Lingual Papillae Research Articles

The Innervation of the Fetal Buffalo Tongue

, 1998).Since the lingual papillae of the domestic ruminant show an extreme variety of shape, webelieved it interesting to extend the study to innervation of the lingual papillae of the waterbuffalo, analyzing the morphological, structural, and immunohistochemical characteristicsof the gustatory papillae.MATERIALS AND METHODSThe study was performed on 25 water buffalo fetuses of both sexes, from 5 to 60cm inlength, and originating from a local slaughter-house. The fetus length, from the sinciput tothe tail root, was utilized as a development index. The fetuses were subdivided into fivegroups in relation to their length (5–7cm; 10–15cm; 20–25cm; 30–35cm; 40–60cm). Foreach group samples were prepared for scanning electron microscopy (SEM), transmissionelectron microscopy (TEM), light microscopy (LM), and the simple immunohistochemicalmethod.The TEM samples were treated by the usual techniques.

Role of IGFBPs in the morphogenesis of lingual papillae

The expression of insulin-like growth factor binding proteins (IGFBPs) during the morphogenesis of lingual papillae of mice was examined by in situ hybridization. Among seven mouse IGFBPs, IGFBP-1, -6, and -7 mRNAs were not expressed in the tongue tissue. At E12, though no papillae have formed yet, IGFBP-2, -4, and -5 were expressed in the entire tongue epithelium. At E14, fungiform papillae appeared in the anterior region and circumvallate papillae were distinguished in the posterior region. Strong expression of IGFBP-5 was observed in the apical region of both fungiform and circumvallate papillae. At this stage, the epithelial elevation of filiform papillae was not clear; but IGFBP-5 was expressed in the apex. At E15, foliate papillae were distinguished and IGFBP-5 was expressed in the dorsal epithelium of ridges. In filiform papillae, IGFBP-3 was expressed in the core of the connective tissue. At E17, the expression of IGFBP-5 disappeared from the apical region of fungiform, filiform, foliate, and circumvallate papillae, whereas that of IGFBP-2 remained. This finding suggests that IGFBP-5 and -2 function to cause evagination of the epithelium into a raised structure. In the epithelium of trenches of foliate and circumvallate papillae, strong expression of IGFBP-4 was observed at E15 and E17. As previously suggested from a study on postnatal mice (Suzuki et al. J Comp Neurol 2005;482:74-84), IGFBP-4 acts in the epithelial invagination to form the trenches, grooves, or furrows of lingual papillae during development.

Scanning Electron Microscopic Structure of the Lingual Papillae of the Common Opossum (Didelphis marsupialis)

The mammalian tongue has evolved for specialized functions in different species. The structure of its papillae tells about the animal's diet, habit, and taxonomy. The opossum has four kinds of lingual papillae (filiform, conical, fungiform, vallate). Scanning electron microscopy of the external features, connective tissue cores, and corrosion casts of the microvasculature show the filiform papillae have a spearhead-like main process and spiny accessory processes around the apical part of the main process. The shape and number of both processes depend on their position on the tongue. On the apex, the main processes have shovel-like capillary networks and the accessory processes have small conical networks. On the lingual radix, the processes have small capillary loops. In the patch region, conical papillae have capillaries arranged as a full sail curving posteriorly. The fungiform papillae are scattered among the filiform papillae and have capillary baskets beneath each taste bud. Giant fungiform papillae on the tongue tip are three to four times larger than the ones on the lingual body. Capillaries of giant papillae form a fan-shaped network. The opossum has three vallate papillae arranged in a triangle. Their tops have secondary capillary loops but not their lateral surfaces. Mucosal folds on the posterolateral border have irregular, fingerlike projections with cylindrical capillary networks. These findings and the structure of the rest of the masticatory apparatus suggest the lingual papillae of opossum have kept their ancestral carnivorous features but also developed the herbivore characteristics of other marsupials.

