Extensive use of vasodilator agents and functional echocardiography to monitor extremely-low-birth-weight infants in Japan.

National surveys were conducted in Japan to assess the current practices for circulatory management of extremely-low-birth-weight infants (ELBWIs) in acute phases. Approximately 80 and 100 institutions were surveyed in 2006 and 2011, respectively. Echocardiography was identified as an important diagnostic tool at 95% of the surveyed institutions. Furthermore, 74% of the institutions survey in 2011 used vasodilator agents. In 2011, the mean velocity of circumferential fiber shortening (mVcfc) and left ventricular end-systolic wall stress (ESWS) were used by 60% of the surveyed institutions to evaluate the relationship between afterload of the left ventricle and left ventricular contractility. Overall, the data collected from these national surveys clarified the current practices for circulatory management of ELBWIs in Japan, particularly the use of echocardiography and cardiovascular agents, including catecholamines and vasodilators.

Improvement of gemcitabine sensitivity of p53-mutated pancreatic cancer MiaPaCa-2 cells by RUNX2 depletion-mediated augmentation of TAp73-dependent cell death.

Pancreatic cancer exhibits the worst prognostic outcome among human cancers. Recently, we have described that depletion of RUNX2 enhances gemcitabine (GEM) sensitivity of p53-deficient pancreatic cancer AsPC-1 cells through the activation of TAp63-mediated cell death pathway. These findings raised a question whether RUNX2 silencing could also improve GEM efficacy on pancreatic cancer cells bearing p53 mutation. In the present study, we have extended our study to p53-mutated pancreatic cancer MiaPaCa-2 cells. Based on our current results, MiaPaCa-2 cells were much more resistant to GEM as compared with p53-proficient pancreatic cancer SW1990 cells, and there existed a clear inverse relationship between the expression levels of TAp73 and RUNX2 in response to GEM. Forced expression of TAp73α in MiaPaCa-2 cells significantly promoted cell cycle arrest and/or cell death, indicating that a large amount of TAp73 might induce cell death even in the presence of mutant p53. Consistent with this notion, overexpression of TAp73α stimulated luciferase activity driven by p53/TAp73-target gene promoters in MiaPaCa-2 cells. Similar to AsPC-1 cells, small interfering RNA-mediated knockdown of RUNX2 remarkably enhanced GEM sensitivity of MiPaCa-2 cells. Under our experimental conditions, TAp73 further accumulated in RUNX2-depleted MiaPaCa-2 cells exposed to GEM relative to GEM-treated non-silencing control cells. As expected, silencing of p73 reduced GEM sensitivity of MiPaCa-2 cells. Moreover, GEM-mediated Tyr phosphorylation level of TAp73 was much more elevated in RUNX2-depleted MiaPaCa-2 cells. Collectively, our present findings strongly suggest that knockdown of RUNX2 contributes to a prominent enhancement of GEM sensitivity of p53-mutated pancreatic cancer cells through the activation of TAp73-mediated cell death pathway, and also provides a promising strategy for the treatment of patients with pancreatic cancer bearing p53 mutation.

Silencing of RUNX2 enhances gemcitabine sensitivity of p53-deficient human pancreatic cancer AsPC-1 cells through the stimulation of TAp63-mediated cell death.

