Identifying the degree of major histocompatibility complex matching in genetically unrelated dogs with the use of microsatellite markers.

BACKGROUND: The dog has served as an important experimental model for biomedical research such as transplantation and developing immunosuppressive agents. Although major histocompatibility complex (MHC) in dogs is a dominant factor of graft rejection, it has not been well investigated in dogs compared with human. For that reason, imprecise cross-matching or time-consuming sequence-based typing methods have generally been used to choose specific donor and recipient pairs. Investigation of matching distribution of MHC in dogs with the use of simple and accurate methods would be beneficial for biomedical researchers. The aim of this study was to identify the diversity of dog leukocyte antigen (DLA) types in genetically unrelated dogs by means of microsatellite markers. METHODS: Thirty-three Beagle and Shih-Tzu dogs, which were negative in cross-matching, were chosen. The genomic DNA was isolated from peripheral blood leukocytes, and highly polymorphic short tandem repeats located in MHC class I and II were amplified with the use of specific primers. RESULTS: Among all of the dogs, MHC matching groups, including class I full match-class II full match (M-M), class I full match-class II haplo match (M-H), class I haplo match-class II full match (H-M), class I haplo match-class II haplo match (H-H) groups, were ∼1.55%, 0.39%, 1.94%, and 6.59%, respectively. MHC class I nonmatch-class II nonmatch (U-U) groups were 58.14% of the total dogs. CONCLUSIONS: Because differences of histocompatibility between donor and recipient leads to various allograft rejections, knowledge of the distribution of MHC matching in unrelated dogs would be helpful in designing studies and to get more accurate results from experiments using dog transplantation models.

Acute rejection after swine leukocyte antigen-matched kidney allo-transplantation in cloned miniature pigs with different mitochondrial DNA-encoded minor histocompatibility antigen.

INTRODUCTION: Graft rejection remains a major cause of morbidity and mortality following renal transplantation. One of the main determinants of success after renal transplantation is histocompatibility between donor and recipient. Most of the research on this topic has addressed human leukocyte antigen (HLA), but the roles played by minor histocompatibility antigens (mHAgs), such as mitochondrially transmitted antigens, are poorly understood. In this study, we evaluated immune responses induced by minor antigens originating from mitochondrial DNA (mtDNA) in a large animal model. METHODS: To characterize whole swine leukocyte antigen (SLA) allele in 8 cloned pigs, we performed SLA genotyping for SLA-1, SLA-2, SLA-3, SLA-DQB1, and SLA-DRB1 as well as the hypervariable region 1 (HV1) of mtDNA. Renal transplantation was performed using SLA-matched pigs with different mtDNA as well as SLA-mismatched cloned animals. Cytokine profiling was performed by incubating peripheral leukocytes with cellular components from SLA-matched different mtDNA and SLA-mismatched cells to evaluate mtDNA-mediated immune response. RESULTS: SLA types were confirmed to be identical, but mtDNA sequences of HV1 varied among cloned pigs. Rejection episodes in the SLA-matched group with different mtDNA were similar to those in the SLA-mismatched group; that is, plasma creatinine and BUN levels were increased and mononuclear cell infiltration was observed in perivascular regions in the matched and SLA-mismatched groups. Furthermore, in vitro studies showed interleukin (IL)-1ß expression to be elevated in SLA-matched and SLA-mismatched groups. CONCLUSION: Cloned pigs are a useful preclinical model to evaluate the immunogenicity of mtDNA encoding minor antigens. The mtDNA originating from nongenomic DNA induced cell-mediated immune rejection after kidney transplantation.

Disparate hypervariable region-1 of mitochondrial DNA did not induce skin allograft rejection in cloned porcine models.

INTRODUCTION: Alloantigen recognition in skin transplantation is the bane for surgeons. Several studies have mainly focused on the immunogenicity of major histocompatibility (MHC) antigens and H-Y minor histocompatibility antigens. However, the roles of the mitochondrial DNA (mtDNA) encorded miHA have not been identified. Therefore, we sought to address the antigenicity of the hypervariable region 1 (HV-1) of mtDNA in skin transplantation using cloned pig models. METHODS: Swine leukocyte antigen and HV-1 of mtDNA were analyzed using sequencing methods. Skin transplantation was performed between MHC-matched, mtDNA-mismatched cloned miniature pigs. Full-thickness skin was grafted between cloned pigs without any immunosuppressants. The grafted tissues were observed for 3 months and evaluated histologically. RESULTS: The cloned pigs shared identical MHC but mtDNA mismatched at 9 positions. Skin grafts between the cloned pigs were accepted and hair growth maintained, whereas MHC-mismatched grafts showed acute rejection within 7 days after transplantation and were replaced by hairless scar tissue. CONCLUSIONS: HV-1 disparate skin grafts were not recognized as alloantigenic by MHC-matched cloned pigs.