Prevalence of Anaemia in Tribal School Children

Tribal population is about 13 per cent of total population of Rajasthan and it is the most unprevileged section of society. Therefore, tribal schools children aged 6-12 years from government schools of villages covered by Kun Primary Health Centre (PHC), Dhariyawad block of Udaipur region, Rajasthan were studied to assess the prevalence of anaemia. A total of 206 children from five schools were examined for clinical signs and symptoms of anaemia. Blood haemoglobin level of clinically anaemic children was analysed using digital haemoglobinometer by cyanmethaemoglobin method. Dietary intake was also noted by 24 hours recall method using food models and standardised bowls set. Nutrient intake was calculated by food composition tables and then compared with RDA (ICMR, 1990). SES was assessed by SES scale of Pareek and Trivedi (1963). Results revealed that 61 per cent children were from low socio-economic status families and remaining 39 per cent from low middle class. Clinical examination showed that 95.1 per cent children were clinically anaemic. Clinical signs such as pale conjuctiva (77.6 %), flat and plate nails (24.3 %), atrophic lingual papillae (12.4 %) and koilonychia. (44.3 %) were observed among the school children. Blood haemoglobin revealed that more than half (60.2 %) of these children were moderately anaemic (7-10 g/dl/), 32.9 per cent were severely anaemic (<7 g/dl) and 0.6 per cent mildly anaemic (10-11.5 g/dl). On the whole, 93.7 per cent children were suffering from different forms of anaemia. The results show high prevalence of anaemia in the tribal school children of Udaipur region suggesting that an intervention to cure and prevent anaemia may be started for school children also.

Lectin Histochemistry of Taste Buds in the Circumvallate Papilla of the Rat

In mammals, taste buds are localized mostly within two regions, i.e. lingual and palatal epithelium. In the lingual epithelium, taste buds are localized in three lingual papillae; fungiform, foliate and circumvallate papillae. In the palatal epithelium, taste buds are present in the nasoincisor papillae and soft palate. In addition, few taste buds are recognized at the laryngeal surface of epiglottis. Taste buds contain 50–80 elongated specialized epithelial cells (taste cells) and proliferative basal cells (progenitor cells). Ultrastructually, taste cells have traditionally been classified into two types of cells; dark and light cells. Dark cells or type I cells have an electron-dense cytoplasm and thought to be supporting cells. Light cells are characterized by the presence of electron-lucent cytoplasm and further subdivided into two types based on the presence of synaptic vesicles. Light cells without apparent synaptic vesicles are called type II cells, while those with synaptic vesicles are termed as type III cells. Progenitor cells are located at the basal portion of the taste buds. There are at least two theories for cell lineage of the taste buds; each type of cells in taste buds has individual progenitor cells (multi cell line theory) and all types of cells are derived from the same progenitor cell (one cell line theory). Several histochemical studies to demonstrate the expression of bioactive substances in specific cell type of taste cells for the analysis of cell lineage of the taste buds. Lectins are useful for histochemistry and have been used in several areas to identify particular cell populations on the base of different ability to bind with high specificity to different carbohydrate epitopes of cell membrane. Although several lectin histochemical studies have been carried out to the taste buds to map the overall distribution of different kinds of lectins in the different location of taste buds in various species (Witt and Reutter, 1988; Witt and Miller, 1992; Zeng et al., 1995; Ohnishi et al., 2000; Kano et al., 2001), few studies are conducted on the relation between lectin binding patterns and specific cell types within the taste buds. The present presentation focused on the lectin binding pattern of rat circumvallate papilla with special references to the relation with type II cells, i.e. α-gustducin-immunoreactive cells. Ulex europaeus agglutinin-I (UEA-I) reacts strongly with α(1, 2)linked fucose. In the taste buds of the circumvallate papilla, UEA-I was found to bind with the cell membrane of taste buds (Figure 1A, compared with Figure 1C). Within the taste buds, UEA-I bound to almost all individual cells in the taste buds. Most blood group H type 2 antigen (AbH) immunoreactive cells co-expressed UEA-I (Figure 1B). In addition to the taste buds, UEA-I binds occasionally to the keratinized layer of the trench wall of the circumvallate papilla and in the lingual epithelium. Although lectin from Lotus tetragonolobus is also specific for L-fucose, it did not, however, bind to the taste bud cells. Peanut agglutinin (PNA) is specific for terminal β-galactose and binds to the membrane of rounded-shaped cells at the basal portion of taste buds. Spindle-shaped cells within the taste buds were devoid of PNA binding. In the trench wall epithelium, PNA strongly labeled the membrane of cells from basal cell layer to granular cell layer. Cells at keratinized cell layer lack PNA bindings. In the lingual epithelium, strong bindings for PNA were also detected from basal cell layer to granular cell layer. Jacalin is purified from jackfruit and specific for D-galactose. Similar to PNA, jacalin binds the membrane of cells located from basal cell layer to granular layer in the trench wall epithelium. Keratinized cells lacked in jacalin binding. Some rounded cells at the basal portion of the taste buds were also found to bind to jacalin. PNA bound to membrane of some rounded-shaped cells that does not react with jacalin. Within the taste buds, some spindle-shaped cells reacted with both wheat germ agglutinin (WGA) and succinyl WGA. The number of cells binding to WGA is slightly smaller than that binding to succinyl WGA. Many cells binding to WGA and succinyl WGA displayed immunoreactivity for α-gustducin; Not all cells lebeled with WGA or succinyl WGA showed α-gustducin immunoreactivity and vise versa. Dolichos bifforus agglutinin (DBA) binds to the mostly apical portion of the spindle-shaped cells within the taste buds of circumvallate papilla. Occasionally, DBA also binds to the cytoplasm of the basal portion of taste buds. The number of cells binding to DBA is low compared to cells binding to WGA or succinyl WGA. Most of cells binding with DBA did not show immunoreactivity for α-gustducin. The present lectin histochemical analysis of taste cells showed that some lectins bind specifically to certain types of cells in the rat circumvallate papilla.

