Concordance of International Regulation of Pediatric Health Research.

OBJECTIVE: To assess the comparability of international ethics principles and practices used in regulating pediatric research as a first step in determining whether reciprocal deference for international ethics review is feasible. Prior studies by the authors focused on other aspects of international health research, such as biobanks and direct-to-participant genomic research. The unique nature of pediatric research and its distinctive regulation by many countries warranted a separate study. STUDY DESIGN: A representative sample of 21 countries was selected, with geographical, ethnic, cultural, political, and economic diversity. A leading expert on pediatric research ethics and law was selected to summarize the ethics review of pediatric research in each country. To ensure the comparability of the responses, a 5-part summary of pediatric research ethics principles in the US was developed by the investigators and distributed to all country representatives. The international experts were asked to assess and describe whether principles in their country and the US were congruent. Results were obtained and compiled in the spring and summer of 2022. RESULTS: Some of the countries varied in their conceptualization or description of one or more ethical principles for pediatric research, but overall, the countries in the study demonstrated a fundamental concordance. CONCLUSIONS: Similar regulation of pediatric research in 21 countries suggests that international reciprocity is a viable strategy.

The Human Genome Organisation (HUGO) and the 2020 COVID-19 pandemic.

This letter is the Human Genome Organisation's summary reaction to the 2020 COVID-19 pandemic. It identifies key areas for genomics research, and areas in which genomic scientists can contribute to a global response to the pandemic. The letter has been reviewed and endorsed by the HUGO Committee on Ethics, Law and Society (CELS) and the HUGO Council.

A real-time PCR-based amelogenin Y allele dropout assessment model in gender typing of degraded DNA samples.

Allelic dropout due to stochastic variation in degraded small quantity DNA appears to be one of the most serious genotyping errors. Most methods require PCR replication to address this problem. The small amounts of valuable samples are often a limitation for such replications. We report a real-time PCR-based amelogonin Y (AMELY) allele dropout estimation model in an AMEL-based gender typing. We examined 915 replicates of AMELY-positive modern male DNA with varying amounts of DNA and humic acid. A male-specific AMEL fragment (AMELy) dropped out in 143 genuine male replicates, leading to gender typing errors. By graphing a scatter plot of the crossing point versus the end cycle fluorescence of the male replicates, a standard graph model for the estimation of the AMELy allele dropout was constructed with the dropout-prone and dropout-free zones. This model was then applied to ancient DNA (aDNA) samples. Nine samples identified as female were found in the dropout-prone zone; with higher DNA concentrations, six were shifted to the dropout-free zone. Among them, two female identifications were converted to male. All the aDNA gender was confirmed by sex-determination region Y marker amplification. Our data suggest that this model could be a basic approach for securing AMELy allele dropout-safe data from the stochastic variation of degraded inhibitory DNA samples.

Secondary Use Provisions in the European Health Data Space Proposal and Policy Recommendations for Korea.

OBJECTIVES: This article explores the secondary use provisions of the European Health Data Space (EHDS), proposed by the European Commission in May 2022, and offers policy recommendations for South Korea. METHODS: The authors analyzed the texts of the EHDS proposal and other documents published by the European Union, as well as surveyed the relevant literature. RESULTS: The EHDS proposal seeks to create new patient rights over electronic health data collected and used for primary care; and establish a data sharing system for the re-use of electronic health data for secondary purposes, including research, the provision of personalized healthcare, and developing healthcare artificial intelligence (AI) applications. These provisions envisage requiring both private and public data holders to share certain types of electronic health data on a mandatory basis with third parties. New government bodies, called health data access bodies, would review data access applications and issue data permits. CONCLUSIONS: The overarching aim of the EHDS proposal is to make electronic health data, which are currently held in the hands of a small number of organizations, available for re-use by third parties to stimulate innovation and research. While it will be very challenging for South Korea to adopt a similar scheme and require private entities to share their proprietary data with third parties, the South Korean government should consider making at least health data collected through publicly funded research more readily available for secondary use.

How isometric are the anatomic femoral tunnel and the anterior tibial tunnel for anterior cruciate ligament reconstruction?

