Showing papers in "The Kobe journal of the medical sciences in 2019"

Antiviral Activity of Cananga odorata Against Hepatitis B Virus.

Abstract: Chronic hepatitis B virus (HBV) infection can lead to liver cirrhosis and hepatocellular carcinoma. Current therapeutic drugs for chronic hepatitis B using pegylated interferons and nucleos(t)ide analogs have limited efficacy. Therefore, the development of novel and safe antivirals is required. Natural products including medicinal plants produce complex and structurally diverse compounds, some of which offer suitable targets for antiviral screening studies. In the present study, we screened various crude extracts from Indonesian plants for anti-HBV activity by determining their effects on the production of extracellular HBV DNA in Hep38.7-Tet cells and HBV entry onto a HBV-susceptible cell line, HepG2-NTCP, with the following results: (1) In Hep38.7-Tet cells, Cananga odorata exhibited the highest anti-HBV activity with a 50% inhibitory concentration (IC50) of 56.5 µg/ml and 50% cytotoxic concentration (CC50) of 540.2 µg/ml (Selectivity Index: 9.6). (2) The treatment of HepG2-NTCP cells with Cassia fistula, C. odorata, and Melastoma malabathricum at concentrations of 100 µg/ml lowered the levels of HBsAg production to 51.2%, 58.0%, and 40.1%, respectively, compared to untreated controls, and IC50 and CC50 values of C. odorata were 142.9 µg/ml and >400 µg/ml. In conclusion, the C. odorata extract could be a good candidate for the development of anti-HBV drugs.

Regulation of Small Intestinal Epithelial Homeostasis by Tsc2-mTORC1 Signaling

Abstract: Mammalian target of rapamycin complex 1 (mTORC1), a protein complex containing the serine/threonine kinase mTOR, integrates various growth stimulating signals. mTORC1 is expressed in intestinal epithelial cells (IECs), whereas the physiological roles of this protein complex in homeostasis of IECs remain virtually unknown. We here generated mice, in which tuberous sclerosis complex 2 (Tsc2), a negative regulator of mTORC1, was specifically ablated in IECs (Tsc2 CKO mice). Ablation of Tsc2 enhanced the phosphorylation of mTORC1 downstream molecules such as ribosomal S6 protein and 4E-BP1 in IECs. Tsc2 CKO mice manifested the enhanced proliferative activity of IECs in intestinal crypts as well as the promoted migration of these cells along the crypt-villus axis. The mutant mice also manifested the increased apoptotic rate of IECs as well as the increased ectopic Paneth cells, which are one of the major differentiated IECs. In addition, in vitro study showed that ablation of Tsc2 promoted the development of intestinal organoids without epidermal growth factor, while mTORC1 inhibitor, rapamycin, diminished this phenotype. Our results thus suggest that Tsc2-mTORC1 signaling regulates the proliferation, migration, and positioning of IECs, and thereby contributes to the proper regulation of intestinal homeostasis.

Role of Hypoxia Inducible Factor-1 Alpha (HIF-1α) in Cytoglobin Expression and Fibroblast Proliferation of Keloids.

Abstract: Background Keloids are characterized by an overabundance of collagen deposition due to elevated activity and proliferation of fibroblasts, which lead to hypoxic conditions. Adaptation to these conditions is regulated by the transcription factor hypoxia inducible factor-1α (HIF-1α). Cytoglobin (Cygb), a reactive oxygen species scavenger, is a target gene of HIF-1α. In our previous study, we showed that Cygb expression in keloid tissue was correlated with HIF-1α expression. However, whether HIF-1α regulates Cygb expression and the proliferation of keloid fibroblasts remained unclear. Therefore, this study aimed to determine the role of HIF-1α in Cygb expression and fibroblast proliferation of keloids. Methods This was an in vitro study using a primary culture of keloid fibroblasts in which ibuprofen was used to inhibit HIF-1α expression. The expression of HIF-1α and Cygb mRNA were analyzed using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) methods, and their protein levels were analyzed using an enzyme-linked immunosorbent assay (ELISA). Fibroblast proliferation was analyzed using a Trypan blue exclusion assay. Results Inhibition of HIF-1α by ibuprofen decreased Cygb mRNA expression but not in all the samples, followed by a decrease in the protein level of Cygb. There was a positive correlation between the HIF-1α protein and Cygb mRNA, probably due to the regulation of Cygb by HIF-1α at the mRNA level, but not the protein level. The proliferation of keloid fibroblasts was significantly decreased and positively correlated with the HIF-1α protein. Conclusion HIF-1α regulates Cygb expression and fibroblast proliferation in keloids.

