Immunohistochemical Localization of Calbindin D28k, Parvalbumin, and Calretinin in the Superior Olivary Complex of Circling Mice.

The circling mouse serves as a hearing loss model. It has spontaneous tmie gene mutations that cause hair cell and cochlear degeneration. However, little is known about the role of the tmie gene in superior olivary complex (SOC) regions, in which sound information from the two ears is integrated and primarily relayed to the nuclei of the lateral lemniscus and inferior colliculus. Several studies have reported that abnormal calcium (Ca2+) homeostasis is associated with the pathology of hearing loss. This study investigated the distribution of Ca2+-binding proteins (CaBPs), such as calbindin D28k, parvalbumin, and calretinin, in the SOC of the circling mouse on postnatal day 16. A comparison of wild-type (+/+), heterozygous (+/cir), and homozygous (cir/cir) mice showed that CaBP immunoreactivity was significantly decreased in the auditory nucleus of the SOC of homozygous (cir/cir) mice. A decline in the CaBPs level in the SOC may be the result of hearing loss through hair cell and cochlear degeneration following tmie gene mutation.

Calcified Pulmonary Nodules Identified in a 350-Year-Old-Joseon Mummy: the First Report on Ancient Pulmonary Tuberculosis from Archaeologically Obtained Pre-modern Korean Samples.

We found calcified pulmonary nodules in a middle-aged female mummy discovered from 350-yr-old Joseon tomb of Korea. In the CT scan, we found six radiopaque nodules in right lung, through the levels of thoracic vertebrae 1 to 6. We also found presumptive pleural adhesions in right thoracic cavity of CT images. We re-confirmed radiological findings by our post-factum dissection on the same mummy. By the differential diagnosis, we speculate that the radiopaque calcification nodules and associated pleural adhesion could have been caused by tuberculosis. This is the first-ever report on the pulmonary tuberculosis identified in archaeologically obtained, pre-modern Korean samples.

Detection Trend of Helminth Eggs in the Strata Soil Samples from Ancient Historic Places of Korea.

For several years, we have conducted a series of studies on the patterns of ancient parasitism prevailing in the soil of rural and urban areas of past Kingdom of Korea. Actually, during our survey of paleoparasitology in archaeological sites of Korean peninsula, numerous ancient parasite eggs were discovered in the samples from the city districts of Hansung (Joseon) and Buyeo (Baikje), the palace moat at Gyeongju (Silla), shell-midden site at Bonghwang-dong (Silla to Joseon), and the reservoir found in Hwawangsansung fortress (Silla). By the paleoparasitological studies, with respect to parasitism in the high-density populations of ancient towns and cities, we have managed to catch glimpses of the patterns prevalent therein: a serious parasitic contamination of the soil in ancient urban areas, but not in rural areas of the past. Our historical research also proposed the plausible mechanism of parasite infection very serious indeed among urban populations in Korean history. Although city dwelling doubtless has accrued significant benefits for people and populations with agriculture, it can be equally supposed that living in such highly populated areas might have facilitated the spread of parasite infection.

V-shaped pits in regions of ancient Baekje kingdom paleoparasitologically confirmed as likely human-waste reservoirs.

In a paleo-parasitological analysis of soil samples obtained from V-shaped pits dating to the ancient Baekje period in Korean history, we discovered Ascaris lumbricoides, Trichuris trichiura, and Clonorchis sinensis eggs. In light of the samples' seriously contaminated state, the V-shaped pits might have served as toilets, cesspits, or dung heaps. For a long period of time, researchers scouring archaeological sites in Korea have had difficulties locating such structures. In this context then, the present report is unique because similar kind of the ancient ruins must become an ideal resource for successful sampling in our forthcoming paleoparasitological studies.

Tracing hepatitis B virus to the 16th century in a Korean mummy.

UNLABELLED: A rare find of a mummified child from the 16th century AD, in Korea, with relatively preserved organs, enabled a search for ancient hepatitis B virus (aHBV) DNA sequences from laparoscopic-derived liver biopsies. Analysis of the complete aHBV genome (3,215 base pairs) revealed a unique HBV genotype C2 (HBV/C2) sequence commonly spread in Southeast Asia, which probably represents an HBV that infected the Joseon Dynasty population in Korea. Comparison of the aHBV sequences with contemporary HBV/C2 DNA sequences revealed distinctive differences along four open reading frames. Genetic diversity between contemporary and recovered aHBV/C2 DNA may be the result of immunologic, environmental, and/or pharmacologic pressures. The calculated time of most recent common ancestor suggests that the Korean HBV sequence origin dates back at least 3,000 years and possibly as long as 100,000 years. This isolate most likely represents the earliest human HBV sequence that colonized Southeast Asia by human migration. CONCLUSION: This study describes the complete sequence of the oldest HBV isolate and the most ancient full viral genome known so far.

