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Introduction: Kawasaki disease (KD), which is the most common multisystem vasculitis with unknown causes in childhood, causes coronary artery lesions (CALs). Treatment with a high dose of intravenous immunoglobulin (IVIG), plus steroids if needed, is the most effective therapy for the acute phase of KD. However, there are some very severe cases who need several times additional treatments and are at risk for CALs. In Japan, there are some scoring systems that initially predict IVIG-resistant patients. However, the problem is that these scoring systems fail in multiethnic populations. The aim of this study is to find universal biomarkers that predict treatment-resistant cases of KD. Methods: The subject was 276 KD patients, including Group 1 (n=214) who needed only 1 st line treatment, Group 2 (n=48) who needed 2 nd line treatment, Group 3 (n=14) who needed 3 rd line treatment or more. Tenascin C (TN-C), Pentraxin 3 (PTX3) and Procalcitonin (PCT) values, which were selected by systematic review, were measured before initial treatment in each group. Results: TN-C; 99.8±41.05 ng/ml in Group 1, 118.0±71.4 ng/ml in Group 2 and 183.0±25.0 ng/ml in Group 3. The TN-C level of Group 3 was significantly higher than that of all the others (p Conclusions: It may be possible to predict treatment-resistant KD cases with high sensitivity and specificity by combining the measurement from the universal biomarkers, TN-C, PTX3 and PCT, before initial treatment.

Background: Left ventricular noncompaction (LVNC) is a hereditary type of cardiomyopathy. Although it is associated with high morbidity and mortality, the related ion channel gene variants in children have not been fully investigated. This study aimed to elucidate the ion channel genetic landscape of LVNC and identify genotype-phenotype correlations in a large Japanese cohort. Methods: We enrolled 206 children with LVNC from 2002 to 2017 in Japan. LVNC was classified as follows: LVNC with congenital heart defects, arrhythmia, dilated phenotype, or normal function. In the enrolled patients, 182 genes associated with cardiomyopathy were screened using next-generation sequencing. Results: We identified 99 pathogenic variants in 40 genes in 87 patients. Of the pathogenic variants, 8.8% were in genes associated with channelopathies, 27% were in sarcomere genes, and 11.5% were in mitochondrial genes. Ion channel gene variants were mostly associated with the arrhythmia classification, whereas sarcomere and mitochondrial gene variants were associated with the dilated phenotype. Echocardiography revealed that the group with ion channel gene variants had almost normal LV ejection fraction and LV diastolic diameter Z scores. Fragmented QRS, old age, and an arrhythmia phenotype were the most significant risk factors for ventricular tachycardia ( P =0.165, 0.0428, and 0.0074, respectively). Moreover, the group with ion channel variants exhibited a greater risk of a higher prevalence of arrhythmias such as ventricular tachycardia, rather than congestive heart failure. Conclusions: This is the first study that focused on genotype-phenotype correlations in a large pediatric LVNC patient cohort with ion channel gene variants that were determined using next-generation sequencing. Ion channel gene variants were strongly correlated with arrhythmia phenotypes. Genetic testing and phenotype specification allow for appropriate medical management of specific LVNC targets.

Cardiomyocytes derived from induced pluripotent stem cells (iPSC) are used to evaluate the function and risk factors of genetic variations linked to congenital and adult heart diseases in human beings. Our purpose in this study was to establish an in vitro cardiac disease model from an immortalized B-cell line stock using patient-specific iPSC-derived cardiomyocytes. Long QT syndrome (LQTS) 3 is caused by a gain-of-function mutation in the SCN5A gene, resulting in a prolonged QT interval as well as the development of early after-depolarizations (EADs). We generated LQT3-iPSC lines from immortalized B-cell lines transformed from peripheral blood obtained from a 24-day-old male (patient #1) and an 11-year-old female (patient #2), both diagnosed with type-3 LQTS due to SCN5A missense mutation (patient #1: exon 28, G4868A, R1623Q; patient #2: exon 20, G3578A, R1193Q). Several control iPSC lines were also created from samples collected from healthy candidates (26–36-year-old male and female). In both the generated patient iPSC lines, mutated genes were not altered after reprogramming, and normal karyotypes were observed. The pluripotency of all these iPSC lines was proven by in vivo teratoma formation and the expression of pluripotent stem cell marker genes and proteins. To evaluate the electrophysiological properties at the multicellular level, we studied the iPSC-derived cardiac cell layers with a microelectrode array mapping system (MED 64, Alpha MED Scientific Inc.). The recorded extracellular electrograms were analyzed to measure the field-potential duration (FPD), which was normalized to account for variations in beating frequency, using Fridericia’s correction (FPDcF, analogous to the QTc interval in the electrocardiogram). FPDcF was observed to be longer in LQT3 patient-derived specimens when compared to that in healthy control specimens (patient #1: P < 0.05, N = 17 and patient #2: P < 0.0001, N = 19 when compared to healthy control N = 39). Moreover, EADs were observed in both LQT3 patient specimens after 15–30 nM of E-4031 was added exclusively to patient samples. The results indicated that LQT3 patient iPSC-derived cardiac cells expressed prolonged QT duration (a common pathophysiological feature of LQTS) and EAD development with marked arrhythmogenicity in our in vitro disease model system.

The incidence of endometrial serous carcinoma (ESC) has been increasing, and ESC is resistant to treatment. We report a patient with ESC who responded to radiotherapy for multiple recurrences. The first recurrence was detected in the vaginal wall and left internal iliac lymph node 5 months after the initial treatment. Concurrent chemoradiotherapy (CCRT) was administered. Radiation was delivered using the intensity modulated radiation therapy technique. The second recurrent tumor was detected in the right internal iliac lymph node after 4 months, and CCRT was conducted. After 4 months, the third recurrence was detected in the right common iliac node, and CCRT was performed. After 8 months, the fourth recurrence was detected in the horizontal portion of the duodenum, and radiotherapy was administered. After 9 months, the fifth recurrence was detected in the vaginal wall. Interstitial brachytherapy was conducted. Grade 2 gastrointestinal injury, nausea and radiodermatitis were observed. During the subsequent 13-month follow-up, there has been no recurrence. Although ESC is resistant to treatment, radiotherapy could be effective in some cases. Even when multiple recurrences occur, radiotherapy may be considered a treatment option if the irradiation level is permissible.