Development of Gustatory Organs and Innervating Sensory Ganglia

Taste function requires neural circuits to transmit gustatory infor-mation from taste receptor cells in taste buds, via afferent nerves tothe soma of ganglion neurons, and through central ganglion proc-esses into the brainstem. During initial formation, the sensoryganglion neurons have a key situation in establishing receptive fieldsby extending neurites bidirectionally, to the peripheral taste organsand to central taste nuclei.Our laboratory is studying functional differentiation of sensoryganglia that innervate the tongue, and morphogenesis andpatterning of the tongue and papilla organs. The lingual ganglia andtaste papillae initially develop independently, but then becomereciprocally dependent as ganglia derive molecular support fromgustatory papillae and the papillae require sensory innervation forgrowth and morphogenesis (Mistretta, 1998). Currently our focus ison the geniculate and trigeminal ganglia, which innervate anteriortongue, and the fungiform papilla taste organs innervated by theseganglia. Geniculate and trigeminal ganglia innervate spatiallycontiguous, but functionally distinct, sensory organs of the fungi-form papilla: the trigeminal neurons innervate lateral papilla epithe-lium and subserve somatosensation and nociception, whereas thegeniculate axons project to central apical papilla epithelium to inner-vate cells that will form taste buds for gustatory sensation (Mistrettaand Hill, 2003).In this brief paper we summarize some recent work on develop-ment of tongue and taste regions, and on early functional pheno-types of the geniculate and trigeminal ganglia.

Jacalin and peanut agglutinin (PNA) bindings in the taste bud cells of the rat: New reliable markers for type IV cells of the rat taste buds

Lectin histochemistry of Jacalin (Artocarpus integrifolia) and peanut agglutinin (PNA), specific lectins for galactosyl (beta-1, 3) N-acetylgalactosamine (galactosyl (beta-1, 3) GalNAc), was applied to the gustatory epithelium of the adult rat. In the ordinary lingual epithelium, Jacalin and PNA labeled the cell membrane from the basal to granular cell layer. They also bound membranes of rounded-cells at the basal portion of taste buds, but the number of PNA labeled cells was smaller than that of Jacalin labeled cells. There was no apparent difference in the binding patterns of Jacalin and PNA among the taste buds of the lingual papillae and those of the palatal epithelium. Occasionally, a few spindle-shaped cells were labeled with Jacalin, but not with PNA. Double labeling of Jacalin and alpha-gustducin, a specific marker for type II cells, revealed that Jacalin-labeled spindle-shaped taste cells were immunonegative for alpha-gustducin. Spindle-shaped cells expressing protein gene product 9.5 (PGP 9.5) immunoreactivity lacked Jacalin labeling. During the development of taste buds in circumvallate papillae, the binding pattern of Jacalin became almost identical from postnatal day 5. The present results indicate that rounded cells at the basal portion of the taste buds cells (type IV cells) bind to Jacalin and PNA, and these lectins are specific markers for type IV cells of the rat taste cells.