Runt-related transcription factor 2 (RUNX2) has been considered to be one of master regulators for osteoblast differentiation and bone formation. Recently, we have described that RUNX2 attenuates p53/TAp73-dependent cell death of human osteosarcoma U2OS cells bearing wild-type p53 in response to adriamycin. In this study, we have asked whether RUNX2 silencing could enhance gemcitabine (GEM) sensitivity of p53-deficient human pancreatic cancer AsPC-1 cells. Under our experimental conditions, GEM treatment increased the expression level of p53 family TAp63, whereas RUNX2 was reduced following GEM exposure, indicating that there exists an inverse relationship between the expression level of TAp63 and RUNX2 following GEM exposure. To assess whether TAp63 could be involved in the regulation of GEM sensitivity of AsPC-1 cells, small interfering RNA-mediated knockdown of TAp63 was performed. As expected, silencing of TAp63 significantly prohibited GEM-dependent cell death as compared with GEM-treated non-silencing cells. As TAp63 was negatively regulated by RUNX2, we sought to examine whether RUNX2 knockdown could enhance the sensitivity to GEM. Expression analysis demonstrated that depletion of RUNX2 apparently stimulates the expression of TAp63, as well as proteolytic cleavage of poly ADP ribose polymerase (PARP) after GEM exposure, and further augmented GEM-mediated induction of p53/TAp63-target genes, such as p21 (WAF1) , PUMA and NOXA, relative to GEM-treated control-transfected cells, implying that RUNX2 has a critical role in the regulation of GEM resistance through the downregulation of TAp63. Notably, ablation of TAp63 gave a decrease in number of γH2AX-positive cells in response to GEM relative to control-transfected cells following GEM exposure. Consistently, GEM-dependent phosphorylation of ataxia telangiectasia-mutated protein was remarkably impaired in TAp63 knockdown cells. Collectively, our present findings strongly suggest that RUNX2-mediated repression of TAp63 contributes at least in part to GEM resistance of AsPC-1 cells, and thus silencing of RUNX2 may be a novel strategy to enhance the efficacy of GEM in p53-deficient pancreatic cancer cells.

[Primary arterial switch operation for complete transposition of the great arteries (type I) of a neonate weighing 1,378 g].

A 2-day-old female baby, delivered by emergent cesarean section at 35 weeks of gestational age with a birth weight of 1,378 g, was referred to our institute for intensive care of heart failure. By echocardiography and cardiac catheterization, the patient was diagnosed with isolated complete transposition of the great arteries. Primary arterial switch operation was performed at 13 days of age. No technical difficulty arose, imposed by the small size of cardiovascular structure. On the 5th postoperative day, surgical repair of intestinal perforation was performed. Convalescence thereafter was uneventful. She returned home on the 64th postoperative day with the body weight of 2,310 g. We conclude that primary arterial switch operation can be a feasible surgical option even in a neonate with very low birth weight.

Congenital pulmonary-systemic collateral vein without obstructed left atrial egress is associated with conotruncal anomalies.

We present a case of congenital pulmonary systemic collateral vein associated with truncus arteriosus. Pulmonary systemic collateral vein with nonobstructed left atrial egress is different from those with obstructed left atrial egress in that it is functionally redundant. Including this case, 8 patients among 33 reported cases with pulmonary systemic collateral veins have had nonobstructed left atrial egress. Association with conotruncal anomalies in 4 of these 8 patients, as well as the reported finding that neural crest cells are distributed not only in the conotruncus but also in the cardinal vein, indicates that neural crest cells play roles in remodeling of the systemic venous system.

Identification of early-responsive genes associated with the hypersensitive response to tobacco mosaic virus and characterization of a WRKY-type transcription factor in tobacco plants.

In order to identify genes that are involved in the defense reaction against pathogen attack, we screened for examples that are regulated during the hypersensitive response (HR) to infection by tobacco mosaic virus (TMV) of tobacco ( Nicotiana tabacum cv. Xanthi nc) carrying the N gene, which confers resistance to TMV. Among seven genes initially identified by fluorescent differential display, one clone was further characterized because its transcripts accumulated rapidly and transiently after the onset of HR. Its full-length cDNA of 1346 bp encoded a polypeptide consisting of 258 amino acids. The deduced protein contained a single WRKY domain, a Cys(2)His(2) zinc-finger motif and a leucine-zipper motif, showing high similarity to WIZZ, a member of the family of WRKY transcription factors in tobacco. The gene was thus designated TIZZ. A GFP-TIZZ fusion protein was found to localize to the nucleus upon introduction into epidermal cells of onion. Bacterially expressed TIZZ was able to bind to the W-box (TTGAC) element that is recognized by other WRKY proteins, but transactivation assays showed it to be unable to activate reporter gene expression by itself. TIZZ transcripts were induced in TMV-infected nahG transgenic tobacco plants, in which salicylic acid fails to accumulate. Neither exogenously applied salicylic acid nor mechanical wounding induced TIZZ transcript accumulation. These results indicate the presence of salicylic acid-independent pathways for HR signal transduction, in which a novel type of WRKY protein(s) may play a critical role for the activation of defense.