Background Kawasaki disease (KD), a systemic vasculitis, is suspected to be related to abnormalities in innate immunity. Based on the important role of IL-1 signaling in innate immunity, we investigated the effects of an anti-IL-1β antibody using a Candida albicans water-soluble fraction (CAWS)-induced mouse model of KD. Methods CAWS (0.5 mg/mouse) was injected intraperitoneally into 5-week-old DBA/2 mice on five consecutive days. An anti-Murine IL-1β antibody (01BSUR) was administered at various doses (2.5, 5.0, and 10.0 mg/kg) and time points (2 days before, same day, and 2, 5, 7, and 14 days after CAWS administration). After 4 weeks, vasculitis in the aortic root was investigated histologically. Cytokines including IL-1β, -6, -10, and TNF-α were also measured. Results Groups administered 01BSUR at all doses showed a significant reduction in the area of vasculitis. In addition, 01BSUR inhibited vasculitis until 7 days after CAWS administration. In the analysis of various time points, the level of IL-6 was lower in all groups compared to the CAWS only group, but the levels of IL-1β, TNFα, and IL-10 were lower when 01BSUR was administered before CAWS. On the other hand, TNFα and IL-10 levels were restored when 01BSUR was administered after CAWS, suggesting that 01BSUR may have additional effects beyond blocking IL-1β signaling. Conclusions The anti-IL-1β antibody significantly attenuated CAWS-induced vasculitis. The mechanism of inhibiting vasculitis is thought to include inhibition of the IL-1β pathway and additional effects beyond blocking IL-1β signaling.

BACKGROUND Tenascin-C (TN-C) is an extracellular matrix glycoprotein that is heavily upregulated at sites of inflammation. We conducted a retrospective study to assess the utility of TN-C as a novel biomarker to predict the risk of developing coronary artery lesions (CAL) and resistance to intravenous immunoglobulin (IVIG) in patients with Kawasaki disease (KD).Methods and Results:We collected blood samples of 111 KD patients (IVIG-responder: 89, IVIG-resistant: 22; CAL: 8) and 23 healthy controls, and measured the serum levels of TN-C. TN-C levels on admission were significantly higher in patients than in healthy controls and in patients during convalescence after IVIG administration (69.6 vs. 20.4 vs. 39.7 ng/ml, respectively; P

Lissencephaly is a type of cortex convolution malformation caused by a neuronal migration disorder in early fetal development. It is characterized by a thick cortex and agyria (absence of gyri) or pachygyria (broad gyri) on the brain surface. Clinical manifestations include severe mental retardation and intractable epilepsy. Here, we report a four months old boy with lissencephaly started focal seizures in a patient aged 3 months who was treated with 2 anticonvulsants. Spasms and regression of psychomotor development were present from 6 months of age. West syndrome was diagnosed on the basis of the types of seizures and electroencephalographic findings (hypsarrhythmia). Treatment with ACTH was started in an attempt to control the spasms. Decreases in convulsive seizures and recovery of regressed psychomotor development were subsequently noted. However, convulsive seizures recurred when the ACTH was tapered. The epileptic seizures accompanying lissencephaly thus appear to be intractable.

Hypertrophic pyloric stenosis (HPS) is the principal disease to consider in neonates presenting with frequent projectile vomiting and poor weight gain. Ramstedt pyloromyotomy is commonly used for the surgical treatment of HPS. The present study investigated the efficacy of nonsurgical medical treatment using intravenous administration of atropine and the examined the clinical course and results of ultrasonography and a contrast upper gastrointestinal series. A 34-day-old girl was admitted with chief complaints of projectile vomiting and poor weight gain. HPS was diagnosed on the basis of the clinical course and results of imaging studies. After intravenous administration of atropine, projectile vomiting resolved and weight increased without complications. On imaging studies, barium introduced into the stomach by tube rapidly entered the duodenum after atropine administration. Ultrasonography initially showed no reductions in hypertrophic muscle in the pyloric region, but gradual reductions were identified in subsequent months.

Background: Kawasaki disease (KD) is a systemic vasculitis and the leading cause of acquired heart disease. There are some cases that show unresponsiveness to initial intravenous immunoglobulin (IVIG) and require addition treatment. High incidence of coronary artery lesions (CAL) is seen in unresponsive cases. Pentraxin 3 (PTX3) is produced at the site of vascular inflammation, and used as a new biomarker for vasculitis. The aim of this study is to explore the application of PTX3 to KD. Methods: 128 patients with KD are enrolled. Blood samples are collected at before IVIG and 1, 3, 6 month later from the onset of KD. PTX3 values are compared with IVIG unresponsive scoring system by Kobayashi et al. (Circulation, 2006). Results: Mean values of PTX3 before IVIG and 1, 3, 6 month were respectively 25.1*, 7.1*, 3.8, 3.6 ng/ml (asterisk shows statistical significance with age-matched control: 3.6). Mean values of PTX3 in unresponsiveness (n=20) and responsiveness (n=108) at before IVIG and 1, 3, 6 month were 46.8* vs. 20.9, 9.1* vs. 6.7, 4.2 vs. 3.7, 3.9 vs. 3.5 ng/ml (asterisk shows statistical significance between two groups). There is statistical positive correlation between PTX3 and score points by Kobayashi et al. (r=0.602, p=0.000). According to the statistical analysis, the area under the ROC curve (AUC) was 0.87 and sensitivity and specificity of PTX3 as IVIG unresponsiveness prediction were 90 and 81 %, if cut off value was set as 28 ng/ml. Conclusions: 1. vasculitis continues at least 1 month after onset of disease. 2. PTX3 can be a candidate biomarker for prediction of unresponsiveness in patients with KD.

The electrophysiology of the heart changes dramatically during development. These changes in electrical properties are caused by changes in the number (density) and types of ion channels, with resultant changes in the magnitude and kinetic properties of the various types of ionic currents. The alterations in the ion channels are resposible for the changes in the resting potential and characteristics of the action potentials. In this review, we focused on the developmental changes of the L-type Ca2+ channel (especially its regulation via β-adrenoceptor, and muscarinic acetylcholine agonsist, tyrosine kinase) and the T-type Ca2+ channel. As mentioned in this review, the immature heart differs from the adult heart in both size and function. Therefore, we have to pay attention to the characteristics of the immature heart when we treat neonates and children.

ObjectivesWe used myocardial fractional flow reserve (FFRmyo) and coronary flow reserve (CFR) to estimate cut-off values for assessment of the functional severity of coronary stenosis and myocardial ischemia, and we tested the usefulness of coronary blood hemodynamic measurements before and after plain old balloon angioplasty (POBA) and coronary artery bypass graft surgery (CABG).BackgroundFractional flow reserve and CFR are useful for assessing the functional severity of coronary artery stenosis, coronary microvascular dysfunction, and myocardial ischemia during cardiac catheterization in adults. However, there have been no reports on the use of these measurements in children with Kawasaki disease (KD).MethodsThe study group included 128 patients with 314 coronary branches. The subjects were classified into three groups: normal coronary group, with 206 branches; abnormal coronary artery without ischemia group, with 58 branches; and ischemia group, with 50 branches.ResultsIn each branch, CFR and FFRmyowere significantly lower in the ischemia group than in the other groups. Cut-off values for assessing the functional severity of coronary stenosis and CFR were approximately equal to those obtained for adults (CFR

