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To determine whether temporal subtraction (TS) CT obtained with non-rigid image registration improves detection of various bone metastases during serial clinical follow-up examinations by numerous radiologists. Six board-certified radiologists retrospectively scrutinized CT images for patients with history of malignancy sequentially. These radiologists selected 50 positive and 50 negative subjects with and without bone metastases, respectively. Furthermore, for each subject, they selected a pair of previous and current CT images satisfying predefined criteria by consensus. Previous images were non-rigidly transformed to match current images and subtracted from current images to automatically generate TS images. Subsequently, 18 radiologists independently interpreted the 100 CT image pairs to identify bone metastases, both without and with TS images, with each interpretation separated from the other by an interval of at least 30 days. Jackknife free-response receiver operating characteristics (JAFROC) analysis was conducted to assess observer performance. Compared with interpretation without TS images, interpretation with TS images was associated with a significantly higher mean figure of merit (0.710 vs. 0.658; JAFROC analysis, P = 0.0027). Mean sensitivity at lesion-based was significantly higher for interpretation with TS compared with that without TS (46.1% vs. 33.9%; P = 0.003). Mean false positive count per subject was also significantly higher for interpretation with TS than for that without TS (0.28 vs. 0.15; P

To develop and evaluate a deep learning-based algorithm (DLA) for automatic detection of bone metastases on CT.This retrospective study included CT scans acquired at a single institution between 2009 and 2019. Positive scans with bone metastases and negative scans without bone metastasis were collected to train the DLA. Another 50 positive and 50 negative scans were collected separately from the training dataset and were divided into validation and test datasets at a 2:3 ratio. The clinical efficacy of the DLA was evaluated in an observer study with board-certified radiologists. Jackknife alternative free-response receiver operating characteristic analysis was used to evaluate observer performance.A total of 269 positive scans including 1375 bone metastases and 463 negative scans were collected for the training dataset. The number of lesions identified in the validation and test datasets was 49 and 75, respectively. The DLA achieved a sensitivity of 89.8% (44 of 49) with 0.775 false positives per case for the validation dataset and 82.7% (62 of 75) with 0.617 false positives per case for the test dataset. With the DLA, the overall performance of nine radiologists with reference to the weighted alternative free-response receiver operating characteristic figure of merit improved from 0.746 to 0.899 (p.001). Furthermore, the mean interpretation time per case decreased from 168 to 85 s (p = .004).With the aid of the algorithm, the overall performance of radiologists in bone metastases detection improved, and the interpretation time decreased at the same time.• A deep learning-based algorithm for automatic detection of bone metastases on CT was developed. • In the observer study, overall performance of radiologists in bone metastases detection improved significantly with the aid of the algorithm. • Radiologists' interpretation time decreased at the same time.

To compare observer performance of detecting bone metastases between bone scintigraphy, including planar scan and single-photon emission computed tomography, and computed tomography (CT) temporal subtraction (TS). Data on 60 patients with cancer who had undergone CT (previous and current) and bone scintigraphy were collected. Previous CT images were registered to the current ones by large deformation diffeomorphic metric mapping; the registered previous images were subtracted from the current ones to produce TS. Definitive diagnosis of bone metastases was determined by consensus between two radiologists. Twelve readers independently interpreted the following pairs of examinations: NM-pair, previous and current CTs and bone scintigraphy, and TS-pair, previous and current CTs and TS. The readers assigned likelihood levels to suspected bone metastases for diagnosis. Sensitivity, number of false positives per patient (FPP), and reading time for each pair of examinations were analysed for evaluating observer performance by performing the Wilcoxon signed-rank test. Figure-of-merit (FOM) was calculated using jackknife alternative free-response receiver operating characteristic analysis. The sensitivity of TS was significantly higher than that of bone scintigraphy (54.3% vs. 41.3%, p = 0.006). FPP with TS was significantly higher than that with bone scintigraphy (0.189 vs. 0.0722, p = 0.003). FOM of TS tended to be better than that of bone scintigraphy (0.742 vs. 0.691, p = 0.070). Sensitivity of TS in detecting bone metastasis was significantly higher than that of bone scintigraphy, but still limited to 54%. TS might be superior to bone scintigraphy for early detection of bone metastasis. • Computed tomography temporal subtraction was helpful in early detection of bone metastases. • Sensitivity for bone metastasis was higher for computed tomography temporal subtraction than for bone scintigraphy. • Figure-of-merit of computed tomography temporal subtraction was better than that of bone scintigraphy.

