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

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.

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

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.

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

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.

Alterations in Spontaneous Movement, Corticosterone, and Cytokines in Mice Exposed to 835 MHz Radiofrequency Radiation / 체질인류학회지

Although commercialization of mobile phones has raised much concerns about the effects of radiofrequency radiation on the human body, few experimental studies have been conducted on the effects of radiofrequency radiation on physiological homeostasis, immune and inflammatory responses. Therefore, we presently investigated the effect of 835 MHz radiofrequency radiation on spontaneous wheel exercise, hormone and cytokines levels in the plasm of mice. Mice were divided into 4 groups as control, exercise, radiofrequency radiation, radiofrequency radiation & exercise group. The body weight, corticosterone and blood cytokine levels were checked for 10 weeks. Followed by the exposure to radiofrequency radiation for 6 hours a day, the more increase in body weight was observed in the radiofrequency radiation & exercise group than in the spontaneous exercise group. When the amount of spontaneous exercise was measured for 10 weeks, the amount of exercise was increased in the both control and spontaneous exercise group, while the amount of exercise was decreased in the radiofrequency radiation group. To determine whether the homeostasis, immune and inflammatory responses are indirectly affected by radiofrequency radiation exposure, IL-1β, IL-6, IL-12 (p70), TNF-α, IFNγ, and GM-CSF were measured by ELISA kit, respectively. As a result, the blood levels of IL-6, IL-12 (p70) and TNF-α in the spontaneous exercise group were higher than that of control group, and each cytokine levels in the radiofrequency radiation & exercise group were lower than that of control group. However, the corticosterone, IL-1β, IFNγ and GM-CSF didn't show statistically significant differences in all groups. It has been confirmed that exposure to high frequency electromagnetic waves for a long time can affect the amount of exercise, body weight, and some inflammatory cytokines such as IL-6, IL-12 (p70) and TNF-α.

The distribution of calbindin-D28k, parvalbumin, and calretinin immunoreactivity in the inferior colliculus of circling mouse / 대한해부학회지

The circling mice with tmie gene mutation are known as an animal deafness model, which showed hyperactive circling movement. Recently, the reinvestigation of circling mouse was performed to check the inner ear pathology as a main lesion of early hearing loss. In this trial, the inner ear organs were not so damaged to cause the hearing deficit of circling (cir/cir) mouse at 18 postnatal day (P18) though auditory brainstem response data indicated hearing loss of cir/cir mice at P18. Thus, another mechanism may be correlated with the early hearing loss of cir/cir mice at P18. Hearing loss in the early life can disrupt the ascending and descending information to inferior colliculus (IC) as integration site. There were many reports that hearing loss could result in the changes in Ca²⁺ concentration by either cochlear ablation or genetic defect. However, little was known to be reported about the correlation between the pathology of IC and Ca²⁺ changes in circling mice. Therefore, the present study investigated the distribution of calcium-binding proteins (CaBPs), calbindin-D28k, parvalbumin, and calretinin immunoreactivity (IR) in the IC to compare among wild-type (+/+), heterozygous (+/cir), and homozygous (cir/cir) mice by immunohistochemistry. The decreases of CaBPs IR in cir/cir were statistically significant in the neurons as well as neuropil of IC. Thus, this study proposed overall distributional alteration of CaBPs IR in the IC caused by early hearing defect and might be helpful to elucidate the pathology of central auditory disorder related with Ca²⁺ metabolism.

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.

Bone tumors in pre-modern skulls from human skeletal series of Joseon Dynasty / 대한해부학회지

To date, there are still very few reports on benign-tumor cases based on East Asian skeletal series, even though other regions and continents have been well represented. In our study on the Joseon Human Skeletal Series, we identified benign bone tumors in two skeletons (cases Nos. 75 and 96). Our radiological analyses showed both cases to be homogeneous sclerotic bone masses aligned with the cranial vault suture. In a subsequent series of differential diagnoses, we determined both cases to be osteoma, the most common bone-tumor type reported for archaeological samples. Our study is the osteoarchaeological basis for this, the first-ever report on benign bone neoplasm in a pre-modern East Asian population.