Revealing Joseon period People's single nucleotide polymorphism associated with lactase gene by ancient DNA analysis of human remains from archaeological sites in Korea.

Lactase non-persistence (LNP), one of the causes of lactose intolerance, is related to lactase gene associated single nucleotide polymorphisms (SNPs). Since the frequency of LNP varies by ethnic group and country, the research to reveal the presence or absence of LNP for specific people has been conducted worldwide. However, in East Asia, the study of lactase gene associated SNPs have not been sufficiently examined so far using ancient human specimens from archaeological sites. In our study of Joseon period human remains (n=14), we successfully revealed genetic information of lactase gene associated SNPs (rs1679771596, rs41525747, rs4988236, rs4988235, rs41380347, rs869051967, rs145946881 and rs182549), further confirming that as for eight SNPs, the pre-modern Korean people had a lactase non-persistent genotype. Our report contributes to the establishment of LNP associated SNP analysis technique that can be useful in forthcoming studies on human bones and mummy samples from East Asian archaeological sites.

Effect of the calcineurin inhibitor FK506 on K+-Cl- cotransporter 2 expression in the mouse hippocampus after kainic acid-induced status epilepticus.

Calcineurin (CaN)-mediated excitotoxicity impairs γ-aminobutyric acid (GABA) transmission and induces neuronal apoptosis. Ca(2+)-dependent K(+)-Cl(-) cotransporter 2 (KCC2) participates in GABAergic inhibitory transmission. However, the mechanism by which CaN mediates GABA receptor-mediated KCC2 in seizures is not fully understood. In the present study, we investigated the altered expression of KCC2 and the effects of the CaN inhibitor FK506 on KCC2 expression in the mouse hippocampus following kainic acid (KA) treatment. FK506 was injected twice 24 h and 30 min before KA treatment and then mice were treated with KA and killed 2 days later. FK506 had anticonvulsant effect on KA-induced seizure activities. CaN cleavage was evident in the hippocampus 24 h after KA treatment. FK506 pretreatment blocked the truncation of CaN in the KA-treated hippocampus. Cresyl violet and TUNEL staining showed that FK506 prevented KA-induced hippocampal cell death. In particular, Western blot analysis showed that KCC2 expression was time dependent, with a peak at 6 h and a return to decreased levels at 48 h, whereas FK506 pretreatment inhibited the KA-induced decrease in KCC2 expression in the hippocampus. Immunofluorescence showed that FK506 pretreatment protected the loss of inhibitory GABAergic KCC2-expressing neurons following KA treatment. Taken together, these results provide evidence that altered KCC2 expression may be associated with Ca(2+)-mediated seizure activity and indicate that neuron-specific KCC2 may be involved in neuroprotection after seizures.

Tendon impingement of the extensor digiti minimi: clinical cases series and cadaveric study.

The authors describe two unique clinical cases of closed extensor digiti minimi (EDM) tendon injuries after hyperflexion of the wrist with full finger flexion and one case of chronic tenosynovitis around the EDM tendon. All three cases were thought to be related to the bifurcation of the EDM tendon and synovial septum. Subsequently, variations in EDM tendons were investigated in 49 cadaveric hands with a focus on patterns of tendon bifurcation and their relationships with the surrounding synovial sheath. The EDM tendon was found to be bifurcated in 74% (n = 36) of hands and all of these hands contained a synovial septum. In 9 (25%) hands, the EDM tendon bifurcated proximal to the retinaculum, in 15 (42%), it bifurcated distal to the retinaculum, and in the other 12 hands (33%), the tendon bifurcated at the retinacular level. In 6 of the 15 hands with an infraretinacular bifurcation, the tendon was found to impinge on the synovial septum during passive flexion of the wrist with full finger flexion, and the mean distance between the synovial septum and the bifurcation point in these specimens was 0.6 cm (range, 0.4-0.7 cm), which was differed significantly from hands not showing impingement (P = 0.01). This study shows that distal bifurcation of the EDM tendon may lead to tendon impingement on the septum and suggests that this is a potential etiology of chronic tenosynovitis of the fifth compartment and of acute closed tendon injuries.

