Arterial supply of the human soft palate.

PURPOSE: Palatal surgery for snoring or obstructive sleep apnea is commonly performed; however, the vascular anatomy of the soft palate is not well described. The purpose of this study was to evaluate the vascular anatomy of the soft palate. METHODS: This study was performed on 22 adult cadaveric heads. All specimens were prepared with vascular injections using red liquid silicone through the common carotid artery. The palate was then harvested and decolorized, and the vascular anatomy was studied. RESULTS: Of 22 specimens, 20 had sufficient uptake of the silicone dye. The anterior and posterior branches of the ascending palatine artery were the main feeding vessels of the soft palate; however, they were not always present simultaneously. The anterior branch alone was identified 25% of the time (5/20), while the posterior branch alone was found in 35% (7/20) of specimens. Both anterior and posterior branches were simultaneously present in 40% (8/20) of cases. Mean diameter of the anterior branch (0.73 ± 0.09 mm) was significantly greater than that of the posterior branch (0.48 ± 0.08 mm, p < 0.001). Mean vertical distance from the palatal arch to the posterior branch was 9.3 ± 2.4 mm. CONCLUSIONS: The soft palate is supplied by the anterior and posterior branches of the ascending palatine artery; however, the arteries were not always present simultaneously. In cases where the anterior branch is absent and the posterior branches terminate close to the uvula, injury to the primary blood supply to the palate might occur more frequently during surgery.

[Hong Seok-hoo's translation of "New Edition of Physiology Textbook"(1906) and its meanings].

Hong Seok-hoo, who took charge of Jejungwon, was successful in translating Jiro Tsuboi's book titled "New Edition of Physiology Textbook (1897)" in Japanese and publishing it with a title of "New Edition of Physiology Textbook" in 1906. Jiro Tsuboi, the original author of that book, was a doctor having majored in Hygienics in Germany and was also known to have done pioneering work in Hygienics and Occupational and Environmental Medicine in Japan. At that time, he wrote that book for the purpose of teaching his students at Ordinary Middle School and Normal School. Therefore, it was not intended as a Physiology textbook for medical students, but an introductory book explaining Physiology with a wide range of subjects including hygienic matters in a broader sense. Hong Seok-hoo made an almost complete translation of the "New Edition of Physiology Textbook." While editing the book, however, he changed some of the most Japanese-style contents to meet the Korean conditions then, and made up for some insufficient contents with reference to the original author's other books. Although it was not included in an original version of that book, he also compiled a physiology dictionary in order to help Korean readers acquire medical terms in a more systematic way. Just like other textbooks of Jejungwon, the "New Edition of Physiology Textbook" was also put into Korean only. Hong Seok-hoo accepted Japanese-style medical terms, but also changed some of them or coined new words, considering the Korean circumstances then. He seemed to do so in an effort to introduce Western medicine in a more independent way while overcoming his limitations of translation. In particular, this book criticized that a long-term use of cosmetics might cause a serious lead poisoning from a Christian viewpoint, saying that a God-created human body should be kept intact as it is. In addition, in the course of reediting premodern books, the term "Lord" was changed into "God," which is considered a kind of fusion between traditional values and missionary medicine. While translating books, Jejungwon could put such fusion into practice because it was a hospital established under the banner of the propagation of Christianity. Besides the "New Edition of Physiology Textbook," at least five physiology textbooks were also translated into Korean in the last years of Daehan Empire for the purpose of teaching students modern subjects like Physiology, Health and Hygienics in educational institutions including Boseong School, Hwimun School and Soongsil School. On the other hand, the "New Edition of Physiology Textbook" was first translated at the end of Daehan Empire in order to foster more professional doctors in medical schools compared to those schools. In this respect, by translating the "New Edition of Physiology Textbook," Jejungwon can be considered as playing a pioneering role in translating Physiology textbooks in the late Daehan Empire.

Timetable for upper eyelid development in staged human embryos and fetuses.