To assess whether temporal subtraction (TS) images of brain CT improve the detection of suspected brain infarctions. Study protocols were approved by our institutional review board, and informed consent was waived because of the retrospective nature of this study. Forty-two sets of brain CT images of 41 patients, each consisting of a pair of brain CT images scanned at two time points (previous and current) between January 2011 and November 2016, were collected for an observer performance study. The 42 sets consisted of 23 cases with a total of 77 newly developed brain infarcts or hyperdense artery signs confirmed by two radiologists who referred to additional clinical information and 19 negative control cases. To create TS images, the previous images were registered to the current images by partly using a non-rigid registration algorithm and then subtracted. Fourteen radiologists independently interpreted the images to identify the lesions with and without TS images with an interval of over 4 weeks. A figure of merit (FOM) was calculated along with the jackknife alternative free-response receiver-operating characteristic analysis. Sensitivity, number of false positives per case (FPC) and reading time were analyzed by the Wilcoxon signed-rank test. The mean FOM increased from 0.528 to 0.737 with TS images (p < 0.0001). The mean sensitivity and FPC improved from 26.5% and 0.243 to 56.0% and 0.153 (p < 0.0001 and p = 0.239), respectively. The mean reading time was 173 s without TS and 170 s with TS (p = 0.925). The detectability of suspected brain infarctions was significantly improved with TS CT images. • Although it is established that MRI is superior to CT in the detection of strokes, the first choice of modality for suspected stroke patients is often CT. • An observer performance study with 14 radiologists was performed to evaluate whether temporal subtraction images derived from a non-rigid transformation algorithm can significantly improve the detectability of newly developed brain infarcts on CT. • Temporal subtraction images were shown to significantly improve the detectability of newly developed brain infarcts on CT.

Purpose To determine the improvement of radiologist efficiency and performance in the detection of bone metastases at serial follow-up computed tomography (CT) by using a temporal subtraction (TS) technique based on an advanced nonrigid image registration algorithm. Materials and Methods This retrospective study was approved by the institutional review board, and informed consent was waived. CT image pairs (previous and current scans of the torso) in 60 patients with cancer (primary lesion location: prostate, n = 14; breast, n = 16; lung, n = 20; liver, n = 10) were included. These consisted of 30 positive cases with a total of 65 bone metastases depicted only on current images and confirmed by two radiologists who had access to additional imaging examinations and clinical courses and 30 matched negative control cases (no bone metastases). Previous CT images were semiautomatically registered to current CT images by the algorithm, and TS images were created. Seven radiologists independently interpreted CT image pairs to identify newly developed bone metastases without and with TS images with an interval of at least 30 days. Jackknife free-response receiver operating characteristics (JAFROC) analysis was conducted to assess observer performance. Reading time was recorded, and usefulness was evaluated with subjective scores of 1-5, with 5 being extremely useful and 1 being useless. Significance of these values was tested with the Wilcoxon signed-rank test. Results The subtraction images depicted various types of bone metastases (osteolytic, n = 28; osteoblastic, n = 26; mixed osteolytic and blastic, n = 11) as temporal changes. The average reading time was significantly reduced (384.3 vs 286.8 seconds; Wilcoxon signed rank test, P = .028). The average figure-of-merit value increased from 0.758 to 0.835; however, this difference was not significant (JAFROC analysis, P = .092). The subjective usefulness survey response showed a median score of 5 for use of the technique (range, 3-5). Conclusion TS images obtained from serial CT scans using nonrigid registration successfully depicted newly developed bone metastases and showed promise for their efficient detection. © RSNA, 2017 Online supplemental material is available for this article.

Temporal subtraction of CT (TS) images improves detection of newly developed bone metastases (BM). We sought to determine whether TS improves detection of BM by radiology residents as well. We performed an observer study using a previously reported dataset, consisting of 60 oncology patients, each with previous and current CT images. TS images were calculated using in-house software. Four residents independently interpreted twice the 60 sets of CT images, without and with TS. They identified BM by marking suspicious lesions likely to be BM. Lesion-based sensitivity and number of false positives per patient were calculated. Figure-of-merit (FOM) was calculated. Detectability of BM, with and without TS, was compared between radiology residents and board-certified radiologists, as published previously. FOM of residents significantly improved by implementing TS (p value