PURPOSE: The purpose of this study was to evaluate the isometry of an anatomic femoral tunnel and anterior tibial tunnel positions. METHODS: Tibial tunnels were made at 2 different locations in 10 cadaveric knees: the conventional tunnel and a more anterior position. Three-dimensional computed tomography (CT) scanning was then performed at 0°, 30°, 60°, 90°, and 120°. After removal of the anterior cruciate ligament from its femoral attachment, the 2 different femoral tunnels were marked at (1) the vertical femoral tunnel point and (2) the anatomic femoral tunnel point. After scans were repeated for coordinate transformation, the change in length between the tunnels was calculated with imaging software (OsiriX, version 3.2; Apple, Cupertino, CA) and the center of rotation for the femoral tunnels was calculated with a least squares fitting algorithm. RESULTS: The conventional tibial tunnel-vertical femoral tunnel combination showed the least excursion as knee flexion angle changed. The vertical femoral tunnel combination groups showed a trend toward increasing length as the knee flexion angle increased. In contrast, the anatomic femoral tunnel combination groups displayed a trend toward decreased length with increasing knee flexion. At less than 30° of flexion, the tibial anterior-anatomic femoral tunnel showed the least excursion. CONCLUSIONS: The anatomic femoral tunnel was nonisometric, and the differences in isometry for each tunnel type were explained primarily by differences in relations between the centers of rotation of tunnels and tunnel position. When a femoral anatomic tunnel is chosen for anterior cruciate ligament reconstruction, the anterior tibial tunnel offers greater isometric benefits than the conventional tibial tunnel, especially in near full extension. CLINICAL RELEVANCE: The distance between anatomic femoral and tibial tunnels is greatest in full extension and decreases with flexion. This would result in graft laxity. The surgeon should give consideration to a more anterior tibial tunnel position, which shows less excursion in early flexion.

Streamlining ethics review for international health research.

Single-site review means protection and efficiency.

A western Eurasian male is found in 2000-year-old elite Xiongnu cemetery in Northeast Mongolia.

We analyzed mitochondrial DNA (mtDNA), Y-chromosome single nucleotide polymorphisms (Y-SNP), and autosomal short tandem repeats (STR) of three skeletons found in a 2,000-year-old Xiongnu elite cemetery in Duurlig Nars of Northeast Mongolia. This study is one of the first reports of the detailed genetic analysis of ancient human remains using the three types of genetic markers. The DNA analyses revealed that one subject was an ancient male skeleton with maternal U2e1 and paternal R1a1 haplogroups. This is the first genetic evidence that a male of distinctive Indo-European lineages (R1a1) was present in the Xiongnu of Mongolia. This might indicate an Indo-European migration into Northeast Asia 2,000 years ago. Other specimens are a female with mtDNA haplogroup D4 and a male with Y-SNP haplogroup C3 and mtDNA haplogroup D4. Those haplogroups are common in Northeast Asia. There was no close kinship among them. The genetic evidence of U2e1 and R1a1 may help to clarify the migration patterns of Indo-Europeans and ancient East-West contacts of the Xiongnu Empire. Artifacts in the tombs suggested that the Xiongnu had a system of the social stratification. The West Eurasian male might show the racial tolerance of the Xiongnu Empire and some insight into the Xiongnu society.

Erratum to: Milk Containing BF-7 Enhances the Learning and Memory, Attention, and Mathematical Ability of Normal Persons.

[This corrects the article DOI: 10.5851/kosfa.2009.29.2.278.].

Immunohistochemical study on the distribution of glycogen synthase kinase 3alpha in the central nervous system of SOD1(G93A) transgenic mice.

OBJECTIVE: To identify glycogen synthase kinase (GSK) 3alpha expression in a mouse model of familial amyotrophic lateral sclerosis (ALS), we investigated the changes of GSK3alpha in the central nervous system of SOD1(G93A) transgenic mice by immunohistochemistry. METHODS: We used 12 SOD1(G93A) transgenic and ten wild-type (wt) SOD1 transgenic mice bred by 'The Jackson Laboratory' under the strain designations B6SJL-TgN (SOD1(G93A)) 1 Gur/J and B6SJL-TgN (SOD1) 2 Gur/J, respectively. Immunohistochemistry was performed in accordance with the free-floating method described earlier. RESULTS: In symptomatic transgenic mice, GSK3alpha-immunoreactive astrocytes were detected in the spinal cord, brainstem and cerebellum of symptomatic SOD1(G93A) transgenic mice. In contrast to symptomatic mice, no GSK3alpha-immunoreactive astrocytes were observed in any brain region of wtSOD1 and pre-symptomatic mice, and the number and intensity of stained cells were not different at the age of 8 and 13 weeks. DISCUSSION: These results provide the first evidence that GSK3alpha-immunoreactive astrocytes were found in the CNS of SOD1(G93A) transgenic mice after clinical symptoms, suggesting a possible role in the pathologic process of ALS. However, the mechanisms underlying the increased immunoreactivity for GSK3alpha and the functional implications require elucidation.

