Identification of fetal blood stains by radioimmunoassay of α1-fetoprotein

Summary Radioimmunoassay of ₁-fetoprotein(AFP) for medico-legal identification of fetal blood stains using a commercial kit is described. The AFP content in fetal blood stains on filter paper ranged from 21 – 320 ng/9 mm². The protein was detected in stains of adult blood and retroplacental blood in only negligible amounts. Aging of the blood stains did not influence the values up to 1 month. The method is simple and sensitive enough for application to medico-legal-practice.     

Formation and decay of the E1′ center and of its precursor

The E₁′ center has been used for ESR dating of quartz with assuming that the signal intensity increases with natural radiation dose as those of other ESR signals do. However, this simple assumption is not necessarily correct. Formation and decay of the E₁′ center are closely related with its precursor, diamagnetic oxygen vacancies. Gamma ray of large dose (>100 kGy) creates oxygen vacancies giving little dose rate effect, which, therefore, might be useful for dating of granites and high dose dosimetry.     

Radiolabeling of the biologically active peptide 163–171 of human interleukin-1β

N-succinimidyl[2,3-³H]propionate was used for the radiolabeling of the biologically active peptide fragment 163–171 of human interleukin-1β (VQGEESNDK). Suitable reaction conditions were studied to obtain useful labeling of the molecule. A mixture of mono- (70%) and bi- (30%) propionyl derivatives was obtained with a total ³H specific activity of 87 Ci/mmol of peptide. The conditions for an efficient chromatographic separation of labeled peptide from unreacted reagents and by-products were established. The labeled peptide maintained the same biological activity as that of the corresponding unlabeled molecule, indicating that the labeling procedure did not alter the biological characteristics of the peptide. This thus allows the use of the radiolabeled peptide for receptor binding studies.     

Reproducibility of volume and densitometric measures of emphysema on repeat computed tomography with an interval of 1?week

Objectives

The reproducibilities of CT lung volume and densitometric measures of emphysema were assessed over 1?week. The influence of breathhold on reproducibility was assessed.

Methods

HRCT was performed on 44 subjects at inspiration on two visits with a 7-day interval. CT lung volume, relative area below -950HU (RA950-raw), and 15th percentile density (PD15-raw) were computed. Volume correction was used to obtain RA950-adj and PD15-adj. Reproducibilities between visits were assessed using concordance correlation coefficient (CCC) and repeatability coefficient (RC). Reproducibilities were compared between raw and adjusted measures. Differences between visits were computed for volume and density measures. Correlations were computed for density differences versus volume difference. Subgroup analysis was performed using a 0.25?L volume difference threshold.

Results

High CCC were observed for all measures in full group (CCC?>?0.97). Reproducibilities of volume (RC?=?0.67?L), RA950-raw (RC?=?2.3%), and PD15-raw (RC?=?10.6HU) were observed. Volume correction significantly improved PD15 (RC?=?3.6HU) but not RA950 (RC?=?1.7%). RA950-raw and PD15-raw had significantly better RC in <0.25?L subgroup than ??0.25?L. Significant correlations with volume were observed for RA950-raw and PD15-raw (R ²?>?0.71), but not RA950-adj or PD15-adj (R ²?Conclusions Good breathhold and RA950 reproducibilities were achieved. PD15 was less reproducible but improved with volume correction or superior breathhold reproduction.

Key Points

? Good breath-hold reproducibility is achievable between multiple CT examinations. ? Reproducibility of densitometric measures may be improved by statistical volume correction. ? Volume correction may result in decreased signal. ? Densitometric reproducibility may also be improved by achieving good breath-hold reproduction. ? Careful consideration of signal and noise is necessary in reproducibility assessment.     

Disruption of Hif-1α enhances cytotoxic effects of metformin in murine squamous cell carcinoma

Purpose: In the present study, we investigated whether the disruption of the Hif-1α gene affects the sensitivity of SCC VII cells to metformin and also if metformin functions as a radiosensitizer using murine squamous cell carcinoma (SCC VII) cells.

