Structure–function relations of primate lower incisors: a study of the deformation of Macaca mulatta dentition using electronic speckle pattern interferometry (ESPI)

Teeth adopt a variety of different morphologies, each of which is presumably optimized for performing specific functions during feeding. It is generally agreed that the enamel cap is a crucial element in controlling the mechanical behavior of mammalian teeth under load. Incisors are particularly interesting in terms of structure–function relations, as their role in feeding is that of the ‘first bite’. However, little is known how incisor cap morphology is related to tooth deformation. In the present paper we examine the mechanical behavior of mandibular central incisors in the cercopithecine primate Macaca mulatta under loads similar to those encountered during ingestion. We map three‐dimensional displacements on the labial surface of the crown as it is compressed, using electronic speckle pattern interferometry (ESPI), an optical metrology method. In addition, micro‐computed tomography is used to obtain data regarding the morphology of the enamel cap, which in the M. mulatta lower incisors exhibits missing or very little enamel on the lingual face. The results showed that although compressed along a longitudinal axis, deformation in the incisors mostly occurred in the lingual direction and orthogonal to the direction of the applied load. Both isolated, embedded teeth and teeth in the mandible showed considerable lingual deformation. Incisor deformation in the mandible was generally greater, reflecting the additional freedom of movement enabled by the supporting structures. We show that the association with adjacent teeth in the arch is significant for the behavior of the tooth under load. Finally, loading two teeth simultaneously in the mandible showed that they work as one functional unit. We suggest that these results demonstrate the importance of enamel cap morphology in directing deformation behavior; an ability stemming from the stiffness of the enamel cap overlying the more pliable dentin.     

Application of monoclonal antibodies in functional and comparative investigations of heavy-chain immunoglobulins in new world camelids

Of the three immunoglobulin G (IgG) isotypes described to occur in camelids, IgG2 and IgG3 are distinct in that they do not incorporate light chains. These heavy-chain antibodies (HCAbs) constitute approximately 50% of the IgG in llama serum and as much as 75% of the IgG in camel serum. We have produced isotype-specific mouse monoclonal antibodies (MAbs) in order to investigate the roles of HCAbs in camelid immunity. Seventeen stable hybridomas were cloned, and three MAbs that were specific for epitopes on the gamma chains of llama IgG1, IgG2, or IgG3 were characterized in detail. Affinity chromatography revealed that each MAb bound its isotype in solution in llama serum. The antibodies bound to the corresponding alpaca IgGs, to guanaco IgG1 and IgG2, and to camel IgG1. Interestingly, anti-IgG2 MAbs bound three heavy-chain species in llama serum, confirming the presence of three IgG2 subisotypes. Two IgG2 subisotypes were detected in alpaca and guanaco sera. The MAbs detected llama serum IgGs when they were bound to antigen in enzyme-linked immunosorbent assays and were used to discern among isotypes induced during infection with a parasitic nematode. Diseased animals, infected with Parelaphostrongylus tenuis, did not produce antigen-specific HCAbs; rather, they produced the conventional isotype, IgG1, exclusively. Our data document the utility of these MAbs in functional and physiologic investigations of the immune systems of New World camelids.     

Application of Monoclonal Antibodies in Functional and Comparative Investigations of Heavy-Chain Immunoglobulins in New World Camelids

Of the three immunoglobulin G (IgG) isotypes described to occur in camelids, IgG2 and IgG3 are distinct in that they do not incorporate light chains. These heavy-chain antibodies (HCAbs) constitute approximately 50% of the IgG in llama serum and as much as 75% of the IgG in camel serum. We have produced isotype-specific mouse monoclonal antibodies (MAbs) in order to investigate the roles of HCAbs in camelid immunity. Seventeen stable hybridomas were cloned, and three MAbs that were specific for epitopes on the γ chains of llama IgG1, IgG2, or IgG3 were characterized in detail. Affinity chromatography revealed that each MAb bound its isotype in solution in llama serum. The antibodies bound to the corresponding alpaca IgGs, to guanaco IgG1 and IgG2, and to camel IgG1. Interestingly, anti-IgG2 MAbs bound three heavy-chain species in llama serum, confirming the presence of three IgG2 subisotypes. Two IgG2 subisotypes were detected in alpaca and guanaco sera. The MAbs detected llama serum IgGs when they were bound to antigen in enzyme-linked immunosorbent assays and were used to discern among isotypes induced during infection with a parasitic nematode. Diseased animals, infected with Parelaphostrongylus tenuis, did not produce antigen-specific HCAbs; rather, they produced the conventional isotype, IgG1, exclusively. Our data document the utility of these MAbs in functional and physiologic investigations of the immune systems of New World camelids.     