We have developed a novel medical device software program that is intended to improve radiologists’ performance with regard to the detection of bone metastases on CT images. To establish the protocol of the clinical performance evaluation for regulatory approval, the feasibility of nonclinical study (NCS) and clinical study (CS) conforming to the principles of GCP was compared. There were two main issues with establishing the protocol in Japan. Firstly, retrospective case collection was not accepted for CS due to concerns regarding selection bias. However, it is difficult to prospectively collect cases for use in an observer study due to the wide variety of bone metastases cases required. Secondly, when setting the reference standard (RS) for bone metastases, the utilization of any patient information was not accepted for NCS due to concerns regarding an inappropriate RS setting. However, it was predicted that it would be difficult to set accurate RSs only using CT images. Consequently, following discussion with a Japanese regulatory body, the protocol was established as an NCS using retrospectively collected CT images. Furthermore, we agreed to use anonymized clinical images to set the RS and to rescue the test results by reviewing the RS after the observer study, hence ensuring reliability. An NCS was a feasible method of evaluating the novel medical device software while also complying with quality management systems. By using real-world clinical image databases, this clinical performance evaluation method may be both effective and efficient as a path to regulatory approval of certain medical device software.

In our previous study, we developed a computer-aided diagnosis (CADx) system using imaging findings annotated by radiologists. The system, however, requires radiologists to input many imaging findings. In order to reduce such an interaction of radiologists, we further developed a CADx system using derived imaging findings based on calculated image features, in which the system only requires few user operations. The purpose of this study is to check whether calculated image features (CFT) or derived imaging findings (DFD) can represent information in imaging findings annotated by radiologists (AFD). We calculate 2282 image features and derive 39 imaging findings by using information on a nodule position and its type (solid or ground-glass). These image features are categorized into shape features, texture features and imaging findings-specific features. Each imaging finding is derived based on each corresponding classifier using random forest. To check whether CFT or DFD can represent information in AFD, under an assumption that the accuracies of classifiers are the same if information included in input is the same, we constructed classifiers by using various types of information (CTT, DFD and AFD) and compared accuracies on an inferred diagnosis of a nodule. We employ SVM with RBF kernel as classifier to infer a diagnosis name. Accuracies of classifiers using DFD, CFT, AFD and CFT $$+$$ AFD were 0.613, 0.577, 0.773 and 0.790, respectively. Concordance rates between DFD and AFD of shape findings, texture findings and surrounding findings were 0.644, 0.871 and 0.768, respectively. The results suggest that CFT and AFD are similar information and CFT represent only a portion of AFD. Particularly, CFT did not contain shape information in AFD. In order to decrease an interaction of radiologists, a development of a method which overcomes these problems is necessary.

Although hospital information systems (HIS) have recently become more widely used, data stored in HIS databases are not fully utilized in clinical research. One major reason is the lack of suitable tools to utilize the stored data for clinical research. Patient eligibility criteria for clinical studies often consist of multiple conditions incorporating logical operations across multiple data tables. These are too complex to perform using the existing HIS interfaces, which were originally designed for clinical business rather than clinical research. Moreover, these criteria sometimes require importing patient lists from external data sources not stored in the HIS databases. Certainly end-user computing (EUC) is effective in satisfying these demands, but in reality, EUC is too difficult for many clinical researchers to master because it requires them to understand the complicated composition of the data tables in the HIS database. Thus, we have developed a search system to meet these needs using user-friendly interfaces. Since the objective of a clinical study is to prove a relationship between a certain purported cause and the outcome, target patients must be selected based on several items of clinical information. Patient eligibility criteria for clinical studies are usually complex, consisting of multiple conditions combined with logical "AND", "OR" and "NOT" operations as well as parentheses. It is a challenge to design user interfaces that can specify logical operations and parentheses among multiple search conditions. Our search system is designed to support set operations between the lists of search results as well as functions for loading and saving multiple lists, instead of logical operations and parentheses among search conditions. The system was developed using a middleware package for EUC, and was released in our institute. In this study, both the proposed search system and search interfaces for our institutional HIS were evaluated using application forms submitted to the Informatics Department requesting search and export of data stored in the HIS database. The requests were processed using both systems, and the results were recorded and analyzed for evaluation. Our search system fulfilled a significantly larger number of requests than the HIS interfaces (paired t-test, p

Quality assurance (QA) guidelines for medical display systems in Japan, JESRA X-0093, were published in August 2005 and have been used in many medical fields to maintain image quality on medical displays. This report offers detailed explanations of terms and testing methodologies in the guidelines, taking into account users with little knowledge of display technology. The management grade classifications, luminance meters, test patterns, and evaluation methods for executing the QA are supplementally described based on the technical background of related things. In addition, the validity of the evaluation methods and judgment criteria for uniformity and contrast response tests were examined in some experiments. The experimental results of the contrast response indicated that some cases presented inadequate display contrast even if the contrast responses were set within ± 15% of the standard acceptable range for grade 1. The luminance responses of displays used in two computed tomography systems (CTs) and one magnetic resonance imaging system (MRI) were also measured, and the results indicated that their responses with conventional gamma responses were problematic for comparing images with those of medical displays.