Harris lines observed in human skeletons of Joseon Dynasty, Korea / 대한해부학회지

The Harris line (HL), caused by bone-growth arrest and manifesting on X-rays as a radiopaque transverse line in the metaphysis of the long bones, is an indicator reflecting stress conditions such as disease or malnutrition. HL frequency has been assumed to differ between pre-modern and modern societies, as reflective of increased caloric intake and overall nutritional improvements attendant on industrialization. To determine if such a change occurred in Korea, in the present study we compared the respective HL statuses in medieval Joseon and modern Korean population samples. HLs were found in 39.4% (28/71) of the Joseon Koreans. Whereas only 27.5% (11/40) of the males showed an HL, fully 54.8% (17/31) of the females exhibited it. Notably, HLs were observed in only 16.4% (35/213) of the modern Koreans; more remarkably still, the HL rate was almost the same between the sexes, 16.7% (20/120) for the males and 16.1% (15/93) for the females. The HL frequency was much higher in the Joseon Koreans than in their modern counterparts, reflecting the improvement of nutritional status that had been achieved in the course of South Korea's modernization. This HL-frequency decrease was much more obvious in the female populations. The higher HL frequency among the Joseon females might reflect the relatively poor nutritional condition of females in pre-modern Korean society.

Forensic Anthropological Study on Saw Marks Appearing on the Tibiae of a Joseon Skeleton / 체질인류학회지

Tomb with lime-soil mixture barrier (LSMB) was constructed by the people from upper class people of Joseon Dynasty. The coffin of LSMB was surrounded by hard concrete barriers, being successfully protected from outside invasions until the archaeological excavation begins. The human remains were extremely preserved well, providing important information on the health and illness of the people of Joseon dynasty. Recent investigation into human skeletons from LSMB in Yong-in city was another forensic anthropological case that was very meaningful to our research. During the examination on the elderly Joseon female bones, we discovered unusual saw-marks on the shaft of both tibiae. We could not find any osteological evidences suggestive of healed bone process. Considering archaeological and anthropological findings altogether, the occurrence time of saw-marks was considered to be perimortem period. However, as for why such a saw mark was made on the tibiae, we did not get any information about it. We expect that this report facilitate other researcher to do explore the usefulness of forensic anthropology examination on the similar human skeleton cases identified in various archaeological ruins.

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.

Evidence of Periostitis in Joseon Dynasty Skeletons / 체질인류학회지

Periostitis is one of the human diseases commonly encountered in archaeological samples. It is known to be an important health indicator for paleopathologists examining skeletal remains. In our recent study on a Joseon skeletal series (n=101), non-specific, primary periostitis was observed only in five individuals (#4, #29, #137, #175, and #290). Notably, there were no secondary periostitis-suggestive signs (e.g. syphilis), except for those caused by fractures (#33 and #41). As this is the inaugural Korean-skeletal-series report on periostitis, the results presented in these pages should prove significant to interested paleopathologists.

Long bone fractures identified in the Joseon Dynasty human skeletons of Korea / 대한해부학회지

Fracture is one of the pathological signs most frequently encountered in archaeologically obtained bones. To expand the paleopathological knowledge on traumatic injuries, it is desirable to secure data on long bone fractures from as wide a geographic and temporal range as possible. We present, for the first time, evidence of long bone fractures in a 16th-18th century Joseon skeletal series (n=96). In this study, we found 3 Colles' fractures of the radius in 2 individual cases. The pattern of fractures was unique. Although previous reports show that the ulna is broken more often than the radius, ulnar fracture associated with fending off a blunt attack was rare in our series (1/7 cases). Transverse fractures, typically caused by intentional violence, were also very rare (1/7 cases) in this study. These results may reflect the relatively tranquil lives of the Joseon people in 16th-18th century Korea. We also found post-fracture complications such as deformations, bone length shortening, and osteomyelitis. The present study would be of interest to medical scientists in related fields because it is one of the few studies conducted on long bone fractures among pre-modern societies in East Asian countries, thus far.

Vertebral osteophyte of pre-modern Korean skeletons from Joseon tombs / 대한해부학회지

Spinal osteophytic changes are known to be affected by differences in age, sex, population, and mechanical stress. We examined Joseon skeletons (n=87) to obtain vertebral osteophytosis data on a pre-modern Korean population. The mean osteophytic value (MOV) of vertebrae increased in the cervical-thoracic-lumbar order. More severe osteophytosis was found in the vertebrae (C5, T9, T10, and L4) farthest from the line of gravity, while the general pattern of vertebral osteophytosis appeared similar to those of previous reports on other skeletal series. More severe osteophytes were much more common in the males, possibly due to their engaging in more strenuous physical labor than that of females. We also observed MOV patterns seemingly unique to the Joseon people, and findings not typically reported in previous studies. Although a full explanation of the factors contributing to vertebral-osteophytic development in Joseon Koreans will require further studies, the present results are meaningful to anatomists and anthropologists interested in osteophytic patterns occurring in an East Asian population.

