Evaluation of two semi-supervised learning methods and their combination for automatic classification of bone marrow cells.

Differential bone marrow (BM) cell counting is an important test for the diagnosis of various hematological diseases. However, it is difficult to accurately classify BM cells due to non-uniformity and the lack of reproducibility of differential counting. Therefore, automatic classification systems have been developed in which deep learning is used. These systems requires large and accurately labeled datasets for training. To overcome this, we used semi-supervised learning (SSL), in which learning proceeds while labeling. We used three methods: self-training (ST), active learning (AL), and a combination of these methods, and attempted to automatically classify 16 types of BM cell images. ST involves data verification, as in AL, before adding them to the training dataset (confirmed self-training: CST). After 25 rounds of CST, AL, and CST + AL, the initial number of training data increased from 425 to 40,518; 3682; and 47,843, respectively. Accuracies for the test data of 50 images for each cell type were 0.944, 0.941, and 0.976, respectively. Data added with CST or AL showed some imbalances between classes, while CST + AL exhibited fewer imbalances. We suggest that CST + AL, when combined with two SSL methods, is efficient in increasing training data for the development of automatic BM cells classification systems.

Difference in left atrial myocardial dynamics during reservoir phase between hypertrophic cardiomyopathy and hypertensive heart determined using three-dimensional speckle tracking echocardiography.

We aimed to investigate left atrial (LA) myocardial dynamics during reservoir phase using three-dimensional speckle-tracking echocardiography (3DSTE) focusing on its longitudinal-circumferential relationship in patients with left ventricular (LV) hypertrophy and clarifying the difference in LA myocardial reservoir dynamics between hypertrophic cardiomyopathy (HCM) and hypertension with LV hypertrophy (HT-LVH). We studied 4 age-matched groups consisting of 27 patients with HCM, 16 with HT-LVH, 22 hypertensive patients without LV hypertrophy (HT), and 18 normal controls. Using 3DSTE, we measured LA global longitudinal strain (LA-LSR), global circumferential strain (LA-CSR), and global area strain (LA-ASR) during the reservoir phase, as well as LV global longitudinal strain (LV-LS), global circumferential strain (LV-CS), and global area strain (LV-AS). LA-LSR was significantly lower in the HCM and HT-LVH groups than in the controls, but there was no significant difference between the HCM and HT-LVH groups. LA-CSR and LA-ASR were significantly lower in the HCM group than in the other three groups, among which no significant difference was detected. In all subjects, LA-LSR was significantly correlated with LV-LS but not with LV-CS. LA-CSR was correlated with neither LV-LS nor LV-CS. In conclusion, both longitudinal and circumferential LA myocardial expansion during reservoir phase were reduced in HCM, while only the longitudinal one was reduced in HT-LVH. Reduction of LA circumferential expansion may reflect a more serious and intrinsic impairment of LA myocardial distensibility in HCM. Measuring LA-CSR and LA-ASR using 3DSTE would contribute to a more accurate understanding of LA reservoir function abnormality in HCM.

Prognostic value of an echocardiographic index reflecting right ventricular operating stiffness in patients with heart failure.

PURPOSE: We recently reported a noninvasive method for the assessment of right ventricular (RV) operating stiffness that is obtained by dividing the atrial-systolic descent of the pulmonary artery-RV pressure gradient (PRPGDAC) derived from the pulmonary regurgitant velocity by the tricuspid annular plane movement during atrial contraction (TAPMAC). Here, we investigated whether this parameter of RV operating stiffness, PRPGDAC/TAPMAC, is useful for predicting the prognosis of patients with heart failure (HF). METHODS: We retrospectively included 127 hospitalized patients with HF who underwent an echocardiographic examination immediately pre-discharge. The PRPGDAC/TAPMAC was measured in addition to standard echocardiographic parameters. Patients were followed until 2 years post-discharge. The endpoint was the composite of cardiac death, readmission for acute decompensation, and increased diuretic dose due to worsening HF. RESULTS: 58 patients (46%) experienced the endpoint during follow-up. Univariable and multivariable Cox regression analyses demonstrated that the PRPGDAC/TAPMAC was associated with the endpoint. In a Kaplan-Meier analysis, the event rate of the greater PRPGDAC/TAPMAC group was significantly higher than that of the lesser PRPGDAC/TAPMAC group. In a sequential Cox analysis for predicting the endpoint's occurrence, the addition of PRPGDAC/TAPMAC to the model including age, sex, NYHA functional classification, brain natriuretic peptide level, and several echocardiographic parameters including tricuspid annular plane systolic excursion significantly improved the predictive power for prognosis. CONCLUSION: A completely noninvasive index of RV operating stiffness, PRPGDAC/TAPMAC, was useful for predicting prognoses in patients with HF, and it showed an incremental prognostic value over RV systolic function.

