Yoji Urata

Elevated expression of inducible nitric oxide synthase and inflammatory cytokines in the alveolar macrophages after esophagectomy.

Objective To evaluate the role of inducible nitric oxide (NO) synthase (iNOS) and inflammatory cytokines in alveolar macrophages (AMs) after esophagectomy in the pathogenesis of acute lung injury. Design Prospective, exploratory, open-labeled clinical study. Setting Intensive care unit and operating room in a university hospital. Patients Thirteen patients receiving esophagectomy with carcinoma of the esophagus (postesophagectomy group), ten patients just before the surgery (preoperation group), and seven patients receiving surgery less invasive than esophagectomy (other surgery group) were selected. Interventions Bronchoalveolar lavage fluid (BALF) and blood samples were obtained from study groups. Measurements and Main Results The AMs in the BALF collected from each group were stained immunohistochemically with antibodies against iNOS, interleukin (IL)-6, and IL-8. The intensities of these expressions were determined by semiquantitative cytofluorometric system. NOx (NO2−+NO3−), IL-6, and IL-8 levels in the BALF and plasma were measured concurrently. The expressional intensities of iNOS, IL-6, and IL-8 in AMs obtained from the postesophagectomy group were maximal 24 hrs after the skin incision and significantly more evident than those from other groups. The IL-6, IL-8, and NOx levels in BALF and IL-6 and IL-8 levels in plasma in the postesophagectomy patients were also elevated. The intensities of iNOS and inflammatory cytokines expressions in AMs were closely related to postoperative respiratory failure. Conclusions The activation of topical alveolar macrophages may be involved in the pathogenesis of pulmonary complications in the postoperative period after esophagectomy.

Simultaneous signalling through c-mpl, c-kit and CXCR4 enhances the proliferation and differentiation of human megakaryocyte progenitors: possible roles of the PI3-K, PKC and MAPK pathways

We assessed the effect of signalling through CXCR4 on the proliferation and differentiation of human megakaryocytic progenitor cells (CFU‐Meg) in the presence or absence of stem cell factor (SCF) and/or thrombopoietin (TPO), using peripheral blood‐derived CD34+IL‐6R− cells as a target. TPO alone induced a significant number of CFU‐Meg colonies. Although stromal cell‐derived factor‐1 (SDF‐1) or SCF alone did not support CFU‐Meg colony formation, these factors had a synergistic effect on CFU‐Meg colony formation in the presence of TPO. The combination of SDF‐1, SCF and TPO induced twice as many CFU‐Meg colonies as TPO alone. To investigate the mechanism of this synergistic action, we examined the effects of various protein kinase inhibitors on CFU‐Meg colony formation. LY294002 and GF109203X (inhibitors of PI3‐K and PKC respectively) completely or partially inhibited this synergistic action. In contrast, a MEK inhibitor (PD98059) did not inhibit CFU‐Meg colony formation. It significantly increased the higher ploidy classes (16N to 64N) of megakaryocytes supported by TPO, TPO + SCF, TPO + SDF‐1, and TPO + SCF + SDF‐1, whereas it abolished the effect of SDF‐1 on the increase of higher ploidy classes of megakaryocytes supported by TPO. These results suggest that MAPK may negatively or positively regulate the nuclear maturation of megakaryocytes, known as endomitosis. In the presence of PD98059, proplatelet formation (PPF) was significantly augmented, suggesting that the MAPK pathway may also inhibit the initiation of PPF. In conclusion, simultaneous activation of three signals through c‐mpl, c‐kit and CXCR4 can induce the in vitro proliferation and differentiation of CFU‐Meg, and SDF‐1 is a potentiator of human megakaryocytopoiesis.

Role of human interleukin-9 as a megakaryocyte potentiator in culture

OBJECTIVE This study investigated the effect of interleukin-9 (IL-9) on the proliferation and differentiation of human colony-forming unit megakaryocytic progenitor cells (CFU-Meg). MATERIALS AND METHODS Peripheral blood-derived CD34(+)IL-6R(-) cells were sorted and cultured in the presence of IL-9, erythropoietin (Epo), stem cell factor (SCF), and thrombopoietin (TPO) alone or in combination. The number of pure and mixed megakaryocyte colonies, the size of pure megakaryocyte colonies, the ploidy distribution of megakaryocytes, and proplatelet formation were investigated. RESULTS Apart from TPO, no single factor could support CFU-Meg-derived colony formation, but each two-factor combination among IL-9, Epo, and SCF supported a few CFU-Meg colonies. Interestingly, the combination of Epo+SCF+IL-9 induced four to six times as many CFU-Meg colonies as any of the two-factor combinations. Neutralizing monoclonal antibodies (mAbs) for IL-9 receptor and c-kit completely abolished this synergistic effect. In contrast, addition of neutralizing anti-c-Mpl or anti-CXCR4 Abs did not influence colony formation, indicating that this synergistic effect was independent of TPO or SDF-1. Moreover, the endogenous production of TPO by cultured CD34(+)IL-6R(-) cells in the presence of Epo+SCF+IL-9 was ruled out by reverse transcriptase polymerase chain reaction for TPO mRNA. Interestingly, the combination of TPO, Epo, SCF, and IL-9 supported the largest number of pure and mixed megakaryocyte colonies, suggesting that this combination of cytokines might recruit primitive megakaryocytic as well as multipotential progenitors. This combination also potently enhanced proplatelet formation compared with TPO alone or a combination of Epo, SCF, and IL-9. CONCLUSION This study demonstrated for the first time that human IL-9 can potentiate human megakaryocytopoiesis in the presence of Epo and/or SCF.

