Lymphoscintigraphy for sentinel lymph node mapping in Japanese patients with malignant skin neoplasms of the lower extremities: comparison with previously investigated Japanese lymphatic anatomy.

OBJECTIVES: Lymph nodes (LN) and lymphatic drainage were identified by lymphoscintigraphy using 99(m)Tc-phytate in order to map the sentinel lymph nodes (SLNs) in patients with malignant skin neoplasms of the lower extremities, and to compare the results with an atlas of Japanese lymphatic anatomy. METHODS: Sentinel lymphoscintigraphs of 18 patients with malignant skin neoplasms of the lower extremities (9 men, 9 women; age range 45-84 years, mean age 66 years) were analyzed retrospectively, and the LNs detected were identified as SLNs or secondary nodes. RESULTS: The patterns of lymphatic drainage were divided into three different categories: (1) initial drainage into inguinal LN without visualization of popliteal LNs (inguinal type), (2) initial drainage into popliteal LNs and then into intrapelvic LNs (popliteal type), and (3) initial drainage into both popliteal and inguinal LNs (inguinal and popliteal type). More than half of the cases were the inguinal and popliteal type, as both inguinal and popliteal LNs were identified as SLNs. In the cases in which the hallux and its surrounding area were injected, all were the inguinal type and popliteal LNs were not visualized. In one case, only dynamic images detected lymphatic drainage without visualization of popliteal LNs. In contrast to the previously published literature on Japanese lymphatic anatomy, SLN lymphatic drainage from the skin of the lower extremities was wide and overlapping in many areas. However, in agreement with currently accepted anatomy, only the great saphenous lymphatic vessel drained the skin of the hallux and its surrounding area. The present results suggest that it is important to confirm lymphatic drainage in order to identify SLNs in the lower extremities. CONCLUSIONS: The patterns of lymphatic drainage from the skin of the foot were divided into three different categories. In contrast to previously published Japanese lymphatic anatomy, lymphatic drainage from the skin of the lower extremities was wide and overlapping in many areas. However, only the great saphenous lymphatic vessel drained the skin of the hallux and its surrounding area in agreement with currently accepted Japanese lymphatic anatomy. It is important to confirm lymphatic drainage to identify SLNs in the lower extremities.

Heavy ion beam irradiation regulates the mRNA expression in megakaryocytopoiesis from human hematopoietic stem/progenitor cells.

Heavy ion beams are a high-LET radiation that has greater biological effect than electron beams or X-rays. However, little is known about the effect of heavy ion beams on the proliferation and differentiation of human hematopoietic stem/progenitor cells (HSPCs). The present study examined the effect of heavy ion beams on gene expression in human HSPCs, especially during early stage of megakaryocytopoiesis. Human CD34+ cells were exposed to monoenergetic carbon-ion beams (290 MeV/nucleon, LET = 50 KeV/m) that were generated by an accelerator (Heavy Ion Medical Accelerator in Chiba). The expression of various genes related to early hematopoiesis, megakaryocytopoiesis/erythropoiesis, cytokine receptors and oxidative stress were analyzed by real-time RT-PCR. Friend leukemia virus integration 1, an early hematopoiesis-related gene, showed significantly higher mRNA expression than the control at 6 hr after irradiation. In contrast, no significant differences were observed in almost all of the other early hematopoiesis-related genes, cytokine receptor-coded genes and megakaryocytopoiesis/erythropoiesis-differentiation pathway-related genes, respectively. An analysis of the response of the genes to oxidative stress revealed the expression of heme oxygenase 1 to show a 1.5-fold and 11.9-fold increase from the day 0 control at 24 hr after 0.5 Gy and 2 Gy irradiation, respectively. Similarly, the NAD(P)H dehydrogenase-quinone 1 expression also showed a 22.0-fold and a 21.8-fold increase at 6 hr in comparison to the initial control. These results showed that the heavy ion beams affect megakaryocytopoiesis/ erythropoiesis differentiation of human HSPCs on the gene expression level.