In this study, we examined the development of the upper eyelids to provide a basic understanding of gross anatomical structures and information relative to mechanisms of congenital anomalies in the upper eyelids. We studied the upper eyelids by external and histological observation in 48 human embryos and in fetuses from 5 to 36 weeks postfertilization. The upper eyelid fold began to develop at Stage 18. Upper and lower eyelids fused from the lateral cantus at Stage 22, and fusion was complete by 9 weeks of development. Mesenchymal condensations forming the orbital part of the orbicularis oculi (OO), tarsal plate, and the eyelashes and their appendages, were first seen at Week 9. Definite muscle structures of the upper eyelid, such as the orbital part of the OO and the levator palpebrae superioris and its aponeurosis, and the Müller's muscle were observed at 12 and 14 weeks, respectively. In addition, orbital septum, arterial arcade and orbital fat pad, and tarsal gland (TG) were apparent at 12, 14, and 18 weeks, respectively. Opening of the palpebral fissure was observed at Week 20. In addition, we defined the directional orientation between the levator aponeurosis and orbital septum and the growth pattern of the TG. Our results will be helpful in understanding the normal development of the upper eyelid and the origins of upper eyelid birth defects.

Hoxc8 represses BMP-induced expression of Smad6.

Proper regulation of bone morphogenetic protein (BMP) signaling is critical for correct patterning and morphogenesis of various tissues and organs. A well known feedback mechanism is a BMP-mediated induction of Smad6, an inhibitor of BMP signaling. Hoxc8, one of the Hox family transcription factors, has also been shown to negatively regulate BMP-mediated gene expression. Here we add another level of Hoxc8 regulation on BMP signaling. Our results show that Hoxc8, when over-expressed in C3H10T1/2 or C2C12 cells, suppressed basal Smad6 promoter activity and its mRNA expression. Activation of Smad6 transcription either by BMP2 treatment or Smad1 over-expression was also abolished by Hoxc8. When chromatin was precipitated from mouse embryos with anti-Smad1 or anti-Hoxc8 antibody, Smad6 promoter sequence was enriched, suggesting that Hoxc8 proteins make complexes with Smad1 in the Smad6 promoter region. Yet, a lack of Hox binding motifs in the Smad6 promoter sequence suggests that instead of directly binding to the DNA, Hoxc8 may regulate Smad6 expression via an indirect mechanism. Our results suggest that the Smad6-mediated negative feedback mechanism on BMP signaling may be balanced by the repression of Smad6 expression by Hoxc8.

The third helix of the Hoxc8 homeodomain peptide enhances the efficiency of gene transfer in combination with lipofectamine.

Protein transduction domains (PTDs) have been shown to cross the biological cell membranes efficiently through a receptor and energy independent mechanism. Because of its ease in membrane transducing ability, PTDs could be used as a gene delivery vector. Since we already have shown that purified Hoxc8 homeoprotein has the ability to cross the cellular membrane, we analyzed the possibility of the third helix of the Hoxc8 homeodomain as a useful gene delivery vector. For that purpose, a 16-aa long synthetic oligopeptide Hoxc8 Protein Transduction Domain (HPTD) was chemically synthesized and then tested to see whether the HPTD could form a complex with DNA or not. Gel retardation analysis revealed that the HPTD interacts with plasmid DNA efficiently but failed to transfer the DNA into the cells. However, HPTD can enhance the efficiency of gene transfer in combination with Lipofectamine which doubled the gene transfer rate into COS-7 cells compared with the DNA/Lipofectamine control. An MTT assay indicated that the amount of HPTD used in the complex for the transfection did not show any cytotoxicty in COS-7 cells. The TEM studies showed compact particle formation in the presence of HPTD. These results indicate that the HPTD could be a good candidate adjuvant molecule to enhance the gene transfer efficiency of Lipofectamine in eukaryotic cells.

[Life and activities of Chu Hyun Chik].