Deep Learning-Based Nuclear Lobe Count Method for Differential Count of Neutrophils.

Differentiating neutrophils based on the count of nuclear lobulation is useful for diagnosing various hematological disorders, including megaloblastic anemia, myelodysplastic syndrome, and sepsis. It has been reported that one-fifth of sepsis-infected patients worldwide died between 1990 and 2017. Notably, fewer nuclear-lobed and stab-formed neutrophils develop in the peripheral blood during sepsis. This abnormality can serve as an early diagnostic criterion. However, testing this feature is a complex and time-consuming task that is rife with human error. For this reason, we apply deep learning to automatically differentiate neutrophil and nuclear lobulation counts and report the world's first small-scale pilot. Blood films are prepared using venous peripheral blood taken from four healthy volunteers and are stained with May-Grünwald Giemsa stain. Six-hundred 360 × 363-pixel images of neutrophils having five different nuclear lobulations are automatically captured by Cellavision DM-96, an automatic digital microscope camera. Images are input to an original architecture with five convolutional layers built on a deep learning neural-network platform by Sony, Neural Network Console. The deep learning system distinguishes the four groups (i.e., band-formed, two-, three-, and four- and five- segmented) of neutrophils with up to 99% accuracy, suggesting that neutrophils can be automatically differentiated based on their count of segmented nuclei using deep learning.

Left ventricular global longitudinal strain calculated from manually traced endocardial border lengths utilizing the images for routine ejection fraction measurement by biplane method of disks.

PURPOSE: The purpose of this study was to test whether the fractional change in the endocardial border length between end-diastole and end-systole as manually traced in left ventricular ejection fraction (LVEF) measurement using the biplane method of disks (MOD) was consistent with the global longitudinal strain derived from speckle-tracking echocardiography. METHODS: For 105 patients who underwent echocardiography, two- and four-chamber images with manually traced endocardial lines for LVEF measurement by MOD were stored. LV endocardial lengths at end-diastole and at end-systole were measured on both images to calculate the fractional length changes, which were averaged (GLSMOD). Speckle-tracking analysis was performed to measure global longitudinal strains in the apical two- and four-chamber and long-axis images, and the three values were averaged (GLSSTE) according to the ASE and EACVI guidelines. RESULTS: There was no significant difference between GLSMOD and GLSSTE. GLSMOD correlated well with GLSSTE (r = 0.81, p < 0.001), and there was no fixed bias in the Bland-Altman analysis. The intraclass correlations for the intra- and inter-observer comparisons for GLSSTE were excellent, and those for GLSMOD were adequate. CONCLUSION: The fractional LV endocardial border length change, GLSMOD, showed sufficient agreement with GLSSTE to justify its use as a substitute for the STE-derived global longitudinal strain.

Simple and noninvasive method to estimate right ventricular operating stiffness based on echocardiographic pulmonary regurgitant velocity and tricuspid annular plane movement measurements during atrial contraction.

