Biomedical subjects

Kunio Doi

Publications and source records attributed to Kunio Doi.

At least 19 recordsLinked to original sources

PubMed2007

Usefulness of texture analysis for computerized classification of breast lesions on mammograms.

This work presents the usefulness of texture features in the classification of breast lesions in 5,518 images of regions of interest, which were obtained from the Digital Database for Screening Mammography that included microcalcifications, masses, and normal cases. Sixteen texture features were used, i.e., 13 were based on the spatial gray-level dependence matrix and 3 on the wavelet transform. The nonparametric K-NN classifier was used in the classification stage. The results obtained from receiver operating characteristic analysis indicated that the texture features can be used for separating normal regions and lesions with masses and microcalcifications, yielding the area under the curve (AUC) values of 0.957 and 0.859, respectively. However, the texture features were not very effective for distinguishing between malignant and benign lesions because the AUC was 0.617 for masses and 0.607 for microcalcifications. The study showed that the texture features can be used for the detection of suspicious regions in mammograms.

Breast Neoplasms↗

PubMed2007

Computer-aided diagnosis for improved detection of lung nodules by use of posterior-anterior and lateral chest radiographs.

RATIONALE AND OBJECTIVES: We developed a computerized scheme for detection of lung nodules in the lateral views of chest radiographs, in order to improve the overall performance in combination with the computer-aided diagnostic (CAD) scheme for posterior-anterior (PA) views. MATERIALS AND METHODS: We used 106 pairs of PA and lateral views of chest radiographs (122 lung nodules) for development of the CAD scheme. In the CAD scheme for lateral views, initial candidates of lung nodules were identified by use of a nodule enhancement filter based on the edge gradients. Thirty-four image features extracted from the original and the nodule-enhanced images were used for the rule-based scheme and for artificial neural networks (ANNs) for removal of some false-positive candidates. The computer performance was evaluated with a leave-one-case-out test method for ANNs. For PA views, we used the existing CAD scheme, which was trained with one-half of 924 chest images and then tested with the remaining images. RESULTS: When the CAD scheme was applied only to PA views, the sensitivity in the detection of lung nodules was 70.5%, with 4.9 false positives per image. Although the performance of the computerized scheme for lateral views was relatively low (60.7% sensitivity with 1.7 false positives per image), the overall sensitivity (86.9%) was improved (6.6 false positives per two views), because 20 (16.4%) of the 122 nodules were detected only on lateral views. CONCLUSIONS: The CAD scheme by use of lateral-view images has the potential to improve the overall performance for detection of lung nodules on chest radiographs when combined with a conventional CAD scheme for standard PA views.

Diagnosis, Computer-Assisted↗

PubMed2006

Etoposide induces apoptosis and cell cycle arrest of neuroepithelial cells in a p53-related manner.

We clarified that etoposide (VP-16), a topoisomerase II inhibitor, induced apoptosis in the mouse fetal brain. Apoptotic mechanisms and cell cycle arrest in this system were investigated. Four mg/kg of VP-16 was injected into pregnant mice on day 12 of gestation (GD12). The cell cycle and expression of protein and mRNA of p53 and its transcriptional target genes were examined in the fetal brain. The number of p53- and p21-protein-positive cells peaked at 4 h after treatment (HAT). The expression of p21 mRNA was significantly increased at 4 HAT and 8 HAT. The expression of fas mRNA was significantly increased from 2 to 12 HAT. Significant expression of puma mRNA was observed from 1 HAT to 48 HAT. Flow cytometric analysis revealed that VP-16 induced S-phase accumulation and G2 arrest at 4 and 8 HAT, and VP-16-induced apoptosis was significantly increased from 4 to 24 HAT. In an experiment using BrdU treatment of pregnant mice, the migration of neuroepithelial cells in the fetuses was delayed as compared to the migration of controls, and BrdU-positive signals were observed in some pyknotic cells from 8 to 12 HAT. The present results suggest that VP-16 might induce cell cycle arrest at G2/M phase and apoptosis in a p53-related manner.

Animals↗

PubMed2006

Automated hepatic volumetry for living related liver transplantation at multisection CT.