Chu Hyun Chik was one of those who graduated first from Jejungwon Medical School in 1908, and had carried on an independence movement as well as religious, educational, and social movement both as a doctor and a Christian. He opened the Inje Hospital in Sunch'on, North Pyeongan Province in 1909, and was put in prison on charges of being involved in Incident of '105 People' as he joined in Sinminhoe in which christians (Christians) frrom Gwanseo showed their initiative with 3.1 Movement as a momentum, he started to raise funds for an independence movement mainly in North Pyongan Province, as a councilor of the Ministry of Finance of Shanghai Provisional Government of Korea. After he moved into Andong, Manchuria, he continued to support the spread of an independence movement by connecting Shanghai Provisional Government of Korea with the country. In October, 1919, he came to Sanghai as an exile and lead diverse activities as a member of Shin Han young man party and one of the leading men of Korean Christendom, especially related to An Chang Ho and christians around him and joining in Hungsadan. In 1925 when he returned home, he opened the Dongje Hospital and devoted himself to the developments of religious, educational, and social movement as a president of YMCA, Sunch'on and an executive of a branch of Suyang Donguhoe in Sunch'on. By Incident of Suyang Donguhoe he was put in prison, resisting Japanese Imperialism and died in 1942.

The third helix of the murine Hoxc8 homeodomain facilitates protein transduction in mammalian cells.

Previously, we have demonstrated that purified Hoxc8 homeoprotein has the ability to penetrate the cellular membrane and can be transduced efficiently into COS-7 cells. Moreover, the Hoxc8 protein is able to form a complex with DNA molecules in vitro and helps the DNA be delivered intracellularly, serving as a gene delivery vehicle. Here, we further analyzed the membrane transduction activity of Hoxc8 protein and provide the evidence that the 16 amino acid (a.a.191-206, 2.23 kDa) third helix of murine Hoxc8 protein is an efficient protein transduction domain (PTD). When the 16 amino acid peptide was fused at the carboxyl terminal of enhanced green fluorescence protein (EGFP), the fusion proteins were transduced efficiently into the primary pig fetal fibroblast cells. The transduction efficiency increased in a concentration-dependent manner up to 1 microM, and appeared to plateau above a concentration of 1 microM. When tandem multimers of PTD, EGFP-PTD(2), EGFP-PTD(3), EGFP-PTD(4), and EGFP-PTD(5), were analyzed at 500 nM of concentration, the penetrating efficiency increased in a dose-dependent manner. As the number of PTDs increased, the EGFP signal also increased, although the signal maintained plateau after EGFP-PTD(3). These results indicate that the 16 amino acid third helix is the key element responsible for the membrane transduction activity of Hoxc8 proteins, and further suggest that the small peptide could serve as a therapeutic delivery vehicle for large cargo proteins.

Membrane transducing activity of recombinant Hoxc8 protein and its possible application as a gene delivery vector.

In order to examine whether the Hoxc8 protein can deliver nucleic acid into mammalian cells, we designed several Hoxc8-derived recombinant proteins to be synthesized as glutathione S-transferase (GST) fused forms in E. coli (GST-Hoxc8(1-242), containing a full length of Hoxc8; GST-Hoxc8(152-242), possessing a deletion of the acidic N-terminus of Hoxc8; GST-Hoxc8(149-208), which contained the homeodomain only). After labeling these proteins with Oregon 488, we examined their membrane transduction ability under the fluorescence microscope and verified that all three proteins showed similar transduction efficiency. The ability of the proteins to form in vitro protein-DNA complexes was analyzed on agarose gel; both GST-Hoxc8(1-242) and GST-Hoxc8(149-208) formed complexes. In contrast, the GST-Hoxc8(152-242) protein did not form a complex. The GST-Hoxc8(149-208) protein formed a complex with DNA at a mass ratio of 1ú1 (DNAúprotein), and GST-Hoxc8(1-242) formed a complex at a mass ratio of 1ú5. When the DNA (pDsRed1-C1) and protein complexes were added to culture media containing mammalian cells, the cells uptook the complexes, which was indicated by red fluorescence expression under the fluorescent microscope. These results indicate that recombinant Hoxc8 derivatives that harbor a homeodomain are able to traverse the mammalian cellular membrane. DNA that is bound to the recombinant derivatives can be carried across the membrane as well. This process could be applied in the development of a useful delivery vector for gene therapy in the future.

[Life and medical activities of Yun Ti Wang].