We aimed to describe the development of an inference model for computer-aided diagnosis of lung nodules that could provide valid reasoning for any inferences, thereby improving the interpretability and performance of the system. An automatic construction method was used that considered explanation adequacy and inference accuracy. In addition, we evaluated the usefulness of prior experts’ (radiologists’) knowledge while constructing the models. In total, 179 patients with lung nodules were included and divided into 79 and 100 cases for training and test data, respectively. F-measure and accuracy were used to assess explanation adequacy and inference accuracy, respectively. For F-measure, reasons were defined as proper subsets of Evidence that had a strong influence on the inference result. The inference models were automatically constructed using the Bayesian network and Markov chain Monte Carlo methods, selecting only those models that met the predefined criteria. During model constructions, we examined the effect of including radiologist’s knowledge in the initial Bayesian network models. Performance of the best models in terms of F-measure, accuracy, and evaluation metric were as follows: 0.411, 72.0%, and 0.566, respectively, with prior knowledge, and 0.274, 65.0%, and 0.462, respectively, without prior knowledge. The best models with prior knowledge were then subjectively and independently evaluated by two radiologists using a 5-point scale, with 5, 3, and 1 representing beneficial, appropriate, and detrimental, respectively. The average scores by the two radiologists were 3.97 and 3.76 for the test data, indicating that the proposed computer-aided diagnosis system was acceptable to them. In conclusion, the proposed method incorporating radiologists’ knowledge could help in eliminating radiologists’ distrust of computer-aided diagnosis and improving its performance.

A Japanese adult female voxel (volume pixel) phantom (JF) was developed using CT images of a healthy female volunteer. The height (152 cm) and weight (44 kg) of JF are smaller than the averages of Japanese adult female. The voxel size of JF is 0:98 × 0:98 × 1mm3. Therefore, the shapes of small or complicated organs, such as thyroid and stomach, are distinctly reproduced. The shapes of female-specific organs, such as ovary and uterus, in JF are also realistically represented compared with those of a previously developed phantom. These results suggest that JF enables the accurate evaluation of organ doses due to diverse exposures to Japanese adult females. Photon-specific absorbed fractions (SAFs) for liver or stomach as target organs and for kidney as a source organ were calculated using JF and compared with those of other phantoms. It was concluded that the SAFs calculated using JF and other phantoms depend on the organ distances from source to target.

Objective The purpose of this study was to investigate the relationship between visual score of emphysema and homology-based emphysema quantification (HEQ) and evaluate whether visual score was accurately predicted by machine learning and HEQ. Materials and methods A total of 115 anonymized computed tomography images from 39 patients were obtained from a public database. Emphysema quantification of these images was performed by measuring the percentage of low-attenuation lung area (LAA%). The following values related to HEQ were obtained: nb0 and nb1. LAA% and HEQ were calculated at various threshold levels ranging from −1000 HU to −700 HU. Spearman’s correlation coefficients between emphysema quantification and visual score were calculated at the various threshold levels. Visual score was predicted by machine learning and emphysema quantification (LAA% or HEQ). Random Forest was used as a machine learning algorithm, and accuracy of prediction was evaluated by leave-one-patient-out cross validation. The difference in the accuracy was assessed using McNemar’s test. Results The correlation coefficients between emphysema quantification and visual score were as follows: LAA% (−950 HU), 0.567; LAA% (−910 HU), 0.654; LAA% (−875 HU), 0.704; nb0 (−950 HU), 0.552; nb0 (−910 HU), 0.629; nb0 (−875 HU), 0.473; nb1 (−950 HU), 0.149; nb1 (−910 HU), 0.519; and nb1 (−875 HU), 0.716. The accuracy of prediction was as follows: LAA%, 55.7% and HEQ, 66.1%. The difference in accuracy was statistically significant (p = 0.0290). Conclusion LAA% and HEQ at −875 HU showed a stronger correlation with visual score than those at −910 or −950 HU. HEQ was more useful than LAA% for predicting visual score.

