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

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.

Periprosthetic Fracture after Hook Plate Fixation in Neer Type II Distal Clavicle Fracture: A Report of 3 Cases

Hook plate fixation is a treatment method for the displaced distal clavicle fracture with favorable results regarding bone union and shoulder function, however possible complications include impingement syndromes, subacrormial erosions, acromial fractures, and periprosthetic fractures. In this report, we observed 3 cases of periprosthetic fracture after hook plate fixation. All cases of periprosthetic fractures were initiated at the medial end screw holes. The causes of these periprosthetic fractures appeared to be the off centered fixation of medial end screws near the anterior or posterior cortex which were specific during operations with hook plates with more than 6 holes and the increased stress on the medial end screw by over-reduced or inferiorly reduced position of the distal end of the clavicle by the hook plate.

Improvement of andropause symptoms by dandelion and rooibos extract complex CRS-10 in aging male

Many aging male suffer various andropause symptoms including loss of physical and mental activities. This study evaluated the putative alleviative effects of CRS-10 dandelion and rooibos extract complex (CRS-10) on the symptoms of andropause. The survival rate of TM3 Leydig cells (TM3 cells) treated with CRS-10 was measured based on typical physiological stress. After daily intake of CRS-10 for 4 weeks, the level of testosterone, physical activity and both the number and activity of sperm in older rats (18 weeks) were measured. Furthermore, thirty males were surveyed with AMS (Aging Males' Symptoms) questionnaire after intake of 400 mg of CRS-10. Overall, CRS-10 protected TM3 cells from serum restriction and oxidative stress via activation of ERK and Akt pathways. The level of testosterone and activation of spermatogenesis in rats were significantly enhanced. In addition, physical locomotion was markedly improved. Daily intake of 400 mg of CRS-10 improved the quality of life among agingmale respondents, according to a clinical survey using the AMS. The results indicate the potential of CRS-10 as a safe and efficacious natural substance for reducing or alleviating andropause symptoms.

HLA-DRB1 Study of DNA from Ancient Human Skeleton by Sequence-based Typing / 체질인류학회지

The analysis of ancient human DNA is increasingly used recently in the study of anthropology and human evolution. Although mitochondrial DNA and Y chromosomal DNA has commonly been the target in the field of human DNA study, HLA analysis of ancient human DNA is extremely rare. This study aimed to develop the PCR method of ancient human DNA for analyzing the sequence of HLA. Authors established a new method for HLA-DRB1 analysis by sequence-based typing. Alleles of HLA-DRB1 were analyzed and typed by sequencing with DNA of ancient human skeletons from Korea and Mongolia 3000-500 years ago. The types of HLA-DRB1 were determined by comparing the sequences with those of HLA database (http://www. ebi.ac.uk/Tools/blast2/nucleotide.html). The alleles of HLA-DRB1 of ancient human DNA from Korea and Mongolia were classified by types. The frequencies of HLA-DRB1 types of Mongolia were also presented according to the geography such as West, Central, East, and North. In summary, our method was successful in the analyzing the type of HLA-DRB1 from DNA of ancient human bones. Authors anticipate that many researchers could do their research in a better way to get the genetic information for the kinship analysis between individuals or communities from ancient human bones.

A Kinship Analysis of Ancient Human Bones and Teeth from Mongolia / 체질인류학회지

The kinship was analyzed genetically on the three 2000 year old ancient human bones and teeth excavated in Mongolia. The samples were processed in a clean room to prevent the contamination from modern human DNA. The DNA extraction and purification was done with ion-exchange column kit (Qiagen G-tip 20G, USA). The PCR was done with purified DNAs from ancient human bones for paternal Y-SNP haplogroup, maternal mtDNA haplogroup, and autosomal short tandem repeats (STR). Two samples belonged to the maternal D major haplogroup, which is one of the most frequent types in the present North East Asia. One of them, showing male genotype, belonged to the paternal C major haplogroup, which is also one of the most frequent types in the present North East Asia. The remaining one belonged to the paternal R major haplogroup, frequent in the present Europe, and the maternal U haplogroup, frequent in the present Europe and East Mediterranean. The repeated results were consistent in the autosomal STR PCR. The STR data were analyzed with DNA-VIEW program (http://www.dna-view.com), which showed no close kinship among the three ancient humans. Our method was successful in the analyzing kinship among ancient human bones, which has been possible in few restricted laboratories in the World. Authors anticipate that many researchers could do their research in a better way to get the genetic information from ancient human bones.

