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Frailty is associated with mortality in maintenance dialysis patients. For incident dialysis patients, we used the clinical frailty scale (CFS) to investigate frailty as related to mortality or hospitalization within 2 years. We retrospectively reviewed the medical records of patients initiating hemodialysis or peritoneal dialysis during 2016-2018. Based on those records, two dialysis nurses independently used a 9-point CFS (1 = "Very fit" to 9 = "Terminally ill") to assess each patient's frailty at dialysis initiation. Patients with a mean CFS value of 5 or higher were classified into the frail group. The 2-year survival rates or hospitalization-free rates after the initiation of dialysis were compared between the frail (mean CFS score ≥ 5) and non-frail (mean CFS score < 5) groups. The analysis included 155 incident dialysis patients with mean age of 66.7 ± 14.1 (71% male). Frailty was inferred for 39 (25%) patients at dialysis initiation. Kaplan-Meier analyses showed that the survival rate and hospitalization-free rate within 2 years were significantly lower in the frail group than in the non-frail group (p < 0.01). Cox proportional hazards regression analyses revealed the CFS score as associated with the occurrence of a composite outcome, independently of age (hazard ratio 1.34, 95% confidence interval 1.04-1.72). Frailty assessment based on clinical judgment using CFS might predict adverse outcomes in dialysis-initiated patients., (© 2022. The Author(s).)

Background: Dysphagia is a growing health problem in aging societies. An observational cohort study targeting community-dwelling populations revealed that 16% of elderly subjects present with dysphagia. There is a need in elderly communities for systematic dysphagia assessment., Objective: This study aimed to verify whether laryngeal elevation in the pharyngeal phase could be measured from the body surface using thin and flexible stretch sensors., Methods: Thirty-two elderly subjects (17 males, 15 females; mean age ± SD: 89.2 ± 6.2 years) with suspected dysphagia underwent a swallowing contrast examination in which seven stretch sensors were attached to the front of the neck. The elongation of the sensors was measured and compared to the laryngeal elevation time values obtained using videofluorography. The sensor signal detected the laryngeal elevation start time, conclusion of the descent of the larynx, and the laryngeal elevation time. The respective laryngeal elevation times obtained using videofluorography and using the sensor were compared using the Bland-Altman method., Results: The laryngeal elevation time was 1.34 ± 0.46 s with the stretch sensor and 1.49 ± 0.56 s with videofluorography. There was a significant positive correlation between the duration obtained by both methods (r = .69, P < .0001). A negative additional significant bias of -0.15 s (95% confidence interval -0.30 to -0.03, P = .046) was noted in the laryngeal elevation time from the videofluorography measurement., Conclusion: Laryngeal elevation time can be measured non-invasively from the neck surface using stretch sensors., (© 2020 John Wiley & Sons Ltd.)

Purpose: Children with cerebral palsy may face difficulties using handheld pointing devices, due to involuntary muscle movements. This study aimed at describing the idea of the new wearable sensor switch and assessing its feasibility as an access solution in a case of mixed-type cerebral palsy. Methods: The study participant was a 17-year-old male with mixed-type cerebral palsy characterized by chorea-athetotic movements and bilateral spasticity with gross motor function classification system level V. He exhibited sudden and irregular involuntary upper limb movements when sitting. Because spastic finger movements limited his ability to use a handheld mouse, he used a trackball near his neck as a pointing device (previous input method). The wearable switch system using a stretchable strain sensor was introduced; the sensor was attached to a groove worn on the dorsal regions of the right hand crossing the proximal interphalangeal and metacarpophalangeal joints of the middle finger (new input method). The switch turned on when the subject flexed his middle finger. Results: The user successfully turned the switch on and typed almost the same numbers of characters per trial compared with the previous input method. The speed of his head movements during typing reduced ( p  < .01), and his sitting posture was nearly upright during computer operation ( p  < .01). No involuntary movement, requiring physical assistance, was observed when using the wearable switch. Conclusion: The new switch system can be a new option for people with difficulty using standard handheld input devices due to paralysis and involuntary muscle movements.Implications for rehabilitationCerebral palsy is a major cause of motor dysfunction and spasticity and dyskinesia in the fingers and upper limbs may prevent children with cerebral palsy from using handheld input devices.Wearable devices may be useful for children with cerebral palsy who have limited access to handheld pointing devices.We developed a new wearable switch to control devices using a flexible stretchable sensor.The wearable switch contributed to the improvement of sitting posture and reduction of neck burden during the typing task at the speed equivalent to that using the previous method in a child with mixed type of cerebral palsy exhibiting choreoathetotic movements and bilateral spasticity.