Purpose: Because of the recent innovative growth in computer technology, digital imaging, and the Internet, we can take advantage of these facilities for education and clinical work in nuclear medicine. We developed a tele-nuclear medicine conference system with electronic mail (e-mail) on the Internet.Methods: Twenty-one physicians (20 radiologists, 1 neurologist), 6 technologists and 2 medical students in six university hospitals (Japan 5, Canada 1), 5 local hospitals in Japan participated in this project. We used digital still cameras (330k pixels) equipped with a floppy disk drive and 10 x optical zoom to digitize images with JPEG compression (640×480 matrix). The images were attached to e-mail messages (containing a brief description of each case). The mail was sent simultaneously to all members on the mailing list. Scintigram and SPECT images as well as other radiological images were sent by e-mail. Reply mails about each case were sent to all members via the mailing list.Results: During a period of 6 months, 18 cases (tumor/infection: 7, bone: 6, cardiovascular: 1, neurology; 3, endocrine: 1) with 144 e-mails (average 5.6/case) were submitted to the conference. The average period of discussion was 15.6 days. The number of attached images was 1 to 9 (average, 4.2/e-mails). JPEG compression rate was 1/10 to 1/20. The quality of the images was good enough for discussion. Some cases required additional images for further discussion.Conclusion: Our tele-nuclear medicine conference with an electronic mailing list and digital camera was simple and low-cost. The conference system was useful for education and clinical work.

We compared early and delayed Tc-99m ECD SPECT scans in 32 SLE patients (Group 1, definite neuropsychiatric disorders; Group 2, minor neurologic symptoms or normal) with those of normal controls by visual inspection and semi-quantitative evaluation. With visual interpretation, 13 out of 14 patients in Group 1 (93%) and 7 out of 18 patients in Group 2 (39%) had diffuse uneven decrease in early scans. Seven patients in Group 2 (39%) who had normal early scans demonstrated focal decrease in the medial frontal lobe in delayed scans. With cerebral region to cerebellar ratios, in early scans, the medial frontal lobe in Group 1 and Group 2 was significantly lower than in normal controls, and lateral frontal lobe and occipital lobes in Group 1 were significantly lower than in normal controls. Nevertheless, in delayed scans, every cortical region except for the parietal lobe in Groups 1 and 2 was significantly lower than in normal controls. The retention rates in all regions in SLE patients were significantly lower than in normal controls. No case showed SPECT improvement on follow-up studies in either group in spite of clinical improvement. Delayed Tc-99m ECD brain SPECT of high sensitivity might be useful in detecting CNS involvement. Although the SPECT findings did not correlate with the neuropsychiatric symptoms, early and delayed Tc-99m ECD SPECT seems to provide useful objective diagnostic information in SLE patients.

Color image consistency has not been accomplished yet except the Digital Imaging and Communication in Medicine (DICOM) Supplement 100 for implementing a color reproduction pipeline and device independent color spaces. Thus, most healthcare enterprises could not check monitor degradation routinely. To ensure color consistency in medical color imaging, monitor color calibration should be introduced. Using simple color calibration device . chromaticity of colors including typical color (Red, Green, Blue, Green and White) are measured as device independent profile connection space value called u’v’ before and after calibration. In addition, clinical color images are displayed and visual differences are observed. In color calibration, monitor brightness level has to be set to quite lower value 80 cd/m 2 according to sRGB standard. As Maximum brightness of most color monitors available currently for medical use have much higher brightness than 80 cd/m 2 , it is not seemed to be appropriate to use 80 cd/m 2 level for calibration. Therefore, we propose that new brightness standard should be introduced while maintaining the color representation in clinical use. To evaluate effects of brightness to chromaticity experimentally, brightness level is changed in two monitors from 80 to 270cd/m 2 and chromaticity value are compared with each brightness levels. As a result, there are no significant differences in chromaticity diagram when brightness levels are changed. In conclusion, chromaticity is close to theoretical value after color calibration. Moreover, chromaticity isn’t moved when brightness is changed. The results indicate optimized reference brightness level for clinical use could be set at high brightness in current monitors .