Yun Ti Wang studied medicine in England, different from other Korean medical doctors in early days. Yun, who entered medical school at Glasgow University in England in March 1919, graduated with a Bachelor of Medicine in October 1925, along with an England medical license. Yun began working as an instructor at Severance Medical College from November 1927, and received Doctor of Medicine from the College of Obstetricians and Gynecologists at Kyoto Imperial University in August 1936. After the Liberation, Yun began working as a faculty member at the medical school at Seoul University, and he also worked as a Chief of the Second Medical Clinic of the school from 1946. Yun made a great effort in order to build an integrated committee, eventually contributing to the launching of Joseon Medical Associates in 1947. He was also elected as a first president at Joseon Obstetrics and Gynecology Associates, which was organized at the same year as the Joseon Medical Associates. Yun entered military as an army surgeon in April 1949 and has worked there until he was appointed as a principal at the Army Medical School in September 1953. His contributions to the development of military medical services include the following: expansion of medical facilities in army, stable system of workforce in military medical service, launching of Medical Aid and establishment of Department of Medical Care, and introduction of new medical technologies in anesthesiology and neurosurgery, etc. The career of Yun can be largely divided into the field of gynecology and military medical services. In the gynecological field, Yun contributed to the settlement of obstetrics in Korea, by taking in charge of the obstetrics class at Severance following medical missionaries. As for the military medical services, he has contributed to the establishment of military medical system as well as to the opening of new academic areas. The impact of his activities on the establishment of military medical services is especially significant, since it was a field that no Korean citizens had access to during the colonization era.

Efficient cellular uptake of recombinant murine Hoxc8 homeoprotein in COS-7 cells.

In order to analyze the self-delivery activity of Hoxc8, recombinant Hoxc8 protein (rHoxc8) was designed to be expressed and purified in E. coli as a glutathione S-transferase and green fluorescent protein-fused form (GST-GFP-Hoxc8). After purification using glutathione sepharose beads, the 82 kDa fusion protein was separated on the SDS-PAGE gel and confirmed by detecting the fluorescence through luminescent image analyzer. When rHoxc8 was added to culture media for 30 h, most of the COS-7 cells contained the fusion proteins, showing green fluorescence under the fluorescent microscope. When the efficiency of cellular uptake was examined after Hoechst staining, almost 100% of the cells exhibited the GFP signal, revealing that rHoxc8 can traverse the cellular membrane of COS-7 cells efficiently, suggesting that the rHoxc8 could be applied in the development of efficient and useful delivery vectors for therapeutic molecules.

[Suh Yang Park, who became a doctor as a son of a butcher].

Dr. Suh Yang Park was born in 1885 as a son of a butcher, which was the lowest class at that time in Korea. However, contact with western missionaries, including Dr. 0. R. Avison, provided him with an understanding of western civilization. After entering Chejungwon Medical School in 1900, Dr. Park learned basic sciences, basic medical sciences like anatomy and physiology, and other Western medical specialties such as internal medicine and surgery. He graduated from medical school in 1908 and received Government Certificate from Home Office, the first in Korea in this field, which granted the right to practice medicine. His certificate number was 4th overall. As both a doctor and a talented musician, Dr. Park actively participated in the social enlightenment movement. He was quite progressive in his time, having surpassed the social limitations as a son of a butcher, as well as actively propagating his knowledge of Western civilization onto others. After graduation, he had served as a professor at the school he graduated from, until he went into exile in Manchuria in 1917 due to the annexation of Korea by Japan in 1910. There, he opened a hospital and provided medical treatment for Koreans. He also established a school for young Koreans, inspiring them with a sense of nationalism. Also, Dr. Park was an active member of various Independence Movement Organizations in Manchuria. Then in 1932, at the time when Japan took control of Manchuria, his school was closed down. As a result, Dr. Park couldn't help but stop his anti-Japanese activities. In 1936, he returned to his homeland and passed away in 1940, just five years before the liberation of Korea from Japanese occupation.

Membraneous localization of Jpk is not essential to exert cytotoxicity in F9 teratocarcinoma cells.