It was recently shown that invasively determined right ventricular (RV) stiffness was more closely related to the prognosis of patients with pulmonary hypertension than RV systolic function. So far, a completely noninvasive method to access RV stiffness has not been reported. We aimed to clarify the clinical usefulness of our new echocardiographic index of RV operating stiffness using atrial-systolic descent of the pulmonary artery-RV pressure gradient derived from pulmonary regurgitant velocity (PRPGDAC) and tricuspid annular plane movement during atrial contraction (TAPMAC). We studied 81 consecutive patients with various cardiac diseases who underwent echocardiography and cardiac catheterization. We measured PRPGDAC and TAPMAC using continuous-wave Doppler and M-mode echocardiography, respectively, and calculated PRPGDAC/TAPMAC. RV end-diastolic pressure (RVEDP) and RV pressure increase during atrial contraction (ΔRVPAC) were invasively measured, and RV volume change during atrial contraction (ΔVAC) was calculated from echocardiographic late-diastolic transtricuspid flow time-velocity integral and tricuspid annular area; thus ΔRVPAC/ΔVAC was used as the standard index for RV operating stiffness. PRPGDAC/TAPMAC well correlated with ΔRVPAC/ΔVAC (r = 0.84, p < 0.001) and RVEDP (r = 0.80, p < 0.001), and the area under the receiver operating characteristic curve to discriminate RVEDP > 12 mmHg was 0.94. Multivariate regression analysis revealed that PRPGDAC/TAPMAC was the single independent determinant of ΔRVPAC/ΔVAC (ß = 0.86, p < 0.001). PRPGDAC/TAPMAC is useful to estimate RV operating stiffness and a good practical indicator of RVEDP.

Right ventricular basal inflow and outflow tract diameters overestimate right ventricular size in subjects with sigmoid-shaped interventricular septum: a study using three-dimensional echocardiography.

Sigmoid-shaped ventricular septum (SS), a frequently encountered minor abnormality in echocardiographic examinations of the elderly, may have some influence on RV shape. We aimed to determine the influence of SS on the accuracy of the 6 RV linear diameter measurements in the light of three-dimensional echocardiographic (3DE) RV volume. The aorto-septal angle (ASA) was measured in the parasternal long-axis view using two-dimensional echocardiography (2DE) as an index of SS in 70 patients without major cardiac abnormalities who were subdivided into 35 with SS (ASA ≤ 120°) and 35 without SS (NSS). We measured RV end-diastolic volume (RVEDV) using 3DE; in addition, using 2DE, we measured basal RV diameter, mid-cavity diameter, longitudinal diameter and end-diastolic area in the apical four-chamber view; proximal RV outflow tract (RVOT) diameter in the parasternal long-axis view; and proximal and distal RVOT diameters in the parasternal short-axis view. RVEDV did not differ between the SS and NSS groups. The SS group had greater basal RV diameter and proximal and distal RVOT diameters than the NSS group. RV mid-cavity diameter, longitudinal diameter, and end-diastolic area did not differ between the groups. Among the 2DE parameters of RV size, RV end-diastolic area was most strongly correlated with RVEDV (r = 0.67), followed by RV mid-cavity diameter (r = 0.58). When SS is present, the echocardiographic basal RV diameter and RVOT diameters overestimate RV size, and the measurement of RV end-diastolic area and mid-cavity diameter more correctly reflect 3D RV volume.

Quantitative distinction of the morphological characteristic of erythrocyte precursor cells with texture analysis using gray level co-occurrence matrix.

BACKGROUND: Morphological characteristics of blood cells are still qualitatively defined. So a texture analysis (Tx) method using gray level co-occurrence matrices (GLCMs; CM-Tx method) was applied to images of erythrocyte precursor cells (EPCs) for quantitatively distinguishing four types of EPC stages: proerythroblast, basophilic erythroblast, polychromatic erythroblast, and orthochromatic erythroblast. METHODS: Fifty-five images of four types of EPCs were downloaded from an atlas uploaded by the Blood Cell Morphology Standardization Subcommittee (BCMSS) of the Japanese Society of Laboratory Hematology (JSLH). Using in-house programs, two types of GLCMs-(R: d=1, θ=0°) and (U: d=1, θ=270°)-and nine types of texture distinction index (TDI) were calculated with images removed outer part of cell. RESULTS: Three binary decision trees were sequentially divided among four types of EPC with the sum average of GLCM (U), the contrast of GLCM (R), and the sum average of GLCM (U). The average concordance rate (sensitivity) of CM-Tx method with the judgments of eleven experts in the BCMSS of the JSLH was 95.8% (87.5-100.0), and the average specificity was 97.6% (92.5-100.0). CONCLUSIONS: The CM-Tx method is an effective tool for quantitative distinction of EPC with their morphological features.