Incidence of unilateral distal vertebral artery aplasia: evaluation by combining basiparallel anatomic scanning-magnetic resonance imaging (BPAS-MRI) and magnetic resonance angiography.

PURPOSE: The incidence of a unilateral aplastic distal vertebral artery (VA) has been reported as 0.2% of cases on cerebral angiography. During our daily magnetic resonance (MR) examinations, however, we frequently encounter MR angiograms (MRAs) that do not demonstrate unilateral VA. The purpose of this study was to calculate the frequency of aplastic unilateral distal VA by MR images performed for asymptomatic people. MATERIALS AND METHODS: Over a time span of 1 year, 237 asymptomatic people (140 men, 97 women; ages 28-67 years, mean 54.4 years) underwent brain MRI during a "brain check-up examination" in our hospital. To identify an aplastic unilateral distal VA, we retrospectively compared three-dimensional time-of-flight MRA with basiparallel anatomic scanning (BPAS)-MRI which was designed for recognition of the arterial outer contour. RESULTS: Aplasia of the unilateral distal VA was confirmed in 11 persons (4.6%). According to our classification, hypoplastic distal VA in 12 (5.1%) and asymptomatic acquired unilateral distal VA occlusion was also proved in 2 (0.8%). CONCLUSION: We found that the frequency of aplastic unilateral VA was 4.6% in asymptomatic people using a combination of MRA and BPAS-MRI for assessment of an intracranial VA.

Inhibition of transforming growth factor-beta, hypoxia-inducible factor-1alpha and vascular endothelial growth factor reduced late rectal injury induced by irradiation.

Tumor hypoxia and angiogenesis associated with malignant progression have been studied widely. The efficacy of angiogenesis inhibition combined with radiotherapy has been demonstrated in cancer treatment. Here, we studied the effect of hypoxia and angiogenesis inhibition on radiation-induced late rectal injury. The rectum of C57BL/6N mice was irradiated locally with a single dose of 25 Gy. Radiation-induced histological changes were examined at 90 days after irradiation by hematoxylin-eosin (H.E.) staining and azan staining. Pimonidazole was administered and its distribution was assayed by immunohistochemistry staining. Expression of transforming growth factor beta1 (TGF-beta1), hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) was assessed on the fibrotic region using real-time PCR and immunohistochemistry. In addition, the effects of TGF-beta, VEGF and HIF-1alpha on radiation-induced injury were investigated by the administration of neutralizing antibody of TGF-beta, antibody of VEGF or YC-1 (3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole) which was developed as an agent for inhibiting HIF-1 expression after irradiation respectively. Fibrosis and uptake of pimonidazole were found 90 days after irradiation. The expression of TGF-beta1, HIF-1alpha and VEGF significantly increased with the formation of fibrosis induced by irradiation compared with unirradiated controls. In addition, treatment of neutralizing antibody of TGF-beta, antibody of VEGF or YC-1 reduced the development of radiation-induced injury. Our results suggested that radiation-induced hypoxia may play an important role in late rectal injury. Although the inhibition of HIF-1alpha and VEGF reduced the radiation induced late injury, the precise mechanism is still unclear.

Placental/umbilical cord blood-derived mesenchymal stem cell-like stromal cells support hematopoietic recovery of X-irradiated human CD34+ cells.

AIMS: The potential of human mesenchymal stem cell-like stroma prepared from placental/umbilical cord blood for hematopoietic regeneration by X-irradiated hematopoietic stem cells is herein assessed. MAIN METHODS: Placental/umbilical cord blood-derived mesenchymal stem cell-like stromal cells were applied to a regenerative ex vivo expansion of X-irradiated human CD34(+) cells in a serum-free liquid culture supplemented with a combination of interleukine-3 plus stem cell factor plus thrombopoietin. KEY FINDINGS: The total number of cells and of lineage-committed myeloid hematopoietic progenitor cells generated in the co-culture of both non-irradiated and X-irradiated cells with stromal cells was significantly higher than those in the stroma-free culture. In addition, the number of CD34(+) cells and CD34(+)/CD38(-) cells, immature hematopoietic stem/progenitor cells also increased more than the stroma-free culture. The stromal cells produced various types of cytokines, although there was little difference between the co-cultures of non-irradiated and X-irradiated cells with stromal cells. Furthermore, when X-irradiated cells came in contact with stromal cells for 16 h before cytokine stimulation, a similar degree of hematopoiesis was observed, thus suggesting the critical role of cell-to-cell interaction. SIGNIFICANCE: The present results showed the potential efficacy of human mesenchymal stem cell-like stroma for hematopoietic regeneration from irradiated hematopoietic stem/progenitor cells.

