Shunji Mimura

Acute respiratory failure associated with intrathoracic masses in neonates

BACKGROUND Intrathoracic masses are uncommon in children. Occasionally, they present with acute respiratory failure in the neonatal period. Although emergency resection usually is the treatment of choice, other modalities are sometimes necessary to stabilize the patient. METHODS Seven neonates with intrathoracic masses were treated. Five had congenital cystic adenomatoid malformations (CCAM), 1 had a mediastinal teratoma, and 1 had a pneumatocele. These cases were reviewed retrospectively. RESULTS Four of the 7 infants had respiratory failure in the neonatal period. A patient with a large mediastinal teratoma and 1 with a CCAM that increased rapidly after presentation underwent emergency operation, relieving respiratory distress. The other 2 large CCAMs presented with severe respiratory distress immediately after birth because of pulmonary hypoplasia. One neonate with a Stocker-I CCAM died after emergency resection. One more recent patient with a Stocker-III CCAM survived after successful treatment with delayed resection, performed 3 days after birth. Nitric oxide (NO), and extracorporeal membrane oxygenation (ECMO) were instituted as supportive care because of profound persistent fetal circulation (PFC). CONCLUSIONS Acute respiratory failure associated with intrathoracic masses in neonates may be managed in 1 of 2 ways. A small mass that increases rapidly should be resected soon after presentation. In neonates with large masses with associated PFC, surgery can be delayed until the patient is stable. ECMO, NO, and high-frequency oscillation (HFO) can be used aggressively for stabilizing such neonates.

Abnormal cerebral neuronal migration in a rat model of intrauterine growth retardation induced by synthetic thromboxane A2

Many reports have associated intrauterine growth retardation (IUGR) with adverse neurological outcome, but the underlying pathology is imperfectly understood. We have developed a new rat model of IUGR using maternal administration of synthetic thromboxane A(2) (STA(2)). In the present study, the effect of this insult on neuronal migration in the rat cerebral cortex was examined. Bromodeoxyuridine (BrdU), a time-specific cell marker was administered intraperitoneally to the mothers on embryonic day (E) 19. At postnatal day (P) 3, P4, P5, and P6, pups were terminally anesthetized and brains were removed. BrdU-labeled cells were detected immunohistochemically and counted in cerebrum, which was divided into the cortical plate (CP), the intermediate zone, and the subventricular/ventricular zone (SVZ+VZ). Numbers of labeled cells in the three areas over time were compared between IUGR and control animals. Numbers of labeled cells in SVZ+VZ were significantly greater in IUGR than in controls at P3, 5, and 6 (P<0.05). In contrast, labeled cells in the CP were significantly less abundant in IUGR animals than in controls at P3, 4, and 6 (P<0.05). We concluded that neuronal migration was delayed in IUGR rats.

Underexpression of neural cell adhesion molecule and neurotrophic factors in rat brain following thromboxane A2-induced intrauterine growth retardation

Intrauterine growth retardation (IUGR) often results in clinical neurodevelopmental disorders. To clarify the influence of uteroplacental insufficiency on central nervous system development, we have created a model of IUGR in rats using maternal administration of synthetic thromboxane A(2). We investigated expression patterns of neural cell adhesion molecule (NCAM) and reelin in this model by semiquantitative competitive polymerase chain reactions. On postnatal day 2, NCAM expression was decreased in rat cerebral cortex, and reelin expression was decreased in hippocampus from levels in controls without maternal thromboxane exposure. No significant differences in NCAM expression were seen in hippocampus, nor did reelin expression differ in cerebral cortex between control and IUGR groups. We also examined expression of two neurotrophic factors, brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3). In cerebral cortex the IUGR group showed less BDNF and NT-3 expression than controls. Delay of neuronal migration and histological changes observed in our IUGR rats may be related to altered expression of these molecules.

Dangerous effects of tin-protoporphyrin plus photoirradiation on neonatal rats

In vivo and in vitro effects of porphyrins (tin-protoporphyrin [SnPP], cobalt-mesoporphyrin, haemin and protoporphyrin) on neonatal rats were investigated. Under photoirradiation a high mortality rate was recognized in SnPP injected rats. None died from the application of SnPP without photoirradiation. In photoirradiated rats the median lethal dose (LD50) value of SnPP was calculated to be about 7.4 μmol/kg body weight. Haemolysis and malonaldehyde formation of red blood cells were induced by SnPP together with photoirradiation. SnPP may be useful in reducing bilirubin levels in severely jaundiced infants under non-photoirradiated conditions or dim light, but prophylactic administration of SnPP to the majority of infants is not recommended.

Plasma and erythrocyte levels of trace elements and related antioxidant enzyme activities in low-birthweight infants during the early postnatal period

To discover the relationship between antioxidant enzyme activities and trace elements in low‐birthweight infants during the early postnatal period, we analysed catalase (CAT), CuZn‐superoxide dismutase (CuZn‐SOD) and glutathione peroxidase (GSH‐Px) activities in erythrocytes, and compared them with Fe, Cu, Zn and Se levels in plasma, and Cu, Zn and Se levels in erythrocytes until 16 wk after birth. Thirteen low‐birthweight infants whose mean birthweight and gestation were 1520 ± 293 g and 32.0 ± 2.8 wk were enrolled in this study. All infants were without chronic complications, well nourished, and predominantly fed standard formula or preterm formula based on cows milk, commercially available in Japan. Cu and Zn levels in erythrocytes did not decline after birth, in contrast to a temporal decrease in plasma Zn. Erythrocyte CAT activity was significantly higher at 16 wk than that at birth or 4 wk of age. Erythrocyte CuZn‐SOD activity did not change throughout the study period. Only Se in plasma and erythrocytes decreased remarkably after birth, which resulted in a significant decline in erythrocyte GSH‐Px activity at 8 and 16 wk (11.2 ± 2.0 and 11.0 ± 1.1 U/g Hb) compared to that at birth (12.4 ± 2.1 U/g Hb).