Recent studies have shown that protein kinase C (PKC) delta is proteolytically activated at the onset of apoptosis induced by DNA-damaging agents, tumor necrosis factor, and anti-Fas antibody. However, the relationship of PKC delta cleavage to induction of apoptosis is unknown. The present studies demonstrate that full-length PKC delta is cleaved at DMQD330N to a catalytically active fragment by the cysteine protease CPP32. The results also demonstrate that overexpression of the catalytic kinase fragment in cells is associated with chromatin condensation, nuclear fragmentation, induction of sub-G1 phase DNA and lethality. By contrast, overexpression of full-length PKC delta or a kinase inactive PKC delta fragment had no detectable effect. The findings suggest that proteolytic activation of PKC delta by a CPP32-like protease contributes to phenotypic changes associated with apoptosis.

These studies demonstrate that treatment of human U-937 cells with ionizing radiation (IR) is associated with activation of a cytoplasmic myelin basic protein (MBP) kinase. Characterization of the kinase by gel filtration and in-gel kinase assays support activation of a 40 kDa protein. Substrate and inhibitor studies further support the induction of protein kinase C (PKC)-like activity. The results of N-terminal amino acid sequencing of the purified protein demonstrate identity of the kinase with an internal region of PKC delta. Immunoblot analysis was used to confirm proteolytic cleavage of intact 78 kDa PKC delta in control cells to the 40 kDa C-terminal fragment after IR exposure. The finding that both IR-induced proteolytic activation of PKC delta and endonucleolytic DNA fragmentation are blocked by Bcl-2 and Bcl-xL supports an association with physiological cell death (PCD). Moreover, cleavage of PKC delta occurs adjacent to aspartic acid at a site (QDN) similar to that involved in proteolytic activation of interleukin-1 beta converting enzyme (ICE). The specific tetrapeptide ICE inhibitor (YVAD) blocked both proteolytic activation of PKC delta and internucleosomal DNA fragmentation in IR-treated cells. These findings demonstrate that PCD is associated with proteolytic activation of PKC delta by an ICE-like protease.

The cellular response to ionizing radiation (IR) includes induction of the c-jun and EGR1 early response genes. The present work has examined potential cytoplasmic signaling cascades that transduce IR-induced signals to the nucleus. The results demonstrate activation of the 40S ribosomal protein S6 kinase, pp90rsk, in human U-937 myeloid leukemia cells. Partial purification of pp90rsk by affinity chromatography demonstrated an increase in S6 peptide phosphorylation when comparing irradiated with control cells. IR-induced activation of pp90rsk was further confirmed in immune-complex kinase assays. In contrast to these findings, there was no detectable induction of pp70S6K. Previous work has demonstrated that mitogen-activated protein kinase activates pp90rsk. The present results further show that IR treatment is associated with induction of mitogen-activated protein kinase activity and that this event is temporally related to activation of pp90rsk and early response gene expression. These findings suggest that activation of the mitogen-activated protein kinase/pp90rsk cascade is involved in the response of cells to IR exposure.

Treatment of human myeloid leukemia cells with 12-O-tetradecanoylphorbol-13-acetate (TPA), an activator of protein kinase C (PKC), is associated with induction of monocytic differentiation. Since PKC can act immediately upstream to the cytoplasmic Raf-1 serine/threonine protein kinase, we studied activation of Raf-1 during induction of the differentiated monocytic phenotype. The results demonstrate that Raf-1 is activated during TPA-induced monocytic differentiation of HL-60 cells. In contrast, there was little effect of TPA on this kinase in an HL-60 variant, designated HL-525, which is resistant to TPA-induced differentiation. Treatment of both HL-60 and HL-525 cells with okadaic acid, an inhibitor of serine/threonine protein phosphatases 1 and 2A, was associated with Raf-1 activation and induction of the monocytic phenotype. Since Raf-1 can activate the mitogen-activated protein (MAP) kinases, we also studied the relationship between MAP kinase activation and monocytic differentiation. Treatment of HL-60, but not HL-525, cells with TPA was associated with increased MAP kinase activity as determined by phosphorylation of myelin basic protein and the c-Jun Y peptide. Okadaic acid-induced differentiation of both HL-60 and HL-525 cells was similarly accompanied by increases in MAP kinase activity. These findings indicated that activation of Raf-1/MAP kinase signaling is associated with induction of a differentiated monocytic phenotype and that okadaic acid bypasses a defect in this cascade in TPA-treated HL-525 cells. While recent studies have shown that HL-525 cells are deficient in PKC beta, the present results demonstrate that PKC beta expression is up-regulated in the HL-525 variant by treatment with retinoic acid. The results also demonstrate that retinoic acid-treated HL-525 cells respond to TPA with activation of Raf-1 and MAP kinase, as well as induction of monocytic differentiation. Taken together, the results indicate that activation of Raf-1/MAP kinase signaling is associated with monocytic differentiation and that stimulation of serine/threonine protein phosphorylation by TPA or okadaic acid is sufficient for reversal of the leukemic HL-60 phenotype.