Immunohistochemical Study on the Distribution of Insulin-like Growth Factor Binding Protein 7 (IGFBP7) in the Central Nervous System of Adult Rats / 체질인류학회지

In the present study, we performed immunohistochemical studies to investigate the detailed distribution of insulin-like growth factor binding protein 7 (IGFBP7) in the central nervous system of adult rats. Twelve adult (4~6 month old) Sprague-Dawley rats were examined in this study. Immunohistochemistry using specific antibodies against IGFBP7 was performed in accordance with the free-floating method. In the present study, IGFBP7 immunoreactivity was observed in the cerebral cortex, hippocampus, brainstem, cerebellum and spinal cord. In the cerebral cortex, heavily stained neurons were seen in layers II-VI. In the hippocampus, pyramidal cells in CA1-3 region were strongly immunoreactive for IGFBP7. Strong immunoreactive neurons were also found in the supraoptic nucleus, paraventricular nucleus, periaqueductal gray and oculomotor nucleus. In the cerebellum, IGFBP7 immunoreactivity was prominent in the Purkinje cells and cerebellar output neurons. IGFBP7-immunoreactive neurons were prominent in the superior vestibular nucleus, cochlear nucleus, trigeminal motor nucleus, nucleus of the trapezoid, and facial nucleus. IGFBP7-immunoreactive neurons were also observed mainly in the anterior horn of the spinal cord. The first demonstration of IGFBP7 localization in the whole brain may provide useful data for the future investigations on the structural and functional properties of IGFBP7.

Protective Effect of HSP90 on Neuronal Cell Death-induced by beta-Amyloid Peptide / 체질인류학회지

In the present study, we determined the protective mechanism of HSP90 against neuronal cell death induced by Abeta. For the evaluation of protective role of HSP90, we used human neuroblastoma SK-N-SH cell lines, examined AlamarBlue assay, Western blot analysis and immunofluorescence assay. Incubation of SK-N-SH cells with Abeta significantly induced neuronal cell death. However, HSP90 induced by mild heat shock could attenuate neuronal apoptosis in Abeta treated condition. To identify the role of HSP90, we determined localization of HSP90 in SK-N-SH cells. Interestingly, HSP90 was increased and localized in mitochondria as treatment of mild heat shock. Also, treatment or increase of HSP90 largely elevated level of Bcl-2 expression, whereas inhibition of HSP90 with HSP90 antisense oligonucleotide significantly decreased Bcl-2 expression. In contrast to Bcl-2, Bax expression was regulated independently by HSP90. Moreover, increase of HSP90 could attenuate collapse of mitochondrial membrane potential induced by Abeta. However, HSP90 antisense oligonucleotide largely increase breakdown of mitochondrial membrane potential induced by Abeta. These data suggest that HSP90 as chaperone protein significantly attenuates neuronal damage and protects neuroanl cells from neurotoxin such as Abeta.