Beta-propeller protein-associated neurodegeneration (BPAN) is one of the neurodegenerative disorders characterized by iron deposition in the brain and is the only known disease in humans to be caused by an aberration in autophagocytosis. Here, we present the case of a 42-year-old woman with BPAN identified by the WDR45 mutation. From early childhood, she was recognized as having global developmental delay, and she frequently sucked her hand, which was considered to be a stereotypical movement. She had a febrile convulsion at 6 months of age but there was no history of epilepsy. The delay in language development was more severe than the delay in motor development; she was able to dress herself, walk unaided, and follow simple instructions until adolescence. After the age of 20, her movement ability rapidly declined. By the time she was 42 years old, she was bedridden and unable to communicate. Brain magnetic resonance imaging (MRI) at 21 years revealed no abnormality except non-specific cerebral atrophy. However, MRI at 39 years revealed abnormalities in the globus pallidus and substantia nigra, with neurodegeneration and iron accumulation in the brain. Genetic analysis for WDR45 revealed that she had a splice site mutation (NM&#95;007075.3: c.830 + 2 T > C) which was previously reported, and a diagnosis of BPAN was confirmed. For specific therapies to be developed for BPAN in the future, it is necessary to establish early diagnosis, including genetic analysis.

Objective: We clarified the asymptomatic deficiency of serum free-carnitine in the severe motor and intellectual disabilities (SMIDs) fed with enteral carnitine-deficient formulas for years, and investigated the adequate method to supply enteral L-carnitine and maintain its normal levels., Methods: In 45 SMIDs who has been fed with carnitine-deficient formulas and/or receiving valproate, the serum free-carnitine levels were examined. To the carnitine deficient cases we introduced L-carnitine and/or a carnitine-supplemented formula to normalize and maintain the serum free-carnitine levels., Results: Thirty-one out of 34 cases (91.2%) fed with carnitine-deficient formulas for years had serum free-carnitine deficiency. Supplement of 15 - 30 mg/kg/day L-carnitine was effective to normalize the serum free-carnitine levels within 3 months. After successful supplementation, smaller dosage of L-carnitine (100 or 300 mg/day) was enough to maintain the normal levels. Replacement of the carnitine-deficient formulas to carnitine-supplemented ones was also useful to normalize the serum free-carnitine levels., Conclusions: Smaller dosage of L-carnitine or a carnitine-supplemented formula is sufficient to normalize and maintain the serum free-carnitine levels in SMIDs fed with carnitine-deficient formulas for years.

The aim of this present study was to establish a new in vitro assay, double artificial membrane permeation assay (DAMPA), to evaluate the human intestinal permeability of drugs. A double artificial membrane with an intracellular compartment was constructed in side-by-side chambers by sandwiching a filter containing buffer solution with impregnated lipophilic filters with dodecane containing 2w/v% phosphatidylcholine. Permeation data of ionic compounds clearly indicated that not only the pH value of the apical solution but also that of the intracellular compartment affected the permeability across the double artificial membrane. DAMPA was performed with 20 compounds at physiological pH (apical; 6.5, intracellular and basal; 7.4). Paracellular and transcellular permeabilities of compounds in human epithelium were estimated based on the characteristics of the paracellular pathway using physicochemical properties of compounds with the Renkin function and the area factor i.e. the difference in the effective surface area between human epithelium and the double artificial membrane, respectively. The human intestinal permeability of each compound was predicted by the sum of estimated transcellular and paracellular permeabilities. Predicted human intestinal permeability was significantly correlated with the fraction of absorbed dose in humans, indicating that DAMPA has the potential to predict oral absorption of drugs in humans., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objective: Clinical severity of alternating hemiplegia of childhood (AHC) is extremely variable. To investigate genotype-phenotype correlations in AHC, we analyzed the clinical information and ATP1A3 mutations in patients with AHC., Methods: Thirty-five Japanese patients who were clinically diagnosed with AHC participated in this study. ATP1A3 mutations were analyzed using Sanger sequencing. Detailed clinical information was collected from family members of patients with AHC and clinicians responsible for their care., Results: Gene analysis revealed 33 patients with de novo heterozygous missense mutations of ATP1A3: Glu815Lys in 12 cases (36%), Asp801Asn in 10 cases (30%), and other missense mutations in 11 cases. Clinical information was compared among the Glu815Lys, Asp801Asn, and other mutation groups. Statistical analysis revealed significant differences in the history of neonatal onset, gross motor level, status epilepticus, and respiratory paralysis in the Glu815Lys group compared with the other groups. In addition, 8 patients who did not receive flunarizine had severe motor deteriorations., Conclusions: The Glu815Lys genotype appears to be associated with the most severe AHC phenotype. Although AHC is not generally seen as a progressive disorder, it should be considered a disorder that deteriorates abruptly or in a stepwise fashion, particularly in patients with the Glu815Lys mutation.

