Establishment of functional imprinting of the H19 gene in human developing placentae.

We have found that the imprinted H19 gene can be expressed either biallelically or monoallelically in the developing human placentae. H19 biallelic expression is confined to the placenta until 10 weeks of gestation, after which it becomes exclusively maternal, and does not affect allele-specificity or levels of IGF2 expression. The promoter region of H19 is hypomethylated at all stages of placental development, while the 3' portion shows progressive methylation of the paternal allele with gestation. Our observations demonstrate that the establishment of functional H19 imprinting occurs during the early development of the placenta and provide an opportunity to understand the mechanism by which the H19 primary imprint is manifested in somatic cells.

Radiation dose reduction method combining the ECG-Edit function and high helical pitch in retrospectively-gated CT angiography.

INTRODUCTION: The purpose of this study was to demonstrate that dose reduction does not compromise image quality when combining high helical pitch (HP) and the ECG-Edit function during low HP retrospectively gated computed tomography angiography (CTA). METHODS: This study made use of a pulsating cardiac phantom (ALPHA 1 VTPC). The heart rate (HR) of the cardiac phantom was changed in five intervals, every 5 beats per minute (bpm), from 40 to 60 bpm. Evaluation of a range of HR was important because data loss might occur when combining a low HR and high HP. We performed retrospectively gated CTA scans five times using a low HP (0.16) and high HP (0.24), for each of the five HR intervals, using a 64-detector row CT scanner. The CT volume dose index (CTDIvol) was recorded from the CT console of each scan. For the images with data loss, data were repaired using the ECG-Edit function. We compared the CTDIvol, estimated cardiac phantom volume, and the visualization of the coronary ladder phantom between HP 0.16, with or without repaired HP 0.24, using the ECG-Edit function. RESULTS: Data loss occurred with a HR of 40 bpm and 45 bpm when using HP 0.24. The CTDIvol was reduced by approximately 33% with HP 0.24 when compared with HP 0.16. There were no significant differences in the mean cardiac motion phantom volume and visualization scores between HP 0.16 and with and without repaired HP 0.24 using the ECG-Edit function (p < 0.05). CONCLUSION: The ECG-Edit function is potential useful for repairing the lost data in patients with a low HR, and when combined with a high HP, it is possible to reduce the radiation dose by approximately 33%. IMPLICATIONS FOR PRACTICE: The ECG-Edit function and high HP may be a viable option in pediatric CTA studies.

Deep learning with convolutional neural network for estimation of the characterisation of coronary plaques: Validation using IB-IVUS.

INTRODUCTION: Deep learning approaches have shown high diagnostic performance in image classifications, such as differentiation of malignant tumors and calcified coronary plaque. However, it is unknown whether deep learning is useful for characterizing coronary plaques without the presence of calcification using coronary computed tomography angiography (CCTA). The purpose of this study was to compare the diagnostic performance of deep learning with a convolutional neural network (CNN) with that of radiologists in the estimation of coronary plaques. METHODS: We retrospectively enrolled 178 patients (191 coronary plaques) who had undergone CCTA and integrated backscatter intravascular ultrasonography (IB-IVUS) studies. IB-IVUS diagnosed 81 fibrous and 110 fatty or fibro-fatty plaques. We manually captured vascular short-axis images of the coronary plaques as Portable Network Graphics (PNG) images (150 × 150 pixels). The display window level and width were 100 and 700 Hounsfield units (HU), respectively. The deep-learning system (CNN; GoogleNet Inception v3) was trained on 153 plaques; its performance was tested on 38 plaques. The area under the curve (AUC) obtained by receiver operating characteristic analysis of the deep learning system and by two board-certified radiologists was compared. RESULTS: With the CNN, the AUC and the 95% confidence interval were 0.83 and 0.69-0.96, respectively; for radiologist 1 they were 0.61 and 0.42-0.80; for radiologist 2 they were 0.68 and 0.51-0.86, respectively. The AUC for CNN was significantly higher than for radiologists 1 (p = 0.04); for radiologist 2 it was not significantly different (p = 0.22). CONCLUSION: DL-CNN performed comparably to radiologists for discrimination between fatty and fibro-fatty plaque on CCTA images. IMPLICATIONS FOR PRACTICE: The diagnostic performance of the CNN and of two radiologists in the assessment of 191 ROIs on CT images of coronary plaques whose type corresponded with their IB-IVUS characterization was comparable.