Development of a sustained-release recombinant human growth hormone formulation.

Recombinant human growth hormone (rhGH) therapy for short stature must be administered as a daily injection because of its poor bioavailability and short half-life. In the present study, a sustained-release formulation of rhGH (SR-rhGH), DA-3003, was prepared using double emulsion solvent evaporation with poly(D,L-lactide-co-glycolide) (PLGA), zinc oxide and hydroxypropyl-beta-cyclodextrin (HPCD) as the release modulator, stabilizer, and aggregation-prevention agent, respectively. After a single administration of DA-3003, the elevated concentration of rhGH in plasma was sustained for 14 days in rats and 28 days in monkeys. The plasma concentration of insulin-like growth factor-1 (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3), which are pharmacodynamic markers of rhGH administration, increased and remained elevated for approximately 28 days in monkeys. Monkeys administered DA-3003 did not develop antibodies to hGH, indicating safety of the SR-rhGH formulation comparable to that observed with daily rhGH injections (Growtropin II). There were no significant differences in efficacy between Growtropin II (daily dose of 5 microg/animal for 14 days) and DA-3003 (weekly dose of 35 microg/animal for 14 days with a dosing interval of a week) in hypophysectomized rats, as assessed by changes in body weight and the width of the tibial growth plate. These results show that a sustained-release rhGH formulation, DA-3003, has the potential to be used safely and efficaciously in a weekly dosing regimen.

Topography and location of the depressor anguli oris muscle with a reference to the mental foramen.

The labiomandibular fold (LMF) is the area of the face that extends from the mouth corner to the mandibular border, and its prominence tends to increase with age. The LMF can be formed by the medial or lateral border of the depressor anguli oris (DAO). The aim of this study was to demonstrate the topographical anatomy between the DAO and mental foramen, thereby providing critical information for the safest and most effective site at which to inject botulinum toxin type A (BTX-A). Thirty-four hemifaces from Korean adult cadavers were dissected. The maximum width between the medial borders of the bilateral DAO, parallel to the intercheilion horizontal line, was 59.9 +/- 4.6 (mean +/- SD) mm below the lower lip. The minimum width between the medial borders of the attachment of bilateral DAO was 29.7 +/- 4.8 mm at the mandibular border. The mental foramen was located in the middle third from the cheilion to the mandibular border in 28 cases (90.3%), and it was mostly confined within the DAO muscle coverage in 21 cases (67.7%). The buccal branch of the facial nerve entered through the middle third of the lateral border of DAO and then distributed. Concomitantly, the marginal mandibular branch of the facial nerve entered through the lower third of the lateral border of DAO in 17 cases (60.7%). These results represent additional reference data for identifying the position of the mental foramen on the facial skin, and will be useful for providing criteria for the most effective site for injecting BTX-A when treating the LMF.

Topography of the outer mandibular symphyseal region with reference to the autogenous bone graft.

The mandibular symphysis is preferred as a donor site for the relatively small grafts needed for the autogeneous bone graft procedure. This study was undertaken to determine the morphology and composition of the cortical and trabecular bone in the mandibular symphyseal region using 35 mandible specimens from Koreans. The topographical patterns through the thickness of the cortical plate and the width of the trabecular bone were observed. In this study, the labial cortical plate of the mandible became thicker from the superior to the inferior aspects (P < 0.05). However, the trabecular bone width exhibited a different distribution pattern compared to the thickness of the labial cortical plate. This observation concerning the cortical and trabecular bones assists in determination of the depth of osteotomy. The results provide useful information on the mandibular symphysis graft prior to dental implant placement. These results will enable the volume of the cortical plate in the mandibular symphyseal region and its proper size, depth, and location to be predicted when removing a graft block.

Branching patterns of the facial nerve and its communication with the auriculotemporal nerve.

