A gamma-ray computed tomography scanner for the quantitative measurement of bone density

A special purpose gamma-ray computed tomography scanner has been developed for the precise measurement of bone density in the distal forearm. Details of the scanner hardware and computer analysis technique are given. Suitable phantoms have been used to test the operation of the scanner, which has been used to measure trabecular and cortical bone density with a precision better than 1%.     

组胺H2受体拮抗剂对骨髓造血重建的影响

用组胺H_2受体拮抗剂(甲氰咪胍或呋喃硝胺)处理正常和亚致死量γ-射线照射小鼠,探讨正常体内造血和再生骨髓中造血重建与组胺受体的关系。发现非毒性剂量的甲氰咪胍对正常小鼠骨髓多能造血干细胞(CFU-s)无抑制作用,但可抑制小鼠体内粒单系祖细胞(CFU-GM)的生长和亚致死量照射后CFU-s产率的恢复。组胺可能与骨髓的再生有关,组胺H_2受体拮抗剂可抑制骨髓的造血重建。     

碲化镉（CdTe）探测器的原理及医学应用

本文介绍了一种新型的化合物半导体探测器——CdTe探测器,它具有灵敏度高,探测效率高、能量分辨率好、可以在室温下使用,以及对湿度不敏感和体积小等优点,并介绍了该探测器在医学应用方面的前景。     

Gamma-ray induction of deoxyribonucleic acid synthesis in temperature-sensitive DNA initiation mutants of Escherichia coli

DNA synthesis was followed after gamma-ray irradiation of several different temperature-sensitive mutants with defects in the initiation process. The results indicate that only dnaA and dnaI mutants show induction of supplementary DNA synthesis after gamma-ray irradiation. The induction of DNA synthesis by gamma-ray irradiation was also shown to be recA+ dependent in the dna5 mutant.     

X线、CT、SPECT、PET在评估小鼠的骨成像中的作用

目的利用各种影像诊断设备对正常小鼠的骨进行成像,观察其在小鼠骨成像中最佳成像参数。方法分别使用X线、CT、SPECT、PET对小鼠的骨进行拍摄成像。结果X线和CT均可以清楚地对小鼠的骨组织成像,而SPECT、PET由于其分辨率和特异性不高,成像较模糊。结论X线和CT检查对小鼠的骨成像明显,对小鼠疾病的观察有重要意义。而SPECT、PET对诊断小鼠的骨疾病意义不是很大。     

Fluorine concentrations in bone biopsy samples determined by proton-induced gamma-ray emission and cyclic neutron activation

Fluorine concentrations in bone biopsy samples taken from the iliac crest of subjects, divided into four groups depending on the length of dialysis treatment, and aluminium levels in blood and bone pathology, in terms of osteoporosis, were determined by two instrumental methods. Proton-induced gamma-ray emission (PIGE), making use of the resonance reaction of¹⁹F(p,αγ)¹⁶O at 872 keV, and cyclic neutron activation analysis (CNAA), using the¹⁹F(n,γ)²⁰F reaction in a reactor irradiation facility, were employed. Rutherford backscattering (RBS) was used to calculate the volume, and, hence, mass of the sample excited in PIGE by determining the major element composition of the samples in order to express results in terms of concentration. From this preliminary investigation, a relationship is suggested between fluorine concentrations in bone and aluminium levels in the system.

     

The enhancement of haemopoietic stem cell recovery in irradiated mice by prior treatment with cyclophosphamide.

Studies are reported of the enhancement of stem cell recovery following whole body irradiation as a result of prior administration of cyclophosphamide. It is shown that the much larger enhancement of regeneration observed for the hosts own surviving stem cells, compared to the regeneration of injected bone marrow stem cells, is due to the different numbers of stem cells initiating the regeneration in conjunction with the time course of stem cell regeneration. The results show that the environmental changes produced by cyclophosphmide greatly enhance haemopoietic recovery even though at the dose used this agent is relatively toxic to stem cells. Furthermore it has been shown that the level of stem cell regeneration is nearly independent of the gamma-ray dose in the range 3-8 gray (300-800 rad). If human bone marrow should respond similarly it follows that regeneration produced by cytotoxic drugs administered prior to radiation embodies a considerable safety factor as far as recovery of the haemopoietic system is concerned.     