Background: In order to clarify the factors causing hyperammonemia and to predict occurrences during treatment with valproic acid (VPA), we investigated the effect of the genetic polymorphism of carbamoyl-phosphate synthase 1 (CPS14217C>A) on susceptibility of hyperammonemia, together with the effect of coadministration of other anticonvulsants., Methods: Seventy-nine patients with epilepsy were enrolled, and five of them had hyperammonemia. Univariate and multivariate logistic regression analyses were performed., Results: The aspartate aminotransferase level in the patients with hyperammonemia was significantly higher than that in those without hyperammonemia. The risk of hyperammonemia was significantly influenced by the number of anticonvulsants concomitantly administered with VPA. Also, the distribution of the CPS14217C>A genotype differed depending on whether the patients had hyperammonemia or not. No significant effects of CPS14217 genotypes and the number of anticonvulsants coadministered with VPA on the serum concentrations of VPA were observed. The multivariate logistic regression analysis showed that the concomitant administration of two or more anticonvulsants with VPA and the heterozygous or homozygous carrier state of the A allele of the CPS14217C>A polymorphism were independent risk factors for developing hyperammonemia., Conclusions: These findings suggested that in epileptic patients undergoing VPA therapy, CPS14217A polymorphism and the number of coadministered anticonvulsants would be considered as risk factors for hyperammonemia, even if the serum VPA concentrations were controlled., (© 2010 The Authors. Pediatrics International © 2010 Japan Pediatric Society.)

We investigate whether the functional near-infrared spectroscopic (fNIRS) signal includes a signal from the changing skin blood flow. During a locomotor task on a treadmill, changes in the hemodynamic response in the front-parietal area of healthy human subjects are simultaneously recorded using an fNIRS imaging system and a laser Doppler tissue blood flow meter. Independent component analysis (ICA) for fNIRS signals is performed. The skin blood flow changes during locomotor tasks on a treadmill. The activated spatial distribution of one of the components separated by ICA reveals an overall increase in fNIRS channels. To evaluate the uniformity of the activated spatial distribution, we define a new statistical value-the coefficient of spatial uniformity (CSU). The CSU value is a highly discriminating value (e.g., 2.82) compared with values of other components (e.g., 1.41, 1.10, 0.96, 0.61, and 0.58). In addition, the independent component signal corresponding to the activated spatial distribution is similar to changes in skin blood flow measured with the laser Doppler tissue blood flow meter. The coefficient of correlation indicates strong correlation. Localized activation areas around the premotor and medial somatosensory cortices are shown more clearly by eliminating the extracted component.

Ornithine transcarbamylase (OTC) deficiency is the most common inborn error of the urea cycle. Although a combination of molecular methods have been used including DNA sequencing of all 10 exons and exon-intron boundaries of OTC gene, only approximately 80% of patients with OTC deficiency are found to have mutations. We report two known and three novel mutations of the OTC gene in five Japanese patients including two neonatal-onset, one late-onset, and two symptomatic female patients. Known nonsense mutations (c.578G>A and c.421C>T) were detected in a neonatal-onset male and a symptomatic female patient, respectively. Mutation analysis revealed two novel mutations including one splice site mutation (c.386+1G>C) in a symptomatic female patient and one missense mutation (c.515T>A) in a late-onset male patient. In the remaining case, which was a neonatal-onset male patient, no mutation was disclosed by direct sequencing of all 10 exons and their flanking intron sequences. Therefore, OTC mRNA in the liver was analyzed by RT-PCR, and remarkably, a 135-nt insertion was detected between exons 5 and 6. Genomic DNA analysis of intron sequences revealed a single nucleotide change at 265 bp downstream from the 3' end of exon 5, which created the novel splice acceptor site. Thereby, a 135-nt exon was created from the central part of an intron sequence. This is the first report of mutation deep in the intronic sequence in the OTC gene. Molecular analysis using genomic DNA and mRNA will increase the mutation detection ratio in the OTC gene.