Genetic Evaluation of ALADIN Gene in Early-Onset Achalasia and Alacrima Patients

BACKGROUND/AIMS: ALADIN gene has been known to cause achalasia, alacrima, adrenal abnormalities and a progressive neurological syndrome. A considerable proportion of achalasia patients has been known to show alacrima (decreased secretion of tear). However, the genetic mechanism between achalasia and alacrima has not been defined yet. We postulated that ALADIN gene may be involved in the occurrence of early-onset achalasia; thus, we investigated the correlation of ALADIN gene in early-onset achalasia patients. METHODS: From 1989 to 2007, patients who were diagnosed as primary achalasia before age 35 were enrolled. All of the enrolled patients were asked for (1) blood sampling for DNA, (2) Shirmer test and (3) dysphagia questionnaires. RESULTS: The ALADIN gene in exon 1, 2, 10, 11 and 12 from 19 patients was investigated (M:F = 12:7). The mean age of patients at diagnosis was 27 +/- 5 (15-35) years old. Eight out of 19 (42%) showed alacrima by the positive Shirmer test. In spite of thorough exam in the genetic study, there was no definite abnormal genetic finding in this study. CONCLUSIONS: A considerable number of achalasia patients showed alacrima. Due to the limitation of this study, it is difficult to conclude that early-onset achalasia may have significant correlations with the ALADIN gene.

Anthropological Study on Human Skeletons from Joseon Tomb, for Confirming Ryu Ja-Gwang, a Famous Historical Figure of Medieval Korea / 체질인류학회지

Anthropological study on human skeletons from Joseon tomb, for confirming Ryu Ja-Gwang, a famous historical figure of medieval Korea. Ryu Ja-Gwang was the famous figure of Joseon Dynasty who achieved great successes in royal court. According to the records in Joseon Dynasty, he tried to hide his own tomb after death under the apprehension that the political enemies might do harm to his corpse. In 1974, the descendants of Younggwang Ryu clan discovered a Joseon tomb that was lost for the past several hundred years. Since they suspected if the tomb might be their legendary ancestor's, anthropological studies were asked to be done on the skeletons collected from the tomb. In our study, the estimated stature of a male from the tomb was 160.5+/-3.8 cm. The age at death was estimated to be 58.6+/-5.2. He seems to have been suffered from diffuse idiopathic skeletal hyperostosis (DISH), the disease prevalent among the people taking high-caloric diets. Since nuclear DNA could not be amplified in this case, the personal identification of the dead person could not be successfully traced by molecular techniques. Taken together, we could not conclude that the skeletons were those of Ryu Ja-Gwang because the radiocarbon dating, age at death or height data could not be well matched with those described in historical documents. Even so, this study could be significant to the related researchers because it was one of the first trials to identify debatable historical figure using various anthropological techniques. Considering that the anthropological studies on famous historical people have been performed briskly, our study could be the basis for the similar studies in Korea, performed in near future.

Decreased Immunoreactivities and Functions of the Chloride Transporters, KCC2 and NKCC1, in the Lateral Superior Olive Neurons of Circling Mice

OBJECTIVES: We tested the possibility of differential expression and function of the potassium-chloride (KCC2) and sodium-potassium-2 chloride (NKCC1) co-transporters in the lateral superior olive (LSO) of heterozygous (+/cir) or homozygous (cir/cir) mice. METHODS: Mice pups aged from postnatal (P) day 9 to 16 were used. Tails from mice were cut for DNA typing. For Immunohistochemical analysis, rabbit polyclonal anti-KCC2 or rabbit polyclonal anti-NKCC1 was used and the density of immunolabelings was evaluated using the NIH image program. For functional analysis, whole cell voltage clamp technique was used in brain stem slices and the changes of reversal potentials were evaluated at various membrane potentials. RESULTS: Immunohistochemical analysis revealed both KCC2 and NKCC1 immunoreactivities were more prominent in heterozygous (+/cir) than homozygous (cir/cir) mice on P day 16. In P9-P12 heterozygous (+/cir) mice, the reversal potential (Egly) of glycine-induced currents was shifted to a more negative potential by 50 microM bumetanide, a known NKCC1 blocker, and the negatively shifted Egly was restored by additional application of 1 mM furosemide, a KCC2 blocker (-58.9+/-2.6 mV to -66.0+/-1.5 mV [bumetanide], -66.0+/-1.5 mV to -59.8+/-2.8 mV [furosemide+bumetanide], n=11). However, only bumetanide was weakly, but significantly effective (-60.1+/-2.9 mV to -62.7+/-2.6 mV [bumetanide], -62.7+/-2.6 mV to -62.1+/-2.5 mV [furosemide+bumetanide], n=7) in P9-P12 homozygous (cir/cir) mice. CONCLUSION: The less prominent immunoreactivities and weak or absent responses to bumetanide or furosemide suggest impaired function or delayed development of both transporters in homozygous (cir/cir) mice.