The LSR (label switch router), which uses label switching technology, performs packet transmission by a label transmission module that uses fixed-length labels, in addition to by an ordinary network transmission module. It realizes high-speed and high-performance packet transmission by making correspondence with IP packets and fixed-length labels, and by transmitting them using the label transmission module. The basic part of unicast label distribution protocol used by LSR has been specified by IETF (Internet Engineering Task Force). As for multicast label switching technology, various protocols (FANP [5], TAG [7], IPSOFACT [11]) have been proposed, and they have been compared by Ooms and colleagues [13]. However, there has been no study that evaluated the effectiveness of multicast label switching technology by comparing two cases, one that uses label switching technology for multicast packet transmission and another that does not. In this article, we evaluate the effectiveness of multicast label switching technology by designing and implementing FANP, a protocol for multicast label switching, by measuring LSP setup time, which is overhead of LSR compared with ordinary routers, and by measuring throughput for the case with label switching technology and for the case without it. LSP setup time was about 40 ms per LSR, which shows that overhead of label switching technology is small. Also, we verified that the throughput is higher when we used the label switching technology than when we did not use it, and it is effective in routers that reproduce multicast packets, such as backbone routers with many multicast receivers. © 2004 Wiley Periodicals, Inc. Electron Comm Jpn Pt 1, 87(7): 66–74, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecja.1189

To evaluate the potency of levosimendan, a newly developed cardiotonic agent, as a phosphodiesterase-3 inhibitor, we examined its effects on the L-type Ca 2+ current ( I Ca,L ) in single human atrial cells using the whole-cell voltage-clamp method. Levosimendan significantly increased I Ca,L in a concentration-dependent manner ( E max , 139.0±1.8%; EC 50 , 54±3.6 nM). The increase in I Ca,L induced by 1 μM levosimendan was significantly greater in human atrial cells (136.7±11.0%, n =8) than in rabbit atrial cells (23.5±3.5%, n =6) (depolarization to +10 mV in each case). In rat atrial and ventricular cells, I Ca,L was unaffected by 1–10 μM levosimendan. These results indicate that the selective phosphodiesterase-3 inhibitor levosimendan increases cardiac-cell I Ca,L significantly more strongly in human than in rabbit and rat. It seems likely that the positive inotropic effect of levosimendan on the human myocardium depends on an increase in I Ca,L that is modulated by adenosine 3′5′-cyclic monophosphate (cAMP)-dependent phosphorylation.

Label-switching technology enables high performance and flexible layer-3 packet forwarding based on the fixed-length label information that is mapped to the layer-3 packet stream. A label-switching router (LSR) forwards layer-3 packets based on their layer-3 address information or their label information that is mapped to the layer-3 address information. Two label-mapping policies have been proposed. One is traffic driven mapping, where the label is mapped for a layer-3 packet stream of each host-pair according to the actual packet arrival. The other is topology driven mapping, where the label is mapped in advance for a layer-3 packet stream toward the same destination network, regardless of actual packet arrival to the LSR. This paper evaluates the required number of labels under each of these two label-mapping policies using real backbone traffic traces. The evaluation shows that both label-mapping policies require a large number of labels. In order to reduce the required number of labels, we propose a label-mapping policy that is a combination of the two label-mapping policies above. This is traffic-driven label mapping for the packet stream toward the same destination network. The evaluation shows that the proposed label-mapping policy requires only about one-tenth as many labels as the traffic-driven label mapping for the host-pair packet stream and the topology-driven label mapping for the destination-network packet stream.

Postnatal changes in the slow Ca2+ current (ICa(L)) were investigated in freshly isolated ventricular myocytes from neonatal (1-7 days old) and adult (2-4 months old) rats, using whole-cell voltage clamp and single-channel recordings. The membrane capacitance (mean ± SEM) averaged 23.2 ± 0.5 pF in neonates (n = 163) and 140 ± 4.1 pF in adults (n = 143). ICa(L) was measured as the peak inward current at a test potential of +10 mV (or +20 mV) by applying a 300-ms pulse from a holding potential of -40 mV; 1.8 mM Ca2+ was used as charge carrier. The basal ICa(L) density was 6.7 ± 0.2 pA/pF in neonatal and 7.8 ± 0.2 pA/pF in adult cells (p < 0.05). The time course of inactivation of the fast component (at +10 ms) was significantly longer in the neonatal (10.7 ± 1.4 ms) than in the adult (6.6 ± 0.4 ms) cells (p < 0.05). Ryanodine (10 muM) significantly increased this value to 18.0 ± 1.9 in neonate (n = 8) and to 17.7 ± 2.0 in adult (n = 9). For steady-state inactivation, the half-inactivation potential (Vh) was not changed in either group. For steady-state activation, Vh was 5.1 mV in the neonatal (n = 6) and -7.9 mV in the adult cells (n = 7). Single-channel recordings revealed that long openings (mode-2 behavior) were occasionally observed in the neonatal cells (11 events from 1080 traces / 11 cells), but not in the adult cells (400 traces / 4 cells). Slope conductance was 24 pS in both the neonatal and adult cells. Results in rat ventricular myocytes suggest the following: (i) the peak Ca2+ current density is already well developed in the neonatal period (being about 85% of the adult value); (ii) the fast component of inactivation is slower in neonates than in adults; and (iii) naturally occurring long openings are occasionally observed in the neonatal stage but not in the adult. Thus, the L-type Ca2+ channels of the neonate were slightly lower in density, were inactivated more slowly, and occasionally exhibited mode-2 behavior as compared with those of the adult.Key words: Ca2+ channels; neonatal heart cells; whole-cell voltage clamp; single-channel recordings; long openings of Ca2+ channels.

Postnatal changes in the slow Ca 2+ current (ICa(L)) were investigated in freshly isolated ventricular myocytes from neonatal (1-7 days old) and adult (2-4 months old) rats, using whole-cell voltage clamp and single-channel recordings. The membrane capacitance (mean ± SEM) averaged 23.2 ± 0.5 pF in neonates (n = 163) and 140 ± 4.1 pF in adults (n = 143). ICa(L) was measured as the peak inward current at a test potential of +10 mV (or +20 mV) by applying a 300-ms pulse from a holding potential of -40 mV; 1.8 mM Ca 2+ was used as charge carrier. The basal ICa(L) density was 6.7 ± 0.2 pA/pF in neonatal and 7.8 ± 0.2 pA/pF in adult cells (p < 0.05). The time course of inactivation of the fast component (at +10 ms) was significantly longer in the neonatal (10.7 ± 1.4 ms) than in the adult (6.6 ± 0.4 ms) cells (p < 0.05). Ryanodine (10 μM) significantly increased this value to 18.0 ± 1.9 in neonate ( n = 8) and to 17.7 ± 2.0 in adult (n = 9). For steady-state inactivation, the half-inactivation potential ( Vh) was not changed in either group. For steady-state activation, Vh was 5.1 mV in the neonatal (n = 6) and -7.9 mV in the adult cells (n = 7). Single-channel recordings revealed that long openings (mode-2 behavior) were occasionally observed in the neonatal cells (11 events from 1080 traces / 11 cells), but not in the adult cells (400 trace s / 4 cells). Slope conductance was 24 pS in both the neonatal and adult cells. Results in rat ventricular myocytes suggest the following: ( i) the peak Ca 2+ current density is already well developed in the neonatal period (being about 85% of the adult value); ( ii) the fast component of inactivation is slower in neonates than in adults; and (iii) naturally occurring long openings are occasionally observed in the neonatal stage but not in the adult. Thus, the L-type Ca 2+ channels of the neonate were slightly lower in density, were inactivated more slowly, and occasionally exhibited mode-2 behavior as compared with those of the adult. 881