Sotos syndrome is caused by the haploinsufficiency of the NSD1 gene located in 5q35. More than 70% of the Japanese cases carry microdeletions encompassing of this gene, while point mutations are common in Caucasians. Only 15 familial cases of Sotos syndrome have been reported and all cases shown to have not microdeletions but point mutations. We identified the first Japanese familial case (mother and 3 children). They carry the same mutation at splice donor site of intron 13 (IVS13+1G>A), which results in the in-frame skipping of exon 13. This is also the first familial case caused by the mutation of the splice donor site. Each member of this family showed variable phenotypes and mental development. The present report will contribute to further understanding of genotype-phenotype correlation in Sotos syndrome.

Hypoxia-ischemia (HI) causes injury to oligodendrocytes (OLs), cells which create the myelin sheath in the developing brain. OLs pass successively through progenitor and immature stages during differentiation into mature OLs. Only the OLs in the progenitors stage can express the platelet-derived growth factor-a receptor (PDGF-R(alpha)) so that its expression is one of the cellular markers of OL progenitors. Activation of PDGF-R(alpha) results in OL proliferation, but not OL differentiation. To study the response of OL progenitors after neonatal HI brain injury, we investigated the expression of PDGF-R(alpha) in a neonatal rat stroke model (combination of left common carotid artery ligation and exposure to 8% O2 for 2 h). In the injured cerebral cortex, PDGF-R(alpha) mRNA levels increased significantly (p<0.01) with a peak at 0.5 h after HI insult, and returned to baseline levels within 48 h post-injury. PDGF-R(alpha) protein levels increased significantly at 72-96 h (p<0.05) and then returned to basal levels. Immunohistochemistry showed clear staining of PDGF-R(alpha) only in the injured cerebral cortex at 72 h after HI insult. In contrast, no staining was observed in the cortex of sham-operated controls. These results indicate that the expression of PDGF-R(alpha) increases rapidly and transiently only in the injured cerebral cortex after HI insult and may play a protective role through modulating the glial differentiation under the condition of cellular damage in the developing brain.

Background: Periventricular cysts are not rare findings in neonates. However, they are sometimes associated with serious clinical complications, such as congenital viral infections and anomalies., Methods: We performed a retrospective follow-up study on newborns who had periventricular cysts on routine cranial ultrasound examination., Results: We followed 13 infants (three preterm) with periventricular cysts. Ten had single or multiple germinolysis cysts and the remaining three had choroid plexus cysts. All infants had various kinds of underlying complications, including congenital viral infection (two with cytomegalovirus and one with rubella),Sotos syndrome (n = 4), intrauterine growth retardation (n = 5), large-for-dates(n = 4), congenital heart disease (n = 1),myelomeningocele (n = 1) and other minor anomalies. All cases of germinolysis except for one developed a neurodevelopmental abnormality and/or delay. In contrast,all three cases with choroid plexus cysts appeared to develop well,despite the underlying complications., Conclusions: Germinolysis cysts seem to be associated with systemic diseases and should be treated as a high-risk sign for impaired neurological development.

We diagnosed Pallister-Mosaic syndrome (PMS) in a 4-month-old female infant. In addition to the presence of non-specific anomalies, involving anorectal, finger and ear anomalies, characteristic cranio-facial features and irregular skin lesions that appeared after age 2 months suggested the possibility of genetic mosaicism, PMS in particular. Fluorescence in situ hybridization technique revealed an extra copy of chromosome 12p; i (12p) in 30% of cultured skin fibroblasts. When focal skin lesions accompany neurodevelopmental disabilities in early infancy, genetic analysis for mosaicism should be considered for differential diagnosis. Significantly, we describe several phenotypic features and neuroimaging findings of the PMS in the present case, which have not been described in previous reports. The neuroimaging abnormalities we encountered, such as polymicrogyria, speculating congenital brain anomaly, may explain the severe motor and intellectual disabilities of PMS.

Spinal muscular atrophy (SMA) is one of the most common autosomal recessive disorders characterized by degeneration of anterior horn cells in the spinal cord, and leads to progressive muscular weakness and atrophy. At least three SMA-related genes have been identified: SMN1, NAIP and p44t. We analyzed these genes in 32 SMA patients and found that the SMN1 gene was deleted in 30 of 32 patients (94 %), irrespective of clinical type. The NAIP gene was deleted in 6 patients and its deletion rate was higher in type I patients than that in type U or V. Further, in type I patients lacking the NAIP gene, deterioration in their respiratory function is more rapid than in those type I patients retaining the NAIP gene. Since complete p44t deletion was observed in only 3 patients, the correlation between the p44t deletion and severity of SMA remained ambiguous. We concluded that the NAIP deletion was closely related to the clinical severity of SMA and was a predictive marker of SMA prognosis, while the SMN1 deletion did not correlate with clinical severity.