Distribution of keratins, vimentin, and actin in the testis of two South American camelids: vicuna (Vicugna vicugna) and llama (Lama glama). An immunohistochemical study

The purpose of the present study was to investigate the pattern of distribution of cytokeratins, vimentin and muscular actin in the testis of vicuna (Vicugna vicugna) and llama (Lama glama) two species of camelids native of the Andean high plateau of South America. Testicular biopsies of four vicunas and five llamas were used. Animals were healthy breeders. The tissues were processed by standard immunohistochemistry with antipancytokeratinAE1/AE3, antikeratin 18 (K 18), CAM 5.2 (antikeratin 5, 18, and 19), antivimentin, and smooth-muscle-specific antiactin antibodies to track the cytoskeletal pattern of testicular cells. Using AE1/AE3 antibody the immunostaining was found in the epithelial lining of tubuli recti and rete testis. The reaction was relatively stronger in the apical cytoplasm of epithelial cells. The testicular cells of the two species showed no reaction to K 18 and CAM 5.2 antibodies. Antivimentin antibody stained the basal cytoplasm of the Sertoli cells, the Leydig cells, and the epithelial lining of tubuli recti and rete testis. In the last two structures the immunostain was relatively more intense in the basal cytoplasm of epithelial cells. Antiactin antibody stained the peritubular cells and the muscle cells of the lamina propria oftubuli recti and rete testis. The presence in these species of only some keratins found in man, its coexpression with vimentin in epithelial lining of tubuli recti and rete testis and the peritubule organization, so different from other ungulates may reflect a differential adaptation of the cytoskeleton to particular reproductive strategies.     

Effect of Down syndrome on the dimensions of dental crowns and tissues

Abnormal growth in Down syndrome (DS) is reflected by variable reduction in size and simplification in form of many physical traits. This study aimed to compare the thickness of enamel and dentine in deciduous and permanent mandibular incisor teeth between DS and non‐DS individuals and to clarify how these tissues contribute to altered tooth size in DS. Sample groups comprised 61 mandibular incisors (29 permanent and 32 deciduous) from DS individuals and 55 mandibular incisors (29 permanent and 26 deciduous) from non‐DS individuals. Maximum mesiodistal and labiolingual crown dimensions were measured initially, then the crowns were sectioned midsagittally and photographed using a stereomicroscope. Linear measurements of enamel and dentine thickness were obtained on the labial and lingual surfaces of the crowns, together with enamel and dentine–pulp areas and lengths of the dentino‐enamel junction. Reduced permanent crown size in DS was associated with a reduction in both enamel and dentine thickness. After adjustments were made for tooth size, DS permanent incisors had significantly thinner enamel than non‐DS permanent teeth. The DS permanent teeth also exhibited significant differences in shape and greater variability in dimensions than the non‐DS permanent teeth. Crown dimensions of deciduous incisors were similar in size or larger in DS compared with non‐DS deciduous teeth. Enamel and dentine thicknesses of the deciduous teeth were similar in DS and non‐DS individuals. The findings indicate that growth retardation in DS reduces both enamel and dentine deposition in the permanent incisors but not in the earlier‐forming deciduous predecessors. The results are also consistent with the concept of amplified developmental instability for dental traits in DS. Am. J. Hum. Biol. 13:690–698, 2001. © 2001 Wiley‐Liss, Inc.     