Placental/umbilical cord blood-derived mesenchymal stem cell-like stromal cells support hematopoietic recovery of X-irradiated human CD34+ cells.

AIMS: The potential of human mesenchymal stem cell-like stroma prepared from placental/umbilical cord blood for hematopoietic regeneration by X-irradiated hematopoietic stem cells is herein assessed. MAIN METHODS: Placental/umbilical cord blood-derived mesenchymal stem cell-like stromal cells were applied to a regenerative ex vivo expansion of X-irradiated human CD34+ cells in a serum-free liquid culture supplemented with a combination of interleukine-3 plus stem cell factor plus thrombopoietin. KEY FINDINGS: The total number of cells and of lineage-committed myeloid hematopoietic progenitor cells generated in the co-culture of both non-irradiated and X-irradiated cells with stromal cells was significantly higher than those in the stroma-free culture. In addition, the number of CD34+ cells and CD34+/CD38− cells, immature hematopoietic stem/progenitor cells also increased more than the stroma-free culture. The stromal cells produced various types of cytokines, although there was little difference between the co-cultures of non-irradiated and X-irradiated cells with stromal cells. Furthermore, when X-irradiated cells came in contact with stromal cells for 16 h before cytokine stimulation, a similar degree of hematopoiesis was observed, thus suggesting the critical role of cell-to-cell interaction. SIGNIFICANCE: The present results showed the potential efficacy of human mesenchymal stem cell-like stroma for hematopoietic regeneration from irradiated hematopoietic stem/progenitor cells.

Bolus injection of contrast agents with various iodine concentrations and delivery rates for intracranial three-dimensional CT angiography: evaluation of intracranial arteriovenous contrast using a multidetector-row CT scanner.

PURPOSE: We evaluated the difference in computed tomography (CT) attenuation values of the intracranial arterial and venous systems among the various contrast injection protocols (higher iodine delivery rate or higher concentration of the agent) on the source images of intracranial three-dimensional CT angiography (3D-CTA) using a multidetector-row CT (MDCT) scanner. MATERIALS AND METHODS: We used 100 ml of iopamidol 300 at an injection rate of 3.0 ml/s, 100 ml of iopamidol 300 at an injection rate of 3.7 ml/s, and 80 ml of iopamidol 370 at an injection rate of 3.0 ml/s. There were 10 patients in each group. Attenuation values of the bilateral internal carotid arteries (ICAs), basilar artery trunk, bilateral cavernous sinuses (CSs), and Galenic vein were measured quantitatively on the axial CT angiographic source images obtained by four-channel MDCT. RESULTS: Injection of the high-concentration contrast with a higher iodine-delivery rate achieved good arteriovenous contrast at the cavernous portion. With the same rate of iodine delivery, injection of the intermediate concentrate agent increased the CT value of not only the ICAs but also the CSs. CONCLUSION: High-concentration contrast could increase ICA attenuation without intracavernous attenuation gain during the "first-pass" phase.

Effects of a 2-step culture with cytokine combinations on megakaryocytopoiesis and thrombopoiesis from carbon-ion beam-irradiated human hematopoietic stem/progenitor cells.

