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The influx of microplastics (MPs) into the Arctic Ocean poses a collective risk, particularly with pronounced sea ice depletion due to global warming. A total of 73 replicate sediment samples were collected at different depths (38 to 79 m) from Chukchi and the Beaufort Seas at 8 stations in the Arctic region during the R/V Mirai cruise (MR22-06C) from August to September 2022. The average concentration of MPs is 79.25 ± 31.08 items/kg d.w. Fibrous MPs of 0-1 mm size range are predominant, with blue being the most prevalent colour. Polymer characterization identified polyethylene (PE) as the predominant polymer. Arctic Ocean regions face heightened health risks from the coexistence of MPs and harmful additives, amplifying concerns over plastic pollution. The alarming surge in MPs within Arctic sediment underscores the urgent need for a proactive, collaborative approach to mitigate this environmental threat and its far-reaching impacts., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background and Aim: The diagnostic accuracy for cholangiocarcinoma (CCA) is inadequate, necessitating the exploration of novel diagnostic approaches. Protoporphyrin IX (Pp IX), a metabolic product of 5-aminolevulinic acid (5-ALA), emits red fluorescence upon blue light exposure. Because it accumulates selectively in cancer cells, photodynamic diagnosis using 5-ALA (5-ALA-PDD) has been integrated into clinical practice for diverse cancer types. Nevertheless, there is currently no device capable of capturing Pp IX-derived fluorescence for real-time 5-ALA-PDD within the biliary tract, largely due to challenges in device miniaturization., Methods: To investigate the feasibility of real-time 5ALA-PDD in CCA, we developed two essential components of the cholangioscopy system: a small-diameter flexible camera and a light guide for emitting blue light. We evaluated the detectability of Pp IX fluorescence using these devices in experimental gels and animal models., Results: Our camera and light guide were smoothly inserted into the lumen of existing cholangioscopes. Incorporating a long-pass filter at the camera tip enabled efficient detection of red fluorescence without significantly impacting white-light observation. The integration of these devices facilitated clear visualization of red fluorescence from gels containing Pp IX at concentrations of 5 μM or higher. Additionally, when observing subcutaneous human CCA tumor models in nude mice treated with 5-ALA, we successfully demonstrated distinct red fluorescence from Pp IX accumulation in tumors compared to peritumoral subcutaneous areas., Conclusion: The integration of our device combination holds promise for real-time 5-ALA-PDD in human CCA, potentially enhancing the diagnostic accuracy for this complex condition., Competing Interests: Declaration of competing interest Shiho Furudate and Yuki Kodama are employees of Japan Lifeline Co., Ltd., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

The Fukushima Daiichi Nuclear Power Plant accident caused a radioactive contamination of deposited radionuclides, including 137 Cs, on the land surface. Cesium-137 deposited on the land surface was strongly adsorbed on soil particles and was then washed off through soil erosion. Trends of temporal variation of 137 Cs wash-off varied greatly depending on land use. Therefore, it is important to reflect the characteristics of 137 Cs migration processes in each land use to clarify the long-term fate of 137 Cs. In this study, a 30-year simulation of environmental fate of 137 Cs was conducted using a distributed radiocesium prediction model, taking into account the characteristics of the 137 Cs behavior in each land use. Overall, in the Abukuma River basin, the 137 Cs transported into the ocean for 30 years was estimated to correspond to 4.6 % of the initial deposition in the basin, and the effective half-life of 137 Cs deposited in the basin was estimated to be 3.7 years shorter (by 11.6 %) than its physical half-life. These results suggested that 137 Cs deposited from the accident could still remain for decades. Based on the analysis of the 137 Cs behavior in land use, in 2011, the contribution of 137 Cs export to the ocean from urban lands was estimated to correspond to 70 % of the total 137 Cs export. Meanwhile, from 2012 to 2040, the contribution of 137 Cs export from agricultural lands was estimated to correspond to 75 % of the total 137 Cs export. The reduction ratios excluding radioactive decay of 137 Cs remained in areas with and without human activities for 30 years after the accident, defined as the ratios of the total outflow to the initial deposition, were estimated to be 11.5 %-17.7 % and 0.4 %-1.4 %, respectively. These results suggested that human activities enhance the reduction of 137 Cs remaining in land in the past and future., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Objectives: Social distancing due to the coronavirus disease 2019 crisis can exacerbate inactivity in older adults. Novel approaches for older adults must be designed to improve their activity and maintain their health. This study examined the effect of nudge-based behavioral interventions on health-promoting activities in older adults in Japan., Design: Two-arm, participant-blinded randomized controlled trial., Setting and Participants: Japanese continuing care retirement community residents (n = 99, median age 82 years, 73% women) INTERVENTION: Two-step nudge-based behavioral intervention promoting tablet usage., Methods: We enrolled participants from an ongoing Internet of Things project in a retirement community in Japan. For the health promotion program, tablet computers were installed in a common area for participants to receive information about their health. The intervention group received a 1-time loss-emphasized nudge (first step), followed by asking questions about when they planned to use it again (second step). The control group used the tablet computers without being asked those questions. The main outcome was the participants' mean daily tablet activity every 4 weeks for the next 16 weeks., Results: Ninety-nine individuals were randomly assigned to the intervention or control group. The rate ratios for tablet use were significantly higher in the intervention group in the second and third periods. The subgroup analysis showed that these effects were largely attributable to men., Conclusions and Implications: Nudge-based interventions can be effective in promoting activities for older adults, especially older men. The finding of this study indicates a possible intervention to engage people who are socially isolated., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The recent influx of microplastics into the Arctic Ocean may increase environmental stress on the western Arctic marine ecosystem, which is experiencing significant sea-ice loss due to global warming. Quantitative data on microplastics in the western Arctic Ocean are very limited, and the microplastic budget of the water column is completely unknown. To fill in gaps in our knowledge of Arctic microplastics, we observed surface concentrations (number of particles per unit volume of seawater) of meso- and microplastics using a neuston net, and we observed wind speeds and significant wave heights in the Chukchi Sea, Bering Strait, and Bering Sea. From these observations, we estimated the total number (particle inventory) and mass (mass inventory) of microplastics in the entire water column by taking into account the effect of vertical mixing. The particle inventory of microplastics in the Chukchi Sea ranged from 0 to 18,815 pieces km -2 with a mean and standard deviation of 5236 ± 6127 pieces km -2 . The mass inventory ranged from 0 to 445 g km -2 with a mean and standard deviation of 124 ± 145 g km -2 . Mean particle inventories for the Chukchi Sea were one-thirtieth of those for the Arctic Ocean on the Atlantic side and less than one-tenth of the average for the global ocean, suggesting that the Chukchi Sea is less polluted. However, the annual flux of microplastics from the Pacific Ocean into the Chukchi Sea, estimated from microplastic concentrations in the Bering Strait, was about 5.5 times greater than the total amount of microplastic in the entire Chukchi Sea water. This suggests that microplastic inflows from the Pacific Ocean are accumulating in large amounts in reservoirs other than the Chukchi Sea water (e.g., sea ice and seafloor sediments) or in the downstream regions of the Pacific-origin water., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Introduction: In Japan, patients with coronavirus disease 2019 (COVID-19) who do not require medical intervention are provided care in recovery accommodation facilities (RAFs). However, some patients may require hospitalization if their symptoms become more severe during their stay. We conducted an observational study using epidemiological data of patients with COVID-19 admitted to RAFs in Tokyo., Methods: This was an observational cohort study using data from COVID-19 patients admitted to one of the RAFs in Tokyo from December 2020 to November 2021. Admissions to the facilities were limited to patients with asymptomatic or mild COVID-19 with no underlying disease or at least stable underlying disease at the time of admission. Patients were hospitalized when they required oxygen administration or when they had, or persistent fever, or severe respiratory symptoms. We evaluated the association between hospitalization and the risk factors for hospitalization using a Cox regression model., Results: The number of patients with COVID-19 admitted to the RAF was 6176. The number of hospitalized patients was 393 (6.4%), and the median length of stay was 5.50 days (IQR: 4.50, 6.50). In the Cox regression analysis, the hazard ratio increased with age and was significantly higher among patients aged >60 years (HR = 10.23, 95% CI: 6.72-15.57) than those in other age groups. This trend is similar to that observed in the sensitivity analysis., Conclusion: Patients with diabetes, the elderly, obesity, and medications for gout and psychiatric diseases may be at a high risk of hospitalization. In particular, an age over 60 years was strongly associated with hospitalization., (Copyright © 2022 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Concentrations of 129 I and 127 I in kelps (Saccharina) collected from coastal waters off northern Japan were monitored from 2007 to 2019. During the 2007-2008 test operation of the Rokkasho nuclear fuel reprocessing plant, 129 I discharge from the plant increased, and the 129 I concentration and 129 I/ 127 I atom ratio in the kelps reached maxima of 42 μBq/g-dry and 264 × 10 -11 , respectively. By 2009, both had decreased by one order of magnitude. After the Fukushima Dai-ichi Nuclear Power Plant accident in 2011, the 129 I concentration and 129 I/ 127 I atom ratio in the kelps increased to 2.24 μBq/g-dry and 11.6 × 10 -11 , respectively. After 2012, the ratio in kelps decreased to (2.1-8.9) × 10 -11 , which is almost the same as the seawater value off Aomori Prefecture before the test operation. The 129 I/ 127 I atom ratio in kelps thus represents the ambient seawater ratio during the growth period of the kelps., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Concentration ratios (CRs), expressed by dividing 137 Cs activity in seawater by that in marine biota (mainly fish), were obtained from the monitoring of 137 Cs in coastal areas around Japan between 1984 and 2016. Before the TEPCO Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident (1984-2010), mean CRs of 137 Cs, mainly from global fallout (i.e. CR GF ), were almost constant for each species throughout the monitoring period, but were different among species, while the values for several species were dependent on their length (i.e. CR GF-SIZE ). Thus, CR GF and CR GF-SIZE values for 29 of marketable species are given here as references for conditions where marine biota are in approximate equilibrium (or steady state) with their host water with respect to 137 Cs activities in the marine environment. After the FDNPP accident (2011-2016), the impact of the accident has been sustained in eastern Japan waters as indicated by apparent CRs (CR a s) which are being used here as indicators of disequilibrium between organisms and their host water. The recession rates of this disequilibrium (the effective CR a half-lives) ranged from 100 to 1100 days. The identified distinct variation was due to the sample locations, even for the same species, because of the change in 137 Cs activity concentrations in their host water and diet preference differences. Variation among species, even those captured from the same area, was mainly due to diet differences as well as metabolic-physiological differences in 137 Cs retention. Thus, our results from >30 years of systematically monitoring have helped quantify the recession rates of post-FDNPP disequilibrium of 137 Cs in biota for assessment of how long term is required from contaminated condition by underlying spatial, inter- and intra-species factors., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Objectives: A discrepancy in self-reported and performance-based physical functioning levels is often observed among older adults. We investigated the association of discrepancy in self-reported and performance-based physical functioning levels with risk of future falls among community-dwelling older adults., Design: Prospective cohort study., Setting: Two communities in Fukushima Prefecture, Japan., Participants: 1379 older adults who took part in the yearly health checkup in both 2009 and 2010., Measures: The performance-based and self-reported physical functioning levels were evaluated by the Timed Up and Go test and the Short-Form 12 Health Survey (Japanese version) physical functioning subscale, respectively. We divided the participants into 4 groups based on the combinations of low or high performance-based and self-reported physical functioning groups, which were classified by age- and sex-specific reference values. The main outcome was the occurrence of any falls within the 1-year follow-up period, assessed using a self-reported questionnaire., Results: A total of 22% of the participants reported the occurrence of a fall during the follow-up period. In multivariable logistic regression analysis, the adjusted odds ratios of the high self-reported and low performance-based, low self-reported and high performance-based, and low self-reported and low performance-based physical functioning groups were 1.10 (95% confidence interval [CI], 0.67-1.82), 1.76 (95% CI, 1.17-2.66), and 1.80 (95% CI, 1.11-2.90), respectively, compared with the high self-reported and high performance-based physical functioning group., Conclusions: Our findings suggest that the discrepancy as high performance-based but low self-reported physical functioning level is associated with an increased risk of future falls in older adults aged 65-89 years. Clinicians should carefully assess older adults whose subjective perception of their physical functioning capacity is lower than those in similar age and sex groups, even if their actual physical functioning appears to be objectively high., (Copyright © 2018 AMDA – The Society for Post-Acute and Long-Term Care Medicine. Published by Elsevier Inc. All rights reserved.)