This study examines the anatomic relationships and variability of the facial nerve trunk and its branches, with emphasis on the intraparotid connections between the divisions. Microdissections were performed on 30 Korean half-heads, and the facial nerve trunks and branches were exposed. The average depth of the stylomastoid foramen from the skin surface was 21.0 +/- 3.1 mm, and the distance between the stylomastoid foramen and the bifurcation of the temporofacial (upper) and cervicofacial (lower) divisions was 13.0 +/- 2.8 mm. In 26 of 30 dissections (86.7%), the facial nerve trunk bifurcated into two main divisions, and a trifurcation pattern was seen in the other four cases (13.3%). According to the origin of the buccal branches, we classified the branching patterns of the facial nerve into four categories. In type I (13.8% of cases), the buccal branches arose from the two main divisions of the trunk but not from other branches of the facial nerve. In type II (44.8% of cases), the buccal branches arising from the two main divisions were interconnected with the zygomatic branch. In type III (17.3% of cases), the marginal mandibular branch sent nerve twigs to the buccal branch, which originated from the upper and lower divisions. In type IV (17.3% of cases), the nerve twigs from the zygomatic and marginal mandibular branches merged to the buccal branch arising from the two main divisions. Communications between the facial and auriculotemporal nerve branches, which are known as "communicating auriculotemporal nerves," were observed in 28 of the 30 cases (93.3%). Familiarity with these common variations in the facial anatomy provides useful information for the surgeon in careful dissection, preservation of the facial nerve, and complete removal of the tumors in parotidectomies.

Anatomical study concerning the origin and course of the pectoral branch of the thoracoacromial trunk for the pectoralis major flap.

The patterns of the feeding vessels to each muscle determine the extent of their safe transposition and the muscle's value as a pedicled flap in reconstructive surgery. This study aimed to demonstrate the point of origin and the intra- and submuscular course of the pectoral branch of the thoracoacromial trunk (TAT) for pectoralis major (PM) flap surgery. Seventy sides of the PM were dissected based on a clinical reference line that has been used for several decades. The branching point of the TAT from the axillary artery was located lateral to the midclavicular line on the right-sided specimens (100%) and medial to the midclavicular line on the left sides (86%). The branching patterns of the pectoral branch to the PM muscle from the TAT were classified into three types. In type I the pectoral branches originated directly from the TAT (55 cases, 78.6%). In type II (11 cases, 15.7%) and type III (4 cases, 5.7%) the pectoral branch divided from the medial and lateral pedicle of the TAT, respectively. The course of the pectoral branch from the TAT in the PM was categorized into three patterns according to the degree of proximity to the midclavicular line. In 49 cases (70%), the pectoral branch in the PM ran within 1 cm of the midclavicular line. The other cases ran 2 cm (20 cases, 29%) and 3 cm (1 case, 1%) from the midclavicular line, respectively. These results provide topographic data of the pectoral branch based on anatomical landmarks, and will be useful in surgical planning as well as the procedure for PM flap surgery.

Topographic anatomy of the inferior wall of the maxillary sinus in Koreans.

Knowledge of the relationship between the root apex and the inferior wall of the maxillary sinus are crucial for diagnosing and treating a sinus pathosis as well as in assisting in dental implantation. Therefore, identifying the proximity between the root apex and the inferior wall of the sinus and clarifying the cortical thickness of the inferior wall of the sinus is essential for determining the topography of a spreading dental infection into the maxillary sinus. Accordingly, knowledge of the topography between the root apex and the inferior wall of maxillary sinus is important for diagnosing and planning dental implantation, endodontic procedures, and orthodontic treatment. This study was undertaken to clarify the morphological and clinical characteristics of the maxillary sinus, particularly the inferior wall of the sinus in Koreans, and to identify the relationship between the inferior wall of the maxillary sinus and the roots of the maxillary teeth. Twenty-four sides of the maxillae of hemi-sectioned Korean heads were used in this study. All specimens were decalcificated and sectioned coronally. On the sectioned specimens, 21 items were measured using an image analyzing system. The distances between the each root apex and the inferior wall of the maxillary sinus were measured. The distance from the root apex to the inferior wall of the sinus was the shortest in the second molar area and the longest in the first premolar area. The thickness of the cortical plate of the inferior wall of the maxillary sinus was thinnest in the first premolar area but it was thickest in the second premolar area. The vertical relationship between the inferior wall and the roots of the maxillary molars was classified into five types. Type I (the inferior wall of the sinus located above the level connecting the buccal and lingual root apices) dominated (54.5% in the first molar area, 52.4% in the second molar area). The horizontal relationship between the inferior wall of the sinus and the root apex was classified into three types. Type 2 (the alveolar recess of the inferior wall of the sinus was located between the buccal and lingual roots) was most common (80% in the first and second molar area). Overall, this study demonstrated the many anatomical characteristics and determined the relationships between the maxillary sinus and their surrounding structures. These findings may have an impact on the clinical management of patients.

The discomallear ligament and the anterior ligament of malleus: an anatomic study in human adults and fetuses.