FISH chromosome analysis of plutonium workers from the Sellafield nuclear facility

Chromosome analysis using a single-color FISH technique to paint three pairs of chromosomes was undertaken on a group of 46 retired plutonium workers with assessed bone marrow doses >60 mSv, 34 of whom were categorized as having robust dosimetry and 12 for whom internal doses were considered less reliable. Comparisons were made with a group of 34 workers with negligible radiation exposure and a group of 34 workers with similar recorded external gamma-ray doses but negligible internal dose. The simple translocation frequency of 17.65 +/- 1.96 x 10(-3) per genome equivalent for the 34 plutonium workers with robust dosimetry was significantly increased in comparison with that of 10.06 +/- 1.16 x 10(-3) per genome equivalent for the unirradiated control group (P = <0.001) and that of 13.55 +/- 1.43 x 10(-3) per genome equivalent for the group with similar external gamma-ray exposure (P = 0.012). Thus, although in vitro studies have indicated that the majority of alpha-particle-irradiated cells suffer complex non-transmissible chromosome damage, in vivo a significant proportion survive with simple exchanges that can be passed on to descendant cells. In contrast, the three groups demonstrated no significant differences in stable complex aberrations. No evidence of an increase in dicentrics or unstable complex aberrations associated with plutonium exposure was observed, and it can therefore be assumed that there is little, if any, ongoing irradiation of mature lymphocytes. The translocation frequency of 12.08 +/- 1.92 x 10(-3) per genome equivalent for the group of 12 plutonium workers with less reliable internal dosimetry could adequately be accounted for by age and external dose and indicates that the internal bone marrow doses are likely to have been overestimated. Cytogenetic analysis can therefore make a valuable contribution to the validation of internal doses from plutonium deposition.     

Chemical Elements of Human Bone Tissue Investigated by Nuclear Analytical and Related Methods

The mass fractions (in milligrams per kilogram given on a dry mass basis) of 69 chemical elements in the intact rib bone of 84 apparently healthy 15- to 58-year-old citizens (38 females and 46 males) of a nonindustrial region were investigated using five instrumental analytical methods: neutron activation analysis with high-resolution spectrometry of short- and of long-lived radionuclides, particle-induced gamma-ray emission spectrometry, inductively coupled plasma atomic emission, and mass spectrometry. The mass fractions of chemical elements were measured for rib bone of both males and females, taken separately and together. The present results indicate that rib bone can be used as exposure monitors in occupational medicine and environmental health studies to assess, or indicate, the body burden of Al, B, Ba, Be, Ca, Cr, F, Li, Mg, Na, Ni, P, Pb, rare-earth elements (REEs), Sr, and Zn. Our data show an exponential increase with the age of the content of REEs in the rib of people living in an ecologically safe region.     

Data for the Reference Man: skeleton content of chemical elements

This study was undertaken to provide reference values of chemical element mass fractions in intact bone of Reference (European Caucasian) Man/Woman. The rib bone samples investigated were obtained from autopsies of 84 apparently healthy 15–58-year-old citizens (38 females and 46 males) of a non-industrial region in the Central European part of Russia who had suffered sudden death. The mass fractions (mg/kg given on a wet mass basis) of 69 elements in these bone samples were measured by using neutron activation analysis with high-resolution spectrometry of short-lived and long-lived radionuclides, particle-induced gamma-ray emission, inductively coupled plasma atomic emission spectrometry, and inductively coupled plasma mass spectrometry including necessary quality control measures. Using published and measured data, mass fraction values of the 79 elements for the rib bone have been derived. Based on accepted rib to skeleton mass fractions and reference values of skeleton mass for Reference Man, the elemental burdens in the skeleton were estimated. These results may provide a representative bases for establishing related reference values for the Russian Reference Man/Woman and for revising and adding current reference values for the International Commission on Radiological Protection. The data presented will also be very valuable for many other applications in radiation protection, radiotherapy radiation dosimetry, and other scientific fields.     

Quantitative analysis of micro-CT imaging and histopathological signatures of experimental arthritis in rats

Micro-computed tomographic (micro-CT) imaging provides a unique opportunity to capture 3-D architectural information in bone samples. In this study of pathological joint changes in a rat model of adjuvant-induced arthritis (AA), quantitative analysis of bone volume and roughness were performed by micro-CT imaging and compared with histopathology methods and paw swelling measurement. Micro-CT imaging of excised rat hind paws (n = 10) stored in formalin consisted of approximately 600 30-mum slices acquired on a 512 x 512 image matrix with isotropic resolution. Following imaging, the joints were scored from H&E stained sections for cartilage/bone erosion, pannus development, inflammation, and synovial hyperplasia. From micro-CT images, quantitative analysis of absolute bone volumes and bone roughness was performed. Bone erosion in the rat AA model is substantial, leading to a significant decline in tarsal volume (27%). The result of the custom bone roughness measurement indicated a 55% increase in surface roughness. Histological and paw volume analyses also demonstrated severe arthritic disease as compared to controls. Statistical analyses indicate correlations among bone volume, roughness, histology, and paw volume. These data demonstrate that the destructive progression of disease in a rat AA model can be quantified using 3-D micro-CT image analysis, which allows assessment of arthritic disease status and efficacy of experimental therapeutic agents.     