To evaluate whether the continuous treatment of two cytokine combinations is effective in megakaryocytopoiesis and thrombopoiesis in hematopoietic stem/progenitor cells exposed to heavy ion beams, the effects of a 2-step culture by a combination of recombinant human interleukin-3 (IL-3) + stem cell factor (SCF) + thrombopoietin (TPO), which just slightly protected against carbon-ion beam-induced damages, and a combination of IL-3 + TPO, which selectively stimulated the differentiation of the hematopoietic stem/progenitor cells to megakaryocytes and platelets, were examined. CD34(+)-hematopoietic stem/progenitor cells isolated from the human placental and umbilical cord blood were exposed to carbon-ion beams (LET = 50 keV/microm) at 2 Gy. These cells were cultured under three cytokine conditions. The number of megakaryocytes, platelets and hematopoietic progenitors were assessed using a flow cytometer and a clonogenic assay at 14 and 21 days after irradiation, respectively. However, the efficacy of each 2-step culture was equal or lower than that of using the IL-3 + SCF + TPO combination alone and the 2-step culture could not induce megakaryocytes and platelets from hematopoietic stem/progenitor cells exposed to high LET-radiation such as carbon-ion beams. Therefore, additional cytokines and/or hematopoietic promoting compounds might be required to overcome damage to hematopoietic cells by high LET radiation.

Hypoxia expression in radiation-induced late rectal injury.

Tumor hypoxia and angiogenesis have been studied extensively. However, the relation between normal tissue injury and hypoxia is still unclear. In this study, we investigated the effect of hypoxia on radiation-induced late rectal injury in mice. The rectum of C57BL/6N mice was irradiated locally with a single dose of 25 Gy and the following experiments were performed including hematoxylin-eosin (H. E.) staining, azan staining, real-time PCR, immunohistochemistry and immunofluorescene. Radiation-induced fibrotic changes were observed from 14 days and reached the peak 30 days after irradiation. The expression of transforming growth factor beta1 (TGF-beta1), hypoxia-inducible factor-1alpha (HIF-1alpha), vascular endothelial growth factor (VEGF) and endothelial cell marker CD31 increased significantly with the formation of fibrosis induced by irradiation compared with unirradiated control. In addition, the maximum expression of TGF-beta1, HIF-1alpha and VEGF was found at 14, 30 and 90 days after irradiation, respectively. The temporal changes of cytokines were consistent with the dynamic change of fibrosis. Our data suggests that late normal tissue injury involved various cytokines including hypoxia-induced angiogenic cytokines. These results may have important implications in the understanding of radiation-induced late normal tissue injury.

Individual differences in the radiosensitivity of hematopoietic progenitor cells detected in steady-state human peripheral blood.

The aim of this study is to evaluate the individual differences in radiosensitivity of lineage-committed myeloid hematopoietic progenitors, colony-forming cells (CFC), detected in steady-state human peripheral blood (PB). Mononuclear cells were prepared from the buffy-coat of 30 individuals PB, and were assayed for CFC by semi-solid culture supplemented with cytokines. X irradiation was performed in the range of 0.5-4 Gy at a dose rate of about 80 cGy/min. The mean number of hematopoietic progenitor cells is 5866 alpha 3408 in 1 ml of buffy-coat, suggesting that the erythroid progenitor cells are the major population. The total CFC radiosensitivity parameter D(0) and n value are 1.18 alpha 0.24 and 1.89 alpha 0.98, respectively. Using a linear regression analysis, a statistically significant correlation is observed between the D(0) value and the surviving fraction at 4 Gy (r = 0.611 p < 0.001). Furthermore, we evaluate the relationship between individual radiosensitivity and the level of antioxidants, plasma uric acid, plasma bilirubin, and intracellular glutathione. No statistically significant correlations are observed, however, between the D(0) parameter and the level of antioxidants, plasma uric acid, plasma bilirubin, and intracellular glutathione. The present study demonstrates that there are large individual differences in the radiosensitivity of hematopoietic progenitor cells as detected in steady-state human PB. These differences demonstrate almost no correlation with plasma or intracellular antioxidants. The prediction of individual differences in radiosensitivity of CFC can only be measured by 4 Gy irradiation.

[A case of direct carotid cavernous fistula, whom three-dimensional computed tomographic angiography was helpful for decision making of selective coil embolization].