Immunohistochemical Study on the Distribution of Kv1.2 in Aged Rat Brain / 체질인류학회지

In the present study, we demonstrated age-related changes in Kv1.2 immunoreactivity in the rat brain for the first time. Twelve adult (4~6 month old) and 15 aged (20~29 month old) Sprague-Dawley rats were examined in this study. Immunohistochemistry was performed in accordance with the free-floating method, and densitometric measurement using a NIH image program (Scion Image) determined the staining density. In the cerebral cortex of aged rats, there was a significant increase in the number of Kv1.2-immunoreactive neurons in the cingulate cortex, infralimbic cortex and piriform cortex, compare to adult rats. In the hippocampal CA1-3 regions, moderate Kv1.2 immunoreactivity was found in the cell bodies and processes of some medium to large-sized neurons in aged rats. The intensity was increased in the cell bodies of Kv1.2-positive neurons in the amygdala of aged rats, whereas the number of immunoreactive neurons was not significantly increased. It was noteworthy that age-related changes in Kv1.2-immunoreactive neurons were prominent in the facial nuclei, raphe magnus nuclei, and pontine and medullary reticular formation. Although the present study has not addressed multiple mechanisms contributing to neuronal degeneration during aging, the first demonstration of age-related changes in Kv1.2 immnuoreactivity may offer a comprehensive understanding of the pathophysiology of aging and age-related neurodegenerative diseases such as Alzheimer's disease.

Differential Expression of Neuronal Death-related Factors in Aged Rat Cerebellum / 체질인류학회지

In the present study, we investigated the expression of apoptosis-associated proteins in the cerebellum of aged rats: IGF-I receptor (IGF-IR), nitrotyrosine (NT), p53, key pro-apoptotic gene ICH-1 (caspase-2), c-Fos and Bcl-2 family members (Bcl-2 and Bax). Twelve adult (4~6 month old) and 15 aged (24~29 month old) Sprague-Dawley rats were examined in this study. We performed immunohistochemical staining, in situ hybridization and densitometric measurement using a NIH image program (Scion Image) to determine the staining density. In adult rats, there were no immunoreactivities for insulin-like growth factor-I receptor (IGF-IR), nitrotyrosine (NT) or p53 in any region of cerebellum. However, IGF-IR immunoreactivity was found in some Purkinje cells in aged rat cerebellum. The prominent staining of NT or p53 was also localized in the Purkinje cell layer in aged rats. A high density of ICH-1 (caspase-2) immunoreactivity was observed in the molecular and Purkinje cell layers in aged rats. Immunoreactivity for c-Fos was significantly decreased in the granule cells in aged rats. Positive signal for bcl-2 was significantly decreased in the Purkinje cells and granule cells of aged rats. The most intense staining for Bax was observed in the soma of Purkinje cells of adult rats. However, Bax immunoreactivity was not changed in any layers in the cerebellar cortex of aged rats. In conclusion, this study provides the first morphological data concerning the differential regulation of apoptosisrelated genes in rat cerebellum during aging.

Comparison between Morphological Sex and Genotype Sex of Uzbekistan Ancient Bones Using Improved Amelogenin PCR Amplication Method / 체질인류학회지

Determination of male and female is important in anthropology, archeology and forensic science. This study was designed to compare genotype sex of improved amelogenin PCR amplication method with morphological sex of ancient human bones. Sixty human skulls which lived from the Bronze Age to twenties centuries and excavated in Uzbekistan were used in this study. Morphological sex was determined by Uzbekistan scientist, and genotype sex was determined by improved amelogenin PCR amplication developed in this study. Among 20 morphological males, 13 samples (65%) were genotypical male. Among 40 morphological females, 20 samples (50%) were genotypical male. In conclusion, morphological method might be inadequate for sex determination of ancient bones. The improved amelogenin PCR method will be useful in sex determination of ancient bones.

Immunohistochemical Study on the Distribution of Glycogen Synthase Kinase (GSK) 3beta in the Central Nervous System of SOD1G93A Transgenic Mice / 체질인류학회지

In the present study, we investigated influences of glycogen synthase kinase (GSK) 3beta on the development and/or progression of amyotrophic lateral sclerosis (ALS). We used transgenic mice expressing a human Cu/Zn superoxide dismutase mutant (SOD1G93A) as an in vivo model of ALS and examined expressional changes of GSK3beta immunohistochemically in the spinal cord, brain stem and cerebellum. With these experiments we demonstrate that the neurons in these regions of symptomatic SOD1G93A transgenic mice showed increased GSK3beta immunoreactivities compared with wild-type SOD1 transgenic mice. In contrast to symptomatic SOD1G93A transgenic mice, few GSK3beta immunoreactivity changes were detected in 8w- and 13w-old presymptomatic SOD1G93A transgenic mice. These data suggest the possibility that GSK3 functions as a modulating factor of apoptosis-related alterations in ALS and that GSK3beta exert differential functions in the development and/or progression of ALS. But the exact functional significances of these changes require further elucidation.