Novel echocardiographic method to assess left ventricular chamber stiffness and elevated end-diastolic pressure based on time-velocity integral measurements of pulmonary venous and transmitral flows.

Aims: The detection of increased left ventricular (LV) chamber stiffness may play an important role in assessing cardiac patients with potential but not overt heart failure. A non-invasive method to estimate it is not established. We investigated whether the echocardiographic backward/forward flow volume ratio from the left atrium (LA) during atrial contraction reflects the LV chamber stiffness. Methods and results: We studied 62 patients who underwent cardiac catheterization and measured their left ventricular end-diastolic pressure (LVEDP) and pressure increase during atrial contraction (ΔPa) from the LV pressure waveform. Using the echocardiographic biplane method of disks, we measured the LV volume change during atrial contraction indexed to the body surface area (ΔVa), and ΔPa/ΔVa was calculated as a standard for the LV operating chamber stiffness. Using pulsed Doppler echocardiography, we measured the time-velocity integral (TVI) of the backward pulmonary venous (PV) flow during atrial contraction (IPVA) and the ratio of IPVA to the PV flow TVI throughout a cardiac cycle (FPVA). We also measured the TVI of the atrial systolic forward transmitral flow (IA) and the ratio of the IA to the transmitral TVI during a cardiac cycle (FA) and calculated IPVA/IA and FPVA/FA. IPVA/IA and FPVA/FA were well correlated with ΔPa/ΔVa (r = 0.79 and r = 0.81) and LVEDP (r = 0.73 and r = 0.77). The areas under the ROC curve to discriminate LVEDP >18 mmHg were 0.90 for IPVA/IA and 0.93 for FPVA/FA. Conclusion: The FPVA/FA, the backward/forward flow volume ratio from the LA during atrial contraction, is useful for non-invasive assessments of LV chamber stiffness and elevated LVEDP.

Correction to: Altered oscillation of Doppler-derived renal and renal interlobar venous flow velocities in hypertensive and diabetic patients.

In the original publication of this paper the legend of Fig. 1 should read as: Fig. 1 Pulsed Doppler flow velocity recordings of the aorta (a), right renal artery (b), right renal interlobar artery (c), inferior vena cava (d), right renal vein (e), and right renal interlobar vein (f). PSV peak systolic velocity, EDV end-diastolic velocity, V MAX maximum velocity, V MIN minimum velocity.

Quantitative Distinction of Morphological Characteristics of White Blood Cells in Peripheral Blood Through Texture Analysis Using Gray Level Co-Occurrence Matrix.

BACKGROUND: Texture features are valuable clues for skilled technicians to differentiate peripheral blood (PB) white blood cells (WBCs). Some studies have tried to distinguish WBCs automatically by using texture analysis. However, no study so far has applied a gray level co-occurrence matrix (GLCM) to images of PB WBCs. Here, we developed a new GLCM method, called the CM-Tx method, for automatically distinguishing PB WBCs. METHODS: We used a total of 199 images of six different types of PB WBCs, taken from PB smears of 12 healthy volunteers, as objective standard images for the analysis. The six types were band form neutrophil, segmented form neutrophil, eosinophil, basophil, lymphocyte, and monocyte. Using in-house FORTRAN programs, three types of GLCM (R: distance (d) = 1, direction (θ) = 0°), (U: d = 1, θ = 270°) and (AE: d = 1, θ = 15° x q: q = 0, ..., 23), the mean intensity (MI) of each image and nine different texture distinction indexes (TDIs) for each GLCM were calculated. Then, a threshold value (TV) for distinguishing the type of PB WBC was selected from the dot plots of all TDIs and the MI. RESULTS: In total, we made 1,194 GLCMs. Using the selected TVs of the TDI, four sequential binary divisions could distinguish five types of PB WBCs. First, monocytes were distinguished (sensitivity 100%, specificity 100%, p < 0.0001) with the TV of the inverse difference moment of the GLCM (U). Then, segmented and band form neutrophils were distinguished from the remaining (100%, 99%, p < 0.0001) with the TV of the contrast of the GLCM (AE). Next, lymphocytes were distinguished (100%, 98%, p < 0.0001) with the TV of the entropy of the GLCM (AE). Finally, basophils were distinguished (82.4%, 100%, p < 0.0001) from eosinophils with the TV of the summed entropy of the GLCM (R). Band form neutrophils could not be distinguished from segmented form neutrophils. The average sensitivity of the CM-Tx method for the five types was 95.6%, and its average specificity was 99.3%. CONCLUSIONS: The CM-Tx method can distinguish five types of PB WBCs by using numerical differences only in texture futures quantified with GLCM. However, some other method was needed to distinguish the band and segmented form neutrophils from each other.