Flash visual evoked potential (Flash-VEP) is easily recorded in preterm infants. However, its clinical application has not been established due to its great variability in response. Our longitudinal studies on the two components of the N1 wave facilitated peak definition and established normal ranges that are clinically valuable. The N1a (early component of the N1) peak latency decreases at about 4.6 msec/week between 30 and 40 weeks postmenstrual age. A flash-VEP study in the preterm period enables us to observe the neuronal development in the human visual system that normally proceeds in utero. Flash-VEP analyses on preterm infants demonstrated that the decrease in the N1a peak latency reflects the progress of myelination in the visual pathway according to the developmental program irrespective of preterm birth. The developmental changes of the N1 wave configuration reflect the maturation of the neuronal networks in the visual cortex, which is accelerated by extrauterine visual experience. Using improved methodology and peak denomination that we proposed, flash-VEP can be applied to preterm infants safely, and should provide us with neuro-developmental information of the human cerebrum.

Caspases are believed to play a key role in the delayed neuronal cell death observed in the rat brain after hypoxic-ischemic (HI) insult. Caspase inhibitors have been developed as antiapoptotic agents. Hippocampal damage after HI insult is strongly related to tissue temperature, and systemic hypothermia has been introduced clinically for brain protection. In this study, we examined the effects of a caspase inhibitor and systemic hypothermia on neuronal protection in the developing rat brain. Postnatal d 7 rat pups were subjected to the Rice model of hypoxia for 1 h. Systemic hypothermia was induced with a water bath at 29 degrees C. Before HI insult, a pan-caspase inhibitor, boc-aspartyl-(OMe)-fluoromethyl-ketone (BAF), was injected into the cerebral ventricle. The ipsilateral hippocampus was subjected to caspase assays and histologic assessment. The HI group at 37 degrees C (HI-37 degrees C) showed a peak of caspase-3 activity 16 h after insult. This activity was significantly reduced in the presence of BAF or hypothermia (HI-29 degrees C group, p < 0.05) or by the combination of HI-29 degrees C + BAF (p < 0.01 versus HI-37 degrees C). The number of neuronal cells in the ipsilateral hippocampal CA1 region in the HI-37 degrees C group was significantly decreased (62.9% versus control). The number of neuronal cells was maintained in the HI-37 degrees C + BAF group (82.7%), the HI-29 degrees C group (78.7%), and the combination group (95.2%) (p < 0.05 versus HI-37 degrees C). A combination of systemic hypothermia and BAF produced a strong protective effect against neuronal damage in the developing rat brain, along with a reduction in caspase-3 activity.

Heat shock proteins (HSPs) are immediately expressed in neuronal and glial cells under various stressful conditions and play a protective role through molecular chaperones. We investigated the characteristics of the induction manner of heme oxygenase-1 (HO-1) and HSP70 in rat C6 glioblastoma cells. In heat treatment (42 degrees C for 30 min), C6 cells expressed high level of HO-1 and HSP70 mRNAs within 30-60 min, and their proteins at 3 hrs. Heat-induced expressions of HSPs mRNAs were completely inhibited with actinomycin D, suggesting the transcriptional regulation. Oxygen-glucose deprivation (OGD), cystine-free (inhibition of synthesis of glutathione), cyto-toxic (ethanol, sodium butyrate) treatments resulted in different expression manners between HO-1 and HSP70, which suggested that HO-1 and HSP70 play different protective roles against a variety kind of stressful conditions in glial cells. C6 cells can differentiate toward both astrocyte and oligodendrocyte directions. Treatment with dibutyryl cyclic AMP (cAMP) induces expression of glial fibrillary acidic protein (GFAP), a marker of astrocytes, and treatment with retinoic acid (RA) induces expression of myelin proteolipid protein (PLP), a marker of oligodendrocytes, respectively. Heat treatment before the initiation of differentiation by RA reduced the RA-induced expression of PLP mRNA profoundly, but not in GFAP mRNA level induced by cAMP. Heat treatment after the initiation of differentiation by cAMP or RA accelerated the expression of GFAP or PLP mRNAs. Astroglial differentiation by cAMP reduced the heat-induced expressions of HSPs mRNAs, but no change with RA pre-treatment. These results suggested that HSPs may modulate the glial differentiation in the developing brain. On the contrary, glial differentiation may give influence on the stress-induced HSPs expression. The timing of stressful damages, resulting in the expression of HSPs, on the developing brain is critically important for the pathogenesis of glial lesion. In the heat-treated C6 cells, the expression of platelet-derived growth factor (PDGF) receptor-alpha mRNA was significantly decreased. HSPs may have ability to induce the glial differentiation in part through down-regulation of the PDGF pathway.