GENETIC ANALYSIS OF ANCIENT CLONORCHIS SINENSIS EGGS ATTAINED FROM GORYEONG MUMMY OF JOSEON DYNASTY PERIOD.

Although Clonorchis sinensis is a parasite that still infects many people in East Asia, its genetics remain largely unknown. We conducted ancient DNA analysis of C. sinensis eggs obtained from a Joseon period mummy newly discovered in South Korea. Clonorchis sinensis DNA was amplified for internal transcribed spacer 1, cytochrome c oxidase subunit 1, and NADH dehydrogenase subunit 2 and 5 genes. The results of BLAST/NCBI showed that the consensus sequences were 98.24 to 100% identical to the modern and ancient C. sinensis sequences reported from Korea, China, Japan, and other Asian countries. Our report helps to fill in the genetic profile of ancient C. sinensis strains that infected East Asian people hundreds of years ago.

A portrait drawing of the 17th century Korean scholar based on craniofacial reconstruction.

As a technique mainly hiring in forensic investigation field to identify the descents, craniofacial reconstruction (CFR) is also used in archaeology to create the faces from ancient or medieval human remains, when there is little information about his/her appearance. Eung-Cheok Ko (1531-1605) was a writer and scholar in the mid Joseon period. In January of 2019, His mummified body was found at Gumi, Kyeonsangbuk-do, Korea. The remains were anthropologically examined, and archaeological CFR was also requested for this case. This report reveals the case's facial reconstruction process and his portrait that is drawn based on the 3-dimensional CFR result.

Revisiting diaphragmatic hernia of Joseon period Korean mummy by three-dimensional liver and heart segmentation and model reconstruction.

A three-dimensional (3D) segmentation and model reconstruction is a specialized tool to reveal spatial interrelationship between multiple internal organs by generating images without overlapping structures. This technique can also be applicable to mummy studies, but related reports have so far been very rare. In this study, we applied 3D segmentation and model reconstruction to computed tomography images of a Korean mummy with congenital diaphragmatic hernia. As originally revealed by the autopsy in 2013, the current 3D reconstruction reveals that the mummy's heart is shifted to the left due to the liver pushing up to thoracic cavity thorough diaphragmatic hernial defect. We can generate 3D images by calling up the data exclusively from mummy's target organs, thus minimizing the confusion of diagnosis that could be caused by overlapping organs.

Glutamate co-transmission from developing medial nucleus of the trapezoid body--lateral superior olive synapses is cochlear dependent in kanamycin-treated rats.

Cochlear dependency of glutamate co-transmission at the medial nucleus of the trapezoid body (MNTB)--the lateral superior olive (LSO) synapses was investigated using developing rats treated with high dose kanamycin. Rats were treated with kanamycin from postnatal day (P) 3 to P8. A scanning electron microscopic study on P9 demonstrated partial cochlear hair cell damage. A whole cell voltage clamp experiment demonstrated the increased glutamatergic portion of postsynaptic currents (PSCs) elicited by MNTB stimulation in P9-P11 kanamycin-treated rats. The enhanced VGLUT3 immunoreactivities (IRs) in kanamycin-treated rats and asymmetric VGLUT3 IRs in the LSO of unilaterally cochlear ablated rats supported the electrophysiologic data. Taken together, it is concluded that glutamate co-transmission is cochlear-dependent and enhanced glutamate co-transmission in kanamycin-treated rats is induced by partial cochlear damage.

A Prospective Validation and Observer Performance Study of a Deep Learning Algorithm for Pathologic Diagnosis of Gastric Tumors in Endoscopic Biopsies.