[A case of Down syndrome with ventricular septal defect and hemodynamical Eisenmenger syndrome].

A 10 month-old infant with Down syndrome having ventricular septal defect and pulmonary hypertension performed cardiac catheterization, which resulted in a slight increase in the pulmonary vascular resistance from 10.8 to 11.3 unit.m2. Lung biopsy findings showed at most an early grade 2 Heath-Edwards classification, and an index of pulmonary vascular disease of 1.1, both of which indicated operability for total correction. He underwent surgical correction and the pulmonary arterial pressure significantly decrease. Although he suffered chylothorax in 5th postoperative day, he did not developed pulmonary hypertension crisis.

Clinicopathological analysis of premature infants treated with artificial surfactant.

OBJECTIVE: Our aim was to obtain new information about the relationship between infant responses to surfactant replacement therapy and histopathological changes in vital organs. STUDY DESIGN: To accomplish this, the autopsy findings and clinical backgrounds of 41 very low birth weight infants (gestational week 25.6 +/- 2.3; birth weight 806.4 +/- 251.6g) who had died after receiving surfactant replacement therapy were reviewed, and those who responded to therapy were compared with those who did not. Responders were infants in whom the required FiO(2) declined by > 20% or mean airway pressure declined by > 20% within six hours of instilling surfactant (n=18); non-responders were infants who did not meet those criteria (n=23). RESULT: Gestational age, birth weight and time at treatment were similar in responders and non-responders, but survival was significantly longer in responders. The incidences of hyaline membrane disease, pulmonary interstitial emphysema, hemorrhagic necrosis and parenchymal degeneration of the liver and kidney were all higher in non-responders, whereas the incidences of bronchopulmonary dysplasia and pneumonia were higher in responders. Prior to treatment, acidosis and hypothermia were significantly more severe in non-responders, and perinatal complications, such as fetal distress and intrauterine infection, were observed more often in non-responders. Substantial degradation of vital organs had already occurred during the early post-natal or intrauterine life of the non-responders, which would be expected to interfere with the clinical response to instilled surfactant. CONCLUSION: It is anticipated that in the future improved monitoring of immature fetuses will be indispensable to improve intrauterine fetal management and to achieve better control over the timing and mode of delivery.

Zebrafish mutations in Gli-mediated hedgehog signaling lead to lens transdifferentiation from the adenohypophysis anlage.

It is known that the earliest lens marker delta-crystallin is expressed abundantly in Rathke's pouch of the chicken, suggesting a close relationship between the cell states of the adenohypophysis (pituitary) anlage and the early lens. We show here that the zebrafish midline mutants you-too (yot) and iguana (igu) develop lenses from the adenohypophysis anlage. The early adenohypophysis anlage of normal zebrafish expresses lim3 and six3 but in yot(ty119) mutants the anterior part of the anlage lacks lim3 expression, and instead produces a crystallin-expressing cell population which develops into a large lens structure expressing beta and gamma-crystallins, but is not associated with retina tissues. Among the zebrafish mutants with midline defects, midline lenses were observed in two mutant alleles of yot and an allele of igu, but not in other mutants (syu, con, smh, dtr, uml, spi and lok). Two yot mutant alleles with midline lenses likely encode dominant negative forms of the Gli2 protein which will interfere with transcriptional activation by other Gli proteins. The observation argues that overall inhibition of Shh-Gli signaling leads the adenohypophysis anlage to transdifferentiate into lens.

Loss of cadherin-11 adhesion receptor enhances plastic changes in hippocampal synapses and modifies behavioral responses.