Comparative morphological studies on the stereo structure of the lingual papillae of selected primates using scanning electron microscopy

A scanning electron microscope was used to observe the lingual papillae and their connective tissue cores (CTCs) in five primates (tupai, tamarin, crab-eating monkey, mandrill, and human). There were some slender protrusions rising from the top of the filiform papilla in all five types of primate. After removing the epithelium the filiform CTC from the tupai, tamarin and crab-eating monkey displayed a U-shaped arrangement of rod-shaped protrusions. The filiform CTC from the crab-eating monkey also had a columnar base. The human filiform CTC consisted of a primary columnar base, numerous short rod-shaped secondary protrusions from its upper periphery, and a few central protrusions. The filiform CTC from the Mandrill was fundamentally similar to that of the human, however, its base was shorter. The fungiform CTC from the tupai was column shaped, with several depressions for taste buds on the top. There were three vallate papillae in the tupai, tamarin, and mandrill, approximately four in the monkey, and between five and twelve in the human. Moderately developed foliate papillae were found in the tamarin, monkey, mandrill and human. The tupai, however, possessed a finger-like lateral organ instead. The lingual root area of the tupai, tamarin, crab-eating monkey and mandrill was relatively small with a smooth surface. Only the human had a tonsil-structure, which was located on the surface of its larger lingual root.

Scanning electron microscopy of lingual papillae in the common shrew, Sorex araneus, L.

The dorsal surface of the tongue of the adult common shrew (Sorex araneus L.) was examined by scanning electron microscopy. As in the other insectivores, three types of lingual papillae were observed: filiform, fungiform and vallate papillae. The filiform papillae represented the most numerous type of lingual papillae. The characteristic feature of the filiform papillae, covering the apex and corpus of the tongue, is the two processes tilted to the root of the tongue. The filiform papillae on the lingual apex are reduced in size and structure. Five to six fungiform papillae are placed symmetrically along the left and right border of the corpus of the tongue. Two large oval vallate papillae are located on the radix of the tongue. The posterior surface of the tongue in common shrew is covered with a smooth mucosa with the openings of the serous glands.

Prion infection of skeletal muscle cells and papillae in the tongue.

The presence of the prion agent in skeletal muscle is thought to be due to the infection of nerve fibers located within the muscle. We report here that the pathological isoform of the prion protein, PrP(Sc), accumulates within skeletal muscle cells, in addition to axons, in the tongue of hamsters following intralingual and intracerebral inoculation of the HY strain of the transmissible mink encephalopathy agent. Localization of PrP(Sc) to the neuromuscular junction suggests that this synapse is a site for prion agent spread between motor axon terminals and muscle cells. Following intracerebral inoculation, the majority of PrP(Sc) in the tongue was found in the lamina propria, where it was associated with sensory nerve fibers in the core of the lingual papillae. PrP(Sc) staining was also identified in the stratified squamous epithelium of the lingual mucosa. These findings indicate that prion infection of skeletal muscle cells and the epithelial layer in the tongue can be established following the spread of the prion agent from nerve terminals and/or axons that innervate the tongue. Our data suggest that ingestion of meat products containing prion-infected tongue could result in human exposure to the prion agent, while sloughing of prion-infected epithelial cells at the mucosal surface of the tongue could be a mechanism for prion agent shedding and subsequent prion transmission in animals.

Effects of ovariectomy and estrogen replacement on rat oral mucosa

Objective: Although a close relationship between the oral discomfort and the menopause is suggested, it is not clear if the alteration of sex steroids would cause any histological or even pathological change in the oral mucosa. The aim of this study was to investigate the possible effect of sex steroid deficiency on the rat oral mucosa as well as the effect of its replacement. Methods: The oral mucosa of ovariectomized rats was compared to that of sham-operated rats both histologically and immunohistochemically. The same evaluation was also performed after hormone replacement therapy for the ovariectomized rats. Results: The ovariectomized animals presented reduced thickness of the mucosal epithelium with an irregular corneal (keratinised) surface. Partial disappearance of lingual papilla with irregular lete-peg was also noted. The corneal (keratinised) layer in the sublingual region was significantly thinner in ovariectomized animals. The results from immunohistochemistry showed the possibility that the turnover period was prolonged in ovariectomized rats. Estrogen replacement could compensate for the reduction in mucosal thickness in the animal model. Conclusions: The overall results suggest that sex steroids have a specific role in the maintenance of normal oral mucosa, and its deficiency possibly leads to the onset of oral discomfort in postmenopausal women.