Diagnostic performance of computed tomography digital subtraction angiography of the lower extremities during haemodialysis in patients with suspected peripheral artery disease.

INTRODUCTION: With intra-arterial digital subtraction angiography (DSA) considered as the gold standard, we compared the diagnostic value of computed tomography angiography (CTA) and computed tomography-digital subtraction angiography (CT-DSA in hemodialysis (HD) patients suspected of having lower limb peripheral artery disease (PAD). METHODS: In this retrospective study, we enrolled 220 HD patients with suspected PAD. CT-DSA images were obtained by subtracting unenhanced images from enhanced images. The research team calculated the area under the curve (AUC), sensitivity, specificity, positive and negative predictive value (PPV, NPV), and recorded the diagnostic accuracy between the CTA and CT-DSA images using the DSA as gold standard. Visual evaluation of calcifications in the peripheral arteries were also compared between CTA and CT-DSA images. RESULTS: At the above-knee level, the CTA AUC [95% confidence interval (CI)] was 0.68 (CI 0.64-0.72), sensitivity and specificity were 60 and 81%, PPV and NPV were 85 and 53%, and accuracy was 67%. Below the knee, these values were 0.66 (CI 0.62-0.70), 71 and 79%, 79 and 47%, and 66%. For CT-DSA, above-knee, the AUC [95% CI] was 0.88 (CI 0.85-0.91), sensitivity and specificity were 84 and 92%, PPV and NPV were 89 and 97%, and accuracy was 93%. Below the knee, these values were 0.95 (CI 0.93-0.97), 95 and 93%, 96 and 83%, and 93%. The scores for the visualization of calcification in the peripheral arteries was significantly higher for CT-DSA than CTA (p < 0.05). CONCLUSIONS: CT-DSA helps to assess stenotic PAD with high calcification in the lower extremities of HD patients. IMPLICATIONS FOR PRACTICE: On CT-DSA images, the severity of vascular calcification can be assessed for HD patients suspected of PAD of the lower extremities.

The combined application of the contrast-to-noise index and 80 kVp for cardiac CTA scanning before atrial fibrillation ablation reduces radiation dose exposure.

INTRODUCTION: To compare the radiation dose, diagnostic accuracy, and the resultant ablation procedures using 80 and 120-kVp cardiac computed tomography angiography (CCTA) protocols with the same contrast-to-noise ratio in patients scheduled for atrial fibrillation (AF) ablation. METHODS: This retrospective study was performed following institutional review board approval. We divided 140 consecutive patients who had undergone CCTA using a 64-MDCT scanner into two equal groups. Standard deviation (SD) of the CT number was set at 25 Hounsfield units (HU) for the 120-kVp protocol. To facilitate a reduction in radiation dose it was set at 40 HU for the 80 kVp protocol. We compared the two protocols with respect to the radiation dose, the diagnostic accuracy for detecting left atrial appendage (LAA) thrombi, matching for surface registration, and the resultant ablation procedures. RESULTS: At 120 kVp, the dose length product (DLP) was 2.2 times that at 80 kVp (1269.0 vs 559.0 mGy cm, p < 0.01). The diagnostic accuracy for thrombus detection was 100% using both protocols. There was no difference between the two protocols with respect to matching for surface registration. The protocols did not differ with respect to the subsequent time required for the ablation procedures and the ablation fluoroscopy time, and the radiation dose (p = 0.54, 0.33, and 0.32, respectively). CONCLUSION: For the same CNR, the DLP at 80 kVp (559.0 mGy cm) was 56% of that delivered at 120 kVp (1269.0 mGy cm). There was no reduction in diagnostic accuracy. IMPLICATIONS FOR PRACTICE: Maintaining CNR allows for a reduction in the radiation dose without reducing the image quality.