Changes of Tight Junction and Epithelial Cells after Treatment of Cadmium / 체질인류학회지

Cadmium (Cd) affects cell proliferation, differentiation, apoptosis and other cellular activities and can cause numerous molecular lesions that would be relevant to carcinogenesis. The mechanism of adverse effects of Cd has been poorly understood and, especially on the tight junction. Since there is rare information about the effect of Cd on tight junction protein, we here investigated whether Cd can alter the localization of the proteins. This study examined Cd effects on of tight junction (occludin, ZO-1, and ZO-2) using MDCK cell culture. The change of MDCK cell and tight junction was investigated after treatment of cadmium with phase contrast microscopy, TEER, cell viability, Transmission electron microscopy and confocal laser microscopy. After treatment of cadmium, transendothelial electrical resistance decreased with time and concentration dependent manner. AlamarBlue assay revealed that decreased cell viability also decreased with time and concentration dependent manner. The tight junction moved down between intercellular spaces with decreased density and the cellular thickness around cell junctions decreased with increasing concentration and exposure time of CdCl2. The MDCK cells eventually showed cell death with. Confocal laser microscopy revealed that immunofluorescent reaction of occludin, ZO-1 and ZO-2 decreased. Occludin, ZO-1 and ZO-2 were disrupted at tight junction. These data suggest that after treatment of Cd, increased permeability of MDCK cell monolayer increased. This might be accompanied with disruption of occludin, ZO-1 and ZO-2.

Protective Effect of Fibroin BF-7 on Neuronal Cell Death in Alzheimer Model using Amyloid beta Peptide / 대한해부학회지

Factors such as senescence,stress and neurodegenerative diseases,including Alzheimer's disease, contribute to the impairment of organs,especially the brain.They also negatively affect normal brain functions such as memory and cognition.In this study,the neuroprotective role of the natural product BF-7 was examined against A beta - induced neurotoxicity in SK-N-SH human neuronal cells.BF-7 significantly attenuated A beta-induced apoptosis as measured by intracellular calcium levels,accumulation of reactive oxygen species,mitochondrial dysfunction,and caspase activity.These results strongly indicate that BF-7 plays an effective and positive role in the improvement of brain functions,including learning and memory,in our model system for Alzheimer's disease.Thus,BF-7 might be useful for developing strategies to protect the nervous system and improve brain function.

Alpha-tocopherol Prevents H2O2-induced Tight Junction Occludin Disruption in Blood-Brain Barrier / 체질인류학회지

Vitamin E is the most important lipid-soluble antioxidant in humans. Although alpha-tocopherol is suggested that it has protective effect from many diseases, little is known about the prevention of occludin alteration in tight junction of blood-brain barrier (BBB) under pathologic insults producing reactive oxygen species (ROSs). In this study, the effects of alpha-tocopherol on H2O2-induced tight junction occludin were studied. Primary culture of rat brain microvessel endothelial cells was investigated with confocal microscopy, Western blot, and cell viability assay. Alpha-tocopherol had no apparent cytotoxicity up to 2.8 mM. The preincubation with alpha-tocopherol suppressed the H2O2-induced cytotoxicity in Alamar Blue assay and phase contrast microscopy. In confocal laser microscopy and Western blot, H2O2-induced loss of occludin was suppressed by preincubation with alpha-tocopherol. The present findings provide evidence that alpha-tocopherol may be beneficial for cellular protection from pathologic insults. Since alpha-tocopherol was demonstrated to have far fewer adverse effects, it would become a noteworthy nutrient or drug for the treatment of neurodegenerative diseases.