GC/MS and LC/MS-based Tissue Metabolomic Analysis Detected Increased Levels of Antioxidant Metabolites in Colorectal Cancer.

Abstract: Late-stage colorectal cancer is resistant to current treatments. Understanding the biological processes responsible for the development and progression of colorectal cancer could aid the development of new diagnostic and treatment approaches. We used gas chromatography/mass spectrometry and liquid chromatography/mass spectrometry-based metabolomic analysis to measure metabolite levels in pairs of colorectal cancer tissue samples and samples of the adjacent macroscopically normal mucosal tissue from 10 colon cancer patients. Regarding nucleotide metabolomic intermediates, the colorectal cancer tissue contained lower levels of ribulose 5-phosphate and higher levels of xanthine, adenine, and hypoxanthine than the normal tissue. The levels of antioxidant metabolites, such as sulfur-containing amino acids, were also significantly higher in the colorectal cancer tissue. The level of tryptophan was decreased, and the levels of molecules downstream of the tryptophan pathway, such as kynurenine and quinolinic acid, which protect colorectal cancer against the host's immune system and function in de novo nicotinamide adenine dinucleotide synthesis, were increased in the colorectal cancer tissue. The colorectal cancer tissue samples also contained higher levels of lysophospholipids and fatty acids, especially stearic acid and polyunsaturated fatty acids, including arachidonic acid and docosahexaenoic acid. Thus, understanding these cancer-specific alterations could make it possible to detect colorectal cancer early and aid the development of additional treatments for the disease, leading to improvements in colorectal cancer patients' quality of life.

The Influence of Chronotype and Working Condition on Sleep Status and Health Related Quality of Life of Daytime Office Workers.

Abstract: Several companies in Japan introduced early working conditions (including recommendations on early morning work and prohibitions on nighttime overtime work) to decrease the number of long working hours at night. Nevertheless, individuals possess their own chronotype, i.e., their behavioral timing preference-be it morning or evening-that is associated with worker health. The purpose of this study was to investigate the influence of chronotype and working conditions on sleep and health related quality of life (HRQOL) using 126 daytime office workers who were classified as morning or evening type by their Morningness-Eveningness Questionnaire scores. We then compared morning and evening type workers' sleep variables (sleep onset/offset time and total sleep time), sleep quality (using the Japanese version of the Pittsburgh Sleep Quality Index), and HRQOL scores. Additionally, we compared the same sleep variables, sleep quality, and HRQOL scores of each chronotype category of worker under early and normal working conditions. As the results, evening type workers had late sleep onset/offset time, poor sleep quality, and low HRQOL (role-social component) compared to morning type workers. Furthermore, the evening type workers under early working conditions had earlier sleep onset/offset time and poorer sleep quality compared to those workers under normal working conditions. These results suggest that evening type workers in general have poor sleep and low HRQOL and those same workers under early working conditions, in particular, are associated with poor sleep quality. Therefore, in order to optimize worker health, we suggest that working conditions should be taken account of individual chronotypes.

Red LED Light Is Influenced by IL-6 to Promote the Migration Ability of Oral Squamous Cell Carcinoma Cell Line.