Amyloid fibrillation causes serious neurodegenerative diseases and amyloidosis; however, the detailed mechanisms by which the structural states of precursor proteins in a lipid membrane-associated environment contribute to amyloidogenesis still remains to be elucidated. We examined the relationship between structural states of intrinsically-disordered wild-type and mutant α-synuclein (αSN) and amyloidogenesis on two-types of model membranes. Highly-unstructured wild-type αSN (αSN WT ) and a C-terminally-truncated mutant lacking negative charges (αSN 103 ) formed amyloid fibrils on both types of membranes, the model membrane mimicking presynaptic vesicles (Mimic membrane) and the model membrane of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC membrane). Unstructured αSN WT and αSN 103 both bound to Mimic membranes in a helical conformation with similar binding affinity. Promotion and then inhibition of amyloidogenesis of αSN WT were observed as the concentration of Mimic lipids increased. We explain this by the two-state binding model: at lower lipid concentrations, binding of αSN WT to membranes enhances amyloidogenicity by increasing the local concentration of membrane-bound αSN and so promoting amyloid nucleation; at higher lipid concentrations, membrane-bound αSN WT is actually in a sense diluted by increasing the number of model membranes, which blocks amyloid fibrillation due to an insufficient bound population for productive nucleation. Meanwhile, αSN 103 formed amyloid fibrils over the whole concentration of Mimic lipids used here without inhibition, revealing the importance of helical structures for binding affinity and negatively charged unstructured C-terminal region for modulating amyloidogenesis. We propose that membrane binding-induced initial conformations of αSN, its overall charge states, and the population of membrane-bound αSN are key determinants of amyloidogenesis on membranes., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Introduction: Spinal immobilization has been indicated for all blunt trauma patients suspected of having cervical spine injury. However, for traumatic cardiac arrest (TCA) patients, rapid transportation without compromising potentially reversible causes is necessary. Our objective was to investigate the temporal trend of spinal immobilization for TCA patients and to examine the association between spinal immobilization and survival., Methods: We conducted a retrospective cohort study using the Japan Trauma Data Bank 2004-2015 registry data. Our study population consisted of adult blunt TCA patients encountered at the scene of a trauma. The primary outcome was the survival proportion at hospital discharge, and the secondary outcome was the proportion achieving return of spontaneous circulation (ROSC). We examined the association between spinal immobilization and these outcomes using a logistic regression model based on imputed data sets with the multiple imputation method to account for missing data., Results: Among 4313 patients who met the inclusion criteria, 3307 (76.7%) were immobilized. The proportion of patients that underwent spinal immobilization gradually decreased from 82.7% in 2004-2006 to 74.0% in 2013-2015. 1.0% of immobilized and 0.9% of non-immobilized patients had severe cervical spine injury. Spinal immobilization was significantly associated with lower survival at discharge (odds ratio [OR], 0.64; 95% confidence interval [CI], 0.42 to 0.98) and ROSC by admission (OR, 0.48; 95%CI, 0.27 to 0.87). There was no significant sub-group difference of the association between spinal immobilization and survival at discharge by patients with or without cervical spine injury (p for interaction 0.73)., Conclusion: Spinal immobilization is widely used even for blunt TCA patients, even though it is associated with a lower rate of survival at discharge and ROSC by admission. According to these results, we suggest that spinal immobilization should not be routinely recommended for all blunt TCA patients., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objectives: To assess the effect of blood type on survival outcomes and adverse events (AEs) in patients treated with tyrosine kinase inhibitors (TKIs) for metastatic renal cell carcinoma (mRCC)., Materials and Methods: Patients who received TKIs as first-line therapy for mRCC between 2008 and 2015 at our hospital were included in the study (n = 136). Patients were divided into 2 groups based on their blood type as O and non-O. Survival outcomes and AEs were compared according to blood type. Cox regression models were used for univariate and multivariate survival analyses., Results: Of the 136 patients, 34 (25%) and 102 (75%) had O and non-O blood types, respectively. Blood type O was associated with an increased number of disease sites. There were no differences between the 2 groups with respect to other baseline characteristics. The progression-free survival in patients with O and non-O blood types was 12.1 and 11.6 months, respectively; the overall survival was 34.4 and 24.8 months, respectively. On univariate and multivariate analyses, the ABO blood type was not a significant prognostic factor for progression-free survival or overall survival. Furthermore, the incidences of serious AEs were similar in the 2 blood groups., Conclusions: ABO blood type was not associated with survival outcomes or incidences of serious AEs in mRCC patients treated with TKIs. However, blood type O may be associated with an increased number of disease sites., (Copyright © 2017. Published by Elsevier Inc.)

