breast].

A 56-year-old woman noticed soreness and swelling in the right nipple. Two weeks later, she noticed a mass in the outer lower region of the right areolar area, which was excised and the pathology of which was consistent with diffuse large B cell lymphoma (DLBCL). She was admitted when the right nipple mass was noted to be increasing, and was diagnosed as having stage I lymphoma. Her nipple mass was excised, and the pathology was consistent with DLBCL. CHOP therapy was administered three times and she was judged as having complete remission. Malignant lymphoma accounts for 0.15-0.17% of primary breast malignancies. Though the nipple and areolar area seem to be a rare primary site, this should be recognized as a sentinel zone for malignant lymphoma.

[Retrospective analysis of uroepithelial malignancies detected in the renal pelvis and ureter over the past decade at the Jikei University Hospital].

The aim of this study was to investigate recent characteristics and alterations of upper urinary tract cancer based on experience at a single institution over the past decade. Ninety-nine patients with renal pelvic and ureteral cancer resected at the Jikei University Hospital from January 1991 through December 2000 were retrospectively analyzed. Cancer-specific survival by pathologic stage, grade, and various clinical parameters were calculated by the Kaplan-Meier method. Prognostic factors for survival were examined with univariate and multivariate analysis. Cox regression analysis was used for multivariate analysis. Twenty-eight percent of cancers had been detected incidentally without having caused any symptoms. The overall 3-year and 5-year cancer-specific survival rates were 78% and 70%, respectively. The 5-year survival rate was 100% in patients with G1 cancer and 38% in those with G3 cancer. The 5-year survival rate was significantly higher in patients with cancers of lower grade (p = 0.0089), and was also higher in patients with cancers of stage pT1 or lower than in patients with cancers of stage pT2 and higher (p = 0.0038). The survival of patients with recurrence in the bladder was significantly longer than that of patients with recurrence in other organs. Multivariate analysis indicated that patient age and pT were the most important prognostic factors, followed by the presence of symptoms at diagnosis. The incidence of asymptomatic upper urinary tract cancer is increasing at institutions in Japan. We conclude that the cancer grade and stage still have classical predictive value, but that the presence of symptoms at the time of diagnosis is also an important prognostic factor.

[Bilateral pyeloureteritis cystica: a case report].

We report here a case of bilateral pyeloureteritis cystica. A 67-year-old woman was admitted to our hospital with asymptomatic macrohematuria in September 1999. Drip infusion pyelography and enhanced computed tomography demonstrated multiple small, round filling defects in both renal pelvises and ureters. Ureteroscopy and cold punch biopsy were performed, and histological examination revealed pyeloureteritis cystica. This patient was not given adjuvant therapy but was carefully followed up for 3 years and 6 months postoperatively.

[Successful treatment with foscarnet for disseminated varicella-zoster infection after reduced intensity stem cell transplantation in a case of relapsed refractory central nervous system lymphoma].

Visceral disseminated varicella-zoster virus (VZV) infection occurred with acute graft-versus-host disease in a 33-year-old Japanese male with non-Hodgkin lymphoma who had undergone allogeneic stem cell transplantation from an HLA-identical sibling after reduced intensity conditioning chemotherapy. Although ganciclovir and acyclovir treatment was effective temporarily, the number of VZV-DNA copies in the blood remained at a high level, and the hepatitis was prolonged. The patient was treated with foscarnet, which led to improvement of the VZV viremia and the hepatic dysfunction. Foscarnet therapy should be considered for acyclovir-resistant VZV infection in the setting of allogeneic hematopoietic stem cell transplantation.

[Analysis of factors for poor mobilization after administration of recombinant human granulocyte stimulating factor (rHuG-CSF) in healthy donors].

Allogeneic hematopoietic stem cell transplantation is an effective treatment for hematological malignancies. Peripheral blood stem cells (PBSCs) are increasingly used as an alternative to bone marrow for allogeneic transplantation. However, predictive factors for the response to recombinant human granulocyte stimulating factor (rHuG-CSF) in healthy donors have not been extensively studied. We analyzed the side effects, laboratory test results after administration of rHuG-CSF and the factors influencing mobilization of peripheral blood stem cells in 30 healthy donors. Bone pain, fever and headache were observed with high frequency after administration of rHuG-CSF. WBCs and reticulocytes increased, and RBCs and platelets decreased significantly after administration rHuG-CSF. Biochemical examination revealed significant elevations of LDH, CRP, ALP and UA. Univariate analysis showed the age of donors (< 50 vs. > 50, p = 0.041) and the lymphocyte counts before administration of rHuG-CSF (p = 0.032) to be correlated with the number of CD34 positive cells. From a multivariate analysis, the tendency for good mobilization with a twice daily dose of rHuG-CSF (p = 0.065) was observed. The rHuG-CSF schedule may be the most important factor affecting peripheral blood stem cell mobilization and collection in healthy donors.

Antiapoptotic effect of granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and cyclic AMP on human neutrophils: protein synthesis-dependent and protein synthesis-independent mechanisms and the role of the Janus kinase-STAT pathway.