PURPOSE: Gastric cancer remains the leading cause of cancer-related deaths in Northeast Asia. Population-based endoscopic screenings in the region have yielded successful results in early detection of gastric tumors. Endoscopic screening rates are continuously increasing, and there is a need for an automatic computerized diagnostic system to reduce the diagnostic burden. In this study, we developed an algorithm to classify gastric epithelial tumors automatically and assessed its performance in a large series of gastric biopsies and its benefits as an assistance tool. EXPERIMENTAL DESIGN: Using 2,434 whole-slide images, we developed an algorithm based on convolutional neural networks to classify a gastric biopsy image into one of three categories: negative for dysplasia (NFD), tubular adenoma, or carcinoma. The performance of the algorithm was evaluated by using 7,440 biopsy specimens collected prospectively. The impact of algorithm-assisted diagnosis was assessed by six pathologists using 150 gastric biopsy cases. RESULTS: Diagnostic performance evaluated by the AUROC curve in the prospective study was 0.9790 for two-tier classification: negative (NFD) versus positive (all cases except NFD). When limited to epithelial tumors, the sensitivity and specificity were 1.000 and 0.9749. Algorithm-assisted digital image viewer (DV) resulted in 47% reduction in review time per image compared with DV only and 58% decrease to microscopy. CONCLUSIONS: Our algorithm has demonstrated high accuracy in classifying epithelial tumors and its benefits as an assistance tool, which can serve as a potential screening aid system in diagnosing gastric biopsy specimens.

Magnetic resonance imaging performed on a hydrated mummy of medieval Korea.

Previous investigations have shown that magnetic resonance imaging (MRI) can be employed as an efficient non-invasive diagnostic tool in studies on Egyptian mummies. MRI, moreover, because it produces especially clear images of well-hydrated tissue, could be a particularly effective diagnostic option for mummies that still retain humidity within tissues or organs. Therefore, in the present study, we tested MRI on a 17th century mummy, one of the most perfectly preserved 'hydrated mummies' ever found in Korea, in order to determine the quality of images that could be obtained. We found that the diagnostic value of an MRI scan of the hydrated mummy was not inferior to that of a computed tomography scan. The T1- and T2-weighted magnetic resonance (MR) signals showed unique patterns not easily obtained by computed tomography, the resultant MR images revealing the organ specificities clearly. Overall, the quality of the MR images from the hydrated mummy was superb and the scientific value of MRI in the study of hydrated mummies should not be underestimated.

Glutamatergic transmission is sustained at a later period of development of medial nucleus of the trapezoid body-lateral superior olive synapses in circling mice.

Synaptic transmission between the medial nucleus of the trapezoid body (MNTB) and the lateral superior olive (LSO) was investigated in circling mice, an animal model for inherited deafness, using the voltage-clamp technique. In postnatal day 9 (P9) approximately P11 homozygous (cir/cir) circling mice, perfusion with 10 microM DL-APV and 10 microM CNQX reduced the 10 min average of postsynaptic currents (PSCs) to 8.8 +/- 3.0% compared with controls (n = 6). In heterozygous (+/cir) mice in the same age range, the 10 min PSCs average was reduced to 87.5 +/- 3.7% compared with controls (n = 7). In P0 approximately P2 homozygous (cir/cir) and heterozygous (+/cir) mice, the 10 min PSCs averages were 11.0 +/- 2.6% (n = 9) and 84.1 +/- 4.6% (n = 11), respectively. The effects of a glutamate antagonist mixture were almost the same in single fiber stimulation of P9 approximately P11 mice, reducing mean PSCs to 5.2 +/- 3.1% (homozygous (cir/cir) mice, n = 8) and 78.3 +/- 4.3% (heterozygous (+/cir) mice, n = 12). Immunohistochemical study revealed that glycine receptor (GlyR) immunoreactivity in heterozygous (+/cir) mice was more prominent than in homozygous (cir/cir) mice, while immunoreactivities of NR1 and NR2A-type NMDAR of P16 homozygous (cir/cir) mice were more prominent than in heterozygous (+/cir) mice of the same age. No significant difference was found in the immunoreactivity of NR2B-type NMDAR. These results indicate that glutamatergic transmission is sustained at a later period of developing MNTB-LSO synapses in homozygous (cir/cir) mice.

Scanning electron microscope study of ancient parasite eggs recovered from Korean mummies of the Joseon Dynasty.

We have previously shown that parasite eggs have been identified in the coprolites of Korean mummies. These eggs have shed light on parasitic infection patterns in Korean populations living several hundred years ago. We conducted a scanning electron microscopy (SEM) study on ancient Trichuris trichiura, Ascaris lumbricoides, Metagonimus yokogawai, Paragonimus westermani, and Gymnophalloides seoi eggs recovered from Korean mummies of the Joseon Dynasty. We anticipated that the taphonomic conditions of mummification would alter the eggs of certain species but not of others. Our SEM data show that each species of ancient egg exhibited different degrees of preservation. Thus, some of them, for example, M. yokogawai, exhibited a better preservation status than others, suggesting that they should be the first candidates considered when choosing subjects for future paleoparasitological studies.