PURPOSE: To prospectively compare in vivo hepatic automated volumetry with manual volumetry and measured liver volume. MATERIALS AND METHODS: The study was conducted in accordance with the guidelines of the Institutional Review Board of Kumamoto University (Japan). Patient informed consent was obtained. Preoperative multisection computed tomography (CT) was performed in 35 consecutive patients (21 men, 14 women; mean age, 42.8 years; range, 28-72 years) with hepatic disease awaiting living related liver transplantation. The CT scans covered the entire liver at a section thickness of 2.5 mm. Liver volume was estimated by using both the automated and the manual methods. Actual liver weight was obtained for all patients and was converted to hepatic volume on the basis of a predetermined relationship between actual liver weight and volume. Processing time required for both methods was also recorded. Two-tailed paired t test, correlation coefficient, and Bland-Altman tests were used for statistical analyses. RESULTS: Mean liver weight was 881.7 g +/- 249.8 (standard deviation), and mean measured liver volume was 956.00 cm(3) +/- 280.10. Volumetry performed with the automated and manual methods provided liver volumes of 982.99 cm(3) +/- 301.98 and 937.10 cm(3) +/- 301.31, respectively. There was good correlation between measured and estimated volumes obtained with the automated method (r = 0.792, P < .01). The manual and automated methods required 32.8 minutes +/- 6.9 and 4.4 minutes +/- 1.9, respectively. CONCLUSION: The automated method reduced the time required for volumetry of the liver and provided acceptable measurements.

Adult↗

PubMed2006

Gene expression profiles of drug-metabolizing enzymes (DMEs) in rat liver during pregnancy and lactation.

A cDNA microarray analysis was conducted to examine hepatic gene expression profiles in pregnant and lactating F344 rats compared to a virgin control group using an Affymetrix GeneChip system. Of the approximately 16000 gene transcripts interrogated, more than 1000 were significantly modified in their expression when detected either in late pregnancy (19 days of gestation, GD 19, 513 genes upregulated and 579 downregulated) or on the day of delivery (postpartum 0 day, PPD 0, 497 upregulated and 733 downregulated). Particular interest was paid to the gene expression of drug-metabolizing enzymes (DMEs) and nuclear receptors (NRs). Though the expression of a few genes, those for CYP7A1, CYP51 and Sultx3, increased, the expression of a number of genes encoding DMEs (Phase I and Phase II) and NRs decreased during pregnancy and lactation. Changes in the expression of 9 genes encoding DMEs and NRs were confirmed by quantitative real-time PCR. For all 9 genes tested, overall, the results of the microarray and real-time PCR analyses were in agreement. This is the first application of a microarray analysis to the expression profiling of genes encoding DMEs and NRs in the liver of pregnant and lactating rats. When combined with other studies, the present study may provide a basis for investigating the mechanism of toxicity of environmental or other nonphysiologic chemicals to the fetus and mother and drug safety during pregnancy and lactation.

Animals↗

PubMed2006

Diagnostic imaging over the last 50 years: research and development in medical imaging science and technology.

Over the last 50 years, diagnostic imaging has grown from a state of infancy to a high level of maturity. Many new imaging modalities have been developed. However, modern medical imaging includes not only image production but also image processing, computer-aided diagnosis (CAD), image recording and storage, and image transmission, most of which are included in a picture archiving and communication system (PACS). The content of this paper includes a short review of research and development in medical imaging science and technology, which covers (a) diagnostic imaging in the 1950s, (b) the importance of image quality and diagnostic performance, (c) MTF, Wiener spectrum, NEQ and DQE, (d) ROC analysis, (e) analogue imaging systems, (f) digital imaging systems, (g) image processing, (h) computer-aided diagnosis, (i) PACS, (j) 3D imaging and (k) future directions. Although some of the modalities are already very sophisticated, further improvements will be made in image quality for MRI, ultrasound and molecular imaging. The infrastructure of PACS is likely to be improved further in terms of its reliability, speed and capacity. However, CAD is currently still in its infancy, and is likely to be a subject of research for a long time.

Biological Science Disciplines↗

PubMed2006

[Comparison of LCD and CRT monitors for detection of pulmonary nodules and interstitial lung diseases on digital chest radiographs by using receiver operating characteristic analysis].