Abstract: Aim: The purpose of this study was to search for the influence of red LED light of oral squamous cell carcinoma cells (OSCC) Material & Methods: HSC-3 was irradiated with red LED light (630 nm · 3 J). Proliferative capacity was analyzed using WST-1. Transwell migration assay, real-time PCR, and ELISA method were also used to analyze migratory ability. Conclusions: HSC-3 cells irradiated with red LED light showed increased migration ability. Interestingly, we clarified that the expression of Interleukin-6 (IL-6), which promotes the migratory ability of cancer cells, is induced.

Lifestyle of Patients with Alcoholic Liver Disease and Factors Leading to Hospital Readmission: A Prospective Observational Study

Abstract: The objective of this study was to clarify the lifestyle characteristics of patients with alcoholic liver disease (ALD) who were readmitted to the hospital, and to identify the background factors associated with these characteristics. This was a prospective observational study. Over a period of 3 months following hospital discharge, we conducted structured interviews to investigate the following five lifestyle characteristics based on our previous research: dietary intake, alcohol consumption or abstinence, psycho-emotional status, regularity of life habits, adherence to treatment. We also collected data on background factors from medical records and questionnaires. The analysis was performed using conceptual cluster matrices, with participants divided into two groups (at-home recovery and readmission). Lifestyle, health status, and background factors were compared between the two groups. Of the 34 patients with ALD recruited, 21 completed the one-month follow-up and were included in the analysis-14 patients were in the at-home recovery group and 7 in the readmission group. The at-home group's lifestyle was characterized by controlled alcohol consumption, but with maintenance of regular life and eating habits and adherence to treatment. In contrast, irregular eating habits (p=0.006) and the development of irregular life habits or the discontinuation of treatment very quickly after hospital discharge characterized the readmission group's lifestyle. Experiences of loss were a lifestyle-related background factor that was associated with readmission (p=0.017). Based on these findings, supporting patients with ALD in maintaining regular eating habits and taking experiences of loss into consideration would be important in avoiding readmission over the short-term.

Mothers'Breastfeeding-Related Durations and Nursing Management During the Early Postpartum Period in a Mixed Hospital Ward with an Obstetrics Department: A Prospective Observational Study.

Abstract: Purpose This study aimed to quantify duration of in-hospital breastfeeding (BF)-related activities in mothers after childbirth in a mixed ward-type hospital with an obstetrics department in Japan (hereafter, mixed ward). Methods Twenty-two postpartum mothers (primiparous and multiparous) who had vaginal delivery in a mixed ward were instructed to wear radio beacons. A smartphone was placed in the BF room and detected the presence of a nearby beacon, 24 hours daily, measuring the number of hours a mother spent in the nursing room (defined as BF time [hereafter, BF-related duration]). BF-related duration included time spent for BF, diaper changes before and after BF, bottle feeding, and support from nurses. BF-related total hours during postpartum hospitalization were calculated. The effects of parity and each postpartum day number (postpartum days 1-4) on Daily BF-related duration were determined. Results Percentage of total postpartum hospitalization time spent for BF-related duration was 21.6% and 19.9% for primiparous and multiparous mothers, respectively, with no significant difference between groups. BF-related duration was significantly different between parity group and postpartum day, with peaks on postpartum day 4 (387.7 minutes) and day 2 (318.0 minutes) for primiparous and multiparous groups, respectively. (F = 2.813, p = 0.048). Conclusion Mothers spent 20% of their postpartum hospitalization period for BF-related activities. Individual support is necessary, especially for primiparous mothers, who spent more time than multiparous mothers for BF-related activities on postpartum day 4. In a mixed ward, postpartum mothers need a comfortable hospital environment in which midwives can dedicate themselves to mother-child dyad care.

CD44v8-10 and CD44s Are Age-dependently Expressed in Primary Cultured Papillary Thyroid Carcinoma Cells and Are Associated with Cell Proliferation.