Objectives: To describe the registration of systematic review (SR) protocols and examine whether or not registration reduced the outcome reporting bias in high-impact journals., Study Design and Setting: We searched MEDLINE via PubMed to identify SRs of randomized controlled trials of interventions. We included SRs published between August 2009 and June 2015 in the 10 general and internal medicinal journals with the highest impact factors in 2013. We examined the proportion of SR protocol registration and investigated the relationship between registration and outcome reporting bias using multivariable logistic regression., Results: Among the 284 included reviews, 60 (21%) protocols were registered. The proportion of registration increased from 5.6% in 2009 to 27% in 2015 (P for trend <0.001). Protocol registration was not associated with outcome reporting bias (adjusted odds ratio [OR] 0.85, 95% confidence interval [CI] 0.39-1.86). The association between Preferred Reporting Items for Systematic review and Meta-Analysis (PRISMA) adherence and protocol registration was not statistically significant (OR 1.09, 95% CI 0.59-2.01)., Conclusions: Six years after the launch of the PRISMA statement, the proportion of protocol registration in high-impact journals has increased some but remains low. The present study found no evidence suggesting that protocol registration reduced outcome reporting bias., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Aim: The prevalence of cerebral palsy (CP) has not decreased in developed countries over the past 30 years. We examined gestational age-specific trends in the prevalence of CP., Methods: This unselected, population-based study was conducted in Miyazaki prefecture, Japan (10,000 deliveries annually), where 102,999 deliveries were registered between 2001 and 2010. Of these, 312 were stillbirths (⩾22 weeks of gestation), 126 were neonatal deaths (<28 days of birth), and 214 infants were determined to be at risk of CP at peer-review conferences. Survival and neurological damage were compared for two 5-year periods, 2001-2005 and 2006-2010, and infants were classified according to gestational ages., Results: Stillbirths and neonatal deaths decreased significantly during both periods. Likewise, the number of registered high-risk cases of CP decreased by 30.2%, from 126 to 88 cases. After excluding congenital anomalies, the corrected CP prevalence was 1.5 per 1000 (78/51,889) and 1.3 per 1000 (67/51,110), for the two periods, which was not a significant difference. The number of extremely preterm infants (22-25 weeks) did not change over the 10-year period, whereas that of moderately preterm infants (26-36 weeks) increased, and that of term infants significantly decreased (p<0.01). In term infants, asphyxia decreased from 18 to 7 cases (p<0.05)., Conclusions: Perinatal deaths and CP decreased in prevalence during both 5-year periods, and the CP prevalence was 2.1 per 1000 births. Furthermore, fewer term infants were at high risk for CP mainly because of the reduced prevalence of asphyxia., (Copyright © 2015 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

Background: It remains unclear whether performing amniocentesis to detect intra-amniotic infection is useful for improving neonatal outcomes., Aims: To determine the efficacy of amniocentesis on the neonatal outcomes in women exhibiting threatened preterm labor and intact membranes., Study Design: Retrospective cohort study, Subjects: A total of 174 women with threatened preterm labor and intact membranes at 22 to 33 weeks of gestation. Women with obvious clinical chorioamnionitis, multifetal pregnancy and/or major anomalies were excluded., Outcome Measures: Neonatal short- and long-term outcomes, Results: Sixty-seven women underwent amniocentesis (Tap group), while the remaining 107 did not. The prevalence of a positive Gram stain or a positive culture result was 10% in the Tap group. The overall outcomes were not statistically different between the two groups, with the exception of borderline significance (p=0.052) in long-term outcomes, favoring the Tap group. We performed a subgroup analysis focusing on infants born at 22-28 weeks of gestation. Consequently, the Tap group had better neonatal outcomes than the no-Tap group with respect to both short-term (OR 0.19, 95%CI 0.07-0.55) and long-term (OR 0.15, 0.05-0.46) outcomes. A multivariate analysis revealed that after adjusting confounding factors, the gestational age at delivery (OR 0.4, 0.3-0.7) and amniocentesis (OR 0.1, 0.02-0.3) remained significantly different., Conclusions: Amniocentesis is useful for improving neonatal outcomes in infants born at 22-28 weeks of gestation to women exhibiting preterm labor and intact membranes., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Small bowel adenocarcinoma (SBA) is a rare, aggressive malignancy with a poor prognosis, and the mechanisms of carcinogenesis in SBA remain unclear. Our aims were to investigate the molecular mechanisms underlying SBA and to identify treatments by establishing and characterizing an SBA cell line and performing anti-cancer drug screening. SIAC1 cells, established from jejunal SBA, showed epithelial characteristics and formed organoids in 3D culture. SIAC1 cells had a heterozygous β-catenin deletion mutation, resulting in a stable β-catenin protein with enhanced Wnt/β-catenin activity. SIAC1 cells lacked MLH1 and MSH6 expression, and target genes such as TGFBR2 and ACVR2 showed frameshift mutations. Among 10 clinical SBA samples, 2 (20%) had interstitial deletions in β-catenin, expression of mismatch repair protein was aberrant in 4 (40%), and heterozygous frameshift mutations of three target genes were found in all 10 samples. On screening assay using 140 compounds, eribulin significantly inhibited SIAC1 cell growth both in vitro and in vivo by inhibition of the Wnt/β-catenin pathway via enhanced degradation of β-catenin. In conclusion, we established an SBA cell line with molecular characteristics similar to those of clinical SBA samples, including β-catenin deletion and mismatch repair protein deficiency, that will be useful for SBA research. Eribulin might be a candidate for SBA treatment due to its inhibitory effect on Wnt/β-catenin signaling., (Copyright © 2015 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)

Objective: Determine the impact of cesarean section (CS) on neonatal outcome of infants born at 23weeks of gestation., Methods: A retrospective study was performed involving 34 infants born at 23weeks and 91 infants born at 24-26weeks. Indications necessitating delivery were severe pregnancy induced hypertension, non-reassuring fetal heart rate patterns (NRFHRs), or intrauterine infection (IUI). Obstetrical indication for CS included NRFHR and breech presentation. Poor outcome included neonatal death or cerebral palsy. Univariate and multiple logistic analyses were performed to determine the effect of CS for obstetrical indications on poor outcome., Results: The incidence of poor outcome was significantly higher at 23weeks (number of poor outcomes/total number: 22/34) compared to that (31/91) at 24-26weeks (p<0.01). The incidence of a poor outcome was significantly higher at 23weeks for infants having NRFHR (11/16) compared to those at 24-26weeks (15/43, p=0.02). However, the incidence of a poor outcome was similar in infants with IUI (6/10 at 23weeks versus 5/11 at 24-26weeks, p=0.41). Vaginal birth in cases of obstetrical indication for CS at 23weeks was associated with higher risk of a poor outcome (odds ratio: 8.2). In contrast, the risk at 24-26weeks was not higher (OR, 0.8). After adjustment using variables of vaginal birth and IUI, vaginal birth significantly affected poor outcome (OR, 13.0)., Conclusion: Poor neonatal outcome was closely related to the mode of delivery, suggesting that CS for obstetrical indication at 23weeks may improve neonatal outcome., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Objective: We previously showed that acetylcholine receptor (AChR) agonist reduced hypoxic-ischemic brain damage in the newborn rats. To further investigated the interaction between hypoxia and chorinergic anti-inflammatory pathway, we examined the effect of AChR antagonist on brain damage and to see the relation between microglial activation and protective effect of AChR agonist., Study Design: Seven-day-old Wistar rats were divided into 2 groups, one receiving AChR antagonists to see if they have deleterious effects on hypoxic-ischemic brain damage, and the other receiving AChR agonist, carbachol, to investigate the emergence of microglia in the hippocampus. Rats were subjected to left carotid artery ligation followed by 8% hypoxia. Brains were analyzed histologically and immunohistochemically., Results: Antagonists of AChRs significantly enhanced brain damage in 1-h hypoxia-ischemia. In particular, the nicotinic AChR antagonist showed a marked enhancement of brain damage compared to the saline controls (p<0.01). The hippocampal CA1 was most vulnerable to any AChR antagonists, while the cortex was least vulnerable and only responsive to a higher dose of non-selective nAChR antagonist. Carbachol showed significantly less accumulation of microglia in the hippocampus than the saline controls (p<0.01) in hypoxia-ischemia., Conclusion: An AchR-responsive pathway in the brain plays an important role in modifying perinatal brain damage, in which microglial accumulation may be involved., (Copyright © 2012 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