Soft copy reading of digital images has been practiced commonly in the PACS environment. In this study, we compared liquid-crystal display (LCD) and cathode-ray tube (CRT) monitors for detection of pulmonary nodules and interstitial lung diseases on digital chest radiographs by using receiver operating characteristic (ROC) analysis. Digital chest images with a 1000x1000 matrix size and a 8 bit grayscale were displayed on LCD/CRT monitor with 2M pixels in each observer test. Eight and ten radiologists participated in the observer tests for detection of nodules and interstitial diseases, respectively. In each observer test, radiologists marked their confidence levels for diagnosis of pulmonary nodules or interstitial diseases. The detection performance of radiologists was evaluated by ROC analyses. The average Az values (area under the ROC curve) in detecting pulmonary nodules with LCD and CRT monitors were 0.792 and 0.814, respectively. In addition, the average Az values in detecting interstitial diseases with LCD and CRT monitors were 0.951 and 0.953, respectively. There was no statistically significant difference between LCD and CRT for both detection of pulmonary nodules (P=0.522) and interstitial lung diseases (P=0.869). Therefore, we believe that the LCD monitor instead of the CRT monitor can be used for the diagnosis of pulmonary nodules and interstitial lung diseases in digital chest images.

Data Display↗

PubMed2006

Cell cycle progression is required for nuclear migration of neural progenitor cells.

In the developing brain, neural progenitor cells in the ventricular zone (VZ) show a typical migration pattern-interkinetic nuclear migration, in which nuclear position within the VZ is correlated with the cell cycle. However, the mechanisms underlying this regulation remain unclear. To clarify whether the cell cycle progression controls nuclear migration of neural progenitor cells, we determined whether chemically induced cell cycle arrest affected nuclear migration patterns in the VZ. Administration of 5-azacytidine (5AzC) or cyclophosphamide (CP) to pregnant mice induced cell cycle arrest in the fetal neural progenitor cells of the telencephalon: 5AzC induced G2/M-phase arrest, and CP induced S-phase arrest. We used 5-bromo-2'-deoxyuridine (BrdU) labeling to determine the position of the cell in the cell cycle and the nuclei within the VZ at the same time. Cells arrested in G2/M-phase stopped migrating in the inner area of the VZ. Cells arrested in S-phase stopped migrating in the outer area. These results indicate that nuclear position within the VZ was correlated with cell cycle phase, even when the cell cycle was disrupted, and that the nuclei of neural progenitor cells can migrate only when their cell cycle is going. Our results suggest that cell cycle regulators might control the machinery of migration through a common regulatory mechanism.

Animals↗

PubMed2006

Cell cycle and cell death regulation of neural progenitor cells in the 5-azacytidine (5AzC)-treated developing fetal brain.

In the developing brain, neural progenitor cells are susceptible to many extrinsic stresses, including DNA damage. We treated pregnant rats with 5-azacytidine (5AzC), a DNA demethylating and damaging agent, to investigate the cellular responses of the fetal brain, focusing on the regulation of proliferation and cell death. 5AzC first induced the accumulation of cells in abnormal mitosis, G2-phase accumulation, and then apoptosis of the neural progenitor cells. Most of the apoptotic cells were in G1 phase. Cell cycle transition studies suggested that G2/M progression was blocked, after which the cells moved to G1 phase or underwent apoptosis. p53, a key factor for response to DNA damage, and some of its target genes showed increased expression in Western blot and DNA microarray analyses. In 5AzC-treated fetal brains of p53-deficient mice, apoptosis did not occur, although G2/M accumulation was induced. These results suggest that, in the developing brain, apoptosis is p53-dependent but that another mechanism governs the G2/M checkpoint. The G2/M regulator, Cdc2, was activated by dephosphorylation through G2/M accumulation, suggesting accelerated entry into mitosis leading to accumulation of cells showing abnormal mitosis. Furthermore, some cells may have died due to mitotic catastrophe. Throughout brain development, various cell cycle and cell death regulation mechanisms provide neural progenitor cells with options for defense from DNA damage.

Animals↗

PubMed2006

Evaluation of the image quality of temporal subtraction images produced automatically in a PACS environment.

The aim of the study is to evaluate the reliable production of temporal subtraction images in a picture archiving and communication system environment and to establish objective criteria for the evaluation of image quality. A total of 117 temporal subtraction chest images (55 in the upright position, 62 in the supine position) were obtained in five consecutive days. In all of these, we confirmed that there were no interval changes on the original images, and cases with diffuse lung disease were excluded. The temporal subtraction images were classified by three chest radiologists into five levels: 5, excellent; 4, good; 3, acceptable; 2, poor; and 1, very poor. The following were examined: (1) the yield of adequate quality of the temporal subtraction images; (2) whether the temporal subtraction images were obtained in the warping or nonwarping mode; and (3) the correlation of the overall subjective image quality with the relative shift angles, relative shift distances, and the standard deviation of gray levels in the temporal subtraction images. The percentages of acceptable temporal subtraction images were 100% and 66% in the upright and supine positions, respectively. Sixteen (26%) of the 62 supine-position images were made in nonwarping mode, whereas all upright images were made in warping mode. Significant correlations were obtained in the relative shift angle (P < 0.05), relative horizontal shift distance (P < 0.05), and standard deviation of gray levels (P < 0.0001). Temporal subtraction images with acceptable image quality were obtained in the upright position. The objective criteria may be useful for the evaluation of image quality.