Abstract: Papillary thyroid carcinoma (PTC) is the most common endocrine malignancy, arising from follicular cells, and accounts for more than 80% of all thyroid malignant tumors. Although age is the strongest prognostic factor of PTC, and various cut-off ages (40-55 years) were suggested in previous studies, the molecular mechanisms causing age-related changes of PTC cell proliferation remain unclear. CD44 is a major cell surface receptor for hyaluronate and is known as a cancer stem cell marker. However, the association between CD44 and PTC is still unknown. Therefore, we determined the proliferation of primary cultured cells obtained from patients with PTC, and the CD44 mRNA expression profile to elucidate age-related association of CD44 with PTC. The results showed that cell proliferation was significantly decreased according to age. We also found that CD44v8-10 and CD44 splice variants were expressed dominantly in patients with PTC. Moreover, the CD44v8-10/CD44s mRNA expression ratio was significantly increased according to age, and there was a significant negative correlation between this expression ratio and cell proliferation. Our findings suggest that the CD44v8-10/CD44s expression ratio in PTC cells is useful for screening for aggressive PTC and may provide clinically valuable information.

Association Between HBx Variations and Development of Severe Liver Disease Among Indonesian Patients.

Abstract: Multi-site mutations in the hepatitis B virus (HBV) X gene are often found in patients with advanced liver diseases such as liver cirrhosis and hepatocellular carcinoma. It has been reported that modifications in the X protein play crucial roles in the development of HBV-related severe liver disease. However, the prevalence of genetic variations in Indonesian strains has not been systematically assessed. In this study, we sought to investigate the profile of nonsynonymous mutations in the X gene. Overall, 114 Indonesian HBV strains, including 12 in-house samples, were retrieved from GenBank. The mutation frequency in the X gene was compared among strains obtained from patients with chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. The mutation frequencies of the domain and basal core promoter regions were significantly greater in advanced liver diseases compared with chronic hepatitis. In addition, the double mutation K130M/V131I and the triple mutation N88V/K130M/V131I were associated with a 2.5 times higher risk of advanced liver disease. However, the roles of two novel X gene mutations (A12S/T and L16F/P) on hepatocarcinogenesis are unclear relative to wild-type X gene. In conclusion, the development of multi-site mutations in the X gene may represent a strategy by which HBV can escape immune surveillance and thus contribute to hepatocarcinogenesis, even though the biological roles of some variants remain unclear.

Spinal Muscular Atrophy: New Screening System with Real-Time mCOP-PCR and PCR-RFLP for SMN1 Deletion.

Abstract: Background Spinal Muscular Atrophy (SMA) is a common autosomal recessive neuromuscular disorder characterized by degeneration or loss of lower motor neurons. More than 95% of SMA patients show homozygous deletion for the survival motor neuron 1 (SMN1) gene. For the screening of SMN1 deletion, it is necessary to differentiate SMN1 from its highly homologous gene, SMN2. We developed a modified competitive oligonucleotide priming-PCR (mCOP-PCR) method using dried blood spot (DBS)-DNA, in which SMN1 and SMN2-specific PCR products are detected with gel-electrophoresis. Next, we added a targeted pre-amplification step prior to the mCOP-PCR step, to avoid unexpected, non-specific amplification. The pre-amplification step enabled us to combine mCOP-PCR and real-time PCR. In this study, we combined real-time mCOP-PCR and PCR-restriction fragment length polymorphism (PCR-RFLP) to develop a new screening system for detection of SMN1 deletion. Methods DBS samples of the subjects were stored at room temperature for a period of less than one year. Each subject had already been genotyped by the first PCR-RFLP using fresh blood DNA. SMN1/SMN2 exon 7 was collectively amplified using conventional PCR (targeted pre-amplification), the products of which were then used as a template in the real-time PCR with mCOP-primer sets. To confirm the results, the pre-amplified products were subject to the second PCR-RFLP. Results The real-time mCOP-PCR separately amplified SMN1 and SMN2 exon7, and clearly demonstrated SMN1 deletion in an SMA patient. The results of the real-time mCOP-PCR using DBS-DNA were completely consistent with those of the first and second PCR-RFLP analysis. Conclusion In our new system for detection of SMN1 deletion, real-time mCOP-PCR rapidly proved the presence or absence of SMN1 and SMN2, and the results were easily tested by PCR-RFLP. This solid genotyping system will be useful for SMA screening.