Background & Aims: Some clinical findings have suggested that systemic metabolic disorders accelerate in vivo tumor progression. Deregulation of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway is implicated in both metabolic dysfunction and carcinogenesis in humans; however, it remains unknown whether the altered metabolic status caused by abnormal activation of the pathway is linked to the protumorigenic effect., Methods: We established hepatocyte-specific Pik3ca transgenic (Tg) mice harboring N1068fs*4 mutation., Results: The Tg mice exhibited hepatic steatosis and tumor development. PPARγ-dependent lipogenesis was accelerated in the Tg liver, and the abnormal profile of accumulated fatty acid (FA) composition was observed in the tumors of Tg livers. In addition, the Akt/mTOR pathway was highly activated in the tumors, and in turn, the expression of tumor suppressor genes including Pten, Xpo4, and Dlc1 decreased. Interestingly, we found that the suppression of those genes and the enhanced in vitro colony formation were induced in the immortalized hepatocytes by the treatment with oleic acid (OA), which is one of the FAs that accumulated in tumors., Conclusions: Our data suggest that the unusual FA accumulation has a possible role in promoting in vivo hepato-tumorigenesis under constitutive activation of the PI3K pathway. The Pik3ca Tg mice might help to elucidate molecular mechanisms by which metabolic dysfunction contributes to in vivo tumor progression., (Copyright © 2011 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Background: Due to circulatory instability, premature infants require volume loads, catecholamines and steroid supplementation to improve mortality and neurodevelopmental outcome. However, a complete quantitative analysis concerning the relationship between supplementation and gestational age, especially in infants born at 22 to 24 weeks of gestation, is lacking., Aim: To investigate whether less mature infants need higher doses of catecholamine, volume loads and steroid, and whether those who require higher doses have poorer outcome., Study Design: A retrospective, observational study was performed at a tertiary center in a university setting. Among the consecutive 221 premature infants born at 22 to 28 weeks of gestation, we selected 108 infants who had no apparent pathological conditions other than prematurity. Catecholamines, volume loads and steroid, given to attain sufficient blood pressure and urinary output, were quantitatively analyzed during the first 24 hours., Outcome Measures: Quantity of catecholamines, volume expanders and steroid supplementation as a function of gestational age and childhood outcome at 2 years., Results: Catecholamines and volume loads were increased in a step-wise manner with decreasing gestational age. Intact survival rate was significantly lower in infants born before 25 weeks of gestation compared with the more mature infants. Among infants born at 22 to 24 weeks' gestation, catecholamine and volume load increased significantly in poor outcome infants compared with good outcome infants., Conclusions: From a developmental viewpoint, progressively larger doses of catecholamine, volume expanders and corticosteroid are required to stabilize circulatory adaptation to neonatal life in infants between 22 to 28 weeks of gestation., (2010 Elsevier Ltd. All rights reserved.)

Receptor tyrosine kinases (RTKs) are involved in oncogenesis and disease progression for many cancers. Inhibitors targeting them are vigorously developed and some of them are tested in the clinical setting. Amplifications of certain RTKs (c-Met, FGFR2 and ErbB2) have been associated with human gastric cancer progression. According to our genome-wide scans of genetic lesions in 34 gastric cancer cell lines using high-density single-nucleotide polymorphism genotyping microarrays, we confirmed that the c-met locus was amplified in four gastric cancer cell lines (Hs746T, MKN45, NUGC4 and SNU5). It was reported that somatic mutation is occasionally detected in tumor samples of a certain type of cancer with gene amplification. Previous reports showed gastric cancers harbored mutations of FGFR2 and ErbB2, but c-Met oncogenic mutation had not yet been reported. We performed mutational analysis of the cytoplasmic domains of c-Met using the genome DNA of the gastric cancer cell lines, and found that Hs746T cells had a splice site mutation of exon 14. By cDNA sequencing and Western blotting, we showed that the mutation caused juxtamembrane domain deletion. Previously, this mutation had been detected only in lung cancer specimens and this deletion resulted in the loss of Cbl E3-ligase binding causing decreased ubiquitination and delayed down-regulation. In conclusion, four gastric cancer cell lines harbored amplification of c-met locus, and among them, Hs746T had a putative oncogenic mutation with amplification. This information will be useful for screening of inhibitors targeting gastric cancer with c-Met aberration., (2010 Elsevier Inc. All rights reserved.)

Background: Cases suggestive of non-acidemia related cerebral palsy (CP) are likely misdiagnosed as acidemia related CP because of the presence of nonreasuring fetal heart rate (FHR) patterns., Aims: Our purpose was to compare intrapartum FHR patterns between the cases of neurological damage and the cases without disability after severe metabolic acidemia and neonatal encephalopathy, and also to compare the FHR patterns between cases with CP due to asphyxia and cases with CP of other etiology in infants born after 34 weeks., Study Design: From 1998 to 2003, our peer review conferences determined 136 infants with high-risk factors for neurological impairment in the unselected 65,197 live births. High-risk infants were chosen according to our criteria. Among them 58 were eligible infants because they were born at > or =34 weeks of gestation and also had legible FHR traces., Outcome Measures: Incidence of nonreassuring FHR patterns., Results: Fifteen infants were acidemia related and 43 were non-acidemia related high-risk infants. Ten of the 15 acidemia infants developed CP and all had shown bradycardia > or =13 min with a nadir <80 bpm. In the 43 non-acidemia infants, 35 had CP, mental retardation, epilepsy, or hearing loss and 74% (26/35) of them had shown nonreassuring FHR patterns. Incidence of severe bradycardia was significantly elevated in the acidemia related CP compared with acidotic infants without disability, and those with non-acidemia related CP., Conclusions: Even in infants with non-acidemia related CNS impairments, who were born at > or =34 weeks of gestation, 74% had shown intrapartum nonreassuring FHR patterns.

Background: Some of the perinatal deaths are preventable., Aims: To examine with accuracy perinatal deaths and their clinically relevant conditions to further reduce perinatal mortality rates., Study Design: Population-based study., Subjects: 356 perinatal deaths in Miyazaki Prefecture from 1998 to 2005., Outcome Measures: Causes and clinically associated risk factors of perinatal deaths in infants with congenital abnormalities and those of the non-malformed infants., Methods: We performed a population-based study of 87,593 deliveries in Miyazaki from 1998 to 2005, where 356 perinatal deaths were reported. We also held peer-review audit conference twice a year to investigate causes and clinically associated risk factors of the perinatal deaths, where at least 7 obstetricians and 7 neonatologists congregated., Results: Our perinatal mortality rate was 4.1/1000. 99% of the neonatal deaths and 85% of the stillbirths were examined by the peer-review audit conferences to validate the accuracy of causes. Three fourths were non-malformed perinatal deaths, in which stillbirths represented twice the number of neonatal deaths. Prematurity is the major factor attributable to neonatal deaths. Half of stillbirths were unexplainable but associated with overt or subtle fetal growth restriction. Intrapartum asphyxia after 32 weeks of gestation resulted in 10% of perinatal deaths. The audit conferences concluded that 13% (28/222) of the non-malformed infants had a potential of avoiding death., Conclusion: In the advanced region of perinatal medicine in Japan, we still have room to improve perinatal mortality. Most prevalent factors were fetal growth restriction, intrapartum asphyxia after 32 weeks of gestation, and sudden fetal deaths of undiagnosed diabetes near-term.

Background: Intrauterine bacterial infection is important as a high risk factor associated with subsequent brain damage of the newborn., Aims: To see if mature fetuses require both hypoxia and intrauterine infection to lead to cerebral palsy, while premature fetuses need infection alone., Study Design: A retrospective cohort study., Subjects: 230 singleton live-born infants of 22 to 32 weeks of gestation, exposed to intrauterine infection during labor and delivery, from 1995 to 2002., Methods: Mortality and incidence of cerebral palsy at 2 years old were compared among the 3 groups; immature (n=89, 22-27 weeks), premature (n=73, 28-33 weeks) and mature (n=68, >34 weeks). The relationship between cerebral palsy and fetal pH values was examined., Outcome Measures: Mortality and cerebral palsy., Results: Mortality and cerebral palsy were significantly decreased with advancing gestation. Mortality was significantly decreased after 28 weeks of gestation while cerebral palsy was significantly decreased after 34 weeks of gestation. Acidosis was associated with cerebral palsy in mature infants, but not in less mature infants., Conclusions: : Premature infants were more susceptible to intrauterine infection to cause death or cerebral palsy than mature infants. Mature infants may require exposures to both infection and hypoxia but less mature infants need infection alone to cause cerebral palsy, suggesting different pathogenesis during the developmental stage.

Background: It is still controversial that intrapartum exposure to magnesium may or may not reduce brain damage in premature infants in human and animal models., Aims: We investigated the effect of hypoxemia alone under magnesium exposure on fetal cardiovascular changes in chronically catheterized goat fetuses., Study Design: We performed a 3-day experimental protocol with control (10% glucose) on day 1, recovery on day 2, and magnesium on day 3. Magnesium sulfate was directly infused to fetuses in a bolus dose of 270 mg/kg followed by 80 mg/kg/h. Hypoxemia was induced by maternal inhalation of nitrogen gas on day 1 and on day 3. Cerebral blood flow was measured by colored microsphere techniques. Repeated measure ANOVA and Bonferroni's/Dunn's test were used for comparison., Subjects: Six Japanese Saanen goats at 0.85 gestation., Outcome Measures: Fetal heart rate, blood pressure, and cerebral blood flow., Results: Ionized magnesium concentrations were significantly increased. Fetal PO2 decreased significantly from 30 mmHg to 14 mmHg without acidemia. Magnesium exposure significantly attenuated hypoxemia-induced bradycardia but did not affect blood pressure. Hypoxemia significantly increased fetal brain blood flow from the pre-hypoxic levels on day 1. Magnesium exposure further increased hypoxemia-induced brain blood flow on day 3, but statistical significance was limited to the cerebral cortex., Conclusion: In near-term, initially healthy goat fetuses, brain blood flow during acute hypoxemia was significantly increased with magnesium sulfate exposure.