Materials and methods: Cultured SCC VII and SCC VII Hif-1α-deficient cells were incubated with metformin under glucose-free and/or hypoxia-mimetic conditions and cell viabilities were measured. Tumor-bearing mice were continuously given 5-bromo-2’-deoxyuridine (BrdU) to label all proliferating cells. Tumor-bearing mice were then subjected to γ-ray irradiation after the metformin treatment. Immediately after irradiation, cells were isolated from some tumors and incubated with a cytokinesis blocker. The responses of quiescent and total (=?proliferating?+?quiescent) cell populations were assessed based on the frequency of micronuclei using immunofluorescence staining for BrdU.

Results: The disruption of Hif-1α increased the sensitivity of SCC VII cells to metformin in glucose-free medium. Metformin-induced decreases in the percentage of dead cells in the presence of CoCl₂ were partially reduced when Hif-1α was disrupted. In vivo, metformin increased the radiosensitivity of SCC VII Hif-1α-deficient cells.

Conclusion: The combination of disruption of Hif-1α and metformin effectively enhanced the radiosensitivity of SCC VII cells.     

BOLD-MRI of breast invasive ductal carcinoma: correlation of R2* value and the expression of HIF-1α

Objective

To explore the reliability and feasibility of blood oxygenation level-dependent-based functional magnetic resonance imaging (BOLD-fMRI) to depict hypoxia in breast invasive ductal carcinoma.

Methods

A total of 103 women with 104 invasive ductal carcinomas (IDCs) underwent breast BOLD-fMRI at 3.0 T. Histological specimens were analysed for tumour size, grade, axillary lymph nodes and expression of oestrogen receptors, progesterone receptors, human epidermal growth factor receptor 2, p53, Ki-67 and hypoxia inducible factor 1α (HIF-1α). The distribution and reliability of R2* were analysed. Correlations of the R2* value with the prognostic factors and HIF-1α were respectively analysed.

Results

The R2* map of IDC demonstrated a relatively heterogeneous signal. The mean R2* value was (53.4?±?18.2) Hz. The Shapiro–Wilk test (W?=?0.971, P?=?0.020) suggested that the sample did not follow a normal distribution. The inter-rater and intrarater correlation coefficient was 0.967 and 0.959, respectively. The R2* values of IDCs were significantly lower in patients without axillary lymph nodes metastasis. The R2* value had a weak correlation with Ki67 expression (r?=?0.208, P?=?0.038). The mean R2* value correlated moderately with the level of HIF-1α (r?=?0.516, P?=?0.000).

Conclusion

BOLD-fMRI is a simple and non-invasive technique that yields hypoxia information on breast invasive ductal carcinomas.

Key Points

? Blood oxygenation level-dependent-based MRI can be used to assess tumour hypoxia. ? BOLD-fMRI shows characteristic features of breast invasive ductal carcinoma. ? R2* values of BOLD-fMRI correlate with hypoxia inducible factor 1α.     

Effect of Reducing Chemotherapy Drug Doses on the Level of Index of Liver Fibrosis 、TGF-β1 and Recent Curative Effect of HPC

目的 探索减少经导管动脉化疗栓塞术(TACE)中化疗药剂量对原发性肝癌(PHC)患者肝纤维化指标、血清转化生长因子-β1( TGF-β1)水平及短期疗效的影响.资料与方法 36例不可手术切除的PHC患者接受超选择性TACE,按入选标准随机分为两组,A组(n=15,小剂量组)给予小剂量化疗药物;B组(n=21,常规剂量组)给予常规剂量化疗药物.结果 两组患者治疗前后血清学指标比较,常规剂量组术后三项纤维化指标透明质酸(HA)、人Ⅲ型前胶原(hPC-Ⅲ)、Ⅳ-C型胶原(Ⅳ-C)及TGF-β1水平均明显高于手术前(P＜0.01),层粘连蛋白(LN)水平较术前升高(P＜0.05);小剂量组术后四项肝纤维化指标较术前无显著性差异(P＞0.05),TGF-β1水平较术前升高(P＜0.05);近期疗效比较两组无显著性差异(P＞0.05).结论 TACE治疗PHC,适当减少化疗药物剂量可减轻肝纤维化的发生,有利于保护肝功能,而并不影响到患者近期疗效.     

Prognostic value of HIF-1α expression during fractionated irradiation

Background and purpose

Hypoxia and reoxygenation are important determinants of outcome after radiotherapy. HIF-1α is a key molecule involved in cellular response to hypoxia. HIF-1α expression levels have been shown to change after irradiation. The objective of the present study was to explore the prognostic value of HIF-1α expression during fractionated irradiation.