Spontaneous neutrophil apoptosis during culture was delayed by granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF), or dibutyryl-cyclic adenosine monophosphate (cAMP), whereas apoptosis was accelerated by cycloheximide or actinomycin D. G-CSF-mediated antiapoptosis was completely abolished by cycloheximide or actinomycin D, whereas GM-CSF-mediated antiapoptosis was not completely abolished by these inhibitors. Antiapoptosis induced by dibutyryl-cAMP was highly resistant to cycloheximide, and that induced by benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone was unaffected by cycloheximide. G-CSF- and GM-CSF-mediated antiapoptosis and phosphorylation of signal transducer and activator of transcription 3 (STAT3) and STAT5 were inhibited by AG490, an inhibitor of Janus kinase. The level of Mcl-1 protein was not associated with neutrophil apoptosis. The results suggest that (a) neutrophil survival in the resting state is primarily regulated by the constitutive synthesis of antiapoptotic proteins; (b) the prevention of spontaneous apoptosis is mediated through the protein synthesis-dependent and/or protein synthesis-independent mechanisms according to the stimuli used; and (c) the Janus kinase-STAT pathway is involved in G-CSF- and GM-CSF-mediated antiapoptosis.

Infliximab for the treatment of severe steroid refractory acute graft-versus-host disease in three patients after allogeneic hematopoietic transplantation.

Acute graft-versus-host disease (aGVHD) is a serious complication of allogeneic peripheral blood stem cell transplantation (PBSCT). Patients with severe aGVHD not responding to treatment with steroids have a poor prognosis. We treated three patients with severe aGVHD refractory to steroids with infliximab. Patients (MDS 1, NHL 1, ALL 1) developed grade II-IV GVHD at a median of 13 days (range 9-17) after non-myeloablative PBSCT (HLA mismatched). All patients had received treatment with high-dose steroids for a median of 7 days (range 7-10) in addition to mycophenolate mofetil (MMF) (one). Infliximab was given in 3 weekly doses of 5 mg/kg. In one of three patients a partial resolution of diarrhea and minor improvement of skin were observed. One patient died with refractory GVHD. Infliximab is apparently an effective drug for the treatment of aGVHD, but can be more effective at doses of 5 mg/kg or higher and/or by administering it repeatedly every week.

Expression of the inhibitor of apoptosis (IAP) family members in human neutrophils: up-regulation of cIAP2 by granulocyte colony-stimulating factor and overexpression of cIAP2 in chronic neutrophilic leukemia.

Human neutrophils were found to express members of the inhibitor of apoptosis (IAP) family, namely cellular IAP1 (cIAP1), cIAP2, and X-linked IAP. Among these members, cIAP2 expression was selectively up-regulated by stimulation with granulocyte colony-stimulating factor (G-CSF), but not with granulocyte-macrophage CSF. The increased expression of cIAP2 mRNA was detected as early as 30 minutes after in vitro stimulation with G-CSF, and the elevated level of cIAP2 protein was detected at 1 hour. The elevated level of cIAP2 protein was also detected in peripheral blood neutrophils obtained from healthy donors receiving G-CSF administration. G-CSF-induced up-regulation of cIAP2 mRNA and protein, phosphorylation of signal transducer and activator of transcription 3 (STAT3), and the antiapoptotic effects were inhibited by pretreatment of cells with AG490, a specific inhibitor of Janus kinase 2 (JAK2). Mature neutrophils from a patient with chronic neutrophilic leukemia exhibited remarkable overexpression of cIAP2 mRNA and prolongation of survival, whereas cIAP2 mRNA expression and survival in mature neutrophils from patients with chronic myelogenous leukemia were essentially similar to those in normal neutrophils. These findings suggest that cIAP2 expression is up-regulated by G-CSF through activation of the JAK2-STAT3 pathway, and increased expression of cIAP2 protein may contribute to G-CSF-mediated antiapoptosis. In addition, overexpression of cIAP2 may be partly responsible for sustained neutrophilia at least in some cases of chronic neutrophilic leukemia.

[Fatal septic shock caused by transrectal needle biopsy of the prostate; a case report].

A 46-year-old man refer to us because of hemospermia. The prostatic gland was normal in size and consistency at rectal examination. Serum prostate specific antigen was 7.04 ng/ml. Magnetic resonance imaging showed an area of low signal intensity on T2-weighted images in the left peripheral gland, possibly indicative of carcinoma. Transrectal prostate biopsy was performed after intravenous administration of piperacillin. He developed chills and fever (39 degrees C) the next morning following biopsy. He was taken unconscious into the hospital where a diagnosis of septic shock caused by Escherichia coli was made. Five days later, he died. His general condition deteriorated notwithstanding intensive treatment. Postmortem blood cultures were positive for a piperacillin resistant Escherichia coli. Histological examination of the biopsies showed a benign prostatic hyperplasia. Autopsy showed diffuse tissue damage in the heart, lung, liver and kidneys. The prostate had numerous microabscesses. Currently, transrectal prostate biopsy is considered a generally reliable procedure to detect adenocarcinoma of the prostate. Our case seems to the sixth case report of fatal complications.

Renal metastasis of thyroid carcinoma.

We report here the sixth known case of metastatic renal tumor of thyroid carcinoma. In 1999, a 37-year-old man was referred to us with a left renal mass found incidentally by ultrasound during an annual check-up. Computed tomography (CT) revealed a well-defined, hyperdense mass 3 cm in diameter in the middle of the left kidney. Left radical nephrectomy was performed under the preoperative diagnosis of renal cell carcinoma. Histologically, the tumor was metastatic of typical papillary thyroid carcinoma.