We reported the case of an 84-year-old female suffering from chemosis and exophthalmos. Carotid arteriograms revealed a direct carotid-cavernous fistula (CCF) but could not clearly show the fistula point because of its high-flow nature. We then performed three-dimensional computed tomographic angiography (3D-CTA) using a multi detector-row CT (MDCT) scanner. Multi-planar reformatted CT images distinctly revealed a right carotid aneurysm at the cavernous portion and a shunting point between the aneurysm and the cavernous sinus. Based on this information, we opted to performa transvenous coil embolization to treat this patient. Successful selective coil embolization was performed at the fistulous point was done with a smaller number of coils than ordinarily required in sinus packing. Performing 3D-CTA by using a MDCT was helpful in decision making for the selective coil embolization to treat direct CCF.

Severe damage of human megakaryocytopoiesis and thrombopoiesis by heavy-ion beam radiation.

Heavy ions have a unique efficacy for tumor control in radiotherapy. To clarify the effects of heavy-ion beams on hematopoietic stem/progenitor cells, the effects of carbon-ion beams on megakaryocytopoiesis and thrombopoiesis in CD34(+) cells derived from human placental and umbilical cord blood were investigated. The cells were exposed to carbon-ion beams (LET = 50 keV/microm) and then were treated with thrombopoietin (TPO) alone or TPO plus other cytokines. Megakaryocytic progenitor cells, such as megakaryocyte colony-forming units (CFU-Meg), were far more sensitive to carbon-ion beams than to X rays, and no restoration of carbon-ion beam-irradiated CFU-Meg by treatment with any cytokine combination was observed. However, total cell expansion in liquid culture was not different after either carbon-ion beam or X irradiation of CD34(+) cells. The activation of gamma-H2AX, a marker of DNA double strand-breaks (DSBs), was promoted by the cytokine treatment in X-irradiated CD34(+) cells but not in carbon-ion-irradiated cells. These results showed that carbon-ion beams inflicted severe damage on megakaryocytopoiesis and thrombopoiesis and that a better combination of cytokines and other agents may be needed to stimulate the recovery of hematopoietic cells and repair this damage.

Clinical outcome of stereotactic body radiotherapy of 54 Gy in nine fractions for patients with localized lung tumor using a custom-made immobilization system.

PURPOSE: The aim of this study was to investigate the clinical outcome of stereotactic body radiotherapy (SBRT) of 54 Gy in nine fractions for patients with localized lung tumor using a custom-made immobilization system. METHODS AND MATERIALS: The subjects were 19 patients who had localized lung tumor (11 primaries, 8 metastases) between May 2003 and October 2005. Treatment was conducted on 19 lung tumors by fixed multiple noncoplanar conformal beams with a standard linear accelerator. The isocentric dose was 54 Gy in nine fractions. The median overall treatment time was 15 days (range 11-22 days). All patients were immobilized by a thermo-shell and a custom-made headrest during the treatment. RESULTS: The crude local tumor control rate was 95% during the follow-up of 9.4-39.5 (median 17.7) months. In-field recurrence was noted in only one patient at the last follow-up. The Kaplan-Meier overall survival rate at 2 years was 89.5%. Grade 1 radiation pneumonia and grade 1 radiation fibrosis were observed in 12 of the 19 patients. Treatment-related severe early and late complications were not observed in this series. CONCLUSION: The stereotactic body radiotherapy of 54 Gy in nine fractions achieved acceptable tumor control without any severe complications. The results suggest that SBRT can be one of the alternatives for patients with localized lung tumors.

Colonization and differentiation of transplanted embryonic stem cells in the irradiated intestine of mice.