Nested PCR Amplification Secures DNA Template Quality and Quantity in Real-time mCOP-PCR Screening for SMA.

Abstract: Background Spinal Muscular Atrophy (SMA) is a common autosomal recessive disorder caused by SMN1 gene deletion. SMA has been considered an incurable disease. However, a newly-developed antisense oligonucleotide drug, nusinersen, brings about a good outcome to SMA patients in the clinical trials. Now, a screening for SMA is required for early diagnosis and early treatment so as to give a better clinical outcome to the patients. We have invented a new technology, mCOP-PCR, for SMA screening using dried blood spot (DBS) on the filter paper. One of the problems encountered in SMA screening is poor quality and quantity of DNA extracted from DBS. Methods DNA was extracted from DBS of six individuals. Fresh blood DNA of each individual had already been genotyped using PCR/RFLP. The fragments including the sequence of SMN1/SMN2 exon 7 were pre-amplified with conventional PCR. To determine which pre-amplified product is a better template for the real-time mCOP-PCR, we did pre-amplification with a single PCR or pre-amplification with a nested PCR. Results The real-time mCOP-PCR using pre-amplified products with a single PCR brought about ambiguous results in some SMN1-carrying individuals. However, the results of real-time mCOP-PCR following pre-amplification with a nested PCR were completely matched with those of PCR-RFLP. Conclusion In our study on the real-time mCOP-PCR screening system for SMA, a nested PCR secured the DNA template quality and quantity, leading to unambiguous results of SMA screening.

Newborn Screening for Spinal Muscular Atrophy: DNA Preparation from Dried Blood Spot and DNA Polymerase Selection in PCR

Abstract: Background Polymerase chain reaction (PCR) analysis using DNA from dried blood spot (DBS) samples on filter paper is a critical technique for spinal muscular atrophy (SMA) newborn screening. However, DNA extraction from DBS is time-consuming, and elimination of PCR inhibitors from DBS is almost impossible. Methods Exon 7 of the two homologous SMA-related genes, survival motor neuron (SMN) 1 and SMN2, of five SMA patients and five controls were amplified by PCR with a punched-out circle of the DBS paper. Two types of DNA preparation methods were tested; DNA-extraction (extracted DNA was added in a PCR tube) and non-DNA-extraction (a punched-out DBS circle was placed in a PCR tube). As for the DNA polymerases, two different enzymes were compared; TaKaRa Ex Taq™ and KOD FX Neo™. To test the diagnostic quality of PCR products, RFLP (Restriction fragment length polymorphism) analysis with DraI digestion was performed, differentiating SMN1 and SMN2. Results In PCR using extracted DNA, sufficient amplification was achieved with TaKaRa Ex Taq™ and KOD FX Neo™, and there was no significant difference in amplification efficiency between them. In direct PCR with a punched-out DBS circle, sufficient amplification was achieved when KOD FX Neo™ polymerase was used, while there was no amplification with TaKaRa Ex Taq™. RFLP analysis of the direct PCR products with KOD FX Neo™ clearly separated SMN1 and SMN2 sequences and proved the presence of both of SMN1 and SMN2 in controls, and only SMN2 in SMA patients, suggesting that the direct PCR products with KOD FX Neo™ were of sufficient diagnostic quality for SMA testing. Conclusion Direct PCR with DNA polymerases like KOD FX NeoTM has potential to be widely used in SMA newborn screening in the near future as it obviates the DNA extraction process from DBS and can precisely amplify the target sequences in spite of the presence of PCR inhibitors.

The Novel Lipid-Lowering Drug D-47 Ameliorates Hepatic Steatosis and Promotes Brown/Beige-Like Change of White Adipose Tissue in db/db Mice.