Materials and methods

Six human squamous cell carcinoma models xenografted in nude mice were analysed. Tumours were excised after 3, 5 and 10 fractions. HIF-1α expression was quantified by western blot. For comparative analysis, previously published data on local tumour control data and pimonidazole hypoxic fraction was used.

Results

HIF-1α expression in untreated tumours exhibited intertumoural heterogeneity and did not correlate with pimonidazole hypoxic fraction. During fractionated irradiation the majority of tumour models exhibited a decrease in HIF-1α expression, whereas in UT-SCC-5 no change was observed. Neither kinetics nor expression levels during fractionated irradiation correlated with local tumour control.

Conclusion

Our data do not support the use of HIF-1α determined during treatment as a biomarker to predict outcome after fractionated irradiation.     

Inhalation of [123I]alpha1-protease inhibitor: toward a new therapeutic concept of alpha1-protease inhibitor deficiency?

The alpha1-protease inhibitor (alpha1-Pi) is separated from human serum and is therefore extremely expensive. Because only 2%-3% concentrates in the lung after intravenous administration, inhalational therapy for alpha1-Pi deficiency would seem likely to be better. The aims of this study were therefore to determine the pattern of deposition of inhaled alpha1-Pi labeled with 123I and measure the amount deposited in the lungs. METHODS: Eighteen patients with congenital severe alpha1-Pi deficiency were enrolled in the study. The low-specific-activity 123I-labeled alpha1-Pi aerosol (median particle size +/- SD, 3.9 +/- 2.5 microm) was generated by an air pressure-driven nebulizer. The patients inhaled for an average of 23.6 +/- 8.9 min. Static scintigrams in two projections were acquired immediately after (T1) and 1 (T2), 4 (T3), and 24 h (T4) after inhalation. The patients were divided into the following three groups according to their forced expiratory volume in 1 s (FEV1): group I, < or =40% of predicted normal (n = 8); group II, 40% < FEV1 < or = 60% of predicted normal (n = 4); group III, >60% of predicted normal (n = 6). RESULTS: The absolute percentage uptake values of alpha1-Pi in group I were 12.4 for T1, 7.3 for T2, 4.6 for T3, and 1.2 for T4; in group II the values were 13.0, 9.6, 6.2, and 2.0, respectively; and in group III, 14.6, 11.4, 6.5, and 3.6, respectively. Differences between the groups were generally statistically significant. Between T1 and T2, the probability value was <0.05 for group I versus group II, <0.006 for group I versus group III, and <0.39 for group II versus group III. Between T1 and T3, the probability value was <0.29 for group I versus group II, <0.22 for group I versus group III, and <0.94 for group II versus group III. Retention (between T1 and T4) was also dependent on the grade of the disease: P < 0.2 for group I versus group II, P < 0.001 for group I versus group III, and P < 0.02 for group II versus group III. Grading of the uptake pattern by three independent experienced investigators (87% agreement) revealed a peripheral deposition that was group dependent. We found that greater peripheral deposition corresponded with lower lung functional impairment: P < 0.5 for group I versus group II, P < 0.01 for group I versus group III, and P < 0.08 for group II versus group III. Degradation also corresponded with functional impairment: P < 0.05 for group I versus group II, P < 0.006 for group I versus group III, and P < 0.3 for group II versus group III. CONCLUSION: The results of this study show that sufficient amounts of alpha1-Pi can be deposited in the periphery of the lung by inhalation at least in patients with low-grade disease. Inhalation of alpha1-Pi may thus represent a new and more convenient route of drug administration.     