转矩作用对微植体矫治力系内收上前牙影响的三维有限元研究

目的研究转矩力的作用对于微植体矫治力系内收上前牙的影响。方法利用MIMICS软件和有限元软件ABAQUS,建立微植体矫治力系的有限元模型,并在该模型上计算分析有无转矩及转矩作用区域对中切牙和侧切牙移动趋势的影响。结果不加转矩时,中切牙和侧切牙都表现出舌向倾斜的趋势。在100g·cm转矩力的作用下,随着转矩作用区域的增大,中切牙的唇向移动趋势逐渐减小,侧切牙由舌向移动逐渐转为唇向移动。结论转矩能够有效改变牙齿的移动方式,如果转矩从中切牙之间开始向两侧远中方向逐渐减少,效果会更好。     

Detection of anti-Brucella antibodies in llama (Llama glama)

Seven llamas were immunized with killed Brucella abortus S1119.3 cells and bled sequentially, resulting in 64 samples. An eighth llama was kept as a negative control. In addition, 299 llama and 2075 alpaca sera, submitted for diagnostic testing, were included. Sera from all llamas were tested by the buffered antigen plate agglutination test, the complement fixation test, and the indirect enzyme immunoassays using smooth and rough lipopolysaccharides. A competitive enzyme immunoassay and fluorescence polarization assays were also performed. The sensitivity values for llama sera ranged from 92.2% to 100% and the specificity values from 89.6% to 100%. No alpacas were immunized. The specificity values for alpaca sera ranged from 94.8% to 100% specific although some sera gave an 'agglutination like' reaction after about 10 minutes of incubation. The complement fixation test could not be used, as 31% of the sera were anticomplementary and 4% were false positive.     

Structure of the upper teeth of the filefish, Stephanolepis cirrhifer

The teeth of the filefish were investigated with light and scanning electron microscopy. Anterior and posterior rows of pleurodont teeth are present in the maxilla. The teeth in the anterior row possess a large labial and small lingual surface and the teeth in the posterior row possess a small labial and large lingual surface. The enameloid consists of three layers-the outer, middle, and inner layer. The outer layer consists of fine, parallel crystallites oriented perpendicular to the surface. They are similar to the crystallites in enamel derived from ectoderm. In the middle layer the crystallites form parallel bundles that are oriented in the same direction as that of the crystallites in the outer layer. In the inner layer bundles of crystallites interlace with each other. The tubular structures and the terminal branches of the dentinal tubules are located among the crystallites in the inner layer. In the dentine numerous dentinal tubules radiate from the pulp cavity toward the periphery and are seen to curve slightly in an S-shaped course. At the occlusal tip the alternating parabolic layers are seen and these are considered to be mineralized lines. The openings of the dentinal tubules are round or oval, surrounded by interlacing fibrils. The teeth are tightly fixed to the jawbone by bundles of fibrils. At the lower part of the lingual surface of the teeth in the anterior row and the labial surface of the teeth in the posterior row the bundles of fibrils start at the dentine and some fibrils run through connective tissue, while others terminate in projections of the jawbones.(ABSTRACT TRUNCATED AT 250 WORDS)     