Clinical relevance of cagPAI intactness in Helicobacter pylori isolates from Vietnam.

The purpose of this paper is to investigate the relationship between clinical outcome and the intactness of cagPAI in Helicobacter pylori strains from Vietnam. The presence or absence of 30 cagPAI genes was investigated by polymerase chain reaction (PCR) and dot-blotting. H. pylori-induced interleukin-8 secretion and hummingbird phenotype, and H. pylori adhesion to gastric epithelial cells were examined. The serum concentration of pepsinogen 1, pepsinogen 2, and gastrin was also measured in all patients. cagPAI was present in all 103 Vietnamese H. pylori isolates, of which 91 had intact cagPAI and 12 contained only a part of cagPAI. Infection with the partial cagPAI strains was less likely to be associated with peptic ulcer and chronic gastric mucosal inflammation than infection with strains possessing intact cagPAI. The partial cagPAI strains lacked almost all ability to induce interleukin-8 secretion and the hummingbird phenotype in gastric cells. Their adhesion to epithelial cells was significantly decreased in comparison with intact cagPAI strains. Moreover, for the first time, we found an association between cagPAI status and the serum concentration of pepsinogens 1 and 2 in infected patients. H. pylori strains with internal deletion within cagPAI are less virulent and, thus, less likely to be associated with severe clinical outcomes.

Regulation of epidermal growth factor receptor by activated H-ras and V-myc oncogenes in mouse Balb/3T3 cells: possible roles of AP-1.

We previously reported that introduction of H-ras oncogene decreases the epidermal growth factor (EGF) binding activity to cell surface EGF receptor in mouse Balb/3T3. In this study, we have further isolated four H-ras transfectants, four v-myc transfectants and three both H-ras and v-myc (H-ras/v-myc) transfectants of mouse Balb/3T3 cells. In comparison with introduction of v-myc alone or both H-ras and v-myc oncogene, introduction of H-ras alone resulted in a loss of [125I]EGF binding activity to the cell surface EGF receptor. RT-PCR analysis also showed much lower levels of EGF receptor gene expression in H-ras transfectants compared to that of parental untransformed cells (Balb-Neo1), v-myc and H-ras/v-myc transfectants. Our results demonstrated the activated binding of a transcription factor, Stat1 p84/p91, which directly interacts with EGF receptor, to c-sis-inducible element (SIE) in both v-myc and H-rs/v-myc transfectants, but not in H-ras transfectants. Among transcription factors which we have analysed, activator protein 1 (AP-1) but not SP-1 was modulated by H-ras. Gel shift assays demonstrated the mobility pattern of TPA-responsive element (TRE) binding complex with AP-1 derived from H-ras transfectants migrated faster than those from Balb-Neo1, v-myc and H-ras/v-myc. Expression of c-Jun and Fra-1 was increased more than threefold in H-ras transfectants compared with Balb-Neo1, v-myc and H-ras/v-myc transfectants, but that of c-Fos, Jun B and SP-1 was unchanged. Both transient and permanent expression of H-ras enhanced AP-1 activity in mouse cells, but further co-introduction of dominant negative c-jun mutant encoding a transcriptionally inactive product inhibited the H-ras dependent AP-1 induction. Transfection of the dominant negative c-jun mutant also restored down-regulation of EGF binding by activated H-ras oncogene. Down-regulation of EGf receptor by activated H-ras and the possible involvement of a transcription factor, AP-1 will be discussed.

Effects of intensive lipid lowering by low-density lipoprotein apheresis on regression of coronary atherosclerosis in patients with familial hypercholesterolemia: Japan Low-density Lipoprotein Apheresis Coronary Atherosclerosis Prospective Study (L-CAPS).