Application of [11C]SA4503 to selection of novel σ1 selective agonists

IntroductionThe σ₁ ligands are considered to be a new class of potential therapeutic agents for several types of central nervous system disorder. Carbon-11-labeled 1-[2-(3,4-dimethoxyphenyl)ethyl]-4-(3-phenylpropyl)piperazine ([¹¹C]SA4503) was shown to be a promising PET ligand for mapping σ₁ receptors, and was applied to measure receptor occupancy with several therapeutic drugs in the living human brain. In this study, we applied this technique for preclinical in vivo screening of novel σ₁ selective agonists.MethodsSix newly synthesized piperazine derivatives containing arylalkylamine groups and cyclohexylamine derivatives containing phenyl groups were selected and tested for their in vivo σ₁ receptor binding with [¹¹C]SA4503. The test compounds were administered by intravenous co-injection or oral administration. The in vivo receptor binding of [¹¹C]SA4503 was evaluated by a tissue dissection method at a single time point.ResultsOur in vivo screen identified the most promising candidate of novel σ₁ agonist in the piperazine derivatives. Some correlations between in vitro affinity and in vivo receptor blocking rate were observed when considering oral bioavailability. In vivo receptor blocking of piperazine derivatives after oral administration may be predictable by simple co-injection study.ConclusionLigand selection with [¹¹C]SA4503 by the in vivo receptor binding assay was performed successfully. This technique is a practical and high-throughput method that can directly evaluate blood–brain barrier permeability, receptor binding, and bioavailability of drug candidates at the same time.     

Measurement of the G-value for 1·5 keV X-rays

Summary

Although there are several theoretical predictions of the dependence of the G-value on X-ray energy, measurements have not been made below ≈ 7 keV. Using a ferrous sulfate solution modified by the addition of benzoic acid, we have measured the relative G-values for Al_k characteristic X-rays (1·5 keV), ²³⁸Pu α-particles (3·7 MeV), ⁶⁰Co (1·17 MeV) and ¹³⁷Cs (0·66 MeV) γ-rays. This modified ferrous sulfate solution gave a 4-fold increase in sensitivity relative to the conventional solution, making measurements with the Al_k X-rays feasible. The relative ferrous-to-ferric conversions as a function of dose were similar for the two γ-ray energies, yielding G-values of 1·62 and 1·59 µmol J^?1 for the ⁶⁰Co and ¹³⁷Cs radiations, respectively. The α-particle G-value was 0·52 µmol J^?1, or 31 per cent of that for the ⁶⁰Co γ-rays, in good agreement with previous measurements. The Al_k X-rays had a G-value of 0·92 µmol J^?1 or 57 per cent of that of the ⁶⁰Co radiation. This G-value for the 1·5 keV X-rays is within 20 per cent of the values predicted by current theories, and theoretical values are within the error range of our measurement. The consistency between the experimental value reported here and theoretical G-values for ultrasoft X-rays should be valuable for models of radiation action on biological systems.     

Anti-inflammatory effect of low-dose X-irradiation and the involvement of a TGF-β 1 -induced down-regulation of leukocyte/endothelial cell adhesion

Purpose : Low-dose radiotherapy (LD-RT) is known to exert an anti-inflammatory effect, but the underlying radiobiological and immunological mechanisms remain elusive. In recent studies, we observed a reduced adhesion of peripheral blood mononuclear cells (PBMC) to endothelial cells (EC) after LD-RT (0.3-0.7 Gy). This shows that this treatment affects the initial steps of the inflammatory response. To explore the role of inflammatory mediators in this process, we investigated the expression of Transforming growth factor β 1 (TGF- β 1) and Interleukin 6 (IL-6) after LD-RT. Materials and Methods : EC were grown to subconfluence and irradiated with single-dose LD-RT. Twenty-hours after irradiation, EC were treated with IL-1 β for 4 h and then incubated with peripheral blood mononuclear cells (PBMC). Adherent PBMC were counted when using light microscopy. Expression of the cytokines TGF- β 1 and IL-6 was measured by ELISA, and mRNA levels were analysed by the RNAse-protection assay (RPA). Surface expression of E-selectin was quantified by flow cytometry. Results : A relative minimum of adhesion was observed after LD-RT between 0.3 and 0.7 Gy. This was paralleled by an expression maximum of TGF- β 1 and IL-6, as shown by protein and mRNA levels, respectively. Neutralization of TGF- β 1 by monoclonal antibodies, but not of IL-6, increased PBMC adhesion to EC nearly to control levels. In addition, fluorescence activated cell sorter (FACS) analysis of irradiated EC demonstrated a down-regulation of E-selectin in the same dose range. Conclusion : Low-dose X-irradiation between 0.3 and 0.7 Gy induced a relative maximum of TGF- β 1 production by stimulated EC. This results in a down-regulation of leukocyte/PBMC adhesion and may contribute to the anti-inflammatory effect of LD-RT.     