Background Myocardial ischemia and myocardial infarction are the most serious complications of coronary artery lesions in children with Kawasaki disease (KD). Therefore, early detection and treatment of myocardial ischemia in patients with KD is essential. We studied the effectiveness of percutaneous transluminal coronary angioplasty (PTCA) in patients with silent myocardial ischemia detected by dobutamine stress 99m Tc myocardial scintigraphy (TMS), body surface mapping (BMS), and signal-averaged ECG late potentials (ELP). Methods and Results Eight of 76 asymptomatic patients with a coronary stenosis>25% and a positive dobutamine stress test were considered to have silent myocardial ischemia. All eight patients had>95% stenoses demonstrated by coronary angiography (CAG) just before PTCA. After PTCA, CAG showed that all of the coronary artery stenoses had been reduced to Conclusions PTCA effectively dilates stenotic coronary arteries in children with KD. Moreover, dobutamine stress TMS, BMS, and ELP are useful for detecting silent myocardial ischemia and estimating the effectiveness of PTCA. Furthermore, IVUS is useful for evaluating the severity of coronary artery lesions before and after PTCA in patients with KD.

The electrophysiological effect of levosimendan, a novel Ca(2+)-sensitizing positive inotropic agent and vasodilator, was examined on rat mesenteric arterial myocytes using the patch clamp technique. Resting potential was significantly hyperpolarized with levosimendan, with an EC50 of 2.9 microM and maximal effect (19.5 +/- 3.5 mV; n = 12) at 10 microM. Levosimendan (10 microM) significantly increased the whole-cell outward current. The currents intersected close to the calculated EK (-84 mV), suggesting that the activated current was a K+ current. Hyperpolarization and stimulation of K+ current by levosimendan were not prevented by 30 microM H-7 (a non-specific inhibitor of protein kinases) and 100 nM charybdotoxin (a blocker of Ca(2+)-activated K+ channels), but were abolished by 10 microM glibenclamide. In single-channel current recording in open cell-attached patches, two types of K+ channels were observed having conductances of 26 and 154 pS. The 154 pS channels were not affected by levosimendan and glibenclamide. The 26 pS channels were evoked in one-fourth of the patches when 10 microM levosimendan (and 0.1 mM UDP) was added (at -60 mV) and channel activity was abolished by glibenclamide. The mean open probability of the 26 pS channels was 0.094 +/- 0.017 (n = 9), and the mean open time (at -60 mV) was 6.6 ms in the presence of UDP and levosimendan. Although significant hyperpolarization (4.7 +/- 1.5 mV, n = 8) was observed at 1 microM levosimendan, the same concentration did not affect Ca2+ channel currents (n = 10). In summary, levosimendan hyperpolarized the arterial myocytes, probably through activation of a glibenclamide-sensitive K+ channel. This mechanism may contribute to the vasodilating action of levosimendan.

This paper describes an internetworking architecture and related protocol overview based on routers that have asynchronous transfer mode (ATM) cell switching capability in addition to conventional Internet protocol (IP) packet forwarding. The proposed architecture can provide high-throughput and low-latency switched paths for individual application flows or a group of application flows while retaining current router-based internetworking architecture. The proposed router is able to establish the switched path based on the characteristics of flows, e.g., arrival of a data packet with specific upper layer protocols or arrival of more than a certain amount of data packets in a predetermined period, as well as by the reception of an IP-layer resource reservation request, such as resource reservation protocol (RSVP). One important feature that is provided by the proposed router is interoperability with the emerging ATM network platform specified by the ATM Forum and the telecommunications sector of the International Telecommunications Union (ITU-T). The proposed routers can be interconnected with each other over the point-to-point synchronous optical network link as well as over the ATM network platform, which provides permanent virtual channel, virtual path, or switched virtual channel (SVC) services. That enables network carriers to provide Internet/intranet services as well as others, such as telephony, ATM/time division multiplexing leased line, or native ATM SVC services.

Aims: We evaluated the systolic and diastolic functions of both ventricles from the early neonatal period to adolescence using color tissue Doppler imaging and 2‑dimensional tissue tracking echocardiography. Methods: We examined 100 healthy children (aged 1―5 days n=20; 1 month n=20; 1 year n=20; 6―7 years n=20; and 12―13 years n=20. Blood flow velocities in the mitral and tricuspid valves (E were obtained with pulsed Doppler imaging and longitudinal systolic (Sand early diastolic (Epeak velocities at the mid free wall segment of both ventricles were obtained with color tissue Doppler imaging. For longitudinal strain imaging systolic peak values were obtained at the same position. In addition peak systolic radial strain was obtained from a short‑axis view of the left ventricle using the tissue tracking method. Th eE Eratio was calculated. Results: Regarding systolic indices Sincreased during development and stabilized at 6 to 7 years and longitudinal strain reached values of the 12‑ to 13‑year‑old group at 1 year of age in both ventricles. Like longitudinal strain radial strain in the left ventricle reached values of the 12‑ to 13‑year‑old group at the age of 1 year. Similarly the E Eratio was high at 1 month or younger and decreased by 1 year. Conclusions: Systolic and diastolic variables change markedly from birth to 1 year of age and show only small changes thereafter. (J Nippon Med Sch 2013; 80: 260―267

We report a case of infantile acute monocytic leukemia associated with solid tumors in the skin. Light microscopy showed that the tumor cells were mainly spindle-shaped and arranged in tandem. Immunohistochemical staining showed that fibronectin was detected in the extracellular matrix (and partially on the surface of tumor cells), integrin alpha 5 beta 1 on the cell surface, and vinculin and actin were detected in the cytoplasm of the tumor cells. Electron microscopy showed that the tumor cells had perpendicular transmembranous fibrils and closely situated collections of cytoplasmic microfilaments beneath the cell membrane suggesting fibronexus-like structures. We conclude that the expression of fibronectin, integrin alpha 5 beta 1, vinculin, cytoplasmic actin and fibronexus-like structure in leukemic cells indicate these cells possess an adhesive function. The mechanism of formation of the extramedullary solid tumors in this patients involved these adhesion molecules of the tumor cells.