Usefulness of the Continuous-Wave Doppler-Derived Pulmonary Arterial-Right Ventricular Pressure Gradient Just before Atrial Contraction for the Estimation of Pulmonary Arterial Diastolic and Wedge Pressures.

In our new echocardiographic method, pulmonary regurgitant velocity immediately before right atrial (RA) contraction is used to estimate pulmonary artery diastolic pressure (PADP) and mean PA wedge pressure (MPAWP). Our aim here was to compare the usefulness of this new method with that of the conventional method, which uses pulmonary regurgitant velocity at end diastole. We studied 55 consecutive patients who underwent echocardiography and right-sided heart catheterization. The pulmonary regurgitant velocities just before RA contraction and at end diastole were measured to obtain echocardiographic estimates of PADP (EPADPpreA and EPADPED, respectively) by adding the pressure gradients to the echocardiographically estimated RA pressure. Compared with EPADPED, EPADPpreA correlated better with PADP (r = 0.87) and MPAWP (r = 0.80), and direct fixed biases were detected for EPADPED but not for EPADPpreA. The area under the receiver operating characteristic curve distinguishing patients with MPAWP ≥18 mm Hg was greater for EPADPpreA (0.97) than for E/e' (0.94) and E/A (0.83). EPADPpreA is thus useful in estimating PADP and MPAWP in patients with heart disease.

Altered oscillation of Doppler-derived renal and renal interlobar venous flow velocities in hypertensive and diabetic patients.

BACKGROUND AND PURPOSE: Flow velocity oscillation rate (FVOR) of the renal interlobar vein has been reported to be decreased in patients with urinary obstruction or diabetic nephropathy, and increased in those with hypertension during pregnancy. To clarify the clinical role of the renal interlobar venous FVOR, we investigated the flow velocity patterns of the renal vessels in patients with hypertension (HT) and/or diabetes (DM). METHODS AND RESULTS: Pulsed-wave Doppler sonography was performed in 34 patients: 15 with HT, 10 with DM, and nine with both HT and DM (HT-DM). Each FVOR of the right and left interlobar veins was closely and positively correlated with the ipsilateral interlobar arterial resistive index (RI), especially in the HT group, but not with the estimated glomerular filtration rate. The right interlobar venous FVOR was decreased in the DM and HT-DM groups compared to the HT group. CONCLUSION: The renal interlobar venous FVOR is strongly influenced by the arterial RI in HT patients, and is reduced in DM patients without an obvious relationship with diabetic nephropathy. These findings should be noted for the clinical application of renal interlobar venous flow analysis.

Semi-Automatic Rating Method for Neutrophil Alkaline Phosphatase Activity.

BACKGROUND: The neutrophil alkaline phosphatase (NAP) score is a valuable test for the diagnosis of myeloproliferative neoplasms, but it has still manually rated. Therefore, we developed a semi-automatic rating method using Photoshop® and Image-J, called NAP-PS-IJ. METHODS: Neutrophil alkaline phosphatase staining was conducted with Tomonaga's method to films of peripheral blood taken from three healthy volunteers. At least 30 neutrophils with NAP scores from 0 to 5+ were observed and taken their images. From which the outer part of neutrophil was removed away with Image-J. These were binarized with two different procedures (P1 and P2) using Photoshop® . NAP-positive area (NAP-PA) and granule (NAP-PGC) were measured and counted with Image-J. RESULTS: The NAP-PA in images binarized with P1 significantly (P < 0.05) differed between images with NAP scores from 0 to 3+ (group 1) and those from 4+ to 5+ (group 2). The original images in group 1 were binarized with P2. NAP-PGC of them significantly (P < 0.05) differed among all four NAP score groups. The mean NAP-PGC with NAP-PS-IJ indicated a good correlation (r = 0.92, P < 0.001) to results by human examiners. CONCLUSIONS: The sensitivity and specificity of NAP-PS-IJ were 60% and 92%, which might be considered as a prototypic method for the full-automatic rating NAP score.