On-line preconcentration system for the selective, sensitive and simultaneous determination of chromium species was investigated. Dual mini-columns containing chelating resin were utilized for the speciation and preconcentration of Cr(III) and Cr(VI) in water samples. In this system, Cr(III) was collected on first column packed with iminodiacetate resin. Cr(VI) in the effluent from the first column was reduced to Cr(III), which was collected on the second column packed with iminodiacetate resin. Hydroxyammonium chloride was examined as a potential reducing agent for Cr(VI) to Cr(III). The effects of pH, sample flow rate, column length, and interfering ions on the recoveries of Cr(III) were carefully studied. Five millilitres of a sample solution was introduced into the system. The collected species were then sequentially washed by 1M ammonium acetate, eluted by 2M nitric acid and measured by ICP-AES. The detection limit for Cr(III) and Cr(VI) was 0.08 and 0.15mugl(-1), respectively. The total analysis time was about 9.4min. The developed method was successfully applied to the speciation of chromium in river, tap water and wastewater samples with satisfied results.

Objective: The purpose of this study was to elucidate a role for edaravone, a free radical scavenger 3-methyl-1-phenyl-2-pyrazolin-5-one, in neonatal hypoxic-ischemic brain damage. We determined the level of thiobarbituric acid reactive substances as an index of lipid peroxidation and nitric oxide metabolites as nitric oxide production., Study Design: Seven-day-old Wistar rats were subjected to left common carotid artery ligation followed by 2 hours of 8% oxygen exposure. Then, the rats were administered edaravone (9 mg/kg) or saline solution intraperitoneally. Cerebrospinal fluid was withdrawn just before the rats were killed at 2, 5, 24, and 48 hours after hypoxia, and brains were removed. The thiobarbituric acid reactive substances and nitric oxide metabolites levels were measured in the brain tissue and cerebrospinal fluid, respectively., Results: On the ligated side, edaravone significantly decreased thiobarbituric acid reactive substances levels at 5 and 24 hours after hypoxia, compared with saline group (P < .01). Edaravone significantly decreased the nitric oxide metabolites level in the cerebrospinal fluid only at 5 hours, compared with saline group (P < .01)., Conclusion: Edaravone potently and transiently inhibited lipid peroxidation and the production of nitric oxide in the neonatal rat brain after hypoxic-ischemic insult.

Objective: We examined whether progenitor neural stem cells can differentiate successfully into mature neurons and astrocytes in a rat model of neonatal hypoxic-ischemic encephalopathy., Study Design: Seven-day-old Wistar rats were subjected to hypoxic-ischemic stress. At days 5 to 7 after hypoxic-ischemic stress, 5-bromodeoxyuridine (an early marker of cell proliferation) was injected, and the brains were retrieved at 14, 28, and 42 days after hypoxic-ischemic stress. Immunohistochemical and immunofluorescent studies were carried out for 5-bromodeoxyuridine, neuronal nuclear antigen (a marker protein of matured neuron), and glial fibrillary acidic protein (a protein marker of mature astrocytes)., Results: Only 1% of neuronal nuclear antigen-positive and 4.6% of glial fibrillary acidic protein-positive cells could be detected among the 5-bromodeoxyuridine-immunopositive cells in the peri-infarcted area of the cortex and the striatum, respectively, at 14 days after hypoxic-ischemic stress. There were no such double-staining cells at 28 and 42 days after hypoxic-ischemic stress., Conclusion: The intrinsic ability for neurologic self-repair was limited at the maturation step after hypoxic-ischemic stress in the neonatal rat brain.

Objective: It has been reported that an infarcted area is reduced by the injection of glial cell line-derived neurotrophic factor into brain parenchyma after hypoxic/ischemic insult in neonatal rats. For use of glial cell line-derived neurotrophic factor in humans, we have developed a system for the delivery of a constant supply of glial cell line-derived neurotrophic factor to the brain. The aim of this study was to examine the neuroprotective effect of glial cell line-derived neurotrophic factor with the use of this delivery system., Study Design: Baby hamster kidney cells were transfected with human glial cell line-derived neurotrophic factor complementary DNA, encapsulated in semipermeable hollow fibers, and implanted into the left cerebrum of 12-day-old Wistar rats (glial cell line-derived neurotrophic factor group, 11 rats). Nontransfected baby hamster kidney cells served as controls (control group, 9 rats). Two days after implantation, the rats received a hypoxic/ischemic stress, with a modification of Levine's method. Seven days later the rats were killed, and coronal brain slices were cut 2, 4, 6, 8, and 10 mm from the anterior pole. The cortex, hippocampus, striatum, and thalamus were evaluated for damage severity. The serum concentrations of glial cell line-derived neurotrophic factor were also determined., Results: The left brain hemispheric area was significantly larger; the neuronal damage to each brain region was significantly less, and the serum glial cell line-derived neurotrophic factor concentrations were significantly higher in the glial cell line-derived neurotrophic factor group, compared with the control group., Conclusion: Grafting of encapsulated glial cell line-derived neurotrophic factor-secreting cells is a promising way to protect the neonatal brain from hypoxic/ischemic insult.

Objective: Implantation of encapsulated glial cell line-derived neurotrophic factor-secreting cells into brain parenchyma reduces histological brain damage following hypoxic-ischemic stress in neonatal rats. We examined the effect of glial cell line-derived neurotrophic factors on long-term learning and memory impairment and morphological changes up to 18 weeks after hypoxic-ischemic stress in neonatal rats., Study Design: Baby hamster kidney cells were transfected with expression vector either including (glial cell line-derived neurotrophic factor-hypoxic-ischemic group; n = 10) or not including (control-hypoxic-ischemic group; n = 8) human glial cell line-derived neurotrophic factor cDNA, encapsulated in semipermeable hollow fibers, and implanted into the left brain parenchyma of 7-day-old Wistar rats. Two days after implantation the rats received hypoxic-ischemic stress, and their behavior was then examined in several learning tasks: the 8-arm radial maze, choice reaction time, and water maze tasks, which examine short-term working memory, attention process, and long-term reference memory, respectively. The rats were killed 18 weeks after the hypoxic-ischemic insult for evaluation of brain damage. Two additional control groups were used: the control group (n = 15), which underwent no treatment, and the glial cell line-derived neurotrophic factor group (n = 6), which underwent implantation of the glial cell line-derived neurotrophic factor capsule but did not undergo hypoxic-ischemic stress., Results: The decrease in the size of the cerebral hemisphere was significantly less in the glial cell line-derived neurotrophic factor-hypoxic-ischemic group, compared with the control-hypoxic-ischemic group, and improved performance was observed in all three tasks for the glial cell line-derived neurotrophic factor-hypoxic-ischemic group: for the control-hypoxic-ischemic group versus the glial cell line-derived neurotrophic factor-hypoxic-ischemic group, respectively, in the 8-arm radial maze test, average number of correct choices was 6.2 +/- 0.1 versus 6.9 +/- 0.1 ( P < .01); in the choice reaction time test, average reaction time for a correct response was 2.35 +/- 0.1 seconds versus 1.97 +/- 0.09 seconds ( P < .01); in the water maze test, average swimming length was 1120.0 +/- 95.2 cm versus 841.6 +/- 92.1 cm ( P < .01). All results for the glial cell line-derived neurotrophic factor group were similar to those for the control group., Conclusion: Glial cell line-derived neurotrophic factor treatment is effective in not only reducing brain damage but also inhibiting learning and memory impairment, following hypoxic-ischemic insult in neonatal rats. No adverse effects in learning and memory tests were observed in the glial cell line-derived neurotrophic factor group.