A novel gene, Jopock (Jpk), which was isolated as a trans-acting factor associating with the PSRE of murine Hoxa-7, has been shown to be toxic to both prokaryotic and eukaryotic cells when overexpressed. Here we demonstrate that the overexpression of enhanced green fluorescent protein (EGFP)-tagged Jpk in F9 cells results in the induction of apoptosis, as indicated by phosphatidylserine exposure, DNA fragmentation, and the alteration of mitochondria transmembrane potential. Fluorescence microscopy showed that EGFP-fused Jpk was mainly localized in the endoplasmic reticulum (ER) and a small amount was found in the mitochondria. Deletion mutants with a transmembrane (TM) domain showed a distribution similar to that of EGFP-Jpk, whereas constructs with a deletion of the TM domain localized in the whole cells. Deletion mapping experiments showed that Jpk with an N-terminal part deleted stimulated apoptosis to almost the same extent as that of the wild-type Jpk, indicating that the localization of Jpk in the ER and the TM domain does not appear to be essential for inducing cytotoxicity. Overall, these results suggest that Jpk, particularly the C-terminal part of Jpk and/or 3'UTR, triggers apoptosis through a perturbation of mitochondrial membrane permeabilization.

Dynamic expression pattern of Hoxc8 during mouse early embryogenesis.

The Hoxc8 expression pattern was examined in mouse embryos 7.5-12.5 days postcoitum (dpc) using whole-mount in situ hybridization and RT-PCR. The expression of Hoxc8 started between 7.5 and 8.5 dpc. A strong expression was detected in the ectoderm and mesoderm at 8.5 dpc. At 9.5 dpc, a distinct anterior boundary of Hoxc8 expression was established at the 10th and 16th somites in the neural tube and the paraxial mesoderm, respectively. This staggered expression pattern was maintained throughout the later stages. By 12.5 dpc, the forward progression of the Hoxc8 expression pattern was observed and the stain was weakened. In the ectoderm-derived neural tube, strong Hoxc8 expression was observed in the ventral horn and later in the ventral and mediolateral region of the mantle layer, indicating a possible association with the onset and progression of neural differentiation. In the case of the mesoderm-derivative cells, strong Hoxc8 expression was detected in the sclerotome on the way to the notochord and neural tube and mesonephros, suggesting a role of Hoxc8 in the formation of the vertebrae and ribs and the possible involvement in the differentiation into the kidney.

Early development of the nose in human embryos: a stereomicroscopic and histologic analysis.

OBJECTIVES/HYPOTHESIS: To analyze the morphologic features of the nose in the human embryo from the 4th to 8th developmental week according to Carnegie stage. STUDY DESIGN: Stereomicroscopic and histologic analysis of the morphology of the human embryo. METHODS: A total of 27 cases of embryos, ranging from Carnegie stage 13 to 23, were analyzed. The external morphology was observed with a stereomicroscope, photographed, and analyzed. The histologic features were observed with a light microscope in the horizontally transected specimens stained with hematoxylin-eosin. RESULTS: The nasal placode was observed in stage 13, and it became flat or even concave in stage 14. In stage 15, the nasal pit was formed. In stage 16, the nasal sac and nasal fin were observed. In stage 17, the oronasal membrane was formed by thinning of the nasal fin. In stage 18, the primitive choana was established by a rupture of the oronasal membrane. In stage 19, the lateral palatine process projected vertically below the level of the tongue. The cartilaginous nasal capsule was formed in stage 20. In stage 21, the olfactory area was localized to the upper portion of the lateral nasal wall and the nasal septum. In stage 22, the lateral palatine process developed in a somewhat horizontal orientation. In stage 23, the premaxilla and primitive choana were formed. CONCLUSIONS: The development of the nose is most active from Carnegie stage 13 to 19, which corresponds to the end of the fourth embryonic week to the end of the seventh week. Thus, this period is considered to be the most important period in human nasal embryonic development.

Cloning of TLR3 isoform.

Toll-like receptor (TLR) 3 is a member of the TLR family that confers innate immunity by recognizing viral pathogens. Herein, we report that the TLR3 isoform is expressed on human primary cells and cell lines. This isoform has 2,520 bp cDNAs compared to the 2,712 bp of full cDNA, is produced by deletion of an intron-like sequence within exon 4 and is co-expressed with wild type TLR3 in primary human astrocytes and glioblastoma cell lines. This finding suggests the TLR3 isoform in astrocytes may have a different immunological role for binding ligands during the immune response in brain.