呉共済病院での過去4年間の子宮内膜細胞診2346例中, 偽陽性および偽陰性の誤判定は12例存在した. この誤判定例を, 他施設の細胞検査士16名による追試判定結果と診断別, 経験年数別に比較し, さらに誤判定の原因を細胞学的, 組織学的に検討した.偽陽性は7例 (16%) あり, 内訳は萎縮性内膜 (4例), 臨床情報のなかったIUD挿入例 (1例), 流産後の合胞性内膜炎で絨毛上皮細胞の出現がほとんどない例 (1例), 変性の強い正常内膜 (1例) であった. 偽陰性は5例 (26%) で, 高分化型内膜型腺癌 (4例), 採取部位の不適当な腺棘細胞癌 (1例) であった.追試検査の誤判定率は, 萎縮性内膜21%, IUD挿入例13%, 合胞性内膜炎7%, 正常内膜50%, 高分化型内膜型腺癌44%, 腺棘細胞癌81%であった. 萎縮性内膜では経験年数11年以上の群で偽陽性率が減少する傾向を認めたが, その他は経験年数と誤判定率に関連はなかった.誤判定は, 萎縮性内膜と合胞性内膜炎では再生上皮様異型細胞, IUD挿入例では知識・情報不足, 正常内膜では細胞変性, 腺棘細胞癌では不適切な採取手技に起因し, 高分化型内膜型腺癌と一部の萎縮性内膜では細胞診上判定困難な症例が存在した.

子宮頸癌放射線治療後15年目に発生した子宮体部明細胞腺癌の1例を経験したので, その術前内膜細胞像と病理組織像について検討し, 文献的考察を加えて報告した.患者は66歳女性で, 51歳時子宮頸部扁平上皮癌にて放射線治療を受けている. 治療後15年を経過して不正出血を訴え来院した. 子宮内膜細胞診施行し, 比較的小型で, ラ信トグリーン好性の淡明で豊富な細胞質を持った異型細胞を少数認め, 子宮体部明細胞腺癌と診断した.摘出子宮において腫瘍は体部前壁に不整なポリープ様腫瘤を形成し, 病理組織学的には淡明ないし好酸性の豊富な細胞質を有する異型細胞が乳頭状, 充実性に増殖し, 明細胞腺癌と診断した. 51歳時の子宮頸癌組織には腺腔形成はなく, 癌細胞は充実性増殖, 胞巣形成がみられ, 扁平上皮癌と診断した. 両癌を免疫組織化学的に比較すると, 前者はCEA陰性, ケラチン陽性で, 後者はCEA強陽性, ケラチン強陽性と明らかに差を認めた.

The role of large-conductance voltage-dependent and calcium-activated potassium (BK(Ca)) channels in the regulation of ductus arteriosus (DA) tone is not clear. This study aimed to examine whether BK(Ca) α and β subunits and BK(Ca) currents are present in the rat DA, as well as whether the BK(Ca) channels are involved in O₂-induced ductal constriction. BK(Ca) α and β subunit transcripts (mRNAs) were detected in the DA from premature (19D) and mature (21D) rat fetuses and full-term neonates (NB) by quantitative real-time PCR. The amount of BK(Ca) α mRNAs decreased with advancing development. β1 was the dominant β subunit in the DA, and the amount of β1 mRNAs was greatest in the mature DA. Immunofluorescence staining showed that the majority of BK(Ca) α and β1 proteins were colocated with alpha smooth muscle actin (α-SMA) in the tunica media of the DA in all age groups. The protein expression of the α subunit was greatest in the mature DA, while the expression of the β1 subunit did not differ among all three groups. The 19D and 21D ductus tensions were recorded under various conditions by myograph. The 19D ductus rings exhibited poor O₂ sensitivity and no response to BK(Ca) inhibitor (paxilline) or activator (NS1619). The 21D ductus rings developed significant constriction induced by O₂. Paxilline did not increase the 21D DA tension under either hypoxic or oxygenated conditions. NS1619 dilated the 21D DA only under oxygenated conditions. The recorded BK(Ca) currents were greatest in the 21D DA smooth muscle cells (SMCs) upon using a whole-cell patch clamp. Our study indicated that BK(Ca) channels exist in the DA but are not involved in O₂-induced ductal constriction. Activation of BK(Ca) channels led to vasodilatation in the preconstricted DA induced by O₂, possibly suggesting a way to maintain the patency of DA after birth.

出生時より存在した鼻部腫瘤で発症した先天性白血病の1例を報告した。初診時 (生後4ヵ月) には頭部・背部・大腿部にも腫瘤をみとめ, 血液学的に急性単球性白血病であった。単球性白血病では皮膚浸潤が比較的多いことが知られており, 特に先天性白血病では皮膚結節が初発症状として重要である。近年, 化学療法剤の開発や治療技術の進歩により長期生存例も報告されるようになった。

Background It was investigated whether edaravone, a potent free radical scavenger, would protect against anthracycline-induced cardiotoxicity and prevent cardiac function deterioration. Methods and Results Cultured neonatal rat cardiomyocytes were stimulated by daunorubicin 1 μmol/L either with or without edaravone or superoxide dismutase mimetic Mn (III) tetrakis (1-methyl-4-pyridyl) porphyrin pentachloride (MnTMPyP). Cell viability was estimated by measuring the amount of lactate dehydrogenase (LDH) released into the culture medium. Apoptosis was determined by a caspase-3 activity assay and a histone - DNA complex fragment assay. To investigate whether edaravone interfered with daunorubicin's anti-tumor effect, daunorubicin and edaravone were added to human leukemia K562 cells, and the surviving cells were counted. In addition, edaravone's in vivo effect was evaluated using Sprague - Dawley rats. A total of 15 mg/kg doxorubicin was injected intraperitoneally either with or without simultaneous edaravone injection. Two and 6 weeks after the final injection, left ventricular diastolic diameter and left ventricular fraction shortening were assessed echocardiographically. The LDH assay showed that edaravone significantly inhibited LDH release from cardiac myocytes (p=0.0428). The caspase-3 activity and histone - DNA complex fragment assays demonstrated that edaravone's apoptosis suppression effect was much weaker than that of MnTMPyP. The in vivo study showed that edaravone prevented doxorubicin-induced cardiac deterioration. Finally, edaravone was found to not affect daunorubicin's anticancer effect on K562 cells. Conclusions Edaravone protects cardiomyocytes from anthracycline-induced cardiotoxicity via an anti-necrotic rather than an anti-apoptotic effect. (Circ J 2007; 71: 1815 - 1820)

Background Kawasaki disease (KD) causes coronary artery disease (CAD) in children. In addition, a history of KD is suspected to be a risk factor for the development of atherosclerotic heart disease in the future. Histological senescence changes are a common denominator in atherosclerotic lesions in adults, so the present study investigated whether histological senescence changes had already occurred in KD aneurysm. Methods and Results KD coronary aneurysms and internal mammary arteries retrieved from 5 children with KD (3, 4, 5, 6, and 11 years old, respectively) who underwent coronary artery bypass grafting, as well as giant coronary aneurysm size-reducing operations, were analyzed. Senescence-associated strong β-galactosidase activity was observed in KD aneurysms, but not in the internal mammary arteries. An immunohistochemical analysis of the KD aneurysm using anti-CD31, anti-endothelial nitric oxide synthetase (eNOS), anti-vascular adhesion molecule-1 (VCAM-1), and anti-monocyte chemoattractant protein-1 (MCP-1) showed vascular endothelium CD31 staining, decreased staining of eNOS and strong staining of MCP-1 and VCAM-1. cDNA microarray gene expression profiling revealed increased MCP-1 expression in the KD aneurysm, a finding confirmed by quantitative polymerase chain reaction. Conclusions Histological features of senescence and active remodeling gene expression show that the KD aneurysm is not a silent vasculitis terminal. The future fate of KD aneurysms, including atherosclerosis, should be monitored carefully. (Circ J 2007; 71: 709 - 715)