A novel Ca 2+ -binding protein which is termed S-100P was purified from human placenta with a hydrophobic column followed by an anion exchange column and reverse phase high performance liquid chromatography (HPLC). Molecular mass of the protein was 10 kDa according to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. Using immunoblotting technique, anti-human calcyclin antibodies did not bind to the S-100P. Isoelectric point of S-100P was pI=4.6. S-100P did not formed disulfide-linked dimer. Calcium binding ability was proved by UV difference spectrometry, urea/alkaline gel electrophoresis, and 45 Ca overlay technique. A ninety amino acid sequence of S-100P was determined. It is 49% identical with human S-100β, 38% with human calcyclin, and 37% with human cystic fibrosis antigen.

血液透析患者の血清中にはインドキシル硫酸がアルブミン結合して著明に蓄積している. アルブミン結合物質は通常の血液透析法では殆ど除去されないため, 経口吸着剤AST-120による除去効果に関して検討した.経口吸着剤AST-120を血液透析患者に投与したところ透析前血清インドキシル硫酸濃度は非投与血液透析患者に比較して著明に減少した. しかし, 血清クレアチニン, BUNは有意の低下を示さなかった. また, 掻痒感を訴える血液透析患者にAST-120を投与したところ掻痒感の改善が認められた.AST-120は血液透析患者の血清中に蛋白結合して蓄積しているインドキシル硫酸の濃度を低下させるのに有効であった. これはAST-120が腸内で産生されたインドールを吸着除去することにより肝臓でのインドキシル硫酸の産生を抑制するためと考えられた.

Quinolinic acid was first identified in uremic serum by use of gas chromatography/mass spectrometry. Quantification by selected ion monitoring revealed that the serum concentration of quinolinic acid was markedly increased in chronic hemodialysis patients, and that the acid could be removed by conventional hemodialysis. The serum concentration of quinolinic acid was weakly but significantly correlated with the serum uric acid concentration. Accumulation of quinolinic acid in uremic blood may be involved in the pathogenesis of anemia, suppressed immune system, and uremic encephalopathy.

The stress-activated protein (SAP) kinases are induced by tumor necrosis factor, oncoproteins, and UV light. The present studies demonstrate that ionizing radiation (IR) activates p54 SAP kinase. IR-induced activation of SAP kinase is associated with binding to the SH2/SH3-containing adaptor protein Grb2. This interaction is mediated by the SH3 domains of Grb2 and the proline-rich sequence PPPKIP in the carboxy-terminal region of SAP kinase. We also demonstrated that SAP kinase and the p85 alpha-subunit of phosphatidylinositol (PI) 3-kinase form a complex in irradiated cells. The results indicate that this complex involves binding of the p85 alpha subunit of PI 3-kinase to the SH2 domain of Grb2. The functional role of linking SAP kinase to PI 3-kinase is further supported by the finding that wortmannin, an inhibitor of PI 3-kinase, stimulates SAP kinase activity. These results suggest that the cellular response to IR may include regulation of SAP kinase by a PI 3-kinase-dependent signaling pathway.

Macrophage colony-stimulating factor (M-CSF) is required for the growth and differentiation of mononuclear phagocytes. In the present studies using human monocytes, we show that M-CSF induces interaction of the Grb2 adaptor protein with the focal adhesion kinase pp125FAK. The results demonstrate that tyrosine-phosphorylated pp125FAK directly interacts with the SH2 domain of Grb2. The findings indicate that a pYENV site at Tyr-925 in pp125FAK is responsible for this interaction. We also demonstrate that the Grb2-FAK complex associates with the GTPase dynamin. Dynamin interacts with the SH3 domains of Grb2 and exhibits M-CSF-dependent tyrosine phosphorylation in association with pp125FAK. These findings suggest that M-CSF-induced signaling involves independent Grb2-mediated pathways, one leading to Ras activation and another involving pp125FAK and a GTPase implicated in receptor internalization.