Humans↗

PubMed2006

Computer-aided nodule detection on digital chest radiography: validation test on consecutive T1 cases of resectable lung cancer.

PURPOSE: To evaluate the usefulness of a commercially available computer-assisted diagnosis (CAD) system on operable T1 cases of lung cancer by use of digital chest radiography equipment. MATERIALS AND METHODS: Fifty consecutive patients underwent surgery for primary lung cancer, and 50 normal cases were selected. All cancer cases were histopathologically confirmed T1 cases. All normal individuals were selected on the basis of chest computed tomography (CT) confirmation and were matched with cancer cases in terms of age and gender distributions. All chest radiographs were obtained with one computed radiography or two flat-panel detector systems. Eight radiologists (four chest radiologists and four residents) participated in observer tests and interpreted soft copy images by using an exclusive display system without and with CAD output. When radiologists diagnosed cases as positives, the locations of lesions were recorded on hard copies. The observers' performance was evaluated by receiver operating characteristic analysis. RESULTS: The overall detectability of lung cancer cases with CAD system was 74% (37/50), and the false-positive rate was 2.28 (114/50) false positives per case for normal cases. The mean A(z) value increased significantly from 0.896 without CAD output to 0.923 with CAD output (P = 0.018). The main cause of the improvement in performance is attributable to changes from false negatives without CAD to true positives with CAD (19/31, 61%). Moreover, improvement in the location of the tumor was observed in 1.5 cases, on average, for radiology residents. CONCLUSION: This CAD system for digital chest radiographs is useful in assisting radiologists in the detection of early resectable lung cancer.

Adult↗

PubMed2006

Computer-aided diagnosis for the detection and classification of lung cancers on chest radiographs ROC analysis of radiologists' performance.

RATIONALE AND OBJECTIVES: The aim of the study is to investigate the effect of a computer-aided diagnostic (CAD) scheme on radiologist performance in the detection of lung cancers on chest radiographs. MATERIALS AND METHODS: We combined two independent CAD schemes for the detection and classification of lung nodules into one new CAD scheme by use of a database of 150 chest images, including 108 cases with solitary pulmonary nodules and 42 cases without nodules. For the observer study, we selected 48 chest images, including 24 lung cancers, 12 benign nodules, and 12 cases without nodules, from the database to investigate radiologist performance in the detection of lung cancers. Nine radiologists participated in a receiver operating characteristic (ROC) study in which cases were interpreted first without and then with computer output, which indicated locations of possible lung nodules, together with a five-color scale illustrating the computer-estimated likelihood of malignancy of the detected nodules. RESULTS: Performance of the CAD scheme indicated that sensitivity in detecting lung nodules was 80.6%, with 1.2 false-positive results per image, and sensitivity and specificity for classification of nodules by use of the same database for training and testing the CAD scheme were 87.7% and 66.7%, respectively. Average area under the ROC curve value for detection of lung cancers improved significantly (P = .008) from without (0.724) to with CAD (0.778). CONCLUSION: This type of CAD scheme, which includes two functions, namely detection and classification, can improve radiologist accuracy in the diagnosis of lung cancer.

Clinical Competence↗

PubMed2006

Usefulness of artificial neural network for differential diagnosis of hepatic masses on CT images.