Twenty-five heterozygous familial hypercholesterolemic patients treated with LDL-apheresis and drugs and 11 patients treated with drugs underwent follow-up angiography 2.3 years later. One-hundred thirteen lesions were measured by quantitative angiography. Mean LDL-cholesterol levels during the trial were 140 +/- 34 mg/dl in the apheresis group and 170 +/- 58 mg/dl (P < 0.05) in the control group. The mean changes in minimal lumen diameter of lesions were +0.19 +/- 0.30 mm (improved) in the apheresis group (n = 76) and -0.44 +/- 0.40 mm (worsened) in the control group (n = 37) (P < 0.0001). When progression and regression were defined as a change in minimal lumen diameter of +/- 0.67 mm, in the apheresis group, two (8%) patients had progression, 19 (76%) stayed unchanged and four (16%) had regression, but in the control group seven (64%) patients had progression and four (36%) stayed unchanged. The frequency of regression or no change was significantly higher in the apheresis group than in the control group (P < 0.004). Intensive cholesterol lowering therapy with LDL-apheresis and lipid lowering drugs can achieve a substantial decrease in LDL-cholesterol levels to induce regression of coronary lesions in familial hypercholesterolemic patients with advanced coronary artery disease.

Prolongation of liver allograft survival after interleukin-10 gene transduction 24-48 hours before donation.

BACKGROUND: Interleukin- (IL) 10 may be a potent regulator for controlling of allograft rejection. A single administration of IL-10 is not effective for controlling graft rejection. Gene transfer is an attractive vehicle for prolonging the expression of short-lived proteins. METHODS: Donor or recipient livers were transduced with 1 x 10(10) p.f.u. of replication-deficient adenovirus vectors harboring human IL-10 cDNA (AdCMVhIL-10) via the ileocecal vein before or after rat orthotopic liver transplantation. RESULTS: DA allografts given AdCMVhIL-10 24-48 hr before donation survived for more than 56 days in Lewis recipients, although DA allografts given the adenovirus vector 7 days or 6 hr before, and 3 days after transplantation were rejected within 30 days in recipients. Serum levels of human IL-10 in gene-transferred rats were maximum from day 2 to 7. The serum level of human IL-10 then decreased gradually, and human IL-10 was not detected by ELISA 30 days after gene-transduction. In gene-transduced long-term surviving liver allografts, IL-10 was expressed, and the expression of IL-4 was also up-regulated on posttransplant day 3, despite the expression of Th1 cytokines (IL-2 and interferon-gamma), although in rejected liver allografts, IL-2 and interferon-gamma were expressed without expression of IL-4 and IL-10. CONCLUSIONS: The prolongation of survival of IL-10 cDNA transferred liver allografts might be due to inhibition of the early phase of alloimmune-response by over expression of IL-10, despite the expression of IL-2 and interferon-gamma.

Optimal time for predicting myocardial viability after successful primary angioplasty in acute myocardial infarction: a study using myocardial contrast echocardiography.

This study sought to elucidate serial changes in microvascular integrity during papaverine-induced hyperemia in the risk area for myocardial infarction. In addition, we attempted to determine the optimal time for predicting myocardial viability. Seventy-two patients who underwent serial myocardial contrast echocardiography (MCE) before and shortly after (day 1), 1 day (day 2), and 3 weeks (day 21) after recanalization were studied. In 18 of 72 patients, MCE was performed at baseline and during hyperemia using selective intracoronary infusion of papaverine. Both the peak grayscale ratio (PGSR) within the risk area, and the no- and low-reflow ratio (LR ratio) were analyzed in each stage. Left ventricular regional wall motion (RWM) was determined 6 months after recanalization. The correlation coefficient between PGSR with papaverine on day 1 and that on day 2 was 0.54 (p = 0.02); it was 0.50 (p = 0.04) between day 1 and day 21, and 0.82 (p = 0.001) between day 2 and day 21. On day 1, the correlation coefficient between the LR ratio with papaverine and RWM was 0.60 (p = 0.02), which changed to 0.72 (p = 0.003) on day 2 and 0.54 (p = 0.04) on day 21, respectively. The best time to predict viable myocardium was established on day 2 by receiver operating characteristics curves. ST-segment re-elevation, elapsed time from onset to recanalization, and antecedent angina pectoris were independent factors for PGSR on day 2 using stepwise and multiple linear regression analysis. This study suggests that the optimal time to estimate microvascular integrity for predicting myocardial viability might be 1 day after recanalization, which is neither shortly after recanalization nor during the convalescent stage.