Chromosomal aberrations induced by (12)C6+ ions and 60Co gamma-rays in mouse immature oocytes

The ovaries of Kun-Ming strain mice (3 weeks) were irradiated with different doses of (12)C6+ ion or (60)Co gamma-ray. Chromosomal aberrations were analyzed in metaphase II oocytes at 7 weeks after irradiation. The relative biological effectiveness (RBE) of (12)C6+ ion was calculated with respect to 60Co gamma-ray for the induction of chromosomal aberrations. The (12)C6+ ion and 60Co gamma-ray dose-response relationships for chromosomal aberrations were plotted by linear quadratic models. The data showed that there was a dose-related increase in frequency of chromosomal aberrations in all the treated groups compared to controls. The RBE values for (12)C6+ ions relative to 60Co gamma-rays were 2.49, 2.29, 1.57, 1.42 or 1.32 for the doses of 0.5, 1.0, 2.0, 4.0 or 6.0 Gy, respectively. Moreover, a different distribution of the various types of aberrations has been found for (12)C6+ ion and 60Co gamma-ray irradiations. The dose-response relationships for (12)C6+ ion and 60Co gamma-ray exhibited positive correlations. The results from the present study may be helpful for assessing genetic damage following exposure of immature oocytes to ionizing radiation.     

What do biomarkers tell us about the pathogenesis of ankylosing spondylitis?

Biomarkers may provide information that promotes understanding of prognosis, disease activity, and pathogenesis in ankylosing spondylitis. Biomarkers reflecting disease activity (metallo-proteinase-3) and inflammatory lesions on magnetic resonance imaging predict new bone formation and are ameliorated by anti-tumor necrosis factor therapy, yet this treatment may not prevent new bone formation. Moreover, elevated levels of biomarkers reflecting tissue repair (bone-specific alkaline phosphatase) post-treatment together with magnetic resonance imaging indicates such treatment may even promote repair through new bone formation. Tumor necrosis factor regulation of Dickkopf-1 may constitute a molecular brake that controls osteoblastogenesis through wingless and bone morphogenetic proteins in an established inflammatory lesion in ankylosing spondylitis.     

Imaging of alkaline phosphatase activity in bone tissue

The purpose of this study was to develop a paradigm for quantitative molecular imaging of bone cell activity. We hypothesized the feasibility of non-invasive imaging of the osteoblast enzyme alkaline phosphatase (ALP) using a small imaging molecule in combination with (19)Flourine magnetic resonance spectroscopic imaging ((19)FMRSI). 6, 8-difluoro-4-methylumbelliferyl phosphate (DiFMUP), a fluorinated ALP substrate that is activatable to a fluorescent hydrolysis product was utilized as a prototype small imaging molecule. The molecular structure of DiFMUP includes two Fluorine atoms adjacent to a phosphate group allowing it and its hydrolysis product to be distinguished using (19)Fluorine magnetic resonance spectroscopy ((19)FMRS) and (19)FMRSI. ALP-mediated hydrolysis of DiFMUP was tested on osteoblastic cells and bone tissue, using serial measurements of fluorescence activity. Extracellular activation of DiFMUP on ALP-positive mouse bone precursor cells was observed. Concurringly, DiFMUP was also activated on bone derived from rat tibia. Marked inhibition of the cell and tissue activation of DiFMUP was detected after the addition of the ALP inhibitor levamisole. (19)FMRS and (19)FMRSI were applied for the non-invasive measurement of DiFMUP hydrolysis. (19)FMRS revealed a two-peak spectrum representing DiFMUP with an associated chemical shift for the hydrolysis product. Activation of DiFMUP by ALP yielded a characteristic pharmacokinetic profile, which was quantifiable using non-localized (19)FMRS and enabled the development of a pharmacokinetic model of ALP activity. Application of (19)FMRSI facilitated anatomically accurate, non-invasive imaging of ALP concentration and activity in rat bone. Thus, (19)FMRSI represents a promising approach for the quantitative imaging of bone cell activity during bone formation with potential for both preclinical and clinical applications.     