Objective: There are no established therapies for preventing or rescuing perinatal infection or inflammation-induced perinatal brain damage. We administered dexamethasone (DEX), a synthetic corticosteroid anti-inflammatory drug, to neonatal rats in a model of such damage induced by a combination of lipopolysaccharide (LPS) and hypoxia-ischemia (HI), which produces characteristic histologic and behavioral abnormalities., Study Design: Four hours after the injection of LPS (1 mg/kg, i.p.), 7-day-old Wistar rat pups were subjected to unilateral HI for 1 hour according to Levine's procedure. Injections of 0.5 mg/kg of dexamethasone (DEX-treated group, n = 15) or saline (saline-treated group, n = 15) were given 4 hours before HI. A sham-operated control group received neither LPS nor HI (n = 15). We chose rats of this age because their stage of brain maturation is similar to the human neonate. Over the 7 to 16 weeks after treatment, a choice reaction time (CRT) task was used for assessment of attention processes in each group, an 8-arm radial maze task was used to test short-term memory, and a water maze task was used to test long-term memory. In the CRT task, the reward food was released when the tested animal correctly pressed a lever on the side of an illuminating lamp. The correct and incorrect lever pressings were counted. In the 8-arm radial maze task, rats were allowed to move freely, seeking a reward of food placed at the end of 1 arm. An error was defined as the choice of an arm that had already been visited. In the water maze, rats had to swim to seek a concealed platform as aversive escape motivation. At 19 weeks, the rats were euthanized, the brain was removed, sectioned coronally, and the volume of each part was measured., Results: The striatum, cortex, and hippocampus showed reductions in volume in the saline-treated group (42.7%, 49.2%, and 34.9% decreases compared with the sham-operated controls, respectively), but this was not observed in the DEX-treated group. All learning and memory processes were impaired with the combination of LPS and HI treatment, but these deficits were almost completely prevented by DEX treatment., Conclusion: Dexamethasone is a promising candidate for prevention of infection and inflammation-induced perinatal brain damage. The impact of dexamethasone identifies potential therapeutic pathways once the mechanism of dexamethasone's protection is determined.

Objective: We assessed the long-term effects of perinatal hypoxia-ischemia and endotoxin on attention and short- and long-term memory in neonatal rats with the use of behavioral tasks and brain histologic results., Study Design: Four hours after injections of lipopolysaccharide (1 mg/kg, intraperitoneally) or saline solution, 7-day-old Wistar rat pups were subjected to unilateral hypoxia-ischemia for 1 hour. We studied 4 groups: controls (n = 43 rats), lipopolysaccharide alone (n = 12 rats), hypoxia-ischemia alone (n = 29 rats), and combined lipopolysaccharide + hypoxia-ischemia treatment (n = 34 rats). Seven to 16 weeks after the treatment, we measured attention with a choice reaction time task, short-term memory with an 8-arm radial maze task, and long-term memory with a water maze task. At 19 weeks of age, the brain was removed, fixed, and sectioned coronally; and the volume of each part was measured., Results: A loss of volume in the hippocampus was observed in the lipopolysaccharide, hypoxia-ischemia, and lipopolysaccharide + hypoxia-ischemia groups; a loss of striatum was observed in the hypoxia-ischemia and lipopolysaccharide + hypoxia-ischemia groups, but loss of cortex was observed only in the lipopolysaccharide + hypoxia-ischemia group. The lipopolysaccharide, hypoxia-ischemia, and lipopolysaccharide + hypoxia-ischemia groups showed significantly poorer performance (attention deficit) than controls in the choice reaction time task. Correct choices decreased, and error increased in the lipopolysaccharide + hypoxia-ischemia group compared with the other groups in the radial maze task, which shows short-term memory impairment. Swimming distance was significantly greater in the hypoxia-ischemia and lipopolysaccharide + hypoxia-ischemia groups than in the other 2 groups in the water maze test, which shows long-term memory impairment., Conclusion: Combined lipopolysaccharide and hypoxia-ischemia treatment synergistically induced short-term memory impairment that is associated with loss of cortical volume.

A rat model of neonatal hypoxic-ischemic encephalopathy (Rice's model) was obtained by unilateral ligation of the common carotid artery of 7-day-old rats with hypoxia (exposure to 8% oxygen). To estimate the in vivo intracerebral reducing ability of the mature rats (8 weeks old) of Rice's model, temporal electron paramagnetic resonance (EPR) imaging of the brain of a rat receiving a blood-brain barrier-permeable nitroxide radical, 3-hydroxymethyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl, was performed. In this imaging technique, the decay rate of the EPR signal intensity in a selected region of the brain is indicative of region-specific reducing ability. The effect of neonatal treatment of an antioxidant agent, 3-methyl-1-phenyl-2-pyrazolin-5-one (MCI-186), after a hypoxic-ischemic insult was also tested. It was found that the reducing ability had been depleted in the contralateral hemisphere of Rice's model rats; this depletion was suppressed by administering MCI-186.

Objective: Several studies have shown that abnormal intrapartum fetal heart rate patterns are the results from pre-existing fetal brain damage. We evaluated intrapartum fetal heart rate pattern of cytomegalovirus-infected fetuses and correlated the patterns with neurologic outcomes., Study Design: Between 1991 and 2001, there were 20 cytomegalovirus-infected fetuses. We selected 40 fetuses as control subjects that were matched for gestational age and birth weight. Fetal heart rate was interpreted according to the guidelines of the National Institute for Child and Human Development. The incidence of abnormal fetal heart rate pattern and umbilical blood gases were compared between both groups. We also investigated the factors that contributed to abnormal fetal heart rate pattern in the cytomegalovirus group., Results: Nonreassuring fetal heart rate patterns (prolonged deceleration and recurrent late deceleration) were observed in 8 of 20 fetuses (prolonged deceleration, 7 fetuses; recurrent late deceleration, 1 fetus) in the cytomegalovirus group and in 3 of 41 fetuses (prolonged deceleration, 1 fetus; recurrent late deceleration, 2 fetuses) in the control group (P<.05, Fisher test). Baseline fetal heart rate variability was minimal in 4 of the 7 prolonged deceleration cases in the cytomegalovirus group. Umbilical pH <7.1 was found for 1 fetus in the cytomegalovirus group. The average umbilical arterial pH values were similar in both the groups. In the cytomegalovirus group, there were no differences in the incidence of contributing factors between 8 fetuses with abnormal fetal heart rate pattern (prolonged deceleration and recurrent late deceleration) and 8 fetuses with no change. There were 3 fetuses with cerebral palsy: 2 fetuses in the no change group and 1 fetus in the prolonged deceleration group. Antigenemia was positive exclusively in 4 cases with abnormal fetal heart rate pattern (P<.05)., Conclusion: Cytomegalovirus-infected fetuses are more likely to show abnormal intrapartum fetal heart rate patterns than low-risk control fetuses, which suggests that the perinatal detection of cytomegalovirus is necessary to distinguish hypoxic-ischemic encephalopathy.

Smad4 is a tumor-suppressor gene that is lost or mutated in 50% of pancreatic carcinomas. Smad4 is also an intracellular transmitter of transforming growth factor-beta (TGF-beta) signals. Although its tumor-suppressor function is presumed to reside in its capacity to mediate TGF-beta-induced growth inhibition, there seems to be a Smad4-independent TGF-beta signaling pathway. Here, we succeeded in establishing Smad4 knockdown (S4KD) pancreatic cancer cell lines using stable RNA interference. Smad4 protein expression and TGF-beta-Smad4 signaling were impaired in S4KD cells, and we compared the proteomic changes with TGF-beta stimulation using two-dimensional gel electrophoresis (2-DE) and mass spectrometry. We identified five proteins that were up-regulated and seven proteins that were down-regulated; 10 of them were novel targets for TGF-beta. These proteins function in processes such as cytoskeletal regulation, cell cycle, and oxidative stress. Introducing siRNA-mediated gene silencing into proteomics revealed a novel TGF-beta signal pathway that did not involve Smad4.

Objective: Our purpose was to evaluate the clinical validity of electronic fetal heart rate monitoring to detect fetal acidemia and to evaluate the prevalence of cerebral palsy in unselected low-risk pregnancies., Study Design: We selected two secondary and two tertiary level institutions in which 10,030 infants were born. Among them, 5546 of the pregnancies were low-risk pregnancies by antepartum evaluation. The fetal heart rate patterns were interpreted according to the guidelines of the National Institute of Child Health and Human Development. The correlations between the fetal heart rate pattern and umbilical blood gases and the fetal heart rate pattern and cerebral palsy were studied. Spastic cerebral palsy was diagnosed at > or =1 year by pediatric neurologists. Statistics included unpaired t test, contingency table with chi(2) and Fisher tests, and one-way analysis of variance with Bonferroni/Dunn test., Results: On the basis of the severity of decelerations, frequency of decelerations, and decreased variability, umbilical pH, and Po(2) level were decreased accordingly, and incidence of pH<7.1 was increased. Sensitivity and false-positive rate of nonreassuring fetal heart rate patterns for fetal acidemia were 63% and 89%. There were nine cerebral palsy cases: six of the cases were preexisting asphyxia before monitoring was initiated, two of the cases were cytomegaloviral infections, and one of the cases was a maternal amniotic fluid embolism., Conclusion: In low-risk pregnancies, intrapartum fetal heart rate monitoring was useful to detect fetal acidemia. Cerebral palsy caused by intrapartum asphyxia was restricted to unavoidable accidents under continuous fetal heart rate monitoring.