A 6-year-old boy was hospitalized because of dark feces and facial pallor of 1 weeks duration. Other gastrointestinal symptoms, including vomiting and abdominal pain, were absent, but he felt dizziness when standing and fatigue on effort. Hematologic studies revealed iron-deficiency anemia, and endoscopy showed gastric erosions and a duodenal ulcer. All test results for Helicobacter pylori infection, including H. pylori antigen in stool, anti-H. pylori IgG immunoassay in serum, and the (13)C-urea breath test, were positive. Because an H. pylori-associated gastric ulcer had been diagnosed with endoscopy in the patients father 3 years earlier, father-son transmission was suspected. The patient was treated with triple-agent eradication therapy (proton pump inhibitor [lansoprazol], amoxicillin, and clarithromycin) for 2 weeks. One month after therapy was completed, eradication of H. pylori was confirmed by negative results on the stool antigen test. Peptic ulcer disease can occur in young children, as in this case. The stool antigen test kit is a useful and reliable method that can be used even in preschool children to diagnose H. pylori infection.

Two actin filament-depolymerizing agents, DNase I and cytochalasin D, were used to examine the involvement of the cytoskeleton in the functional interaction between the sulfonylurea receptor (SUR) and the ATP-sensitive K+ (KATP) channels. Isolated rate ventricular cardiomyocytes were studied using open cell-attached patches for single-channel recording. Bath application of DNase I (100 microg/ml) or cytochalasin D (10 microM) stimulated the KATP channel activities (in presence of 30 microM ATP), and these channels became resistant to inhibition by tolbutamide (0.5 mM). After exposure to tolbutamide, the relative NPo value was 0.09 +/- 0.02 in control patches in absence of actin disrupters, and 0.67 +/- 0.22* or 0.65 +/- 0.10*, respectively, in cells treated with DNase I or cytochalasine D (*P0.05 vs. control). The inhibitory action of glibenclamide (10 microM) on the KATP channels was also attenuated by DNase I. Thus, the disruption of the actin cytoskeleton attenuates the ability of SUR to inhibit the opening of KATP channels.

The advent of the mobile wireless Internet has created the need for seamless and secure communication over heterogeneous access networks such as IEEE 802.11, WCDMA, cdma2000, and GPRS. An enterprise user desires to be reachable while outside one's enterprise networks and requires minimum interruption while ensuring that the signaling and data traffic is not compromised during one's movement within the enterprise and between enterprise and external networks. We describe the design, implementation and performance of a Secure Universal Mobility (SUM) architecture. It uses standard protocols, such as SIP and Mobile IP, to support mobility and uses standard virtual private network (VPN) technologies (e.g., IPsec) to support security (authentication and encryption). It uses pre-processing and make-before-break handoff techniques to achieve seamless mobility (i.e., with little interruption to users and user applications) across heterogeneous radio systems. It separates the handlings of initial mobility management and user application signaling messages from user application traffic so that VPNs can be established only when needed, thus reducing the interruptions to users.

In this work, we measure wireless local area network (WLAN) voice performance and capacity. While most WLAN applications today are data centric, the growing popularity of Voice over IP (VoIP) applications and the escalating trend towards convergence with cellular networks will catalyze an increased mix of voice traffic. Since voice applications must compete with each other and with simultaneous data applications for WLAN bandwidth, quantifying voice performance and capacity in the presence of background data traffic is an important issue. We offer a practical investigation into the ability of 802.11b MAC layer to support simultaneous voice and data. We quantify VoIP capacity for standard WLAN networks, indicative of those already in the field, as well as evaluate the practical benefits of implementing backoff control and priority queuing at the access point. Conclusions are drawn based on an extensive set of real-world measurements conducted using off-the-shelf equipment in a commercial testbed.

There are several issues to be resolved when the legacy ATM switch is applied as a forwarding engine for the label switching router (LSR). They are mostly due to inconsistency between the requirements for the current and future Internet and capabilities of most of legacy ATM switches. This paper clarifies issues in constituting ATM-based LSR (ATM-LSR) networks as the Internet backbone, label resource consumption, effect of IP routing loop, support of quality/class of services, and finally desirable solutions for them.

The changes in endothelium-derived vascular regulatory factors during dobutamine (DOB)-induced myocardial ischemia (MI) were investigated in 21 patients with Kawasaki disease aged from 11 months to 18 years. They were classified into an ischemia group (8 patients) and a non-ischemia group (13 patients) based on the results of 99mTc myocardial scintigraphy and DOB stress 99mTc myocardial scintigraphy. In the ischemia group, MI was relatively mild, because there were ischemic changes on the electrocardiogram and no significant symptoms during DOB stress. Catheters were positioned near the orifice of the coronary artery (Ao) and at the coronary sinus (CS). Hemodynamics and the blood concentrations of lactic acid and endothelin-1, as well as NO3-, 6-keto-prostaglandin F1alpha, and thromboxane B2, (which are inactive metabolites of nitric oxide, prostaglandin I2 and thromboxane A2, respectively), were measured at rest and after DOB stress (maximum dose: 30 microg x kg(-1) x min(-1)). The CS/Ao ratio was determined for all parameters. The rate-pressure product, an index of work load, and the cardiac index were significantly increased by DOB stress in both groups. Coronary angiography showed no vasospasm of the epicardial coronary arteries before or after DOB stress in either group. The plasma concentrations of endothelin-1 and 6-keto-prostaglandin F1alpha were significantly increased after DOB stress in the ischemia group, but the serum concentration of NO did not increase. The lack of an increase in NO production during DOB stress may have contributed to the worsening of MI in patients with Kawasaki disease.