Placentation in the alpaca Lama pacos

Reproduction in South American camelids is poorly studied. To extend our knowledge of the development and cellular physiology of the placenta in the alpaca Lama pacos, we have examined specimens from day 150 of pregnancy to term. Morphological investigations using light, transmission and scanning electron microscopy, the histochemical localization of iron, alkaline and acid phosphatase activity, and the immunodetection of placental lactogen hormone were performed. Throughout pregnancy there was a progressive increase in the depths of folds on the uterine mucosa surface together with a thickening of the endometrium. Glandular cells exhibited PAS and acid phosphatase (AcP) positive secretion granules. In the chorion, giant trophoblast polyploid cells gradually became more numerous and larger. Non-giant cells exhibited positive granules for PAS, alkaline phosphatase (AkP) reaction and immunostaining for bovine placental lactogen hormone (PLH). SDS -PAGE electrophoresis and Western blotting procedures also confirmed the presence of a bovine PLH-like glycoprotein in the fetal alpaca placenta. Over the glandular openings, the chorion formed typical areolae, where the trophoblast exhibited AcP and PAS positive reactions. At these sites, the fetal endothelial cells contained iron-storage granules in their cytoplasm. The trophoblast-epithelial interface exhibited a complex microvillous interdigitation, in which an AkP reaction was very prominent. The chorionic capillaries progressively indented adjacent trophoblast cells. These data suggest that although the epitheliochorial alpaca placenta is diffuse, various trophoblast cell types and specialized areas of the maternofetal interface give the placenta micro-regional functions where histiotrophic nutrition, hormone production and molecular exchange are prevalent.     

The Enamel Microstructures of Bovine Mandibular Incisors

Bovine teeth have been considered as an excellent substitute for human teeth for dental research, however, the enamel microstructures of bovine incisors that include arrangements of prisms and interprisms, and their spatial relationships have not been well described. The aim of this study was to investigate the detail enamel microstructures of bovine incisors. Eight bovine mandibular incisors were cut into 77 pieces at eight equal intervals either in the longitudinal direction or in the horizontal direction before each piece had been tangentially cut (parallel to enamel–dentin junction) through the middle of the enamel thickness. All the sectioned surfaces were treated 1 M HCl for 10 sec to expose the prisms and interprisms before observation by scanning electron microscopy. The parallel enamel prisms were located in all the outer enamel, the cervical region and the incisal ridge of the bovine incisors. Most labial inner enamel and the cingulum of lingual inner enamel were composed of the Hunter–Schreger bands with the characteristics of decussating groups of prisms and decussating planes between interprisms and prisms. The interprisms were thicker in the inner enamel than in the outer enamel. Anat Rec, 2012. © 2012 Wiley Periodicals, Inc.     

A method for sampling the stages of amelogenesis on mandibular rat incisors using the molars as a reference for dissection

A method for locating specific stages of amelogenesis on continuously erupting incisors was devised for rats weighing 101 +/- 5 g (n = 32). The technique is based on reflecting reference lines from the mandibular molars as perpendiculars to the labial surface of mandibular incisors. From these reference lines additional measurements are then made along the midline of the labial surface of the incisor in an apical or incisal direction to find the site desired for sampling. Histological studies on 24 decalcified incisors split into segments by using such reference lines and reconstructed by morphometry indicated that a reference line reflected from the contact point between the 2nd and 3rd molars crossed the enamel organ and adjacent enamel at 3,181 +/- 329 microns incisal to the start of the secretory zone of amelogenesis. A reference line from the 2nd and 1st molars crossed the enamel organ and enamel at 1,238 +/- 424 microns incisal to the start of the maturation zone of amelogenesis, while a reference line from the mesial side of the 1st molar crossed the enamel organ and enamel almost exactly where the enamel becomes completely soluble following prolonged decalcification in EDTA. Although reference lines were reproducible within a group of male rats having similar body weights, the linear distance between the apical end of the incisor and the point at which they crossed the tooth increased at a rate of 1 mm per 159 g for rats between 50 and 300 g body weight. This suggests that molars do not maintain a fixed relationship to incisors over time, and extreme care must be taken to standardize an experiment to a specific body weight when using this method.     

A spontaneous mutation: amelogenesis imperfecta with cysts in rats

Amelogenesis imperfecta (AI) is an inherited dental disease of enamel formation in humans, and there are various phenotypes due to the combination of enamel quality and quantity. We encountered four female IGS rats with spontaneous AI including odontogenic cysts in the incisor teeth. Histopathologically, in the incisors of the rats, the enamel organ was disorganized with the remaining enamel matrix residing within the enamel space. The expanding cysts derived from the enamel organ were formed in the periosteal connective tissue on the labial side. At the bottom of the tooth germs, the precursor cells of the epithelial root sheath were arranged regularly and the enamel organs were preserved to the same degree as those of normal rats. In the molar teeth of the affected rats an enamel matrix remained on the neck and crown of the erupted teeth; however, no abnormality was observed at the tooth root. Although an animal model of AI has been developed from mutants of the SHR-SP rat strain, the present cases represent another potential model of the disease because of the differences in the way the enamel matured and the odontogenic cyst formation in the incisors.     