Immunohistochemical study on the distribution of canonical transient receptor potential channels in rat basal ganglia.

In the present study, we examined the localizations of canonical transient receptor potential channels (TRPCs) in rat basal ganglia. The dot-like staining pattern of TRPC5 was observed through the globus pallidus (GP) and caudate-putamen. TRPC7 had a strikingly high level of expression in the neuropil in the GP. In the subthalamic nucleus, strong staining for TRPC5 was observed in the cell bodies, while moderate to high immunoreactivies for TRPC1, TRPC3, TRPC4 and TRPC7 were found in the cell bodies and surrounding neuropil. In the substantia nigra, immunoreactivities for TRPC3 and TRPC7 were prominent in the cell bodies and several processes in the pars compacta and pars reticulata. TRPC6 was expressed in the neuropil, not in the cell bodies. This study may provide useful data for the future investigations on the structural and functional properties of TRPCs.

Age-related changes in glycogen synthase kinase 3beta (GSK3beta) immunoreactivity in the central nervous system of rats.

Although glycogen synthase kinase 3beta (GSK3beta) is emerging as a prominent drug target in the treatment of neurodegenerative diseases such as Alzheimer's disease (AD) and stroke, very little is known about age-related changes in GSK3beta expression and GSK3beta phosphorylation. Therefore, we examined age-related changes in immunoreactivities for GSK3beta and phosphorylated GSK3beta (pGSK3beta) in the central nervous system. In aged rats, there were significant increases in GSK3beta immunoreactivity in the cell bodies and processes of pyramidal cells in most cortical regions. GSK3beta immunoreactivity was also significantly increased in the pyramidal layer of CA1-3 regions, and the granule cell layer of dentate gyrus. Age-related increases were prominent in lateral septal nuclei, compared to the medial septal nuclei. Interestingly, both GSK3beta and pGSK3beta was increased in the prefrontal cortex, while GSK3beta and pGSK3beta was differentially localized in the cerebellar cortex. The first demonstration of age-related alterations in immunoreactivities for GSK3beta and pGSK3beta in the basal forebrain area and cholinergic projection targets may provide useful data for investigating the pathogenesis of age-related neurodegenerative diseases including AD.

ERK1/2 activation attenuates TRAIL-induced apoptosis through the regulation of mitochondria-dependent pathway.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) functions as an extracellular signal, which triggers apoptosis in tumor cells. In order to characterize the molecular events involved in TRAIL cytotoxic signaling, we attempted to determine the role of extracellular signal-regulated kinase 1/2 (ERK1/2), as well as its downstream targets in TRAIL-treated HeLa cells. Here we demonstrate that TRAIL exposure resulted in the activation of ERK1/2, and the elevation of anti-apoptotic Bcl-2 protein levels. ERK1/2 inhibition with PD98059 promoted cell death via the down-regulation of Bcl-2 protein levels, together with increasing mitochondrial damage, including the collapse of mitochondrial membrane potential, the release of cytochrome c from mitochondria to cytoplasm and caspase activity. These results suggest that the ERK1/2 activation is a kind of survival mechanism to struggle against TRAIL-induced stress condition in early stage, via activating cellular defense mechanisms like as the up-regulation of the Bcl-2/Bax ratio, as well as several mitochondrial events.

Biobank Regulation in South Korea.

Like many other countries, South Korea has recognized the importance of biobanks as a tool for medical research and has engaged in two very important tasks to foster biobanking infrastructure: funding biobanks and setting up rules to protect the integrity of biobanks that share potentially sensitive personal information.

The Natural Substance MS-10 Improves and Prevents Menopausal Symptoms, Including Colpoxerosis, in Clinical Research.