Cadherins organize symmetrical junctions between the pre- and postsynaptic membranes in central synapses. One of them, cadherin-11 (cad11), is expressed in the limbic system of the brain, most strongly in the hippocampus. Immunohistochemical studies of the hippocampus showed that cad11 proteins were densely distributed in its synaptic neuropil zones; in cultured hippocampal neurons, their distribution often overlapped with that of synaptophysin, and also occasionally with that of GluR1 at spines. To assess the role of cad11 in synaptic formation and/or function, we analyzed brains of cad11-deficient mice. In these mice, long-term potentiation (LTP) in the CA1 region of the hippocampus was, unexpectedly, enhanced; and the level of LTP saturation was increased. In behavioral tests, the mutant mice showed reduced fear- or anxiety-related responses. These results suggest that the cad11-mediated junctions may modulate synaptic efficacy, confining its dynamic changes to a limited range, or these junctions are required for normal development of synaptic organization in the hippocampus.

Expressed sequence tags from immature female sexual organ of a liverwort, Marchantia polymorpha.

A total of 970 expressed sequence tag (EST) clones were generated from immature female sexual organ of a liverwort, Marchantia polymorpha. The 376 ESTs resulted in 123 redundant groups, thus the total number of unique sequences in the EST set was 717. Database search by BLAST algorithm showed that 302 of the unique sequences shared significant similarities to known nucleotide or amino acid sequences. Six unique sequences showed significant similarities to genes that are involved in flower development and sexual reproduction, such as cynarase, fimbriata-associated protein and S-receptor kinase genes. The remaining unique 415 sequences have no significant similarity with any database-registered genes or proteins. The redundant 123 ESTs implied the presence of gene families and abundant transcripts of unknown identity. Analyses of the coding sequences of 61 unique sequences, which contained no ambiguous bases in the predicted coding regions, highly homologous to known sequences at the amino acid level with a similarity score greater than 400, and with stop codons at similar positions as their possible orthologues, indicated the presence of biased codon usage and higher GC content within the coding sequences (50.4%) than that within 3' flanking sequences (41.9%).

AFP-producing squamous cell carcinoma of the lung in an adolescent.

We report a case of primary lung cancer in a 16-year-old boy. A histologic diagnosis of squamous cell carcinoma was made by bronchoscopic biopsy before surgery. The serum alphafetoprotein (AFP) level was markedly elevated at 193 ng/dl. Preoperative and postoperative evaluation revealed no evidence of scrotal mass. We performed right pneumonectomy with combined resection of the invaded portion of the left atrium under extracorporeal circulation. Despite the rapid improvement in the patient's general condition after surgery, the AFP level continued to increase without a transient decrease and reached 3160 ng/ml on the 23rd postoperative day. When the patient was readmitted because of dyspnea and headache on the 36th postoperative day, hypercalcemia of 13.9 mg/dl was noted, and this was resistant to subsequent treatment. The patient died on the 46th postoperative day.

Blood flow velocities in the cerebral arteries and descending aorta in small-for-dates infants.

Resistive indices (RI) in the anterior cerebral artery (ACA), basilar artery (BA), middle cerebral artery (MCA) and descending aorta (DA) were obtained in 15 small-for-dates (SFD) infants who were, growth retarded because of maternal pregnancy-induced hypertension and in 20 appropriate-for-dates (AFD) infants matched for gestational age between 24 h and 48 h after birth. The RIs in the MCA, ACA and BA were significantly lower, while the RI in the DA was significantly higher, in the SFD infants than in the AFD infants. These changes in RIs in the SFD infants might be similar to the "brain sparing" effect as reported in growth-retarded fetuses.

[Incidence and causes of neonatal seizures in the last 10 years (1981-1990)].