Objective: Our purpose was to determine whether systolic arterial pressure, as predicted from the delay time from the electrocardiogram to pulse oximeter waveform, correlates with directly measured systolic blood pressure., Methods: Six Japanese Saanen goats at 120-125 days' gestation were used. Experiments were conducted in acutely operated animals under anesthesia. We placed catheters in the carotid artery and jugular vein, electrodes on the chest, and a pulse oximeter probe on the tongue. Systolic blood pressure (Psm), fetal heart rate (FHR), delay time (Td), and pulse oximeter waveform were monitored continuously, and the data were stored. Systolic blood pressure was estimated (Pse) by the simple linear equation of Td. Fetal blood pressure was decreased 10% by hydralazine and blood withdrawal. Correlation coefficients and the agreement between Psm and Pse were determined., Results: A significant correlation (P <.01) with good bias values (< 8 mmHg) and precision values (< 5 mmHg) was obtained between Psm and Pse in all animals. There was also a significant correlation between Psm and Pse with high coefficients for hydralazine (r(2) = 0.82 +/- 0.12) and for withdrawal (r(2) = 0.85 +/- 0.15). The correlations became slightly scattered in acidemia (pH < 7.25)., Conclusions: This technique has the potential to provide information on fetal blood pressure during labor and delivery on a beat-by-beat basis, which will aid in the interpretation of electronic FHR recordings.

Objective: To test whether magnesium inhibits apoptosis during hypoxia-ischemia (HI) in the developing brain, we studied the effect of long-term magnesium treatment on an early marker of neuronal damage (loss of microtubule-associated protein-2; MAP-2) and a marker of apoptosis (activated caspase-3) after HI in newborn rats., Methods: Seven-day-old rat pups (n = 107) were exposed to unilateral carotid artery ligation and 2 hours of hypoxia (8% oxygen in 92% nitrogen). Magnesium was administered by a micro-osmotic pump implanted in the back, at an infusion rate of 75 mg/kg per hour for 3 days. Neuronal loss in the cerebral cortex and hippocampus was evaluated by loss of MAP-2 at 24 and 48 hours after HI and visually ranked by a semiquantitative, three-point scale (mild, moderate, and severe). Caspase-3 activation was evaluated similarly in an adjacent section. Area and severity of the damaged lesion were compared between the two staining methods and between magnesium and controls, by chi(2) test and Fisher test., Results: Three-day magnesium administration reduced loss of MAP-2 (from 33 of 50 to 21 of 45, P =.06) and activation of caspase-3 (from 34 of 50 to 21 of 45, P =.04) in the cerebral cortex after HI. There was no significant change in these immunostainings in the hippocampal regions. In the hippocampal CA3, the severity ranked by activated caspase-3 staining was significantly less than that of MAP-2 staining., Conclusion: Long-term, post-HI treatment with magnesium inhibited caspase-3 activation and MAP-2 immunostaining, suggesting that magnesium inhibited apoptotic neuronal death of HI in 7-day-old rats.

An ideal replacement for bone defects is auto-bone tissue, of which there is an ample supply with the required form and with vascularity. Our strategy for generating such bone tissue is as follows. First, bone tissue is induced in muscle by bone morphogenetic protein-2 (BMP-2) with beta-tricalcium phosphate as a carrier to maintain its form in the muscle. Second, the induced bone in the muscle pedicle is grafted to the bone defect to maintain vascularity. In the first experiment, 50 microg of recombinant human BMP-2 (rhBMP-2) was inoculated into the hip abductor muscle of rabbits with beta-tricalcium phosphate under anesthesia. Five weeks after the operation, intramuscular bone formation was observed in all of the samples, and the form and size of the induced bone tissue were identical to those of the carrier. Ten weeks after the operation, the induced bone was partly absorbed. In the second experiment, 50 microg of rhBMP-2 was inoculated in the same manner as previously. Five weeks after the operation, the muscle tissue around the induced bone was incised, leaving just the proximal part as a pedicle. Two or four weeks after the second operation, the induced bone tissue had rich vascularity and no empty lacunae. This indicates the possibility of in vivo bone banking to enable morphologically controlled and vascularized auto-bone grafts.

The transforming growth factor-beta (TGF-beta)-Smad signaling pathway has an important role in carcinogenesis. To study the frequency and mechanism of functional impairment of this pathway in human gastrointestinal cancers, we used a reporter assay to examine the response of 38 cell lines (11 colorectal, 9 pancreatic, 10 gastric, and 8 hepatic cancers) to TGF-beta. We then analyzed TGF-beta type II receptor (T beta RII) gene, immunoblots of Smad4, and restoration of the pathway by rescuing T beta R or Smad. We observed impaired signaling in 91% of colorectal, 67% of pancreatic, and 40% of gastric cancer cell lines, but in none of the hepatic cancer cells. We suggest that this pathway does not function as a tumor suppressor in hepatic carcinogenesis. The impairment is due to inactivation of T beta RII and Smad4 in colorectal and pancreatic cancers. However, because the signal was not recovered by rescuing T beta R or Smad genes in TGF-beta-response-defective gastric cancer cell lines, we suggest that novel molecules or mechanisms are involved in the impaired pathway in some gastric cancers.

Objective: Our purpose was to study the effects of long-term treatment with magnesium sulfate on hypoxic-ischemic brain damage in newborn rats., Study Design: Seven-day-old rat pups (n = 120) were exposed to unilateral carotid artery ligation and 2 hours of hypoxia (8% oxygen in 92% nitrogen). Neuronal loss was evaluated in 4 groups. The loading dose group (n = 25) received a bolus injection of 270 mg/kg magnesium sulfate intraperitoneally. The maintenance group (n = 26) received 3 days of continuous infusion of magnesium sulfate with a micro-osmotic pump at a rate of 72 mg/kg per hour. The loading dose-plus-maintenance group (n = 23) received both. The control group (n = 46) did not receive magnesium. Seven days after the injury the pups were killed and the brains were removed for analysis. Severity of neuronal loss was evaluated in the cerebral cortex and hippocampus and compared among groups by chi2 test., Results: Cyst formation resulting from necrosis was significantly decreased in the maintenance group (15%) and loading dose-plus-maintenance group (9%) but not in the loading dose group (52%) relative to the control group (46%). Severity of neuronal loss in the cerebral cortex and the hippocampus was also significantly improved in the maintenance group and the loading dose-plus-maintenance group but not in the loading dose group. Mortality rates were not different among the groups. Magnesium ion concentrations at 24 hours were significantly decreased from the preinfusion value of 0.49 +/- 0.01 mmol/L in the control group (0.34 +/- 0.03 mmol/L) but remained within normal ranges in the maintenance group (0.46 +/- 0.02 mmol/L with a pump infusing 72 mg/kg per hour and 0.56 +/- 0.05 mmol/L with a pump infusing 24 mg/kg hour). There was an inverse relationship between the maintenance dose and neuronal loss but not between the maintenance dose and mortality rate., Conclusion: Long-term magnesium administration had neuroprotective effects against hypoxia-ischemia in newborn rats.

Objective: We measured fetal plasma concentrations of epinephrine, norepinephrine, and vasopressin during acute hypoxemia in goats and tested whether hypermagnesemia altered these endocrine responses., Methods: Five chronically catheterized goat fetuses at 124-129 days' gestation were used. After 4 hours of infusion (magnesium or vehicle as controls), 30 minutes of hypoxemia was induced by infusing nitrogen gas through a maternal tracheal catheter. Fetal plasma concentrations of epinephrine, norepinephrine, and vasopressin were measured before and during hypoxemia. Both magnesium sulfate and vehicle infusions were performed in each animal. Repeated-measures analysis of variance (ANOVA) and two-way ANOVA with post hoc test were used to determine statistical significance., Results: During hypoxemia, fetal PO(2) decreased significantly from 30 to 14 mmHg with no significant changes in fetal pH or PCO(2) in both groups. Fetal heart rate was reduced significantly by hypoxemia, but to a lesser extent in the magnesium group (change in decrease in fetal heart rate: 41 beats per minute [bpm] in controls versus 26 bpm with magnesium). Mean blood pressure did not change significantly during hypoxemia in both groups. Fetal plasma concentrations of epinephrine, norepinephrine, and vasopressin significantly increased from the prehypoxemic values both with magnesium and in controls. There were no significant differences in these hormone concentrations between magnesium and the controls., Conclusion: Magnesium sulfate had no effect on fetal plasma concentrations of vasopressin, epinephrine, and norepinephrine during acute hypoxemia.

Objective: We investigated the effect of hyperthermic pretreatment 24 hours before hypoxic-ischemic exposure on neuronal cell damage in 7-day-old rats., Methods: Newborn rats were separated on postnatal day 7 into two groups: 1) preheated (those exposed to 2 hours of hyperthermic pretreatment at 42-43C) (n = 29), and 2) nonheated (n = 20). At 24 hours after the hyperthermic stress, rats from both groups were subjected to left carotid artery ligation followed by 2 hours of hypoxia (8% oxygen/92% nitrogen) at 33C. All rats were killed 1 week after hypoxia-ischemia, and brains were extracted for histologic study. A different group of 7-day-old rats (n = 8) was placed in the same hyperthermic environment as mentioned above for 2 hours, and 24 hours after heat exposure brains were extracted for immunohistochemistry of heat-shock protein 70., Results: The total incidence of hypoxic-ischemic brain damage significantly decreased in the preheated group (12 of 25 [48%]) compared with the nonheated group (17 of 20 [85%]; P < .03). The induction of immunoreactive heat-shock protein 70 was observed mainly in glial and vascular endothelial cells and, in a lesser amount, in neuronal cells of the cerebral cortex and hippocampus., Conclusion: Incidence of hypoxic-ischemic brain damage is consistently reduced by 2 hours of hyperthermic pretreatment in 7-day-old rats.