Study on the Mitochondrial Dysfunction by p53 Regulation in Ceramide-induced Neuronal Cell Death / 체질인류학회지

Ceramide induces cell death in a dose- and time-dependent manner in neuroblastoma SK-N-SH cells. To investigate the mechanism of SK-N-SH cell death by C2-ceramide, morphological features and Hoechst 33258 staining were analyzed. In these morphlogic study the cell death by ceramide showed typical apoptotic features, nuclear condensation, fragmentation, and membrane blebbing. Ceramide-induced apoptosis was accompanied by nuclear accumulation of p53. Inhibition of p53 expression with p53 antisense oligonucleotides inhibited apoptosis evoked by ceramide. Also, ceramide induced mitochondrial event, collapse of mitochondrial membrane potential (delta psi m) and interestingly, inhibition of p53 attenuated collapse of mitochondrial membrane potential, suggests that ceramide induces mitochondrial dysfunction through upregulation of p53 expression. These results suggest that ceramide-induced apoptosis is dependent upon increase in cellular p53 levels which play a critical role in the regulation of apoptotic cell death and p53 modulates mitochondrial function such as mitochondrial membrane potential level.

The Effect of BF-7 on the Ischemia-induced Learning and Memory Deficits / 대한해부학회지

Abnormal blood supply to brain, such as ischemia induce neuronal damages, possibly leading to dementia. Such damages should be attenuated by neuroprotective materials which make neurons tolerable against limited blood supply or reinforce of blood. For the animal study, transient middle cerebral artery occlusion was operated with SD rat. To check whether BF-7 attenuated the ischemic damage, BF-7 (10 mg/kg) was oral treated for 7 days once a day. To evaluate the learning and memory of the rat, 8-arm maze test was conducted. For clinical study, Rey Complex Figure Test (RCFT) was used with control group (32 person), 200 mg treated group (33 person) and 400 mg treated group (34 person). Treatment of BF-7 greatly reduced the infarct size. Also the neuronal damages in the hippocampus were significantly diminished. Furthermore, The memory impairment by ischemia was recovered. The clinical test showed that BF-7 greatly enhanced the brain function recognizing and memorizing complex two dimensional figures. Our results implicated that BF-7 plays a positive roles on protecting brain and enhancing brain unction clinically.

Immunohistochemical Study on the Distribution of the Voltage-gated Ion Channels in Gerbil Cerebellum / 체질인류학회지

There is growing evidence that alterations in Ca2+ homeostasis may play a role in processes of brain aging and neurodegeneration. However, few have focused on voltage-gated Ca2+ channel (VGCC) subunits, much less on expression of other voltage-gated ion channels, i.e. voltage-gated K+ (Kv) and Na+ (Nav) channels. In the present study, we have investigated the spatial patterning of VGCCs, Kv1 and Nav channels by immunohistochemistry. This study have shown clearly that the VGCCs, Kv1 and Nav channels have differential distribution in the cerebellum of gerbil, which is used as an ischemia and epilepsy animal model. Immunoreactivities for Cav2.1, Cav1.2 and Cav1.3 were observed in the cell bodies and dendritic branches of Purkinje cells. In particular, Cav1.3 immunoreactivity was most prominent in the cell bodies and dendritic arborizations. A distinct band of punctate immunoreactivity for the Cav2.1, Cav2.2, Cav1.2 and Cav1.3 were observed in cerebellar nuclei. Strong immunoreactivities for Kv1.3, Kv1.4, Kv1.5 and Kv1.6 were observed in the Purkinje cell bodies, whereas Kv1.2 immunoreactivity was found in the basket cell axon plexus and terminal regions around the Purkinje cells. In the cerebellar nuclei, Kv1.2, Kv1.4 and Kv1.6 proteins were clearly detected in the soma of cerebellar output neurons. The most intense staining for Nav1.1 was observed in the granular layer, whereas strong immunoreactivity for Nav1.2 were seen in the Purkinje cell bodies, and extended into their dendrites. The overall results have demonstrated the expression patterns of VGCCs, Kv1 and Nav channels in gerbil cerebellum. Further studies are needed to define changes in other Ca2+ channel types to determine whether any channel changes represent selective loss of specific receptors or of cell loss, and to determine whether changes in Kv and Nav channels are linked to Ca2+ channel changes.