Abstract: D-47 is a newly developed solid dispersion of the arginine salt of (S)-(+)-4-[1-(4-tert-butylphenyl)-2-oxo-pyrrolidin-4-yl]methoxybenzoic acid (S-2E), which inhibits sterol and fatty acid synthesis. D-47 was recently shown to lower the serum level and hepatic content of both triglyceride and cholesterol in a rabbit model of familial hypercholesterolemia. We here investigated the effects of D-47 on dyslipidemia and hepatic steatosis in comparison with those of bezafibrate in the db/db mouse model of obesity. Treatment of db/db mice with D-47 or bezafibrate for 14 days lowered the serum triglyceride concentration without affecting that of cholesterol. D-47, but not bezafibrate, almost completely eliminated lipid droplets in hepatocytes and markedly lowered the triglyceride content of the liver in these mice. The two agents induced similar changes in the hepatic expression of genes including those related to β-oxidation or fatty acid synthesis. D-47 however significantly reduced the mass of white adipose tissue and up-regulated the expression of genes related to energy expenditure, mitochondrial function, fatty acid oxidation or lipolysis in this tissue, indicating that D-47 induced the brown/beige adipocyte-like change in white adipose tissue, whereas bezafibrate had no such effects. Treatment of 3T3-L1 adipocytes with D-47 provoked the expression of genes related to mitochondrial function, fatty acid oxidation or lipolysis. Our data have thus shown that D-47 ameliorated hypertriglyceridemia and hepatic steatosis in an animal model of obesity, and they suggest that this latter effect might be mediated through the change of adipose tissue characteristics.

Neonatal Alloimmune Thrombocytopenia: A Report of Four Cases.

Abstract: Antibodies against fetal platelet alloantigens in maternal blood cause neonatal alloimmune thrombocytopenia (NAIT). We encountered four newborns with NAIT from three women. A woman carried anti-human platelet antigen (HPA)-1a antibody, and vaginally delivered a newborn who had subarachnoid hemorrhage and platelet transfusions. She delivered the second newborn by a cesarean section who had no symptom. The second woman carried anti-human leukocyte antigen-A2 antibody and vaginally delivered a newborn who had no symptom. The third woman with a history of recurrent pregnancy losses carried anti-HPA-4b antibody, and delivered a newborn by a cesarean section who received platelet transfusions and immunoglobulin infusions. Antiplatelet antibody screening may be helpful in women who have a history of blood transfusion, or previous neonates with thrombocytopenia or intracranial hemorrhage.

Fetal Primary Small Bowel Volvulus Associated with Acute Gastric Dilatation Detected by Ultrasonography.

Abstract: Fetal intestinal volvulus is a rare condition, and fetal diagnosis of this disease is still challenging, especially in primary cases not accompanied by other comorbidities, such as intestinal malformations. Herein, we report a case of fetal primary small bowel volvulus associated with acute gastric dilatation detected by ultrasonography. We speculate that the mechanism of acute gastric dilatation in our case was peristatic malfunction of the whole intestine caused by a strangulated ileus resulting from fetal intestinal volvulus. In conclusion, acute gastric dilatation detected by fetal ultrasound can indicate the fetal intestinal volvulus.

Spinal Muscular Atrophy: Advanced Version of Screening System with Real-Time mCOP-PCR and PCR-RFLP for SMN1 Deletion.