Decreased Immunoreactivities and Functions of the Chloride Transporters, KCC2 and NKCC1, in the Lateral Superior Olive Neurons of Circling Mice

OBJECTIVES: We tested the possibility of differential expression and function of the potassium-chloride (KCC2) and sodium-potassium-2 chloride (NKCC1) co-transporters in the lateral superior olive (LSO) of heterozygous (+/cir) or homozygous (cir/cir) mice. METHODS: Mice pups aged from postnatal (P) day 9 to 16 were used. Tails from mice were cut for DNA typing. For Immunohistochemical analysis, rabbit polyclonal anti-KCC2 or rabbit polyclonal anti-NKCC1 was used and the density of immunolabelings was evaluated using the NIH image program. For functional analysis, whole cell voltage clamp technique was used in brain stem slices and the changes of reversal potentials were evaluated at various membrane potentials. RESULTS: Immunohistochemical analysis revealed both KCC2 and NKCC1 immunoreactivities were more prominent in heterozygous (+/cir) than homozygous (cir/cir) mice on P day 16. In P9-P12 heterozygous (+/cir) mice, the reversal potential (Egly) of glycine-induced currents was shifted to a more negative potential by 50 microM bumetanide, a known NKCC1 blocker, and the negatively shifted Egly was restored by additional application of 1 mM furosemide, a KCC2 blocker (-58.9+/-2.6 mV to -66.0+/-1.5 mV [bumetanide], -66.0+/-1.5 mV to -59.8+/-2.8 mV [furosemide+bumetanide], n=11). However, only bumetanide was weakly, but significantly effective (-60.1+/-2.9 mV to -62.7+/-2.6 mV [bumetanide], -62.7+/-2.6 mV to -62.1+/-2.5 mV [furosemide+bumetanide], n=7) in P9-P12 homozygous (cir/cir) mice. CONCLUSION: The less prominent immunoreactivities and weak or absent responses to bumetanide or furosemide suggest impaired function or delayed development of both transporters in homozygous (cir/cir) mice.

Anthropological Study on Ancient Human Skull and Teeth Discovered from Urn Coffin of Proto-Three Kingdoms Period in Korea / 체질인류학회지

Increase in agricultural production or steel manufacturing during Proto-Three Kingdoms Period enabled ancient people to establish well-organized ancient kingdoms in Korean peninsula. A pottery in which human skeletons were buried, the urn coffin, has been regarded as one of the frequently used burial system in ancient Korean society. Among them, the small-sized urn coffin was thought to be used for reburial of adult skeleton or burial for infants and children. In this report, we showed that the human adult bones were buried in a very unique pattern. Based on anthropological study, the dead person buried in the urn coffin died in his/her 30s; and every bone fragment belonged to human skull without exception. The dead person might be a warrior killed in action because there have been reports on exclusive burial of skull in a small-sized, ancient urn coffin in Japan, that was commonly interpreted as the burial of ancient warrior's heads decapitated during ancient battle, and were reburied in the burial ground. Nevertheless, considering that the cervical bones were not found along with the skull bone fragments, the tell tale sign of decapitation, more future studies are needed to confirm the hypothesis. However, even so, it could not be easily denied that very unique funeral customs: burial of human adult skull in a small-sized urn coffin, prevailed in the ancient Korean societies in certain regions.

Construction of Medieval Skeleton Collections with Human Remains from Tombs of Goryeo Dynasty, Korea / 체질인류학회지

Skeletal remains collected from the archaeological fields must be maintained carefully, for being used in scientific studies on the physical characteristics, health status, and pathological disease of the ancient or medieval human populations. Even if Joseon Dynasty Human Sample Collection might be a good example for such studies, few of bone collections predated the Joseon Age (e.g. Goryeo Dynasty) have been established until now, possibly owing to poor preservation condition of archaeological sites in Korea. In this study, we performed anthropological studies on a few cases of Goryeo skeletons, which have been rarely reported by anthropologists in Korea. Judging from the preservation status of bones found in various types of Goryeo graves (e.g. earthen- or stone-chambered tombs), many cases seem to be cremated in accordance with Buddhist funeral rites. Goryeo bone collections must be constructed with the bones identified in the earthen tombs, which were preserved much perfectly than those of any other types of Goryeo tombs.