RATIONALE AND OBJECTIVE: Our purpose in this study is to apply an artificial neural network (ANN) for differential diagnosis of certain hepatic masses on computed tomographic (CT) images and evaluate the effect of ANN output on radiologist diagnostic performance. MATERIALS AND METHODS: We collected 120 cases of hepatic disease. We used a single three-layer feed-forward ANN with a back-propagation algorithm. The ANN is designed to differentiate four hepatic masses (hepatocellular carcinoma, intrahepatic peripheral cholangiocarcinoma, hemangioma, and metastasis) by using nine clinical parameters and 24 radiological findings in dual-phase contrast-enhanced CT images. Thus, the ANN consisted of 33 input units and four output units. Subjective ratings for the 24 radiological findings were provided independently by two attending radiologists. All clinical cases were used for training and testing of the ANN by implementation of a round-robin technique. In the observer test, CT images of all 120 cases (30 cases for each disease) were used. CT images were viewed by seven radiologists first without and then with ANN output. Radiologist performance was evaluated by using receiver operating characteristic (ROC) analysis on a continuous rating scale. RESULTS: Averaged area under the ROC curve for ANN alone was 0.961. The diagnostic performance of seven radiologists increased from 0.888 to 0.934 (P < .02) when they used ANN output. CONCLUSION: The ANN can provide useful output as a second opinion to improve radiologist diagnostic performance in the differential diagnosis of hepatic masses seen on contrast-enhanced CT.

Aged, 80 and over↗

PubMed2006

Improving radiologists' recommendations with computer-aided diagnosis for management of small nodules detected by CT.

RATIONALE AND OBJECTIVES: To evaluate how computer-aided diagnosis (CAD) can improve radiologists' recommendations for management of possible early lung cancers on CT. MATERIALS AND METHODS: Twenty-eight lung cancers and 28 benign lesions were employed. Each group of 28 lesions was classified into subgroups of two sizes (9 between 6 and 10 mm and 19 between 11 and 20 mm) and three patterns (8 with pure ground glass opacity [GGO], 12 with mixed GGO and 8 solid lesions). Sixteen radiologists participated in the observer study, first without and then with CAD. Radiologists' recommendations, including (1) follow-up in 12 months, (2) in 6 months, (3) in 3 months, or (4) biopsy, were compared at three levels of their malignancy probability ratings (low: 1%-33%; medium: 34%-66%; high: 67%-99%) for 896 observations (56 lesions by the 16 radiologists) in the two size subgroups and three patterns. RESULTS: The number of recommendations changed by radiologists by use of CAD was 163 (18%) among all 896 observations. Among these changed recommendations, the fraction showing a beneficial effect from CAD was 68% (111/163), and the fraction showing a beneficial effect regarding biopsy recommendations was 69% (48/70). With CAD, the radiologists' performance regarding biopsy recommendations was significantly improved for 43 lung cancers (31 changed to biopsy versus 12 changed away from biopsy; P = .003) and was also improved for 27 benign lesions (10 changed to biopsy versus 17 changed away from biopsy; P = .18). Most of the cancers with improved recommendations were solid lesions or mixed GGO and relatively large. CONCLUSION: CAD has the potential to improve the appropriateness of radiologists' recommendations for small malignant and benign lesions on CT scans.

Aged↗

PubMed2006

Computer-aided diagnosis scheme for identifying histological classification of clustered microcalcifications by use of follow-up magnification mammograms.

RATIONALE AND OBJECTIVES: Our purpose in this study was to investigate the usefulness of follow-up magnification mammograms (i.e., both current and previous magnification mammograms) in a computer-aided diagnosis (CAD) scheme for identifying the histological classification of clustered microcalcifications. MATERIALS AND METHODS: Our database consisted of current and previous magnification mammograms obtained from 93 patients before and after 3-month follow-up: 11 invasive carcinomas, 19 noninvasive carcinomas of the comedo type, 25 noninvasive carcinomas of the noncomedo type, 23 mastopathies, and 15 fibroadenomas. In our CAD scheme, we extracted five objective features of clustered microcalcifications from each of the current and previous magnification mammograms by taking into account image features that experienced radiologists commonly use to identify histological classifications. These features were then merged by a modified Bayes discriminant function for distinguishing among five histological classifications. For the input of the modified Bayes discriminant function, we used five objective features obtained from the previous magnification mammogram (previous features), five objective features obtained from the current magnification mammogram (current features), and the set of the five previous features and the five current features. RESULTS: The classification accuracies with the five current features were higher than those with the five previous features. These classification accuracies were improved substantially by using the set of the five previous features and the five current features. For the set of the five previous features and the five current features, the classification accuracies of our CAD scheme were 81.8% (9 of 11) for invasive carcinoma, 84.2% (16 of 19) for noninvasive carcinoma of the comedo type, 76.0% (19 of 25) for noninvasive carcinoma of the noncomedo type, 73.9% (17 of 23) for mastopathy, and 86.8% (13 of 15) for fibroadenoma. CONCLUSION: Our CAD scheme with use of follow-up magnification mammograms improved classification performance for mammographic clustered microcalcifications.