Neuropathological changes in the cerebrum of IUGR rat induced by synthetic thromboxane A2

IUGR was induced by maternal administration of synthetic thromboxane A2 (STA2) from the 13th day of gestation. Fetuses and neonates showed a markedly significant weight reduction. In E16 IUGR brain, no pathological abnormalities were found, but morphological changes appeared in the cortical plate of E18 IUGR brain. In E20 IUGR brain, ectopic clusters of differentiating cells cytologically mimicking neuroblasts were found in the neuroepithelial layer, but these abnormal clusters of cells in IUGR brain of late gestation were never observed in PN7. Morphometric analysis of coronal-sectional areas of the brain and cortical plate demonstrated that there were no differences between IUGR rats and controls in E16 and E18. These areas were, however, significantly reduced in E20 and PN7 growth-retarded rats compared with the control. Because the period of STA2 administration coincides with the neuro-developmental stage of cell migration and differentiation, reduction of the uteroplacental blood supply might cause a transient abnormal cytoarchitecture of the cerebral cortex resulting in brain growth retardation.

An Animal Model of Intrauterine Growth Retardation Induced by Synthetic Thromboxane A2

Objective: Intrauterine growth retardation (IUGR) is an important cause of prenatal and neonatal morbidity, and neurologic abnormalities. Although several animal models of IUGR have been developed for scientific investigation, few models approximate the pathophysiology in human fetal growth failure resulting from pregnancy-induced hypertension and preeclampsia. We developed an animal model of IUGR in which fetal growth restriction was induced by administering a synthetic thromboxane A2 analogue (STA2) to the mother. Methods: Timed pregnant Sprague-Dawley rats were used in this study. STA2 was delivered into the peritoneal cavity of the pregnant female at a rate of 20 ng/h from day 13 of pregnancy. The effectiveness of this model was evaluated by monitoring the overall growth of the fetuses and neonates and measuring the weight and biochemical composition of individual organs. Results: Fetuses and neonates from the STA2 group showed a highly significant weight reduction throughout the observation period from day 19 of gestation to postnatal day 7. Weight reduction near and at term exceeded 10% and became more pronounced during the first week after birth. Fetuses on the 20th gestational day exhibited a pattern of growth retardation characteristic of asymmetrical IUGR in which the weight reduction was prominent in the liver with relative sparing of the brain. However, the decrease in brain weight was more than 10%. The protein, DNA, and RNA contents of the liver were lower in the STA2 group. The protein content of the forebrain and brainstem also decreased significantly in the STA2 group compared with the control; however, the DNA content of the forebrain was higher in the STA2 group. Conclusions: This animal model may mimic human IUGR more closely than previous models because the growth restriction is induced in a truly chronic manner.

Protection by L-ascorbic acid against phototoxicity in tin-protoporphyrin-treated suckling rats

The protective effect of free radical scavengers against phototoxicity was investigated with tin-protoporphyrin (SnPP)-treated suckling rats. Six kinds of scavengers (L-ascorbic acid, reduced glutathione, alpha-tocopherol, retinol, uric acid and cystine) were intraperitoneally injected to rats treated with SnPP plus photoirradiation. Among them, L-ascorbic acid was found to be most effective in protecting SnPP-treated rats against phototoxicity. The survival period was markedly prolonged, and the frequency of abnormal behaviors was reduced with the treatment. Lipid peroxidation in vitro with the brain membrane fraction was also suppressed. The other five substances gave only a little antioxidant effect both in vivo and in vitro. The present study shows that L-ascorbic acid may be a promising chemical to prevent the phototoxicity of SnPP.

Cobalt-mesoporphyrin inhibits heme oxygenase activity but it does not induce lipid peroxidation in rat brain membranes during photoirradiation.

We examined the effects of cobalt-mesoporphyrin (CoMP) in vitro. The porphyrin inhibited the activity of rat splenic heme oxygenase but scarcely stimulated peroxidation of lipids in a membrane fraction from rat brain during photoirradiation. The apparent inhibition constant for CoMP was 344 nM. It is suggested that CoMP may be a promising candidate for a chemopreventive of neonatal hyperbilirubinemia that is not associated with phototoxicity.

Rapid Genotyping of 2-bp and 9-bp Deletion Mutations Using the LightCycler Instrument

Instead of sequencing the candidate gene PCR fragment, simple methods such as restriction enzyme digestion after PCR are used to detect known mutation. In some cases, however, such methods cannot be applied and sometimes lead to an ambiguous result. A faster method would be required in case of mass screening or emergency. Recently, an elegant method to detect a known mutation with the LightCycler has been introduced. Rapid real-time PCR is monitored by fluorescent oligonucleotide probes that hybridize the target region of the PCR product. Only when the probes hybridize the template fluorescence resonance energy transfer occurs thereby producing a specific fluorescence emission. Then, by slowly heating the PCR product in the probe-hybridized state, the diminishing fluorescence value gives a melting curve. In the presence of mismatch sequences between the template and the probe, the probe melts off at a lower temperature. Therefore, the melting curve clearly demonstrates its genotype as the differences in Tm. This fluorescence PCR on the LightCycler has displayed its marked ability to detect single nucleotide substitution [1, 2]. However, many inherited diseases are caused by small deletion mutations as well as single nucleotide mutations.