According to some reports, movement of the malleus, resulting from anterior hypertension on the discomallear ligament (DML), could produce aural symptoms related with damage to middle ear structures. The aim of this study was to examine the topographic relationship of the DML and the anterior ligament of malleus (ALM). Four fetuses and 16 adult hemi-sectioned heads were used to determine the anatomic-clinical relevance of DML and ALM in temporomandibular disorder. In fetal specimens, the DML was distinctly interposed between the malleus and the disc of the temporomandibular joint (TMJ), and the ALM had a structure apparently composed of the superior and inferior lamellae, running anteriorly in continuation with the sphenomandibular ligament (SML) through the future petrotympanic fissure (PTF). In all adult specimens, the DML was inserted into the malleus, and it expanded broadly toward the disc and capsular region of the TMJ in a triangular shape and inserted into the disc and capsule of the TMJ. The two-lamellae structure of the ALM was not distinguishable in adult specimens. The overstretched ALM resulted in movement of the malleus in five cases, but similar tension applied to the DML did not cause any movement of the malleus. This result provides an indication of the clinical significance of the ALM, a ligamentous structure continuous with the SML. It is apparent that the ALM has the potential to cause aural symptoms as a result of damage to the middle ear structure.

Topographic anatomy of the mandibular nerve branches distributed on the two heads of the lateral pterygoid.

The purpose of this study was to evaluate the topography of the nerve distribution on the two heads of the lateral pterygoid and to clarify their morphological-functional correlations. The nerve distribution on the lateral pterygoid was studied in 24 hemi-sectioned heads. Both heads of the lateral pterygoid were innervated from the mandibular nerve branches, but with various nerve distribution patterns. The nerves innervating the superior head of the lateral pterygoid originated from the buccal nerve only in 45.8% of cases. In contrast, the nerves innervating the inferior head of the lateral pterygoid originated from both the buccal and mandibular nerve trunk in 58.3% of cases. In the seven distribution categories of the mandibular nerve branches, both the superior and inferior heads of the lateral pterygoid had a common source of nerve innervation in only 20.8% of cases, the buccal nerve. In contrast, in 45.9% of cases, additional nerve twigs from the mandibular nerve trunk were distributed on the inferior head of the lateral pterygoid. In summary, besides the buccal nerve described in anatomy textbooks, the nerve branches that originated directly from the mandibular nerve trunk innervated the lateral pterygoid.

Topographic anatomy of the deep temporal nerves, with references to the superior head of lateral pterygoid.

Commonly, the nerve branches from the anterior mandibular nerve trunk pass between the roof of infratemporal fossa and the superior head of lateral pterygoid. However, varied courses of the mandibular nerve branches can be frequently observed. The purpose of this study was to clarify the positional relationships and the clinical relevance of the course variations of the branches of the anterior mandibular nerve trunk with reference to the surrounding anatomical structures. Thirty-six hemi-sectioned heads were studied in detail. In 20 cases, the posterior deep temporal nerve had a common trunk with the masseteric nerve and was then divided anteriorly (15 cases) or posteriorly (five cases). In 16 cases, the posterior deep temporal nerve arose from the mandibular nerve trunk independently. Based on the branching patterns of the middle deep temporal nerve, type A (one twig of the middle deep temporal nerve) was most frequent and occurred in 41.7%. Similarly, type B (two twigs), type C (three twigs) and type D (four twigs) were observed in 36.1%, 16.7%, and 5.5%, respectively. The twigs of the middle deep temporal nerve, which pierced the muscle fibers of the superior head of lateral pterygoid, were found in 21 cases (58.3%). Cases in which the middle deep temporal nerve pierced through all areas of the superior head were most frequent (56.5%). These results suggest that the piercing patterns of the middle deep temporal nerve show there is a possibility that it may be compressed during the actions of the superior head of lateral pterygoid.

Personal-computer-based three-dimensional reconstruction and simulation of maxillary sinus.

Anatomical descriptions of the maxillary sinus are critical in pathological diagnosis and the treatment planning of surgical procedures. This study was undertaken to develop a new technique for simulating anatomical structures and to clarify the morphological and clinical characteristics of the maxillary sinus. Thirty-three hemi-sectioned Korean heads were used in this study. CT scans and DentaScan reformatted cross-sectional images were taken on all specimens. From the CT images, three-dimensional reconstructed images were made using the V-works program. From the three-dimensional reconstructed images of the maxillary sinus, six categories of maxillary sinus were created, categorized according to their lateral aspects and shapes of the inferior walls. In 55%, a flat inferior wall of the maxillary sinus was observed. All measurements (anterior-posterior length, height, width and volume) of the sinus were larger in males than in females. From the DentaScan reformatted panoramic images, the anterior limit of the maxillary sinus was located in the first premolar area (58%), and the posterior limit was in the third molar and maxillary tuberosity area (94%). We therefore offer a new virtual technique for manipulating three-dimensional reconstructed images easily on a personal computer. On the reconstructed images the three-dimensional morphology could be observed and the anatomical characteristics of the maxillary sinus and surrounding structures could be determined.