Algorithms↗

PubMed2006

Evidence of apoptosis in the subventricular zone and rostral migratory stream in the MPTP mouse model of Parkinson disease.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is commonly used to create animal models of Parkinson disease. There is conflicting evidence on the occurrence of apoptosis induced by MPTP in the mouse substantia nigra pars compacta. We demonstrated that a single acute injection of MPTP induced apoptosis in the subventricular zone (SVZ) and rostral migratory stream (RMS) in the adult C57BL/6 mouse brain. The number of TUNEL-positive cells peaked at 24 hours after injection and decreased thereafter, paralleling the change in the number of cleaved caspase-3-positive cells after MPTP injection. Results of immunohistochemistry and ultrastructural analyses indicated that the majority of apoptotic cells in the SVZ and RMS were migrating neuroblasts (type A cells), whereas a few were astrocytes (type B cells). No apoptosis occurred in transit-amplifying progenitors (type C cells). The decrease in A cell numbers was most marked on day 2 and lasted to day 8 after the administration. A rapid and transient phagocytosis of apoptotic cells by microglial cells was demonstrated to parallel the MPTP-induced apoptosis. The present findings provide new insight into the extensive neurotoxicity of MPTP and may be valuable in reevaluating the MPTP mouse model of Parkinson disease.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

PubMed2006

Image-processing technique for suppressing ribs in chest radiographs by means of massive training artificial neural network (MTANN).

When lung nodules overlap with ribs or clavicles in chest radiographs, it can be difficult for radiologists as well as computer-aided diagnostic (CAD) schemes to detect these nodules. In this paper, we developed an image-processing technique for suppressing the contrast of ribs and clavicles in chest radiographs by means of a multiresolution massive training artificial neural network (MTANN). An MTANN is a highly nonlinear filter that can be trained by use of input chest radiographs and the corresponding "teaching" images. We employed "bone" images obtained by use of a dual-energy subtraction technique as the teaching images. For effective suppression of ribs having various spatial frequencies, we developed a multiresolution MTANN consisting of multiresolution decomposition/composition techniques and three MTANNs for three different-resolution images. After training with input chest radiographs and the corresponding dual-energy bone images, the multiresolution MTANN was able to provide "bone-image-like" images which were similar to the teaching bone images. By subtracting the bone-image-like images from the corresponding chest radiographs, we were able to produce "soft-tissue-image-like" images where ribs and clavicles were substantially suppressed. We used a validation test database consisting of 118 chest radiographs with pulmonary nodules and an independent test database consisting of 136 digitized screen-film chest radiographs with 136 solitary pulmonary nodules collected from 14 medical institutions in this study. When our technique was applied to nontraining chest radiographs, ribs and clavicles in the chest radiographs were suppressed substantially, while the visibility of nodules and lung vessels was maintained. Thus, our image-processing technique for rib suppression by means of a multiresolution MTANN would be potentially useful for radiologists as well as for CAD schemes in detection of lung nodules on chest radiographs.

Algorithms↗

PubMed2006

Repair process of fetal brain after 5-azacytidine-induced damage.

The fetal brain is susceptible to many extrinsic stresses. Some of these stresses induce excessive cell death in the prenatal stage, leading to anomalies in the neonatal brain. However, it is unclear how the developing brain responds to and repairs the prenatal tissue damage. We treated pregnant rats on day 13 of gestation with 5-azacytidine, one of the compounds that induces excessive cell death and inhibits proliferation in neural progenitor cells, to damage the fetal brain, and investigated the repair process up to 60 h after treatment. Histological analysis showed that 5-azacytidine induced strong apoptosis of neural cells. By 60 h, apoptotic cells disappeared and the tissue was repaired, although the telencephalic wall remained thinner than in controls. Flow cytometry analysis showed that the cell cycle distribution also returned to control levels at 60 h, suggesting that the repair process was completed around 60 h. During the repair period, amoeboid microglia infiltrated the brain and ingested the apoptotic cells. These microglial cells were positive for the multiple microglial markers, and mRNAs for the microglia-related cytokines tumor necrosis factor alpha, interleukin 1beta and macrophage colony stimulating factor (M-CSF) were up-regulated. DNA microarray analysis showed the up-regulation of genes relevant to glial cells, inflammation, the extracellular matrix, glycolysis, proliferation and neural development. We show here that the developing brain has the capacity to respond to the damage induced by extrinsic chemical stresses, including changing the expression of numerous genes and the induction of microglia to aid the repair process.

Animals↗