Advanced imaging of the macrostructure and microstructure of bone

Noninvasive and/or nondestructive techniques are capable of providing more macro- or microstructural information about bone than standard bone densitometry. Although the latter provides important information about osteoporotic fracture risk, numerous studies indicate that bone strength is only partially explained by bone mineral density. Quantitative assessment of macro- and microstructural features may improve our ability to estimate bone strength. The methods available for quantitatively assessing macrostructure include (besides conventional radiographs) quantitative computed tomography (QCT) and volumetric quantitative computed tomography (vQCT). Methods for assessing microstructure of trabecular bone noninvasively and/or nondestructively include high-resolution computed tomography (hrCT), micro-computed tomography (muCT), high-resolution magnetic resonance (hrMR), and micromagnetic resonance (muMR). vQCT, hrCT and hrMR are generally applicable in vivo; muCT and muMR are principally applicable in vitro. Although considerable progress has been made in the noninvasive and/or nondestructive imaging of the macro- and microstructure of bone, considerable challenges and dilemmas remain. From a technical perspective, the balance between spatial resolution versus sampling size, or between signal-to-noise versus radiation dose or acquisition time, needs further consideration, as do the trade-offs between the complexity and expense of equipment and the availability and accessibility of the methods. The relative merits of in vitro imaging and its ultrahigh resolution but invasiveness versus those of in vivo imaging and its modest resolution but noninvasiveness also deserve careful attention. From a clinical perspective, the challenges for bone imaging include balancing the relative advantages of simple bone densitometry against the more complex architectural features of bone or, similarly, the deeper research requirements against the broader clinical needs. The considerable potential biological differences between the peripheral appendicular skeleton and the central axial skeleton have to be addressed further. Finally, the relative merits of these sophisticated imaging techniques have to be weighed with respect to their applications as diagnostic procedures requiring high accuracy or reliability on one hand and their monitoring applications requiring high precision or reproducibility on the other.     

Visualizing the “sandwich” structure of osteocytes in their native environment deep in bone in vivo

Osteocytes are the most abundant cells in bone and always the focus of bone research. They are embedded in the highly scattering mineralized bone matrix. Consequently, visualizing osteocytes deep in bone with subcellular resolution poses a major challenge for in vivo bone research. Here we overcome this challenge by demonstrating 3‐photon imaging of osteocytes through the intact mouse skull in vivo. Through broadband transmittance characterization, we establish that the excitation at the 1700‐nm window enables the highest optical transmittance through the skull. Using label‐free third‐harmonic generation (THG) imaging excited at this window, we visualize osteocytes through the whole 140‐μm mouse skull and 155 μm into the brain in vivo. By developing selective labeling technique for the interstitial space, we visualize the “sandwich” structure of osteocytes in their native environment. Our work provides novel imaging methodology for bone research in vivo.      

An Improved Immunostaining and Imaging Methodology to Determine Cell and Protein Distributions within the Bone Environment

Bone is a dynamic tissue that undergoes multiple changes throughout its lifetime. Its maintenance requires a tight regulation between the cells embedded within the bone matrix, and an imbalance among these cells may lead to bone diseases such as osteoporosis. Identifying cell populations and their proteins within bone is necessary for understanding bone biology. Immunolabeling is one approach used to visualize proteins in tissues. Efficient immunolabeling of bone samples often requires decalcification, which may lead to changes in the structural morphology of the bone. Recently, methyl-methacrylate embedding of non-decalcified tissue followed by heat-induced antigen retrieval has been used to process bone sections for immunolabeling. However, this technique is applicable for bone slices below 50-µm thickness while fixed on slides. Additionally, enhancing epitope exposure for immunolabeling is still a challenge. Moreover, imaging bone cells within the bone environment using standard confocal microscopy is difficult. Here we demonstrate for the first time an improved methodology for immunolabeling non-decalcified bone using a testicular hyaluronidase enzyme-based antigen retrieval technique followed by two-photon fluorescence laser microscopy (TPLM) imaging. This procedure allowed us to image key intracellular proteins in bone cells while preserving the structural morphology of the cells and the bone.     

Three‐dimensional intravital imaging in bone research

Intravital imaging has emerged as a novel and efficient tool for visualization of in situ dynamics of cellular behaviors and cell‐microenvironment interactions in live animals, based on desirable microscopy techniques featuring high resolutions, deep imaging and low phototoxicity. Intravital imaging, especially based on multi‐photon microscopy, has been used in bone research for dynamics visualization of a variety of physiological and pathological events at the cellular level, such as bone remodeling, hematopoiesis, immune responses and cancer development, thus, providing guidance for elucidating novel cellular mechanisms in bone biology as well as guidance for new therapies. This review is aimed at interpreting development and advantages of intravital imaging in bone research, and related representative discoveries concerning bone matrices, vessels, and various cells types involved in bone physiologies and pathologies. Finally, current limitations, further refinement, and extended application of intravital imaging in bone research are concluded.      

Effect of Gamma-Ray upon Food Microorganisms

The author has carried out a study concerning a simplified method for the measurement of gamma-ray resistivity of E. coli. This report deals with comparison of each survival curve of 11 species of bacteria and the influence of addition of chemicals on a medium containing E. coli by gamma-ray irradiation by application of this method.