1st Scientific Meeting of the Association of Radiobiologists of the Euratom Countries Rapallo,Italy. 14–16 December 1961

Summary

The response of cultured CHO cells to U.V.L. irradiation during treatment with anisotonic solutions shows that treatment with hypotonic sucrose, NaCl or KCl solutions causes an increase in the cellular U.V.L. sensitivity, while exposure to hypertonic solutions causes a large decrease in U.V.L. sensitivity. Cells exposed to 1·8 M sucrose, NaCl or KCl solutions and given a U.V.L. dose of 252 erg/mm² towards the end of the 20 min solution exposure time have survival levels which are respectively 228, 26, and 23 times higher than the controls, i.e. cells irradiated in phosphate buffered saline.

Cell volume data obtained using a Coulter counter, and nuclear area data of attached cells obtained using an optical microscope with a micrometer reticle, show that cell and nuclear size are related to U.V.L. sensitivity. That is, as cells shrink and the nuclear area decreases, the cells become more U.V.L.-resistant. During hypotonic treatment with 0·1 M NaCl, the cell volume, nuclear area and U.V.L. sensitivity increased in the first 2 to 4 min of exposure time, but at longer exposure times (greater than 3 to 4 min), cell volume, nuclear area and cellular U.V.L. sensitivity decreased. For 0·1 M KCl treatment the cells initially displayed a rapid increase in volume, nuclear area and U.V.L. sensitivity, but at the longer exposure times no decrease in cell and nuclear size were observed, and a slight increase in U.V.L. sensitivity occurred. Changes in U.V.L. sensitivity were related to changes in nuclear size and cell volume; however, calculations showed that during hypertonic treatment there is an ionic effect as well as an osmotic effect. That is, the cellular U.V.L. survival in equal hypertonic concentrations of NaCl or KCl was lower than in the same concentration of sucrose.     

A Technique of “Chimney Nellix” for the Management of Type 1a Endoleak After EVAR

This report presents 3 procedures with visceral “chimney stenting” in conjunction with an endovascular aneurysm sealing (EVAS) device, known as chEVAS, for treatment of type 1a endoleak. It includes the first published chEVAS in a patient with previous fenestrated endovascular aneurysm repair (FEVAR). Cases include an 80-year-old man 8 years after FEVAR for a juxtarenal abdominal aortic aneurysm (AAA); an 85-year-old woman 9 months after endovascular aneurysm repair (EVAR) for a ruptured infrarenal AAA; and an 84-year-old woman 3 months after EVAR for a symptomatic infrarenal AAA. Technical success was achieved in all cases, with 1 postoperative death. The remaining 2 patients had no residual type 1a endoleak at 10 and 14 months respectively.     

Value of the radiolabelled GLP-1 receptor antagonist exendin(9–39) for targeting of GLP-1 receptor-expressing pancreatic tissues in mice and humans

Purpose  

Radiolabelled glucagon-like peptide 1 (GLP-1) receptor agonists have recently been shown to successfully image benign insulinomas in patients. Moreover, it was recently reported that antagonist tracers were superior to agonist tracers for somatostatin and gastrin-releasing peptide receptor targeting of tumours. The present preclinical study determines therefore the value of an established GLP-1 receptor antagonist for the in vitro visualization of GLP-1 receptor-expressing tissues in mice and humans.     

Stabilization of [99mTc]HMPAO—1. Ethanolic preparation

The use of [^99mTc]HMPAO for cerebral blood flow imaging has been hampered by the short useful life time of the labelled radiopharmaceutical. Preparation of [^99mTc]HMPAO in 85% ethanol was found to increase the in vitro stability (92% radiochemical purity at 90 min) in comparison to the aqueous preparation (79% at 90 min). Biological distribution studies in mice indicated similar brain uptake (4.42 ± 1.02 and 4.12 ± 0.82% per gram at 20 min) and brain:blood ratio (0.97 ± 0.27 and 1.03 ± 0.20 at 20 min) of the ethanolic (2 h after preparation) and aqueous (15 min after preparation) formulations of [^99mTc]HMPAO respectively. The improved in vitro stability of [^99mTc]HMPAO prepared in 85% ethanol may prove useful in instances where previously the 30 min time limit precluded its use.