To determine the role of vasoactive peptides such as endothelin (ET), calcitonin gene-related peptide (CGRP) and human atrial natriuretic peptide (hANP) in the regulation of blood pressure in uremic patients, and to determine the effect of various types of dialyzer membranes on hemodialysis (HD)-induced changes in plasma levels of such peptides, plasma ET, CGRP and hANP were measured in HD patients and patients on continuous ambulatory peritoneal dialysis (CAPD). Plasma levels of ET, CGRP, and hANP were significantly higher in HD and CAPD patients than in healthy subjects. There were no significant differences in plasma levels of ET, CGRP, and hANP between hypertensive and normotensive HD patients, and no significant correlation was observed between HD-induced changes in plasma levels and changes in blood pressure. Plasma levels of ET decreased when HD was performed using high-flux membranes, such as polyacrylonitrile (PAN), polymethyl methacrylate (PMMA) and cellulose triacetate (CTA), but did not decrease using a saponified cellulose (SC) membrane. Plasma levels of CGRP decreased in the case of PAN, but increased significantly with PMMA and showed no change with SC and CTA. Plasma levels of hANP decreased in all types of dialyzer membranes due to decreased secretion. These results indicate that the effect of HD on plasma levels of ET and CGRP, but not hANP, depends on the type of dialyzer membrane used.

To determine the diagnostic role of urinary trehalase in chronic glomerular disease, urinary trehalase activity and other urinary markers such as N-acetyl-beta-D-glucosaminidase (NAG), alanine aminopeptidase (AAP), alkaline phosphatase (ALP), gamma-glutamyltranspeptidase (gamma-GTP), lactate dehydrogenase (LDH), lysozyme and beta 2-microglobulin (BMG) were measured in patients with chronic glomerulonephritis, nephrotic syndrome and chronic renal failure. Urinary trehalase activity was significantly increased in chronic glomerular disease, especially nephrotic syndrome, as compared with that in the healthy subjects. The highest incidence of elevated excretion was observed for trehalase with 52% in chronic glomerular disease, followed by NAG. Urinary trehalase activities in the patients were significantly correlated with the urinary levels of protein, NAG and AAP and total score of tubular damage, but not correlated with urinary levels of BMG or lysozyme. In patients with chronic glomerulonephritis and nephrotic syndrome, there was no significant difference in urinary trehalase activities between with and without hematuria. These results indicate that in some patients with chronic glomerular disease, there is tubular involvement as substantiated by elevation of the other urinary enzymes and BMG. Urinary trehalase is elevated more often in these types of disease than other markers of tubular damage.

Serum indoxyl sulphate, which is markedly accumulated in haemodialysis patients, cannot be removed efficiently by haemodialysis due to its albumin-binding property. To determine whether an oral adsorbent (AST-120) can decrease its serum concentration, AST-120 was administered to haemodialysis patients. The patients given AST-120 showed significantly reduced serum concentration of indoxyl sulphate as compared to control haemodialysis patients, even though the serum concentrations of urea nitrogen and creatinine did not decrease significantly in the patients treated with AST-120. The haemodialysis patients with generalised pruritus showed an amelioration of itching after administration of AST-120. These results showed that AST-120 was effective in reducing the serum concentration of albumin-bound indoxyl sulphate in haemodialysis patients by adsorption of indole, a precursor of indoxyl sulphate, in the intestines, and that it relieved itching in haemodialysis patients with generalised pruritus.

Long-term hemodialysis (HD) patients complaining of shoulder joint pain were treated by HD and by push/pull HDF using high-flux synthetic membranes with large pores (Asahi PAN 20CX2) for 2 weeks. The results showed no significant difference in Kt/Vurea between HD and push/pull HDF. Nevertheless, reduction of the plasma beta 2-microglobulin was greater during push/pull HDF than during HD. These findings can be explained by far more convective flux in push/pull HDF than in HD: nearly 30 L during push/pull HDF vs. 3 L during HD. In the present study, there was no alleviation of the shoulder joint pain during HD treatment, whereas marked relief of the symptom was found during push/pull HDF treatment. Since the two treatment modalities differ simply in their efficiency in removing larger molecular weight substances, the joint pain alleviation effected by push/pull HDF could well be ascribed to elimination of an unknown larger molecular weight substance causing this symptom. However, a considerable amount of beta 2-microglobulin was removed both by HD and push/pull HDF. Therefore, the substance causing the joint pain might be larger than beta 2-microglobulin.