Radiation-induced intestinal injury is a common complication in radiotherapy for the cancer located in abdomen or pelvis. However, there is no effective treatment for radiation-induced intestinal injury now. It is therefore important to develop new treatments for radiation-induced intestinal injury. In this study, we investigated whether embryonic stem (ES) cells could be transplanted directly into the radiation-damaged intestine and could colonize and differentiate into the intestinal epithelial cells. The intestines of female nude mice (ICR nu/nu) were irradiated at a single dose of 30 Gy, and were immediately transplanted with male 129/Sv-derived ES cells into the wall of the irradiated intestine by direct injection. The intestine was removed on days 13 to 27 after transplantation. The Y-chromosome DNA of transplanted ES cells in the irradiated intestine was determined by polymerase chain reaction. Colonization and differentiation of transplanted ES cells in the irradiated intestine were analyzed by histological and immunohistochemical methods with antibodies against stage-specific embryonic antigen-1, alpha-smooth muscle actin and cytokeratin AE1/AE3. The cells of donor origin were identified in the intestine of irradiated mice, and intestinal crypt-like structures were observed on day 13 after transplantation. Importantly, we observed that ES cells could differentiate into epithelial cells in the submucosa of irradiated intestine on day 13 and 27 after transplantation. These results suggest that transplanted ES cells could colonize and differentiate in the intestinal intestine. Such a new approach for damaged intestine with transplanted stem cells would be promising.

Attenuation values of the intracranial arterial and venous vessels by bolus injection of various contrast agents: a study with a single-detector helical CT scanner.

PURPOSE: We evaluated the arteriovenous contrast on the source images of intracranial three-dimensional computed tomography (CT) angiography (3D-CTA) using a high-concentration (370 mg I/ml) contrast agent in comparison with intermediate-concentration (300 mg I/ml) contrast. MATERIALS AND METHODS: With a fixed intravenous injection rate and scanning delay, 3D-CTA was performed using a single-detector helical CT scanner in 30 consecutive patients. We used 100 ml of iohexol 300 for 10 patients, 100 ml of iopamidol 300 for 10 patients, and 80 ml of iopamidol 370 for 10 patients. Attenuation values of the bilateral internal carotid arteries, bilateral middle cerebral arteries, basilar artery trunk, bilateral cavernous sinuses, bilateral basal veins, and Galenic vein were measured quantitatively on the axial CT angiographic source images. RESULTS: High-concentration contrast significantly increased the attenuation values of the intracranial arterial system without increasing the attenuation of the venous system. CONCLUSION: High-concentration contrast is helpful for obtaining valuable arteriovenous contrast on source images with intracranial 3D-CTA.

Nuclear accumulation and activation of nuclear factor kappaB after split-dose irradiation in LS174T cells.

Although radiation-induced gene expression has been extensively studied, most of the studies to date have focused on that after single-dose irradiation. As split-dose irradiation, rather than single-dose irradiation, is usual in clinical situations, we investigated the effects of split-dose irradiation on nuclear factor kappaB (NF-kappaB) in the human rectum carcinoma cell line, LS174T. After either single- or split-dose irradiation with a different interval, nuclear localization of NF-kappaB was examined by Western blot and immunofluorescence and its DNA-binding activity was measured by ELISA-based assay. Irradiation-induced NF-kappaB nuclear accumulation and DNA binding activity increased in a dose-dependent manner. The peak of NF-kappaB nuclear accumulation and DNA binding activity was seen 2 to 6 hours after a single dose of 4 Gy irradiation and returned to control levels after 12 hours. In split-dose irradiation, NF-kappaB activity was similar after the first and second doses of 4 Gy irradiation separated by 12 hours. In addition, NF-kappaB activity was decreased by lengthening the interval between irradiation. The cell survival, which was assessed by colony formation assay, showed inverse correlation to this: the surviving fraction was higher after split-dose irradiation than after single-dose irradiation of the same total dose and it increased as the interval between irradiation was lengthened. Thus the present results showed a correlation between NF-kappaB activation and the repair of sublethal damage in split-dose irradiation.

Characteristic features of a high-energy x-ray spectra estimation method based on the Waggener iterative perturbation principle.