Efficacy of triple therapy comprising rabeprazole, amoxicillin and metronidazole for second-line Helicobacter pylori eradication in Japan, and the influence of metronidazole resistance.

BACKGROUND: The widespread use of eradication therapy for Helicobacter pylori in Japan has led to an increase in antibiotic-resistant strains and the problem of re-treatment in cases of eradication failure. AIM: To perform drug sensitivity testing for metronidazole in 92 H. pylori-positive patients who had failed eradication treatment with first-line triple therapy, consisting of a proton pump inhibitor, amoxicillin and clarithromycin, and were administered metronidazole-containing second-line therapy. METHODS: Second-line eradication therapy, consisting of rabeprazole (20 mg b.d.), amoxicillin (750 mg b.d.) and metronidazole (250 mg b.d.), was administered for 1 week and the eradication rates and influence of metronidazole resistance were determined. RESULTS: The eradication rates for rabeprazole-amoxicillin-metronidazole were 88% (81/92) using intention-to-treat analysis and 91% (81/89) using per protocol analysis. The eradication rates were 97% (61/63) for metronidazole-sensitive strains and 82% (18/22) for metronidazole-resistant strains. CONCLUSIONS: As second-line H. pylori eradication treatment in Japan, rabeprazole-amoxicillin-metronidazole triple therapy is effective, even with metronidazole-resistant strains.

Eradication rates of clarithromycin-resistant Helicobacter pylori using either rabeprazole or lansoprazole plus amoxicillin and clarithromycin.

BACKGROUND: The resistance of Helicobacter pylori to clarithromycin has become one of the primary reasons for eradication failure. AIM: To compare the eradication rates of triple therapy using amoxicillin (A), clarithromycin (C) and rabeprazole (R) or lansoprazole (L) against clarithromycin-sensitive and clarithromycin-resistant strains. METHODS: Two hundred and ninety-five patients were randomly divided into four groups and treated for 1 week: 147 cases were treated with RAC, i.e. 49 cases with R20C400 (10 mg R + 750 mg A + 200 mg C, twice daily), 48 cases with R40C400 (20 mg R + 750 mg A + 200 mg C, twice daily) and 50 cases with R40C800 (20 mg R + 750 mg A + 400 mg C, twice daily); 148 cases with treated with LAC (30 mg L + 750 mg A + 200 mg C, twice daily). RESULTS: According to intention-to-treat and per protocol analyses, the eradication rates were 88% and 91% with RAC and 78% and 81% with LAC; the eradication rates with R20C400, R40C400 and R40C800 were 94%, 81% and 86%, respectively, in the intention-to-treat analysis. In addition, the eradication rates for clarithromycin-sensitive strains with RAC and LAC were 98% and 89%, respectively, and for clarithromycin-resistant strains with RAC and LAC were 8.1% and 0%, respectively. CONCLUSIONS: The eradication rate was significantly higher with RAC than LAC. The eradication rate for clarithromycin-resistant strains was low in both groups, and an improved eradication rate could not be achieved by changing the dose of clarithromycin or proton pump inhibitor.

A case of angiocentric immunoproliferative lesions (angiocentric lymphoma) associated with human T-cell lymphotropic virus type 1.