We report the incidence of neonatal seizures in the last 10 years. Between 1981-1990, 28,925 infants were born in the Japanese Red Cross Medical Center. Very low-birth-weight infants (< 1,500 g) were excluded in this study. Seizures were identified in 67 infants (0.23%) during the first 7 days of life. Seizures occurred more frequently in low-birth-weight infants (1,500-2,499 g) than in normal-birth-weight infants (> 2,499 g). Fifty-one infants (0.19%) of 27,276 normal-birth-weight infants and 16 (0.97%) of 1,649 low-birth-weight infants developed seizures. The frequency of seizures was 0.25% during the first 5 years and 0.20% during the second 5 years. In low-birth-weight group, the frequency of seizures decreased form 1.49% to 0.48% during the 5-year periods, and seizures due to post asphyxial hypoxic-ischemic encephalopathy also decreased from 0.14% to 0.06%. The decreased incidence in neonatal seizures during the last 10 years was probably achieved by the progress in intervention of low-birth-weight infants in NICU and by the obstetrical prevention of intrapartum asphyxia.

Development of hepatocellular carcinoma in a man with auto-immune chronic active hepatitis.

A 57 year old man with auto-immune chronic active hepatitis, regularly treated with immunosuppressive therapy, had hepatocellular carcinoma (HCC) 10 years after diagnosis of the hepatitis. Assays of the hepatitis C virus antibodies against capsid and non-structural proteins revealed seronegativity in serial serum samples of this patient stored in the previous 10 years during follow up. The seronegative hepatitis C antibodies excluded hepatitis C virus as the cause of the HCC. The occurrence of HCC in this case suggests the necessity of surveillance for early detection of liver cancer in patients with auto-immune chronic active hepatitis undergoing long-term immunosuppressive therapy.

[Clinical evaluation of imipenem/cilastatin sodium against severe infections complicating hematological disorders and solid tumors].

Imipenem/cilastatin sodium (IPM/CS) was administered to a total of 67 patients with severe infections complicating hematological disorders and solid tumors. Fifty patients are included in the present analysis of efficacy and 64 in that of safety. 1. Out of 31 patients with hematological disorders, responses were excellent in 10 patients, good in 10 patients, and the efficacy rate was 64.5%. Out of 19 patients with solid tumors, responses were excellent in 8 patients, good in 8 patients and the efficacy rate was 84.2%. 2. For patients whose responses to other antibiotics had been poor, the efficacy rate was 59.3% in the group with hematological disorders and 62.5% in the group with solid tumors. 3. The relationship between the neutrophil count and efficacy was studied in the patients with hematological disorders. The efficacy rate for 8 patients whose neutrophil counts were 500/mm3 or less was 75.0%. 4. For the patients with hematological disorders, the efficacy rate for patients from whom causative organisms were isolated was 70.0% and that for patients for whom they were unknown was 61.9%. 5. Adverse reactions were observed in 3 patients and abnormal laboratory test results in 2 patients. However, they were mild and disappeared after discontinuation of this drug. From these results, IPM/CS is considered to be a useful antibiotic for the treatment of severe infections complicating hematological disorders and solid tumors.

A case of Caroli's disease with hepatolithiasis, choledocholithiasis, and cholangiocarcinoma.

A 48-year-old woman with Caroli's disease accompanied by hepatolithiasis, choledocholithiasis, and cholangiocarcinoma is reported. Magnetic resonance imaging was useful for the diagnosis of the cholangiocarcinoma. The relationship between cholangiocarcinoma and hepatolithiasis, choledocholithiasis, and chronic cholangitis in Caroli's disease is discussed.

Cerebral blood flow velocities in the anterior cerebral arteries and basilar artery in hydrocephalus before and after treatment.

We studied Pourcelot's index (PI), which shows cerebral vascular resistance, in the anterior cerebral arteries and basilar artery, and the PI ratio (Pourcelot's index in the anterior cerebral artery/Pourcelot's index in the basilar artery) in 11 measurements of hydrocephalus. The mean values of PI in the anterior cerebral artery, basilar artery, and the PI ratio before treatment were significantly higher than those after treatment and those in normal infants. Before treatment, the mean PI in the anterior cerebral arteries was significantly higher than the mean PI in the basilar artery. All PI ratios increased to 1.00 or more. After treatment and in normal infants, the mean PI in the anterior cerebral arteries was significantly lower than the mean PI in the basilar artery. All PI ratios decreased to less than 1.00. We believe that the PI ratio is useful to evaluate the need or effect of treatment in hydrocephalus.