Abstract: Background Spinal Muscular Atrophy (SMA) is a common autosomal recessive neuromuscular disease characterized by defects of lower motor neurons. More than 95% of SMA patients show homozygous deletion for the survival motor neuron 1 (SMN1) gene. For the screening of SMN1 deletion using dried blood spot (DBS), we developed a new combined system with real-time "modified competitive oligonucleotide priming"-polymerase chain reaction (mCOP-PCR) and PCR restriction fragment length polymorphism (PCR-RFLP). Although our real-time mCOP-PCR method is secured enough to be gene-specific, its amplification efficiency is not as good because the reverse primers carry a nucleotide mismatched with the sequence of the pre-amplified product. The mismatch has consequently been generated in the process of introducing a restriction enzyme site in the pre-amplified products for PCR-RFLP. Method DBS samples of the subjects were stored at room temperature for a period of less than one year. Each subject had already been genotyped by the first PCR-RFLP using fresh blood DNA. SMN1/SMN2 exon 7 was collectively amplified using conventional PCR (targeted pre-amplification). Pre-amplified products were used as template in the real-time mCOP-PCR, and, on the other hand, were digested with DraI enzyme (PCR-RFLP). To improve the amplification efficiency of mCOP-PCR, one nucleotide change was introduced in the original reverse primers (SMN1-COP and SMN2-COP) to eliminate the mismatched nucleotide. Results The real-time mCOP-PCR with a new primer (SMN1-COP-DRA or SMN2-COP-DRA) more rapidly and specifically amplified SMN1 and SMN2, and clearly demonstrated SMN1 deletion in an SMA patient. With the new primers, the amplification efficiencies of real-time mCOP-PCR were improved and the Cq values of SMN1 (+) and SMN2 (+) samples were significantly lowered. Conclusion In the advanced version of our screening system for homozygous SMN1 deletion using DBS, the real-time mCOP-PCR with newly-designed reverse primers demonstrated the presence or absence of SMN1 and SMN2 within a shorter time, and the results were easily tested by PCR-RFLP. This rapid and accurate screening system will be useful for detection of newborn infants with SMA.

Nursing Care Time for Newborns during Hospitalization in a Mixed Hospital Ward with an Obstetrics Department

Abstract: Purpose This study aimed to better understand the nursing care time spent with healthy term newborns from birth to discharge, giving insight into neonatal nursing staff management. Method In total, 30 healthy term newborns in a mixed hospital ward with an Obstetrics Department participated in this study. To measure care time, they had a wireless beacon attached to their cots. This measured how much time the nurses stayed in front of the cot from the time of birth until discharge, 24 hours/day. Collected data were tabulated every 24 hours after birth. Results Seventeen newborns had their data analyzed. The average length of hospital stay for the newborns was 8231.3 minutes. The average nursing care time for the newborns was 533.8 minutes. Nurses provided the highest care time during the first 24 hours after birth (157.6 minutes/24hr). After the first 24, the average nursing care time gradually decreased. The average nursing care time during the first 24 hours after birth was longer than the nursing care time for any other 24-hour periods, with a significant difference (p = 0.001 to 0.046). Conclusion The nursing care time for healthy newborns gradually decreased with the passage of time after birth. Healthy newborns should be treated as individuals, and the number of nursing staff should be adjusted according to the number of newborns in the ward to ensure nursing care quality and to prevent life-threatening events during the first 24 hours after birth.

Study on the Language Formation Process of Very-Low-Birth-Weight Infants in Infancy Using a Formant Analysis.

Abstract: Expressive language development depends on anatomical factors, such as motor control of the tongue and oral cavity needed for vocalization, as well as cognitive aspects for comprehension and speech. The purpose of this study was to examine the differences in expressive language development between normal-birth-weight (NBW) infants and very-low-birth-weight (VLBW) infants in infancy using a formant analysis. We also examined the presence of differences between infants with a normal development and those with a high risk of autism spectrum disorder who were expected to exist among VLBW infants. The participants were 10 NBW infants and 10 VLBW infants 12-15 months of age whose speech had been recorded at intervals of approximately once every 3 months. The recorded speech signal was analyzed using a formant analysis, and changes due to age were observed. One NBW and 3 VLBW infants failed to pass the screening tests (CBCL and M-CHAT) at 24 months of age. The formant frequencies (F1 and F2) of the three groups of infants (NBW, VLBW and CBCL·M-CHAT non-passing infants) were scatter-plotted by age. For the NBW and VLBW infants, the area of the plot increased with age, but there was no significant expansion of the plot area for the CBCL·M-CHAT non-passing infants. The results showed no significant differences in expressive language development between NBW infants at 24 months old and VLBW infants at the corrected age. However, different language developmental patterns were observed in CBCL·M-CHAT non-passing infants, regardless of birth weight, suggesting the importance of screening by acoustic analyses.