Many natural substances were screened to develop nutraceuticals that reduce menopausal symptoms. A complex of Cirsium japonicum var. maackii and Thymus vulgaris extracts, named MS-10, had significant positive effects. Under a low concentration of estrogen, which represents postmenopausal physiological conditions, MS-10 had beneficial effects on estrogen receptor-expressing MCF-7 cells by reversibly enhancing estrogen activity. In addition, in the ovariectomized rat model, changes in bone-specific alkaline phosphatase activity and osteocalcin, as well as low-density lipoprotein cholesterol and triglyceride levels were significantly decreased by MS-10. These results show that MS-10 protected bone health and reduced metabolic disturbances. Furthermore, in a clinical study, all menopausal symptoms, including hot flushes, parenthesis, insomnia, nervousness, melancholia, vertigo, fatigue, rheumatic pain, palpitations, formication, and headache, as well as colpoxerosis, were significantly improved by taking MS-10 for 90 days. Therefore, the evidence supports that MS-10 is an effective natural substance that can safely improve menopausal symptoms, including colpoxerosis.

P53 mediates ceramide-induced apoptosis in SKN-SH cells.

Ceramide induces apoptotic cell death in a dose- and time-dependent manner in neuroblastoma SKN-SH cells. Pretreatment with caspase inhibitors blocks cell death, suggesting that a set of caspase activities including caspase 1, as well as caspase 3, are involved in ceramide-induced apoptosis in SKN-SH cells. Treatment with a caspase inhibitor 3 h after ceramide addition did not inhibit cell death, although caspase activity was substantially reduced. Ceramide-induced apoptosis is accompanied by accumulation of p53 followed by an increase of Bax and decrease of Bcl-2 levels. Inhibition of p53 expression with p53 antisense oligonucleotides inhibits apoptosis and prevents the increase in Bax and decrease in Bcl-2. Furthermore, pretreatment with p53 antisense oligonucleotides markedly inhibits the induction of caspase activity. These results suggest that p53 regulates the ratio Bcl-2/Bax and the expression/activation of caspases during ceramide-induced apoptosis in SKN-SH cells. Caspase inhibition did not alter the expression of p53, Bcl-2 and Bax. Thus ceramide-induced reduction in the Bcl-2/Bax ratio, increase in caspase activity, and apoptosis is dependent upon increases in cellular p53 levels which play a critical role in the regulation of apoptotic cell death.

Baicalein attenuates 6-hydroxydopamine-induced neurotoxicity in SH-SY5Y cells.

It has been suggested that baicalein, a flavonoid obtained from the Scutellaria root, exerts a protective role on neurons against several neuronal insults. However, the protective mechanisms underlying this protective effect remain largely unknown. Our results indicate that baicalein protects SH-SY5Y cells, a dopaminergic neuronal cell line, from 6-hydroxydopamine (6-OHDA)-induced damage by the attenuation of reactive oxygen species (ROS). In order to determine the effects of baicalein on mitochondrial events, mitochondrial membrane potentials (deltapsim) and caspase cascades downstream of mitochondria were assessed. Baicalein inhibited the collapse of deltapsim, suggesting that baicalein reduces the mitochondrial dysfunction associated with 6-OHDA treatment. Baicalein also inhibited caspase-9 and caspase-3 activation, which can be triggered by mitochondrial malfunctions. Furthermore, baicalein induced a significant reduction in the level of phospho-JNK, which is known as an apoptotic mediator in 6-OHDA-induced neuronal cell death. Our results indicate that baicalein protects neurons from the deleterious effects of 6-OHDA via the attenuation of oxidative stress, mitochondrial dysfunction, caspase activity, and JNK activation.

Immunohistochemical study on the distribution of neuronal nitric oxide synthase-immunoreactive neurons in the spinal cord of aged rat.