Background: It is essential to evaluate the renal function for the management of high risk neonates. Color Doppler ultrasound technique can provide a useful information to evaluate the neonatal renal artery blood flow velocities. This study was performed to obtain the normative data of renal blood flow velocities in preterm and fullterm neonates and to compare the renal blood flow velocities with the aortic blood flow velocities. The normal volumetric state of fluid balance and renal function are essential in the management of the sick neonates., Methods: The renal peak systolic blood flow velocity (renal PSFV), the renal mean blood flow velocity (renal MFV), the renal end diastolic blood flow velocity (renal EDFV) and the renal resistance index (renal RI) and also the cardiac output, the aortic peak systolic blood flow velocity (aortic PSFV) and the aortic mean blood flow velocity (aortic MFV) were serially recorded from one to 7 days after birth in 16 preterm and one to 5 days in 23 normal fullterm neonates., Results: The renal PSFV was significantly increased with the postnatal age, and the renal MFV was significantly increased between three and five days of age in the preterm neonates. In the fullterm neonates there was no significant change of the renal PSFV between one and five days of age, but the renal MFV was significantly increased between three and five days of age. There were no statistically significant differences of the renal PSFV, the renal MFV, the renal EDFV and the renal RI between the preterm and the fullterm neonates in the first five days after birth. The cardiac output in the preterm neonates was higher than that in the fullterm neonates: on day 3; 313 +/- 59 vs. 254 +/- 40 ml/kg/min. (p < 0.001) and on day 5; 357 +/- 95 vs. 280 +/- 35 ml/kg/min. (p < 0.01). The renal blood flow velocities were not significantly correlated with the aortic blood flow velocities in the normal fullterm neonates, whereas, in the preterm neonates the renal PSFV correlated with the cardiac output (r = 0.35, p < 0.01), the aortic PSFV (r = 0.45, p < 0.001) and the aortic MFV (r = 0.39, p < 0.01), and the renal MFV also correlated with the aortic PSFV (r = 0.30, p < 0.05) and the aortic MFV (r = 0.32, p < 0.05)., Conclusions: The increased renal flow velocities with the postnatal age in the preterm infants might depend on the increased cardiac output.

In order to clarify the difficulties which handicapped people experienced in the Hanshin-Awaji earthquake disaster, 678 families with handicapped children were studied 1.5 months after the earthquake. All the students who were enrolled in this study were going to some type of school or training classes for handicapped children in January 1995. The study was completed between 1 and 10 March. The questionnaires which were designed for this study consisted of three parts: the difficulties which the families were faced with, their requirements for social and medical services and the symptoms and reactions of their children after the earthquake. A total of 466 answers were obtained from their parents. The kinds of difficulties differed between the families of mentally retarded children and those of the physically handicapped. While many parents with physically handicapped children desired better medical information or materials, many parents with mentally retarded children wanted better care services for their children. Physical and psychological effects of the earthquake were only temporary in most cases. However, some of the handicapped children were still suffering from such reactions as of 10 March. From the results of our study it became evident that a systematic relief program for these handicapped people should be established. The relief program should include the distribution of information regarding medical and social services. Psychological reactions such as panic, excitement and suppression of mental activity were still observed in some handicapped children on 10 March. Further observation will therefore be necessary.

More than 5500 people were killed in the Hanshin-Awaji earthquake disaster. Most of the neonatal intensive care units (NICU) could not offer proper services as a result of the earthquake and the Hyogo Emergency System for Neonates could not function. No one imagined that such a great disaster would occur in their home town, Kobe; however, the devastation of the quake has raised the opportunity to examine the operation of neonatal medicine in an emergency. We sent out questionnaires to perinatologists in the damaged area 2-3 months after the quake. Some hospitals in the demolished area suffered such serious damage that they could not continue work as NICU, and some in the surrounding area had problems in a number of areas, including staff, room capacity and transport methods. Under these difficult circumstances the greatest problem encountered by staff was maintaining fundamental care in NICU, that is to say keeping temperature, formula (nutrition) and prevention of infection techniques operating. In this report we discuss the effects on neonatal medicine of the earthquake in Kobe. Medical staff struggled patiently and made great efforts to recover and maintain the functioning of NICU. We believe this information will be useful for neonatology staff in other areas which may experience an earthquake in the future.