The potential for non-invasive study of mummies: validation of the use of computerized tomography by post factum dissection and histological examination of a 17th century female Korean mummy.

The socio-cultural antipathies of some descendants with regard to invasive examinations of age-old human remains make permission for dissection of Korean mummies of the Joseon Dynasty (1392-1910) difficult to obtain. Overcoming this obstacle necessitated the use of non-invasive techniques, such as multi-detector computerized tomography (MDCT) and endoscopic examination, enabling determination of the preservation status of internal organs of mummies without significantly damaging the mummies themselves. However, MDCT alone cannot clearly differentiate specific mummified organs. Therefore, in much the same way as diagnostic radiologists make their MDCT readings on living patients more reliable by means of comparison with accumulated post-factum data from autopsies or histological studies, examinations of mummies by invasive techniques should not be decried as mere destruction of age-old human remains. Rather, providing that due permission from descendants and/or other relevant authorities can be obtained, dissection and histological examination should be performed whenever opportunities arise. Therefore, in this study, we compared the radiological data acquired from a 17th century mummy with our dissection results for the same subject. As accumulation of this kind of data could be very crucial for correct interpretation of MDCT findings on Korean mummies, we will perform similar trials on other Korean mummies found in forthcoming days if conditions permit.

Gymnophalloides seoi eggs from the stool of a 17th century female mummy found in Hadong, Republic of Korea.

It was previously reported that paleoparasitological clues for parasites infecting humans could be found in the feces of mummies of the Joseon Dynasty (1392-1910) in the Republic of Korea. Here, we report the presence of trematode eggs, including Clonorchis sinensis, Metagonimus yokogawai, and Gymnophalloides seoi (a human parasite known in Korea since 1993) in the feces of a recently excavated female mummy in Hadong, Republic of Korea. This is the first report of the discovery of a G. seoi infection in a human mummy. Since Hadong is currently not an endemic area for G. seoi, we speculate that the parasite might have occurred frequently along coastal areas of the Korean peninsula several hundred years ago and that the endemic areas contracted to, more or less, restricted regions since that time.

Lumbosacral Defects in a 16th-18th-Century Joseon Dynasty Skeletal Series from Korea.

Paleopathological evidence for congenital and degenerative disorders of the lumbosacral vertebrae is informative about ancient individual lifeways and physical conditions. However, very few studies have focused on the paleopathology of the lumbosacral vertebrae in ancient skeletal series from East Asia. One reason for the lack of studies is that skeletal samples from East Asia are typically insufficient in size to represent populations for comparative studies within the continent. Here, we present the first comprehensive analysis of lumbosacral defects in an East Asian human skeletal sample, examining occurrences of spina bifida occulta (SBO), lumbosacral transitional vertebrae (LSTV), and spondylolysis in remains from Joseon tombs dating to the 16-18th centuries in Korea. In this study, we present an alternative methodology for understanding activities of daily life among ancient Koreans through paleopathological analysis.

Post-factum autopsy to confirm cardiac structures visible on computed tomography images of Korean mummies: the radiological basis of paleo-cardiology.

Accurate interpretation of radiological data is important for reliable paleopathological study of mummies. This is especially true for the mummified heart, an anatomically complicated organ that is distorted and displaced due to long-term dehydration and the action of gravity. In the present study, we compared post-factum autopsy results for mummified hearts of differing preservation statuses with corresponding radiological (computed tomography [CT]-image) findings in order to obtain information necessary for accurate radiological diagnosis. We found that the valvular apparatus (especially the aortic valve and chordae tendinae) was easily distinguishable on the CT images of mummies in which more cardiac structures were preserved. We also identified several situations that are known to incur misdiagnosis of cardiac CT images: the presence of pseudo-cavities in the cardiac wall, confusion of the valvular apparatus with cardiac-wall debris, and invisibility of cardiac structures on CT images due to collapse or adhesion. While acknowledging the merits of top-priority CT scanning in non-invasive research, post-factum autopsy also has value as a complementary and confirmatory analysis for enhanced accuracy of diagnoses in paleo-radiological studies.