Establishment of stable melanoma cell line expressing a novel gene, jpk, using a tetracycline-controlled gene expression system.

Jpk, originally isolated as an associating factor with the position-specific regulatory element of Hoxa-7, was found to be toxic to Escherichia coli (1) and to F9 teratocarcinoma cells (2) when transiently transfected and expressed. To investigate the possibility of tumor gene therapy using Jpk, its effect was tested in B16F10 murine melanoma cells. Because Jpk reduces the viability of B16F10 cells when transiently expressed, the Jpk gene was cloned into a tetracycline-controlled gene expression vector, pRetro-On to circumvent the lethal effect in unwanted situations. The retroviral plasmid pRetroJpk purified from the packaging cell was infected into B16F10 melanoma cells and screened in the presence of puromycin. Out of a total of 53 stable clones selected with puromycin, two clones overexpressed Jpk at more than twice the level when induced by doxycycline, a tetracycline-derivative, which implies the amount of the Jpk exhibiting the toxicity is critical. Although these clones control only low levels of Jpk, overexpression of the established melanoma cell line may help us decipher the function of Jpk and apply it as a tumor therapeutic gene in the future.

Timetable for intestinal rotation in staged human embryos and fetuses.

BACKGROUND: The existing data on intestinal rotation during human development are contradictory regarding the timing of major events, and as such an exact timetable for rotation of the intestine in humans is not yet available. METHODS: We studied the initial formation and rotation of the intestine by microdissection and histological observations in 72 human embryos and fetuses at two to 12 weeks postfertilization. The embryos were classified according to the Carnegie staging system. RESULTS: The primordium of the primitive gut was first observed as a yolk sac at stage 5. With the formation of the embryonic foldings, three divisions of the primitive gut (the foregut, midgut, and hindgut) were observed at stage 10. At stage 12, the primitive gut was located on the midline. At stage 15, a 90 degrees counterclockwise rotation of the intestine began. At stage 16, herniation of the intestine into the umbilical cord was not evident in observations of the external form or a transversely sectioned embryo, but was evident in a sagittally sectioned embryo. There was another 90 degrees counterclockwise rotation at stage 20. Reduction of the intestine was a rapid process, since it was still in the cord in fetuses of <40 mm crown-rump length (CRL), and was reduced above 40 mm in general during nine weeks of development. When the intestine returned to the abdominal cavity, the cecum was located in the right lower quadrant (the adult position). CONCLUSIONS: We have developed a standard timetable to describe the rotation of the intestine. The current results will be helpful in studies describing the pathogenesis of some developmental abnormalities in the intestine due to abnormal rotation.

Endotoxin-neutralizing antimicrobial proteins of the human placenta.

Microbial colonization and infection of placental tissues often lead to adverse pregnancy outcomes such as preterm birth, a leading cause of neonatal morbidity and mortality. The fetal membranes of the placenta, a physical and active barrier to microbial invasion, encapsulate the fetus and secure its intrauterine environment. To examine the innate defense system of the human placenta, antimicrobial peptides were isolated from the fetal membranes of human placenta and characterized biochemically. Two salt-resistant antimicrobial host proteins were purified to homogeneity using heparin-affinity and reversed-phase HPLC. Characterization of these proteins revealed that they are identical to histones H2A and H2B. Histones H2A and H2B showed dose-dependent inhibition of the endotoxin activity of LPS and inhibited this activity by binding to and therefore blocking both the core and lipid A moieties of LPS. Consistent with a role for histones in the establishment of placental innate defense, histones H2A and H2B were highly expressed in the cytoplasm of syncytiotrophoblasts and amnion cells, where the histone proteins were localized mainly to the epithelial surface. Furthermore, culturing of amnion-derived WISH cells led to the constitutive release of histone H2B, and histones H2A and H2B contribute to bactericidal activity of amniotic fluid. Our studies suggest that histones H2A and H2B may endow the epithelium of the placenta with an antimicrobial and endotoxin-neutralizing barrier against microorganisms that invade this immune-privileged site.