Emotional and Behavioral Problems of Pre-school Children with Autistic Spectrum Disorder Assessed by the Child Behavior Checklist 1½-5

Abstract: BACKGROUND Children with Autism Spectrum Disorder (ASD) experience a significant disruption in the ability to communicate and interact socially. Early detection is important; the earlier the child is identified with ASD, the faster the intervention can begin. The Child Behavior Checklist (CBCL) is a screening tool capable of identifying various emotional and behavioral disorders. Research data on the use of the CBCL as a screening tool for ASD are still limited in Indonesia. The present study aimed to evaluate the CBCL's ability to detect emotional and behavioral problems in pre-school children with ASD. METHODS A cross sectional study was conducted in Surabaya, Indonesia. Children aged 1½-5 years old diagnosed with ASD, and typically developing (TD) children were enrolled using total sampling. The CBCL/1½-5 questionnaire for parents, consisting of 99 items, was used in this research to evaluate emotional and behavioral problems. Data were analyzed with the Independent-Sample T-test to compare the CBCL/1½-5 scale between ASD and TD children, while the proportion of gender was evaluated by a chi-square test with a 95% confidence interval. RESULTS Fifty children were included in this research, 25 with ASD and 25 TD. Seventy six percent (38) were male and 24% (12) were female. Six of the seven scales were significantly higher in ASD children than TD in the CBCL/1½-5. Children with ASD had higher internalization (p < 0.0001), externalization (p = 0.001), and total problem scores (p < 0.0001) when compared to TD children. CONCLUSION The CBCL/1½-5 can significantly detect the emotional and behavioral problems in pre-school children with ASD.

Anesthetic Management of Transcatheter Aortic Valve Replacement under Extracorporeal Membrane Oxygenation in a Patient with Acute Decompensated Heart Failure: A Case Report.

Abstract: We managed general anesthesia for transcatheter aortic valve replacement (TAVR) under elective extracorporeal membrane oxygenation (ECMO) in a patient with aortic valve stenosis (AS) complicated with acute decompensated heart failure. The patient was an 87-year-old woman with acute heart failure due to severe AS who had been hospitalized. However, her low cardiac output did not improve, and weaned her off catecholamines was difficult, so semi-urgent TAVR was performed. Due to her acute decompensated heart failure complicated by low-left ventricular function, we decided electively to use ECMO for transfemoral TAVR to prevent hemodynamic collapse during induction of anesthesia and surgery, enabling the safe perioperative management of this patient under general anesthesia.

Questionnaire Survey for Bronchial Asthma in Elderly Care Facilities.

Abstract: In developed countries such as North America, the decline in mortality from bronchial asthma has ceased since 2006. The decline in mortality rate is also decreasing in Japan, where about 1,500 asthma deaths have been reported. Among these, elderly people aged 65 years or over account for about 90% of cases. Therefore, the treatment of elderly patients with asthma is an important subject. However, few studies have been conducted on asthma in elderly patients. In this survey, we distributed a questionnaire to 253 elderly care facilities in Kobe, Japan. Ninety facilities responded, and 223 patients in 70 out of 90 facilities were diagnosed with asthma. Dry powder inhaler was the most commonly used dosage form of inhaled corticosteroids. Many facilities have patients who need some assistance during inhalation: only 60% of facilities reported that inhalation is performed accurately. While 31 facilities had patients with a history of hospitalization for asthma attacks, only 14 of these facilities were able to provide appropriate initial treatment. Many facilities have difficulty providing assistance with inhalation to elderly patients whose cognitive function has deteriorated. This survey highlights challenges experienced by care facilities in treating asthma in the elderly.