Overestimation by echocardiography of the peak systolic pressure gradient between the right ventricle and right atrium due to tricuspid regurgitation and the usefulness of the early diastolic transpulmonary valve pressure gradient for estimating pulmonary artery pressure.

We investigated the influence of tricuspid regurgitation (TR) severity on the echocardiographic peak systolic transtricuspid pressure gradient (TRPG) and evaluated the usefulness of the peak early diastolic transpulmonary valve pressure gradient (PRPG) for estimating pulmonary artery (PA) pressure. In 55 consecutive right heart-catheterized patients, we measured the peak systolic right ventricular (RV)-right atrial (RA) pressure gradient (RV-RACATH), peak early diastolic PA-RV pressure gradient (PA-RVCATH), and mean PA pressure (MPAPCATH). Using echocardiography, we obtained the TRPG, PRPG, and an estimate of the mean PA pressure (EMPAP) as the sum of PRPG and the estimated RA pressure, and measured the vena contracta width of TR (VCTR). The difference between the TRPG and RV-RACATH was significantly greater in the very severe TR group (VCTR > 11 mm) than in the mild, moderate, and severe TR groups, and significantly greater in the severe TR group (7 < VCTR ≤ 11 mm) than in the mild TR group. The overestimation of the pressure gradient >10 mmHg by TRPG was not seen in the mild or moderate TR groups, but was observed in the severe and very severe TR groups (22 and 83%, respectively). In the ROC analysis, EMPAP could distinguish patients with MPAPCATH ≥ 25 mmHg with the area under the curve of 0.93, 100% sensitivity, and 87% specificity. In conclusion, TRPG frequently overestimated RV-RACATH when VCTR was >11 mm and sometimes did when VCTR was >7 mm, where EMPAP using PRPG was useful for estimating PA pressure.

Characteristic systolic waveform of left ventricular longitudinal strain rate in patients with hypertrophic cardiomyopathy.

We analyzed the waveform of systolic strain and strain-rate curves to find a characteristic left ventricular (LV) myocardial contraction pattern in patients with hypertrophic cardiomyopathy (HCM), and evaluated the utility of these parameters for the differentiation of HCM and LV hypertrophy secondary to hypertension (HT). From global strain and strain-rate curves in the longitudinal and circumferential directions, the time from mitral valve closure to the peak strains (T-LS and T-CS, respectively) and the peak systolic strain rates (T-LSSR and T-CSSR, respectively) were measured in 34 patients with HCM, 30 patients with HT, and 25 control subjects. The systolic strain-rate waveform was classified into 3 patterns ("V", "W", and "√" pattern). In the HCM group, T-LS was prolonged, but T-LSSR was shortened; consequently, T-LSSR/T-LS ratio was distinctly lower than in the HT and control groups. The "√" pattern of longitudinal strain-rate waveform was more frequently seen in the HCM group (74 %) than in the control (4 %) and HT (20 %) groups. Similar but less distinct results were obtained in the circumferential direction. To differentiate HCM from HT, the sensitivity and specificity of the T-LSSR/T-LS ratio <0.34 and the "√"-shaped longitudinal strain-rate waveform were 85 and 63 %, and 74 and 80 %, respectively. In conclusion, in patients with HCM, a reduced T-LSSR/T-LS ratio and a characteristic "√"-shaped waveform of LV systolic strain rate was seen, especially in the longitudinal direction. The timing and waveform analyses of systolic strain rate may be useful to distinguish between HCM and HT.

A new method to estimate pulmonary vascular resistance using diastolic pulmonary artery-right ventricular pressure gradients derived from continuous-wave Doppler velocity measurements of pulmonary regurgitation.