We report a case of angiocentric immunoproliferative lesions (AILs; angiocentric lymphoma) in the subcutaneous tissue associated with human T-cell lymphotropic virus type 1 (HTLV-1). The patient showed an indolent cutaneous manifestation for 9 years until death. A skin biopsy revealed an angiocentric infiltration of atypical T lymphocytes and a marked coagulative necrosis, which are histologic hallmarks of AILs. Human T-cell lymphotropic virus type 1 antibody was detected by the indirect immunofluorescence test. The proviral integration of HTLV-1 also was confirmed in a DNA extract from paraffin sections using the polymerase chain reaction. The Epstein-Barr virus genome, which has been reported to be associated with AILs, could not be detected by polymerase chain reaction. The present case indicated that HTLV-1 also should be considered one of the pathogenetic factors of AILs.

Effects of lifelong dietary restriction on somatotropes: immunohistochemical and functional aspects.

We investigated the effects of lifelong dietary restriction (DR) on growth hormone (GH)-immunoreactivity and secretory function of somatotropes using a computerized image analysis in tissue sections and a reverse hemolytic plaque assay (RHPA) in dispersed pituitary cells of male F344 rats. DR did not prevent an aging-related reduction in GH immunoreactivity in somatotropes. However, it did augment the mean optical density of GH-immunoreactive area in somatotropes, although it did not alter the immunoreactive area in somatotropes, suggestive of a greater amount of immunoreactive GH in somatotropes of DR rats. DR also increased the percentage of aggregated GH-immunoreactive area in the anterior lobe, indicating a higher cell density of immunoreactive somatotropes. RHPA also confirmed the presence of larger proportions of GH-secreting cells in dispersed cells of DR rats in response to GH-releasing hormone (GHRH) at 24 months, although no change by DR was observed in the estimated amount of GH secreted from individual cells. Our results suggest that lifelong dietary restriction may influence the cytoplasmic GH-content at the steady state, while not modulating the responsiveness of individual somatotropes to GHRH for GH release, and that the major action of DR on somatotropes is an increment in the cell number.

Effects of aging and dietary restriction on mRNA levels of receptors for growth hormone-releasing hormone and somatostatin in the rat pituitary.

Aging impairs and dietary restriction may modulate pituitary response to growth hormone (GH)-releasing hormone (GHRH) and somatostatin (SRIH) for GH secretion. Using the semiquantitative reverse-transcription polymerase chain reaction method, we analyzed the mRNA levels of the GHRH receptor (grfr) and SRIH receptor subtype 2 (sstr2) and subtype 5 (sstr5) in anterior pituitaries of male rats fed ad libitum or 30% dietary restricted. Aging reduced the mRNA levels of these receptors in a slightly different manner. The levels of grfr progressively decreased between 6 and 24 months, whereas those of sstr2 and sstr5 declined after 16 months. Dietary restriction did not diminish the aging-dependent changes, although it slightly augmented the levels of grfr, but not sstr2 and sstr5. The present results suggest that the aging-dependent impairment in pituitary response for GH secretion could result mostly from a decline in grfr rather than relative increase of sstrs. Although DR could slightly enhance the pituitary sensitivity to GHRH, the antiaging action may be minor at the level of gene expression.

Drug combinations with amoxycillin reduce selection of clarithromycin resistance during Helicobacter pylori eradication therapy.

This study investigated the relationship between the drug combinations of the eradication regimens and the prevalence of acquired resistance to clarithromycin. Of 540 patients treated with anti-Helicobacter pylori regimens containing clarithromycin, 55 patients (31 males, mean age, 45.6 years) with failed eradication of H. pylori that was susceptible to clarithromycin before treatment were included. The E test was used to test for susceptibility to clarithromycin (minimum inhibitory concentration (MIC) <1 mg/l). Of the 55 patients, 33 (60.0%) developed clarithromycin resistance after failed eradication. Of the dual therapies, the combination of a proton pump inhibitor (PPI) and clarithromycin resulted in 88.9% (8/9) of the patients acquiring clarithromycic-resistance. With the triple therapies, the percentages of patients acquiring clarithromycin resistant strains after using a PPI+clarithromycin+amoxycillin or a PPI+clarithromycin+metronidazole were 38.7% (12/31) and 90.0% (9/10), respectively (P<0.01). These data suggest that regimens containing amoxycillin may prevent the selection of secondary clarithromycin resistance.