Primary central chondrosarcoma of long bone, limb girdle and trunk: Analysis of 174 cases by numerical scoring on histology.

The aims of this study were: (i) to elucidate clinicopathological characteristics of pcCHS of long bones (L), limb girdles (LG) and trunk (T) in Japan; (ii) to investigate predictive pathological findings for outcome of pcCHS of L, LG and T, objectively; and (iii) to elucidate a discrepancy of grade between biopsy and resected specimens. Clinicopathological profiles of 174 pcCHS (79 male, 95 female), of L, LG, and T were retrieved. For each case, a numerical score was given to 18 pathological findings. The average age was 50.5 years (15–80 years). Frequently involved sites were femur, humerus, pelvis and rib. The 5‐year and 10‐year disease‐specific survival (DSS) rates [follow‐up: 1–258 months (average 65.5)] were 87.0% and 80.4%, respectively. By Cox hazards analysis on pathological findings, age, sex and location, histologically higher grade and older age were unfavorable predictors, and calcification was a favorable predictor in DSS. The histological grade of resected specimen was higher than that of biopsy in 37.7% (26/69 cases). In conclusion, higher histological grade and older age were predictors for poor, but calcification was for good prognosis. Because there was a discrepancy in grade between biopsy and resected specimens, comprehensive evaluation is necessary before definitive operation for pcCHS.

Primary Intra-aortic Epstein-Barr Virus–Positive Large B-Cell Lymphoma Presenting as Aortic Mural Thrombosis: An Entity Distinct From Intravascular Large B-Cell Lymphoma

Intravascular selective growth of neoplastic B lymphocytes is a characteristic finding of intravascular large B-cell lymphoma (IVLBCL). However, because neoplastic B cells of IVLBCL grow merely in the lumina of capillaries or small vessels, primary IVLBCL of the great vessels is considered exceptional. To our knowledge, only 2 primary B-cell lymphomas in the lumina of the vena cava have been reported. However, there has been no report of primary B-cell lymphoma with intra-aortic growth. We describe a novel manifestation of primary Epstein-Barr virus–positive large B-cell lymphoma mainly affecting the lumina of the aorta and its major branches in a 76-year-old man. He had a long-term fever that was refractory to antibiotics and aortic mural thrombosis with visceral embolization. Because he had no detectable mass suggesting a malignancy, it was difficult to diagnose while he was alive. He died without anticancer treatment, and the confirmed diagnosis was made at autopsy.

A long-term survival case of multiple hepatocellular carcinoma with metachronous lymph node metastasis.

A long-term survival case of multiple hepatocellular carcinoma (HCC) with metachronous metastasis to a lymph node is reported. The patient, a 66-year-old woman, had two primary HCC nodules, one each in the left and right hepatic lobes, which were resected. She developed a lymph node lesion and a secondary HCC 45 and 62 months after the first operation, respectively. She has been well for the 7 years since the first operation despite undergoing hepatic resection for HCC twice as well as lymph node resection. Clonal analysis, based on the methylation pattern of the X chromosome-linked androgen receptor gene, suggested that the two primary tumors were multicentric and that the lymph node lesion had arisen by metastasis from the primary tumor in the right hepatic lobe.

Medical tele-education system with superhigh-definition (SHD) image viewer

We have been studying a medical tele-education support system by an individual tutoring system, called CALAT, and a super high definition (SHD) image processing system, called SuperFM-III. Now, we are in a trial operation to use the SuperFM-III for a super high definition image control viewer on the CALAT client side, and have created the courseware of the pathological images. In this paper, we show the concept and the implementation of this system.

EGF binding activity of rat hepatocytes in liver regeneration after partial hepatectomy: Cell cycle analysis by cytofluorometry

Abstract To analyze EGF binding activity in liver regeneration at cell cycle level, we performed the double measurement of nuclear DNA content and cellular bound EGF content in freshly isolated rat hepatocytes from normal and regenerating liver after 67% hepatectomy using autostaging cytofluorometry. The data demonstrated that the bound EGF content of resting cells increased in proportion to their DNA content, while cycling cells had significantly consistently lower bound hEGF throughout cell cycle. These changes are supposed to reflect ‘down-regulation’ of EGF receptor at the cell cycle level, and the data suggested a major role of endogenous EGF in cell proliferation during liver regeneration.