Pulmonary vascular resistance (PVR) is an important hemodynamic parameter in patients with heart failure, especially when the pulmonary arterial pressure is lower due to reduced stroke volume. Several echocardiographic methods to estimate PVR have been proposed, but their applications in patients with organic left-sided heart diseases have been limited. The aim of the present study was to examine the usefulness of our new method to estimate PVR (PVRPR) based on the continuous-wave Doppler velocity measurements of pulmonary regurgitation in these patients. In 43 patients who underwent right heart catheterization, PVRPR was calculated as the difference between the Doppler-derived early- and end-diastolic pulmonary artery (PA)-right ventricular (RV) pressure gradients divided by the cardiac output measured in the left ventricular outflow tract by echocardiography. The PVRPR correlated well with invasive PVR (PVRCATH) (r = 0.81, p < 0.001) without any fixed bias in Bland-Altman analysis. The conventional echocardiographic PVRs showed inadequate correlations with PVRCATH, or a obvious overestimation of PVRCATH. In the receiver operating characteristic analyses to determine the patients with abnormal elevation of PVRCATH (>3 Wood units, WU), the area under the curve was the greatest for PVRPR (0.964) compared to the conventional PVRs (0.649-0.839). PVRPR had 83 % sensitivity and 100 % specificity at the optimal cut-off value of 3.10 WU in identifying patients with PVRCATH >3 WU. Our simple and theoretical PVRPR is useful for the noninvasive estimation of PVR.

Semiautomated Segmentation and Measurement of Cytoplasmic Vacuoles in a Neutrophil With General-Purpose Image Analysis Software.

BACKGROUND: Morphological observation of blood or marrow film is still described nonquantitatively. We developed a semiautomatic method for segmenting vacuoles from the cytoplasm using Photoshop (PS) and Image-J (IJ), called PS-IJ, and measured the relative entire cell area (rECA) and relative areas of vacuoles (rAV) in the cytoplasm of neutrophil with PS-IJ. METHODS: Whole-blood samples were stored at 4°C with ethylenediaminetetraacetate and in two different preserving manners (P1 and P2). Color-tone intensity levels of neutrophil images were semiautomatically compensated using PS, and then vacuole portions were automatically segmented by IJ. The rAV and rECA were measured by counting pixels by IJ. For evaluating the accuracy in segmentations of vacuoles with PS-IJ, the rAV/rECA ratios calculated with results from PS-IJ were compared with those calculated with human eye and IJ (HE-IJ). RESULTS: The rECA and rAV/ in P1 significantly (P < 0.05, P < 0.05) were enlarged and increased, but did not significantly (P = 0.46, P = 0.21) change in P2. The rAV/rECA ratios by PS-IJ were significantly correlated (r = 0.90, P < 0.01) with those by HE-IJ. CONCLUSION: PS-IJ method can successfully segment vacuoles and measure the rAV and rECA, becoming a useful tool for quantitative description of morphological observation of blood and marrow film.

Long-term persistent fetomaternal hemorrhage.

It is not clear that how long the affected fetuses can tolerate fetomaternal hemorrhage (FMH). Incidental serial measurements of the fetal peak systolic velocity of the middle cerebral artery and the retrospective analysis of stocked blood available incidentally indicated that our patient had suffered from FMH for at least 2 weeks prior to delivery.

"Sonocytometry" ­ novel diagnostic method of ultrasonic differentiation of cells in blood flow.

Novel diagnostic method named "sonocytometry", in which streaming blood cell is diagnosed by the reflection of high frequency ultrasound from the cell, is proposed. In the present study, the differentiation of the particle size is performed as a basic study on sonocytometry. Ultrasonic backscatter signal from either 80 or 100 µm diameter polystyrene particles was measured by an ultrasonic transducer with the central frequency of 30 MHz. The spectrum of the reflected signal showed different characteristics according to the particle diameter. Theoretical value of backscatter was calculated by Faran-Hickling model and the correlation coefficient of measured and theoretical value by varying the spherical diameter showed the local maximum value at either 80 or 100 µm diameter. The principle was also validated on the streaming particles in a flow channel. The method successfully classified the particle size. Sonocytometry would be clinically applied for diagnosis of malaria or leukemia.