A 2-month-old girl had focal tonic convulsions. Brain CT showed no abnormalities on admission. Three weeks later she got a severe epileptic status. T 1-weighted MRI demonstrated low intensity areas in the right occipital and left frontal regions, and enhanced CT demonstrated low density areas in the same region. An increase of CMV antibody titer in serum suspected CMV infection associated with brain infarction. Then gamma-globulin was given in addition to PB, CBZ, and VPA, resulting in suppression of seizures. At 9 months of age, she had right tonic hemiconvulsions. An increase of CMV IgM antibody titer showed reactivation of CMV infection. Acyclovir and gamma-globulin were given, and her seizures were controlled. However, she showed progressive motor disability with spastic muscle tonus. CT and MRI showed a severe progressive atrophy of the cerebrum and brain stem.

A human neuroblastoma cell line, GOTO, extends neurite-like processes upon withdrawal of serum from culture medium. This morphological change was accompanied by a 5-fold increase in the neurofilament (NF)-L and a 10-fold increase in the NF-M mRNA levels after 24 h. The addition of a protein kinase inhibitor, H-7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride) also induced the extension of neurite-like processes; however, it did not increase the NF mRNA levels. Thrombin inhibited the extension of neurite-like processes in serum-free culture without affecting the increase in the NF mRNA levels. There was no difference in the number of cells progressing through the S phase between serum-containing and -free cultures at 24 h. This indicates that the increase in the NF mRNA levels upon withdrawal of serum was not preceded by the growth arrest. Treatment with actinomycin D and cycloheximide inhibited the increase in the NF mRNA levels. The half life of the NF gene transcripts was prolonged in the serum-free condition. These results indicate that the serum depletion-induced increase in the NF-L and -M mRNA levels was regulated by both transcriptional and post-transcriptional mechanisms, and the increase in the expression of the NF genes was not simply mediated by growth arrest but controlled by unknown regulator proteins.

Accumulating evidence has demonstrated that protein kinase C (PKC) and protease nexin-1 (PN-1) may be involved in neuronal differentiation including migration, neurite outgrowth, target recognition, and synaptogenesis. We investigated the potential roles of PKC and PN-1 in neurite outgrowth of human neuroblastoma cell line, GOTO. Upon withdrawal of serum GOTO cells extended neurite processes within 3 h and formed fine network of neurites after 24 h. This morphological change was completely inhibited by thrombin and phorbol-12-myristate-13-acetate (PMA). Withdrawal of serum increased the neurofilament (NF)-L and -M mRNA levels and thrombin did not inhibit the effect of withdrawal of serum. A potent PKC inhibitor, H-7 induced neurite outgrowth in the presence of serum, however, it did not increase the NF mRNA levels. Actinomycin D and cycloheximide did not inhibit the initial neurite outgrowth induced by withdrawal of serum, while these inhibited the increase in the NF mRNA levels. Thrombin retracted the serum depletion-induced neurites but did not retract the neurites induced by H-7. The specific activity and subcellular localization of PKC did not differ between GOTO cells cultured in serum-containing and -free media for 12 h. The serine protease inhibitory activity was undetectable in the serum-free conditioned medium of GOTO cells but the PN-1 mRNA was clearly detected by Northern blot analysis to a less extent than glial cells. Withdrawal of serum or treatment with H-7 did not increase the PN-1 mRNA level in GOTO cells, but thrombin increased its level about 7 folds in serum-free condition. These results indicate that the initial neurite outgrowth requires neither new RNA nor protein synthesis, and that PKC negatively regulates neurite outgrowth and thrombin blocks neurite outgrowth through PKC-dependent pathways.

We have studied the expression of smg p25A and synaptophysin in cultured hippocampal neurons isolated from 5-day-old rat brain by an immunocytochemical technique. In a dispersed cell culture seeded on astrocyte monolayers, well-branching neurite proliferation was observed along with age in culture. The synaptophysin immunoreactivity was present in the neuronal cell bodies and neurites at 1 and 5 days in vitro (DIV) and was eventually localized to discrete areas along neurites at 15 DIV while the immunoreactivity in cell bodies became less prominent. On the other hand, the smg p25A immunoreactivity was observed in the neuronal cell bodies and neurites at 1 through 15 DIV. The immunoreactivity for smg p25A or synaptophysin was not observed in astrocytes and this finding was confirmed by an immunoblot analysis. These results indicate that smg p25A as well as synaptophysin is present exclusively in neurons and suggest that these two synapse-associated proteins have different sites of function and different kinetics of synthesis, transport, and/or turnover in cultured hippocampal neurons.