The scanning electron microscopic study of lingual papillae in the silver fox ( Vulpes vulpes fulva, Desmarest, 1820)

The tongues of adult silver foxes were studied using scanning electron microscopy. Five types of lingual papillae were found on the dorsal surface of the tongue. The most numerous papillae were filiform papillae covering the apex and body of the tongue. The filiform papillae on the anterior part of the tongue are divided into 1 main and 10-12 accessory processes. In the posterior part of the body of the tongue the number of accessory processes is reduced. Fungiform papillae are located between the filiform papillae. A cluster of 12 large fungiform papillae was found on the apex of the tongue. Conical papillae are located in the area of the vallate papillae and cover the posterior part of the root of the tongue. Their size increases towards the pharynx, where they are distributed more sparsely. In the silver foxes there were two pairs of vallate papillae. The wall surrounding each papilla and its gustatory trench forms partly connected 6-8 conical papillae. The foliate papillae on both margins of the tongue body are small and consist of 4-5 laminae. The distribution and type of lingual papillae found in the silver fox are similar to those in the other species belonging to the family Canidae.

Morphology of the lingual papillae in the tiger.

The dorsal lingual surfaces of an adult tiger (Panthera tigris altaica) was examined by macroscopical and scanning electron microscopical observations. Filiform, fungiform and vallate papillae were observed. The filiform papillae were distributed over the entire dosal surface of the tongue. The fungiform papillae were present rounded bodies, and more densely distributed on the lingual apex. There were 4 vallate papillae in total on borderline between the lingual body and lingual radix. Each papilla was surrounded by a groove. No foliate papillae were seen on the dorsal surface. Openings of the glandular ducts on the regions of the vallate papillae were found.

The tadpole of Physalaemus fuscomaculatus (Anura: Leptodactylidae), with a description of internal oral morphology

The external morphology and internal oral features of Physalaemus fuscomaculatus (Steindachner, 1864) tadpoles are described and compared with other species in the Physalalemus biligonigerus group (Lynch, 1970). Tadpoles of P. fuscomaculatus have a morphology similar to other species within this group and closely resemble those of P. santafecinus Barrio, 1965 and P. biligonigerus (Cope, 1861). The tadpole of P. fuscomaculatus differs from those of the biligonigerus group by the following characters: i) proportionally shorter tail compared to total length, ii) presence of two narrow ventral gaps on marginal papillae of the oral disc; iii) upper jaw sheath Mshaped; iv) presence of two lingual papillae; v) two to three prepocket papillae, and vi) three to five pustulations anterior to each buccal pocket. This is the first record of Physalaemus fuscomaculatus in southeastern Brazil.

Disruption of sonic hedgehog signaling alters growth and patterning of lingual taste papillae

Taste buds on the anterior part of the tongue develop in conjunction with epithelial–mesenchymal specializations in the form of gustatory (taste) papillae. Sonic hedgehog (Shh) and Bone Morphogenetic Protein 4 (BMP4) are expressed in developing taste papillae, but the roles of these signaling molecules in specification of taste bud progenitors and in papillary morphogenesis are unclear. We show here that BMP4 is not expressed in the early tongue, but is precisely coexpressed with Shh in papillary placodes, which serve as a signaling center for both gustatory and papillary development. To elucidate the role of Shh, we used an in vitro model of mouse fungiform papillary development to determine the effects of two functional inhibitors of Shh signaling: anti-Shh (5E1) antibody and cyclopamine. Cultured E11.5 tongue explants express Shh and BMP4 LacZ in a pattern similar to that of intact embryos, localizing to developing papillary placodes after 2 days in culture. Tongues cultured with 5E1 antibody continue to express these genes in papillary patterns but develop more papillae that are larger and closer together than in controls. Tongues cultured with cyclopamine have a dose-dependent expansion of Shh and BMP4 LacZ expression domains. Both antibody-treated and cyclopamine-treated tongue explants also are smaller than controls. Taken together, these results suggest that, although Shh is not involved in the initial specification of papillary placodes, Shh does play two key roles during pmcry development: (1) as a morphogen that directs cells toward a nonpapillary fate, and (2) as a mitogen, causing expansion of the interplacodal epithelium and underlying mesenchyme.