Despite in vivo studies suggesting an important function for nitric oxide (NO) in the spinal cord in the transmission of pain signals, sympathetic nerve activity and presumably other spinal functions, changes of neuronal NO synthase (nNOS)-containing neurons with aging in the spinal cord has not been investigated. In the present study, we demonstrated for the first time that the number of nNOS-immunoreactive neurons was significantly decreased in the central autonomic nucleus and the superficial dorsal horn of spinal cord in aged rats. Morphologically, the number and length of dendritic branches also seemed to be decreased. Combined with our previous studies, age-related decreases in the number of nNOS-immunoreactive neurons in the central autonomic nucleus and the superficial dorsal horn might be associated with the abnormality of micturition function or pain perception encountered in the elderly. However, the mechanisms underlying the decreased immunoreactivity for nNOS, and the functional implications require elucidation.

Decreased expression of calretinin in the cerebral cortex and hippocampus of SOD1G93A transgenic mice.

In the present study, we investigated the changes of calretinin (CR) expression in the central nervous system of SOD1G93A transgenic mice as an in vivo model of amyotrophic lateral sclerosis (ALS). In wild-type SOD1 (wtSOD1) transgenic mice, many CR-immunoreactive neurons were found in all cortical regions. In the cerebral cortex of SOD1G93A transgenic mice, the number and staining intensity of CR-positive neurons were decreased. In the hippocampal formation, layer-specific alterations in the staining intensity of CR-immunoreactive neurons were observed in the CA1-3 areas and dentate gyrus. In wtSOD1 transgenic mice, CR-immunoreactive neurons with long processes were found in the stratum oriens and stratum radiatum of CA1-3 areas, and heavily stained band-like molecular layer was prominent in the dentate gyrus. CR immunoreactivity was decreased in each layer of CA1-3 areas and dentate gyrus of SOD1G93A transgenic mice. The first demonstration of decreased immunoreactivity for CR in the cerebral cortex and hippocampus of SOD1G93A transgenic mice may provide insights into the pathogenesis of motor neuron degeneration in human ALS although further quantitative studies are needed.

Immunohistochemical study on the distribution of phosphorylated extracellular signal-regulated kinase (ERK) in the central nervous system of SOD1G93A transgenic mice.

In the present study, we performed immunohistochemical studies to investigate the changes of phosphorylated extracellular signal-regulated kinases (pERK) in the central nervous system of SOD1(G93A) transgenic mice. In symptomatic transgenic mice, pERK-immunoreactive astrocytes were detected in the spinal cord, brainstem, central gray and cerebellar nuclei. In contrast to symptomatic mice, no pERK-immunoreactive astrocytes were observed in any brain region of wtSOD1 and presymptomatic mice, and the number and intensity of stained neurons were not different at the age of 8 weeks and 13 weeks. Interestingly, region-specific alterations in pERK immunoreactivity were observed in the hippocampal region and cerebellum. These results provide the first evidence that pERK-immunoreactive astrocytes were found in the CNS of SOD1(G93A) transgenic mice after clinical symptoms, showing a possible consequence of the pathological process of ALS. This study has also demonstrated that pERK increases in the hippocampus and cerebellum, suggesting a role of pERK in an abnormality of cognitive and/or motor function in ALS, respectively. However, the mechanisms underlying the increased immunoreactivity for pERK and the functional implications require elucidation.

Immunohistochemical study on the distribution of the voltage-gated potassium channels in the gerbil cerebellum.

Although there have been many studies on the regional distribution of Kv channels in the rat and mouse cerebellum, there are no reports about Kv channel distribution in the gerbil, which is used as an ischemia animal model. Therefore, we aimed to investigate differences in the spatial patterning of Kv channel alpha-subunit isoforms in the gerbil cerebellum. The greatest concentration of Kv1.2 was found in the basket cell axon plexus and terminal regions around the Purkinje cells. Kv1.1 immunoreactivity was also concentrated in this area although the staining intensity was relatively lower. Both Purkinje cell layer and granular layer were intensely stained with anti-Kv1.3 and Kv1.6 antibodies, whereas immunoreactivities for Kv1.4 and Kv1.5 were detected in the Purkinje cell bodies with much lower intensity in the molecular and granular layers. In the cerebellar nuclei, the cell bodies of cerebellar output neurons showed strong immunoreactivities for Kv1.2, Kv1.4, and Kv1.6 with moderate staining for Kv1.3 and Kv1.5 in the cell bodies. This study on the differential localization patterns of Kv1 channel subunits in the gerbil cerebellum may provide helpful guidelines for correlating current types with particular channels and useful data for the future investigations on the pathological conditions such as ischemia and epilepsy.