Adaptive functional specialisation of architectural design and fibre type characteristics in agonist shoulder flexor muscles of the llama, Lama glama

Like other camelids, llamas (Lama glama) have the natural ability to pace (moving ipsilateral limbs in near synchronicity). But unlike the Old World camelids (bactrian and dromedary camels), they are well adapted for pacing at slower or moderate speeds in high-altitude habitats, having been described as good climbers and used as pack animals for centuries. In order to gain insight into skeletal muscle design and to ascertain its relationship with the llama's characteristic locomotor behaviour, this study examined the correspondence between architecture and fibre types in two agonist muscles involved in shoulder flexion (M. teres major - TM and M. deltoideus, pars scapularis - DS and pars acromialis - DA). Architectural properties were found to be correlated with fibre-type characteristics both in DS (long fibres, low pinnation angle, fast-glycolytic fibre phenotype with abundant IIB fibres, small fibre size, reduced number of capillaries per fibre and low oxidative capacity) and in DA (short fibres, high pinnation angle, slow-oxidative fibre phenotype with numerous type I fibres, very sparse IIB fibres, and larger fibre size, abundant capillaries and high oxidative capacity). This correlation suggests a clear division of labour within the M. deltoideus of the llama, DS being involved in rapid flexion of the shoulder joint during the swing phase of the gait, and DA in joint stabilisation during the stance phase. However, the architectural design of the TM muscle (longer fibres and lower fibre pinnation angle) was not strictly matched with its fibre-type characteristics (very similar to those of the postural DA muscle). This unusual design suggests a dual function of the TM muscle both in active flexion of the shoulder and in passive support of the limb during the stance phase, pulling the forelimb to the trunk. This functional specialisation seems to be well suited to a quadruped species that needs to increase ipsilateral stability of the limb during the support phase of the pacing gait. Compared with other species, llama skeletal muscles are well suited for greater force generation combined with higher fatigue resistance during exercise. These characteristics are interpreted as being of high adaptive value, given the llama's habitat and its use as a pack animal.     

Induction of immune responses and molecular cloning of the heavy chain antibody repertoire of Lama glama

Functional heavy chain immunoglobulins have, so far, only been found in camels and llamas. Antigen-specific fragments of these heavy chain IgGs (V_HH) are of great interest in biotechnology because they are very stable and can be produced at high level by the yeast Saccharomyces cerevisiae. The work described in this paper was conducted to determine whether llamas (Lama glama) are a practical source of antigen-specific V_HH fragments. Llamas were immunised with various types of antigens and the antibody responses were examined during the course of immunisation. Both, conventional and heavy chain IgG antibodies were produced in response to each of the antigens. The heavy chain IgG repertoire displayed a recognition pattern different to that of conventional llama IgGs, resulting in the expansion of the accessible epitope repertoire. Llamas have a lower proportion of heavy chain IgG antibodies in their serum than have camels. To enable the specific and efficient isolation of V_HH genes from peripheral blood B-cells, the long and short-hinge sequences of Lama glama heavy chain IgGs were determined, revealing the presence of a novel subclass of short-hinge heavy chain IgG. Long and short-hinge specific PCR primers were designed to be used in the construction of llama V_HH libraries. We conclude that, using the techniques described, antigen-specific V_HH antibody fragments are readily accessible from the llama, thus providing highly valuable binding molecules for a variety of applications.     