We have redeveloped a high-energy x-ray spectra estimation method reported by Iwasaki et al. [A. Iwasaki, H. Matsutani, M. Kubota, A. Fujimori, K. Suzaki, and Y. Abe, Radiat. Phys. Chem. 67, 81-91 (2003)]. The method is based on the iterative perturbation principle to minimize differences between measured and calculated transmission curves, originally proposed by Waggener et al. [R. G. Waggener, M. M. Blough, J. A. Terry, D. Chen, N. E. Lee, S. Zhang, and W. D. McDavid, Med. Phys. 26, 1269-1278 (1999)]. The method can estimate spectra applicable for media at least from water to lead using only about ten energy bins. Estimating spectra of 4-15 MV x-ray beams from a linear accelerator, we describe characteristic features of the method with regard to parameters including the prespectrum, number of transmission measurements, number of energy bins, energy bin widths, and artifactual bipeaked spectrum production.

Regeneration of megakaryocytopoiesis and thrombopoiesis in vitro from X-irradiated human hematopoietic stem cells.

In the present study, we investigated whether X-irradiated hematopoietic stem cells can be induced to undergo megakaryocytopoiesis and thrombopoiesis in vitro using cytokine combinations that have been demonstrated to be effective for conferring increased survival on irradiated human CD34(+) megakaryocytic progenitor cells (colony-forming unit megakaryocytes; CFU-Meg), such as thrombopoietin (TPO), interleukin 3 (IL3), stem cell factor and FLT3 ligand. Culture of nonirradiated CD34(+) cells in serum-free medium supplemented with multiple cytokine combinations led to an approximately 200- to 600-fold increase in the total cell numbers by day 14 of culture. In contrast, the growth of X-irradiated cells was observed to be one-sixth to one-tenth that of the nonirradiated cultures. Similarly, total megakaryocytes were increased by 50- to 130-fold, while culture of X-irradiated cells yielded one-fourth to one-eighth of the control numbers. At this time, CD41(+) particles, which appeared to be platelets, were produced in the medium harvested from nonirradiated and irradiated cultures. Although radiation suppressed cell growth and megakaryocytopoiesis, there were no significant differences in thrombopoiesis between the two types of culture. These results suggest that X-irradiated CD34(+) cells can be induced to undergo nearly normal terminal maturation through megakaryocytopoiesis and thrombopoiesis by stimulation with appropriate cytokine combinations.

The effects of (-)-epigallocatechin-3-gallate on the proliferation and differentiation of human megakaryocytic progenitor cells.

Epigallocatechin-3-gallate (EGCg) has been widely recognized as a powerful antioxidant and free radical scavenger. The effects of EGCg on the proliferation and differentiation of X-irradiated megakaryocytic progenitor cells (colony-forming unit-megakaryocyte, CFU-Meg) using CD34+ cells prepared from human placental and umbilical cord blood have been shown. In the absence of exogenous thrombopoietin (TPO), no colonies are observed in cultures containing or lacking EGCg (1 nM-100 microM). In the presence of TPO, in contrast, EGCg significantly promotes CFU-Meg-derived colony formations within the 10-100-nM range. A 1.5-fold increase in the total number of CFU-Meg has been counted compared with the control. These favorable effects of EGCg are also observed in the culture of CD34+ cells before and after X irradiation with 2 Gy. Moreover, in order to investigate the function of EGCg promoting megakaryocytopoiesis and thrombopoiesis in ex vivo cultures, both non-irradiated and X-irradiated CD34+ cells are grown in liquid cultures supplemented with TPO. In both cultures, EGCg increases the total number of cells and megakaryocytes. It has been suggested that the favorable effects of EGCg reduce the risk factor from radiation damage in megakaryocytopoiesis.

Theoretical considerations for evaluating the degree of random-periodicity of radiographic noise.

Radiographic noise properties have been evaluated using the Wiener spectrum. However, this approach is not appropriate for periodic noise for two reasons. One is that it takes infinite values at the spatial frequencies of periodic noise. The other is that when adopting a numerical integration, it allows unstable values at each spatial frequency, depending on the integral region. Introducing three types of spectra (W(1), W(2), and W(3)) in connection with the Wiener spectrum, we propose a practical approach to evaluation of periodic noise. Radiographic images sometimes contain noise that is not totally random and not perfectly periodic. Therefore, using the Wiener spectrum and the W(1) spectrum, we also propose two factors for evaluation of the degree of random-periodicity of noise containing periodic signals.