Protective Effect of Green Tea Extract on Neuronal Cell Death in Alzheimer Model using beta-amyloid Peptide / 대한해부학회지

In the present study, we determined whether green tea extract, EGCG protected against beta-amyloid induced neurotoxicity and learning impairment in vitro and in vivo models. Incubation of SK-N-SH cells with Abeta induced activation of caspase-3, mitochondrial dysfunction such as collapse of mitochondrial membrane potential (MMP) and release of cytosolic cytochrome c. Interestingly, pre-treatment of EGCG reduced significantly activation of caspase-3 and increase of mitochondrial damage such as the breakdown of MMP and release of cytochrome c, eventually attenuated the cell death induced by Abeta. These results show that EGCG inhibited caspase activity and blocked mitochondrial damage. For in vivo experiment, ICR mice received vehicle or vehicle plus EGCG (10 mg/kg) i.p. for 3 days. Before 2 days of treatment, 5 microliter of PBS containing 8 nmol of Abeta1-42 were injected into the lateral ventricle. On the 14th day of treatment, animals were applied to passive avoidance test, and after behavial test, animals were sacrificed. Then, morphological techniques were used to determine the extent of neuronal degeneration in the hippocampus. Abeta, but not PBS, injections into hippocampus led to neuronal loss and evidence of widespread apoptosis. EGCG treated animals had significant reductions in the amount of neuronal degeneration, and TUNEL positive cells compared with Abeta alone treated animals. These data suggest that EGCG at therapeutically relevant concentrations, might protect against neuronal degeneration induced by Abeta.

The Activation of ERK1/2 Via Tyrosine Kinase Pathway Attenuates TRAIL-induced Apoptosis in HeLa cell / 체질인류학회지

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) serves as an extracellular signal triggering apoptosis in tumor cells. To characterize the molecular events involved in TRAIL-induced apoptotic signaling, we investigated the role of extracellular signal-regulated kinase 1/2(ERK1/2) in the apoptosis using HeLa cells. Here we show that TRAIL pronounced ERK1/2 activation through a tyrosine kinase-dependent mechanism, subsequently elevated anti-apoptotic Bcl-2 protein levels. Pretreatment with Genistein, an inhibitor of tyrosine kinase, significantly attenuated ERK1/2 activation and enhanced cell death. Moreover, inhibition of ERK1/2 with PD98059 promoted apoptotic cell death through the down-regulation of ERK1/2 activity and Bcl-2 protein levels. Taken together, our results suggest that the activation of ERK1/2 via tyrosine kinase pathway plays a protective role as the mechanism of cellular defense through the up-regulation of Bcl-2 protein levels in TRAIL-induced apoptosis.

Down-regulated Reactive Oxygen Species by Heat Shock Protein 90 in 3-Hydroxykynurenine-induced SKN-SH Cell Death / 체질인류학회지

In this present study, we show that 3HK-induced reactive oxygen species (ROS) accumulation and caspase activation lead to apoptotic cell death. Pretreatment with N-acetylcysteine (NAC), an effective antioxidant, significantly attenuated 3HK-induced apoptosis by way of a reduction of ROS accumulation and caspase activity. SKN-SN cells were protected from 3HK-induced cytotoxicity by heat shock protein (HSP). HSP90 effectively attenuated 3HK-mediated ROS accumulation and apoptosis. In addition, the protective effect of HSP90 was abolished by pretreatment with HSP90 anti-sense oligonucleotide, but not when pretreated with anti-senses for other HSPs. These results suggest that HSP90 protects SKN-SH cells from 3HK-induced cytotoxicity by reducing ROS levels and caspase activity.