Relationships between tooth eruption, occlusion and alveolar bone resorption: cytological and cytochemical studies of bone resorption on rat incisor alveolar bone facing the enamel

The rat labial incisor alveolar bone facing the enamel and bearing the occlusal force was examined by electron microscopy after being compared with the lingual alveolar bone by histological and scanning electron microscopic (SEM) observations. On the labial side, shallow resorptive lacunae were recognized all over the bone surface; these were mainly covered by osteoclasts and some mononuclear cells. The cement line was absent from the bone matrix. On the lingual side, residues of Sharpey's fibers, the bone formation surface and deep resorptive lacunae were observed by SEM. Histologically, bone remodeling areas showing both osteoclasts and active bone-forming osteoblasts on the bone surface, as well as many cement lines in bone matrix, were recognized. Furthermore, electron microscopic and cytochemical studies demonstrated that mononuclear cells located close to osteoclasts displayed osteoblastic characteristics such as alkaline phosphatase activity, a developed Golgi apparatus, and a rough endoplasmic reticulum. These findings indicate that continuous bone resorption occurs on the labial bone surface, while active bone remodeling occurs on the lingual surface. Even on the labial surface, osteoblastic cells close to osteoclasts seem to play an important role in the differentiation and or activation of osteoclasts.     

Non satellite veins in the pelvic limb of the llama (Lama glama)

The objective of this study was to determine the major differences in the venous system of the pelvic limb of the llama (Lama glama) and that of other mammals, including humans. Eight adult llamas, preserved by means of 6% formalin solution at 0°C, were dissected. The venous system was perfused with a solution of 17% coloured industrial gelatin. Two venous systems are recognised, superficial and deep. The veins of the deep system follow the same course as the arteries, except for two important veins located in the femoral, popliteal and crural regions. A more voluminous vein than the femoral originates in a venous tripod in the proximal part of the femoral canal and forms an anastomosis with the femoral vein distally. The popliteal vein gives off an expansion located along the popliteal region. In the leg, it continues with a vein that accompanies the tibial nerve and forms an anastomosis distally with the medial saphenous vein. The described venous distribution represents an interesting complement to the medial saphenous, popliteal and femoral veins, being more important than the last one for its volume. In the thigh it would correspond to the deep femoral vein as in humans and dogs; the homology in the popliteal and leg regions is too difficult because it probably corresponds to the caudal tibial vein of the human, the caudal branch of the medial saphenous vein, and the satellite vein of the tibial nerve in the horse. Anat Rec 256:99–103, 1999. © 1999 Wiley‐Liss, Inc.     

Effects of ascorbate-deficiency on collagen secretion and resorption in cultured mouse incisor germs

The effects of ascorbic acid deficiency on mouse incisors, grown in vitro, has been investigated at the histological and cytological levels. In this model, continuously growing mouse incisors are characterized by the existence of different type of predentin-dentin matrix on its lingual (root-analogue) and labial (crown-analogue) surface and the absence of enamel on the lingual surface. Our observations indicated that ascorbate-deficiency affected the behavior of mouse tooth germs in vitro: odontoblast differentiation was disturbed and morphological evidence for odontoblast-mediated collagen resorption were observed. An abnormal amorphous predentin-dentin matrix existed and the basement membrane was prematurely disrupted. The dentin mineralization, as well as functional differentiation of ameloblasts were strongly hampered. Chronic deficiency led to disorganization of the dental tissues.     

Effects of ascorbate-deficiency on collagen secretion and resorption in cultured mouse incisor germs.

The effects of ascorbic acid deficiency on mouse incisors, grown in vitro, has been investigated at the histological and cytological levels. In this model, continuously growing mouse incisors are characterized by the existence of different type of predentin-dentin matrix on its lingual (root-analogue) and labial (crown-analogue) surface and the absence of enamel on the lingual surface. Our observations indicated that ascorbate-deficiency affected the behavior of mouse tooth germs in vitro: odontoblast differentiation was disturbed and morphological evidence for odontoblast-mediated collagen resorption were observed. An abnormal amorphous predentin-dentin matrix existed and the basement membrane was prematurely disrupted. The dentin mineralization, as well as functional differentiation of ameloblasts were strongly hampered. Chronic deficiency led to disorganization of the dental tissues.