Objectives: Hypoxic-ischemic tolerance can be induced in neonatal rats through hyperthermic preconditioning. The purposes of this study were to determine the interval between hyperthermic preconditioning and a subsequent hypoxic-ischemic insult that would provide optimal neuroprotection against the insult and to examine the relationship between tolerance induction and heat shock protein expression., Study Design: On postnatal day 7 Wistar rat pups were separated into the following 2 groups: a heated group (those exposed to 15 minutes of hyperthermic pretreatment at a brain temperature of 41.5 degrees C-42.0 degrees C) and an unheated control group. At 6, 12, 24, 48, and 72 hours after the hyperthermic stress, rats from both groups were exposed to left carotid artery ligation followed by 2 hours of hypoxia (8% oxygen and 92% nitrogen) at 33 degrees C. Twenty animals from each group were used at each time point. All rats were killed at 1 week after hypoxia-ischemia, at which time the brains were processed and neuronal damage in the cortex and hippocampus was assessed histologically. Another set of 7-day-old rats (n = 30) was studied immunohistochemically at 6, 12, 24, 48, and 72 hours after the same hyperthermic treatment. Expression of 72-kd heat shock protein was measured in neuronal, glial, and vascular endothelial cells., Results: Hyperthermia-induced hypoxic-ischemic tolerance was observed at 6, 12, and 24 hours but not at 48 and 72 hours after hyperthermic preconditioning. Heat shock protein 72 expression in the vascular endothelial cells, rather than in the glial or neuronal cells, was most strongly associated with hypoxic-ischemic tolerance., Conclusion: These findings suggest that heat shock protein 72 in endothelial cells plays an important role in the acquisition of hypoxic-ischemic tolerance at postnatal day 7, a time when maximal angiogenesis occurs and the blood-brain barrier is still immature.

Objective: This study was intended to investigate the temporal changes in heat shock protein 72 expression and microtubule-associated protein 2 disappearance in rat brain at 2 different ages after hypoxic-ischemic insult., Study Design: Both 5-day-old and 14-day-old Wistar rats were subjected to unilateral common carotid artery ligation and hypoxia in 8% oxygen for 2 hours at 33 degrees C. Brain sections were examined sequentially for heat shock protein 72 expression at 0.5, 3, 6, 12, 24, 48, and 72 hours of recovery after hypoxia-ischemia and for microtubule-associated protein 2 disappearance at 0, 24, 48, and 72 hours of recovery and at 7 days of recovery after hypoxia-ischemia. Results of immunohistochemical staining for heat shock protein 72 and microtubule-associated protein 2 were used as markers for detection of early hypoxic-ischemic brain damage. Permanent neuronal damage was assessed with hematoxylin and eosin staining at 7 days after hypoxia., Results: In 5-day-old rats microtubule-associated protein 2 expression was lost as early as 0 hours after hypoxia-ischemia in the cerebral cortex and hippocampus, with a peak at 48 hours after which expression recovered. Expression of heat shock protein 72 was detected in the ligated hemisphere at 0.5 hours after hypoxia-ischemia and peaked at 6 to 24 hours of recovery. In 14-day-old rats microtubule-associated protein 2 was stained in the cortex at 0 hours after hypoxia-ischemia but gradually disappeared in the cerebral cortex and hippocampus after 24 hours of recovery. The expression of heat shock protein 72 was not detected by 6 hours of recovery in the cerebral cortex and by 3 to 12 hours of recovery in the hippocampus, but heat shock protein 72 was persistently expressed in the cortex and hippocampus after 48 hours of recovery. Neuronal damage was significantly less in 5-day-old rats than in 14-day-old rats., Conclusion: In 5-day-old rats hypoxia-ischemia causes earlier changes in heat shock protein 72 and microtubule-associated protein 2 immunostaining results and causes less severe brain damage than in 14-day-old rats.

Destruction of the transcriptional inhibitor IkappaB by the ubiquitin (Ub) system is required for signal-dependent activation of the multifunctional transcriptional factor NF-kappaB, but details of this ubiquitination are largely unknown. We report here that the IkappaBalpha-ubiquitin ligase (IkappaBalpha-E3) is an SCF-like complex containing Skp1, cullin-1, and two homologous F-box/WD40-repeat proteins, betaTrCP1 and betaTrCP2. Intriguingly, all these components are cooperatively recruited to bind to a phosphorylated IkappaBalpha (pIkappaBalpha) produced by tumor necrosis factor-alpha (TNF-alpha) stimulation. IkappaBalpha-E3 bound to pIkappaBalpha catalyzed in vitro ubiquitination of pIkappaBalpha in the presence of ATP, Ub, and E1-activating and E2-conjugating enzymes. Forced expression of betaTrCP1 and betaTrCP2 resulted in dramatic augmentation of the in vitro polyubiquitination activity of IkappaBalpha-E3. These results indicate that the long-sought IkappaBalpha-E3 is an SCF-like complex consisting of multiple proteins which are coordinately assembled during phosphorylation of IkappaBalpha in response to external signals., (Copyright 1999 Academic Press.)

Objective: Our purpose was to study the neuroprotective effect of magnesium sulfate on hypoxicischemic brain damage and how the timing of magnesium administration changes its effect in the newborn rat., Study Design: Seven-day-old rats (n = 91) were exposed to unilateral carotid artery ligation followed by 2 hours of hypoxia (8% oxygen in 92% nitrogen). Magnesium sulfate (270 mg/kg) was intraperitoneally administered 30 minutes before (pretreatment) or 30 minutes after (posttreatment) the 2 hours of hypoxia. In each experiment equivalent amounts of saline solution were administered as controls. Seven days after the injury the rats were killed and the brains were removed for histologic study with hematoxylin and eosin staining. The severity of brain atrophy and neuronal damage was evaluated in the cerebral cortex and hippocampus and compared by chi2 test between the magnesium group and the controls., Results: Ionized magnesium concentrations were significantly increased from 0.52 +/- 0.02 mmol/L to 1.38 +/- 0.07 mmol/L at 0.5 hour after injection and thereafter gradually decreased to 0.73 +/- 0.07 mmol/L at 3.5 hours. Magnesium significantly reduced the incidence of weight loss (0% in magnesium vs 29% in controls) and protected the cerebral cortex from neuronal loss (the incidence of normal, mild, moderate, and severe neuronal loss was 48%, 29%, 5%, and 19% in magnesium and 30%, 5%, 25%, and 40% in controls, respectively) in the pretreatment group. In the posttreatment group magnesium caused increased neuronal damage compared with the controls., Conclusion: Magnesium sulfate has neuroprotective effects against hypoxia-ischemia. This effect is restricted to the pretreatment group in which magnesium sulfate is administered before the insult.

Objective: The aim of this study was to examine the production of 2 types of heat shock protein 70 in the organs of neonatal rats during an episode of mild hypoxic stress that was insufficient to produce histologic changes., Study Design: Seven-day-old rats were subjected to hypoxia (inspired gas of 8% oxygen and 92% nitrogen) at 33 C for 2 hours (n = 5), 3 hours (n = 5), and 4 hours (n = 5). Control rats (n = 5) inspired room air for 4 hours. The brains, hearts, and livers were removed after 4 hours of recovery. The levels of heat shock protein 70 and heat shock cognate protein 70 messenger ribonucleic acid were measured by Northern blot analysis. Arterial pH, Pao 2, PACO 2, and brain temperature were measured before, during, and at 4 hours of hypoxia in another 16 animals. Histologic examinations were carried out in these 16 animals 7 days after hypoxic stress., Results: PaO 2, PACO 2, and brain temperature decreased during the hypoxic stress and returned to prehypoxic values at recovery time. Arterial pH did not change. No histologic changes were observed in any areas of the brain. Heat shock cognate protein 70 messenger ribonucleic acid was normally expressed in the brain, heart, and liver and was further induced after hypoxia in the brain and the heart. There was, however, no additional increase of heat shock cognate protein 70 messenger ribonucleic acid in the liver. there were no increments of the stress-induced form of heat shock protein 70 messenger ribonucleic acid in these organs., Conclusion: Mild hypoxia selectively induced messenger ribonucleic acid of heat shock cognate protein 70, which may play an important role in protecting the brain and the heart against stress.