Label switching technology enables high performance, flexible, layer-3 packet forwarding based on the fixed length label information mapped to the layer-3 packet stream. A Label Switching Router (LSR) forwards layer-3 packets based on their label information mapped to the layer-3 address information as well as their layer-3 address information. This paper evaluates the required number of labels under traffic-driven label mapping policy using the real backbone traffic traces. The evaluation shows that the label mapping policy requires a large number of labels. In order to reduce the required number of labels, we propose a label mapping policy which is a traffic-driven label mapping for the traffic toward the same destination network. The evaluation shows that the proposed label mapping policy requires only about one tenth as many labels compared with the traffic-driven label mapping for the host-pair packet stream,and the topology-driven label mapping for the destination network packet stream.© (1998) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

We investigated the postnatal developmental changes produced by the muscarinic receptor agonist, carbachol, on the L-type Ca2+ current (ICa(L)) in neonatal (aged 5 to 7 days) and adult (aged 2 to 5 months) rat ventricular cells by using the whole-cell voltage clamp technique. Carbachol inhibited the isoproterenol-stimulated ICa(L). The maximal inhibition was 89.3 +/- 4.8% (n = 5) in neonatal cells and 17.7 +/- 7.7% (n = 9) in adult cells. Carbachol inhibited the forskolin-stimulated ICa(L) to almost same extent as the isoproterenol-stimulated ICa(L). In the cells pretreated with pertussis toxin, carbachol failed to inhibit the isoproterenol-stimulated ICa(L), indicating that carbachol produced its effect via a pertussis toxin-sensitive G-protein pathway. The effects of carbachol in adult cells became more pronounced, increasing from 17.7% to 54.8% (n = 11), with the addition of the synthetic inhibitory G-protein alpha subunit (Gi alpha) (1 microM) to the reaction. Conversely, the alpha subunit of another pertussis toxin-sensitive synthetic G-protein (G(o) alpha, 1 microM) failed to mimic the effect of Gi alpha. These results suggest that, in rat ventricular cells, (1) the action of carbachol on ICa(L) showed a marked decrease during development; (2) the decrease in the effect of carbachol in adult cells is in part due to a decrease in the activity of pertussis toxin-sensitive G protein, especially Gi alpha.

Ruthenium red is a well known inhibitor of Ca2+ uptake into mitochondria in vitro. However, its utility as an inhibitor of Ca2+ uptake into mitochondria in vivo or in situ in intact cells is limited because of its inhibitory effects on sarcoplasmic reticulum Ca2+ release channel and other cellular processes. We have synthesized a ruthenium derivative and found it to be an oxygen-bridged dinuclear ruthenium amine complex. It has the same chemical structure as Ru360 reported previously (Emerson, J., Clarke, M. J., Ying, W-L., and Sanadi, D. R. (1993) J. Am. Chem. Soc. 115, 11799-11805). Ru360 has been shown to be a potent inhibitor of Ca2+-stimulated respiration of liver mitochondria in vitro. However, the specificity of Ru360 on Ca2+ uptake into mitochondria in vitro or in intact cells has not been determined. The present study reports in detail the potency, the effectiveness, and the mechanism of inhibition of mitochondrial Ca2+ uptake by Ru360 and its specificity in vitro in isolated mitochondria and in situ in isolated cardiac myocytes. Ru360 was more potent (IC50 = 0.184 nM) than ruthenium red (IC50 = 6.85 nM) in inhibiting Ca2+ uptake into mitochondria. 103Ru360 was found to bind to isolated mitochondria with high affinity (Kd = 0.34 nM, Bmax = 80 fmol/mg of mitochondrial protein). The IC50 of 103Ru360 for the inhibition of Ca2+ uptake into mitochondria was also 0.2 nM, indicating that saturation of a specific binding site is responsible for the inhibition of Ca2+ uptake. Ru360, as high as 10 microM, produced no effect on sarcoplasmic reticulum Ca2+ uptake or release, sarcolemmal Na+/Ca2+ exchange, actomyosin ATPase activity, L-type Ca2+ channel current, cytosolic Ca2+ transients, or cell shortening. 103Ru360 was taken up by isolated myocytes in a time-dependent biphasic manner. Ru360 (10 microM) applied outside intact voltage-clamped ventricular myocytes prevented Ca2+ uptake into mitochondria in situ where the cells were progressively loaded with Ca2+ via sarcolemmal Na+/Ca2+ exchange by depolarization to +110 mV. We conclude that Ru360 specifically blocks Ca2+ uptake into mitochondria and can be used in intact cells.

Yokoshiki et al. (Yokoshiki, H., Sumii, K., Sperelakis, N., 1996. Inhibition of L-type calcium current in rat ventricular cells by the tyrosine kinase inhibitor, genistein and its inactive analog, daidzein. J. Mol. Cell. Cardiol. 28, 807-814) reported that genistein and daidzein inhibited L-type Ca2+ current (I(Ca)(L)) in young rat ventricular cells. Therefore, we investigated the developmental differences in the effect of genistein, an inhibitor of tyrosine kinases, on I(Ca)(L) in freshly-isolated neonatal (3-7 days) and adult (2-5 months) rat ventricular myocytes using whole-cell voltage clamp and single-channel recordings (cell-attached configuration). For whole-cell voltage clamp, I(Ca)(L) was measured as the peak inward current at a test potential of +10 mV by applying a 300 ms pulse from a holding potential of -40 mV. To isolate I(Ca(L), the pipette solution was Cs+-rich and the bath solution was Na+-, K+-free. Ca2+ (1.8 mM) was used as charge carrier. Bath application of 100 microM genistein (sufficient for maximal effect) decreased the basal I(Ca)(L) by 43.3% (n = 27) in neonatal cells and by 30.6% (n = 14) in adult cells (P0.05). In the current/voltage relationships, the potential of peak I(Ca)(L) was shifted to the right by genistein by 8.6 mV in neonatal and by 9.3 mV in adult cells. Genistein produced a shift of the steady-state inactivation curve (to the left) in neonatal cells (from -16.0 +/- 3.9 mV to -26.1 +/- 4.2 mV; P0.05) and in adult cells (-15.9 +/- 3.2 mV to -22.9 +/- 3.3 mV; P0.05); the slope factor was not affected. For single-channel recordings in cell-attached patches, Ca2+ currents were evoked by applying a 150 ms pulse from a holding potential of -40 mV to a test potential of 0 mV. The pipette solution contained 110 mM Ba2+ (as charge carrier), and the bath solution contained 150 mM K+ (to bring resting potential to near zero). Genistein (50 microM) decreased the open probability of the channels from 2.8% to 0.75% (P0.05) in absence of Bay K 8644, and from 24% to 7.9% (P0.05) in presence of Bay K 8644; the mean open time and the slope conductance of the currents were not affected. In conclusion, (1) genistein inhibits the basal I(Ca)(L) in rat ventricular cells and (2) the inhibition of I(Ca)(L) by genistein is greater in immature cells than in adult cells.

Architecture overview of cell switch router (CSR), that is one of actual implementation using label switching paradigm, and the CSR prototype system supporting standard ATM interfaces are described. CSR can operate both with PVC (permanent virtual connection) and with SVC (switched virtual connection) as the VC for cut-through packet forwarding. CSR contains cell switch fabric and IP packet switch fabric to achieve high throughput IP forwarding. IP packets are forwarded either through a cut-through packet transmission, in which packet are forwarded without reassembling IP packet nor IP header processing, or through a conventional hop-by-hop IP packet forwarding. This paper describes and proposes the mechanism to forward the connectionless IP packet flows at the CSR. A CSR prototype system has developed. The CSR prototype system uses PVC and SVC connections to transfer the IP packets. With the CSR prototype system, we can make sure that CSR system could establish the cut-through packet transmission path between the adjacent node with acceptable establishment delay, that was less than few hundred second. The SVC connections for cut- through packet transmission are established on demand using ATM Forum UNI 3.0 or UNI 3.1.