Morphology of the dorsal lingual papillae in the Japanese macaque and Savanna monkey.

The dorsal lingual surfaces of infant Japanese macaque (Macaca fuscata) and adult savanna monkey (Cercopithecus aethiopus) were examined by scanning electron microscopy. Filiform, fungiform, foliate and vallate papillae were found. The filiform papillae were distributed over the entire dorsal surface of the tongue. The fungiform papillae were round in shape, and more densely distributed on the lingual apex. The foliate papillae were seen on the dorsolateral aspect of the tongue. The three vallate papillae were arranged like a triangle with the apex of the triangle directing caudally. Each papilla was surrounded by a groove. The rudiments of the fungiform, foliate and vallate papillae were visible earlier than those of the filiform papillae.

Morphological study of lingual papillae in the fetal cat at mid-gestation, as revealed by light and scanning electron microscopy.

Tongues were removed from five cat fetuses, after approximately 1 month of gestation, for examination by light and scanning electron microscopy. Rudiments of filiform papillae were visible over the entire dorsal surface of the tongue. The epithelium of the dorsal surface of the tongue was of the stratified cuboidal type. No evidence of keratinization was detected anywhere in the entire dorsal lingual epithelium. Rudiments of fungiform papillae were recognizable only at the lingual apex; none were recognizable on other parts of the dorsal surface of the tongue at this stage. By contrast, rudiments of circumvallate and foliate papillae were already distinguishable from the filiform papillae. Differences between these results and those obtained previously in rats and mice are discussed.

Angioarchitectural characteristics of the anterodorsal surface of aging and young rabbit tongue: scanning electron microscopy study

The three-dimensional angioarchitecture of young and aging rabbit tongues were studied using a vascular corrosion cast method. The vascular network of different lingual papillae were observed by scanning electron microscope (SEM). Young animals showed filiform papillae distributed all over considered area, with fungiform ones among them. In comparison to developed specimens, young rabbits demonstrated dense and higher organization of microvascular network.

Phenotypic plasticity of anuran larvae: environmental variables influence body shape and oral morphology in Rana temporaria tadpoles

AbstractEnvironmental variables shaped the morphology of tadpoles of the common frog, Rana temporaria, in various ways at the Pyrenean locality Circo de Piedrafita. Examining only specimens in similar developmental stages, those from small ponds (with higher temperature and higher tadpole density) had lower growth rates, lower relative tail height, lower relative body width and fewer labial keratodonts and keratodont rows. The variation in keratodonts may have been caused by heterochrony related to the slower growth rate. The number of lingual papillae also differed between ponds but was not related to pond size. Higher predator densities caused a higher percentage of damaged tails and a lower relative tail length in specimens with apparently intact tails, probably as a result of incomplete regeneration after mutilations earlier in development.

Scanning electron microscopy study of the tongue and lingual papillae of the california sea lion (Zalophus californianus californianus)

We observed the three-dimensional structures on the external surface and the connective tissue cores (CTCs) of the California sea lion (Zalophus californianus californianus), after exfoliation of the epithelium of the lingual papillae (filiform, fungiform, and vallate papillae), using scanning electron microscopy (SEM) and conventional light microscopy. Macroscopically, the tongue was V-shaped and its apex was rounded. At the posterior area of the tongue, five vallate papillae were arranged in a V shape. In the epithelium, numerous taste buds were distributed on the top of the vallate papillae. On the dorsal surface from the apex to the boundary between the anterior and posterior tongue, filiform papillae were densely distributed. The CTCs of the filiform papillae consisted of a main protrusion (primary core) and many small cores (secondary cores). From the apex to the anterior one-third of the tongue, dome-like fungiform papillae were densely distributed, whereas fewer were located at the posterior two-thirds of the tongue. Several taste buds were found in the epithelium on the fungiform papillae. The size of the filiform papillae gradually increased from the apex to the boundary between the anterior and posterior tongue. At the lingual radix, the conical papillae, which were bigger than any filiform papillae, were densely distributed. The morphological characteristics of the tongue of the California sea lion appear to have been transformed to adapt to an aquatic environment; however, they possess some structures similar to those of land mammals. Anat Rec 267:146–153, 2002. © 2002 Wiley-Liss, Inc.