Effects of dexamethasone (DEX) on the relative abundance of myelin basic protein (MBP), proteolipid protein (PLP) and glial fibrillary acidic protein (GFAP) mRNAs in the developing rat brain were examined. After DEX (1.0 mg/kg body weight) or saline was administered intraperitoneally to 3-day-old rats for 7 consecutive days, wet weight, DNA content and the relative abundance of the glia-specific mRNAs in cerebrum and cerebellum were analyzed at postnatal days (P) 10, 20 and 30. DEX decreased both wet weight and DNA content in cerebellum more profoundly than in cerebrum. The appearance of MBP, PLP and GFAP mRNAs in cerebellum preceded that in cerebrum in the control group. In cerebrum, the relative abundance of MBP and PLP mRNAs was significantly less in the DEX group than that in the control group at P20 and P30. The relative abundance of the GFAP mRNA was significantly less in the DEX group than in the control group at P10 and P20, but there was no significant difference at P30. In cerebellum, a significant decrease in the abundance of MBP, PLP and GFAP mRNAs in the DEX group was observed only at P10 but not at P20 and P30. Our findings indicate that DEX suppresses expression of genes related to glial functions, especially myelination when administered in the early postnatal period.

We have examined the effect of dexamethasone on the metabolism of pulmonary surfactant in normal and hyperoxia-treated rats. The relative abundance of the surfactant-specific apoprotein A (SP-A) mRNA in lung tissues and the contents of disaturated phosphatidylcholine (DSPC) and SP-A were measured in bronchoalveolar lavage fluids and in lung tissues in 4-wk-old rats exposed to room air or greater than 90% oxygen for 7 d with or without simultaneous treatment with dexamethasone (0.5 mg/kg body wt for 7 d). The relative abundance of the SP-A mRNA was marginally increased by hyperoxia (1.3-fold over controls). Dexamethasone increased the relative abundance of the SP-A mRNA to a level comparable to that with hyperoxia treatment (1.5-fold over controls). In lavage fluids, the contents of DSPC and SP-A were increased by 4- and 6-fold over controls by hyperoxia, respectively, but they were increased only by 2-fold by dexamethasone. In lung tissues, the contents of DSPC and SP-A were increased by 3- and 2-fold over controls by hyperoxia, respectively. These values in lung tissues in the air-exposed rats were not significantly increased by dexamethasone. In hyperoxia-treated rats, dexamethasone did not significantly affect the relative abundance of the SP-A mRNA level and the contents of DSPC and SP-A in lavage fluids and lung tissues. These results indicate that mechanisms other than increased synthesis of SP-A are involved in hyperoxia-induced SP-A accumulation and that dexamethasone does not affect the abnormal accumulation of pulmonary surfactant induced by hyperoxia.

Changes in expression and localization of smg p25A, a ras p21-like small GTP-binding protein, in developing rat brain were analyzed in comparison with those of synaptophysin, a well-known synaptic vesicle-specific protein. The smg 25A mRNA was detected in whole brain of rat fetus at 14 days of gestation and its level was increased along with the age and reached the maximum level at postnatal day (P) 20. In postnatal cerebellum, the smg25A mRNA level was also increased age-dependently and the maximum level was observed at P30. Immunoblot analysis with an anti-smg p25A monoclonal antibody (MAb SG-11-7) and an anti-synaptophysin monoclonal antibody (SY 38) showed that expression of both smg p25A and synaptophysin was increased age-dependently in postnatal rat cerebellum. By immunofluorescent cytochemical study with the anti-smg p25A antibody, bright fluorescence was observed in the molecular layer of cerebellum and it was increased in accordance with the cerebellar development. In early postnatal cerebellum, the perikarya of Purkinje cells and the white matter were brightly stained with the antibody, but the fluorescence of these portions was faint in adult cerebellum. The anti-synaptophysin monoclonal antibody also stained the molecular layer of cerebellum but the perikarya of Purkinje cells and the white matter had only a weak immunoreactivity with the antibody irrespective of the age. These results indicate that smg p25A is predominantly present in the nerve terminals and that its amount is increased along with the development of postnatal rat cerebellum. Our results also suggest that smg p25A and synaptophysin have the different kinetics of synthesis, transport, and/or turnover in developing rat cerebellum.