Regulation of L-type Ca2+ channels of vascular smooth muscle (VSM) cells by adenosine 3',5'-cyclic monophosphate (cAMP)-dependent and guanosine 3',5'-cyclic monophosphate (cGMP)-dependent phosphorylation, which requires Mg2+ATP as a phosphate donor, has been reported (T. Ishikawa, J. R. Hume, and K. D. Keef. Circ. Res. 73: 1128-1137, 1993; Z. Xiong, N. Sperelakis, and C. Fenoglio-Preiser. J. Vasc. Res. 31: 271-279, 1994), and regulation by ATP has been demonstrated (Y. Ohya and N. Sperelakis. Circ. Res. 64: 145-154, 1989). However, it has not been elucidated whether the regulation by ATP is mediated by a mechanism that is distinct from phosphorylation. In the present study, we examined the effects of intracellularly perfused ATP on Ca2+ channel currents of VSM cells isolated from rat mesenteric arteries using a whole cell voltage clamp combined with an intracellular perfusion technique. Ba2+ currents (I(Ba)) through Ca2+ channels were evoked by depolarizing pulses from a holding potential of -80 mV with 130 mM Cs+ in the pipette and 100 mM Ba2+ in the bath. The decrease in the ATP concentration (from 5 to 0.1 mM) in the pipette caused a 45 +/- 5% (n = 8) reduction of maximal I(Ba) obtained at +40 mV within 10 min. The dose-response relation between I(Ba) and ATP showed a dissociation constant of 0.53 mM ATP. This concentration is much higher than that usually required for phosphorylation (e.g., few micromolar). Increase in the ATP (from 0.1 to 5 mM) caused an enhancement of maximal I(Ba) by 57 +/- 10% (n = 6), and this enhancement was not prevented in the presence of 30 microM H-7, a nonspecific inhibitor of protein kinases, or 1 microM protein kinase inhibitor, an inhibitor protein of cAMP-dependent protein kinase. These results indicate that slow Ca2+ channels in VSM cells are regulated by intracellular ATP, independently of phosphorylation, implying a direct regulatory action, such as a requirement for ATP binding to the inner surface of the channel, to exhibit activity.

The developmental changes in the isoproterenol stimulation of the L-type calcium current (ICa(L)) were studied in freshly isolated neonatal (3-5-day-old) and adult (2-3-month-old) rat ventricular myocytes using whole-cell voltage clamp (at room temperature). ICa(L) was measured as the peak inward current at a test potential of +10 mV (or +20 mV) by applying a 300 ms pulse from a holding potential of -40 mV. The pipette solution was Cs(+)-rich and Ca(2+)-free. The external solution was Na(+)-free and K(+)-free. Isoproterenol stimulated ICa(L) in a dose-dependent manner. The concentrations of isoproterenol for half-maximal effect were 6.8 nM in neonatal and 13.3 nM in adult. The maximal stimulation of ICa(L) was 147 +/- 14% in neonatal and 97 +/- 7% in adult. The steady-state inactivation curves were not affected by isoproterenol, whereas the steady-state activation curve was shifted to the left in both neonatal and adult. Forskolin (10 microM) increased ICa(L) by 105 +/- 10% in neonatal and 90 +/- 12% in adult. After stimulating ICa(L) by forskolin, the addition of isoproterenol produced a further increase of ICa(L) by 99 +/- 27% in neonatal, but only by 19 +/- 3% in adult. The presence of an inhibitor of cAMP-dependent protein kinase in the pipette did not affect this marked difference between neonatal (87 +/- 23%) and adult (11 +/- 8%). We conclude that, in rat ventricular myocytes, (1) stimulation of ICa(L) by the beta-adrenoceptor agonist, isoproterenol, is already fully developed in the neonatal stage and actually decreases during development; (2) there is evidence for a cAMP-independent stimulation of Ca2+ channels by isoproterenol, and this is greater in neonatal than in adult. We believe that the cAMP-independent pathway is the direct pathway mediated by Gs alpha protein.

Contraction of the heart is regulated by a number of mechanisms, such as neurotransmitters, hormones, autacoids, pH, intracellular ATP, and Ca++ ions. These actions are mediated, at least in part, by actions on the sarcolemmal slow (L-type) Ca++ channels, exerted directly or indirectly. The major mechanisms for the regulation of the slow Ca++ channels of myocardial cells includes the following. cAMP/PK-A phosphorylation stimulates the slow Ca++ channel activity, whereas cGMP/PK-G phosphorylation inhibits. DAG/PK-C phosphorylation and tyrosine kinase phosphorylation are suggested to stimulate the slow Ca++ channel activity. Intracellular application of Gs alpha protein increases the slow Ca++ currents (ICa(L)). Lowering of intracellular ATP inhibits ICa(L). Acidosis and increase in [Ca]i inhibits ICa(L). A number of changes in the Ca++ channels also occur during development and aging. Thus, it appears that the slow Ca++ channel is a complex structure, including perhaps several associated regulatory proteins, which can be regulated by a number of extrinsic and intrinsic factors, and thereby control can be exercised over the force of contraction of the heart.

In 128 children with WPW syndrome, localization estimation was based on the accessory pathway (ACP) localization criteria of the standard 12 lead electrocardiogram for adults proposed by Gallagher et al., Iwa et al., Lindsay et al., and Reddy et al. The rate of consistency of each set of criteria with body surface mapping (BSM) (based on the criteria of Kamakura et al.) was examined along with evaluation of reliability. Among the localization criteria, the most reliable set for children was that developed by Iwa et al. The addition of the precordial lead QRS transitional zone to the criteria established by Iwa et al. was proposed as a means of making the criteria were appropriate for children with WPW syndrome. The consistency rate of localization with this new criterion was added with the results of BSM reached 93% including interpolations. Thus this new localization criterion was considered effective for the estimation of ACP localization in children with WPW syndrome.

A 57-year-old woman with Stage Ia adenosquamous carcinoma arising in a mature cystic teratoma was presented. Pathologically, poorly differentiated squamous cell carcinoma and mucoepidermoid carcinoma were intermingled in the solitary nodule inside of the cyst. She received chemotherapy after operation (total abdominal hysterectomy, bilateral salpingo-oophorectomy, and omentectomy). She is living without evidence of disease 3 years post-treatment. Adenosquamous carcinoma is a rare malignant component in mature cystic teratomas. We reviewed 7 cases of this cancer arising in mature cystic teratomas, including this case. Three cases with Stage Ia were living and well for 9 months to 5 years. Three other cases with Stage Ic, II, and IIIc died of their disease at 6, 13 and 2 postoperative months, respectively. These results might suggest that the prognosis of the tumor is worse in advanced stages than that of squamous cell carcinoma in mature cystic teratomas.