Contribución de la prueba rápida NS1 e IgM al diagnóstico de dengue en Colombia en el periodo pre-zika / Contribution of NS1 and IgM based-rapid tests to dengue diagnosis in Colombia before zika emergence

Objetivo: Se considera que el diagnóstico del dengue es fundamentalmente clínico; sin embargo, las pruebas rápidas basadas en la detección de IgM o NS1/IgM están siendo utilizadas en los servicios de salud. Este estudio determinó la contribución de las pruebas rápidas al diagnóstico de dengue en un área endémica antes de la introducción del virus zika. Metodología: Diseño de corte transversal de pruebas diagnósticas realizado a partir del análisis secundario de un estudio previo en 14 instituciones de salud del Valle del Cauca. Se obtuvo información de 632 participantes con resultados de prueba rápida, diagnóstico clínico y pruebas de referencia ELISA NS1, ELISA IgM y RT-PCR. Se compararon la sensibilidad, especificidad, valores predictivos y razones de verosimilitud del uso solo, en serie, y paralelo de los componentes NS1, IgM, NS1/IgM de la prueba rápida y el diagnóstico clínico con las pruebas Q de Cochran y McNemar para datos pareados. Resultados: La sensibilidad del diagnóstico clínico (61,4% IC95% 56%-66,7%) fue superior a la de las pruebas rápidas (37% IC95% 29,6%-44,7%) (P Conclusión: El diagnóstico clínico tiene una mayor sensibilidad que las pruebas rápidas, pero por si solo no es suficiente para confirmar o descartar dengue. Un resultado positivo en pruebas rápidas en pacientes con diagnóstico clínico de dengue es útil para confirmarlo, pero un resultado negativo no lo descarta.

Objective: Dengue diagnosis is considered to be mainly clinical; however, rapid tests that detect IgM or NS1/IgM are being used in health services. This study assessed the contribution of rapid tests to dengue diagnosis in an endemic area before the emergence of zika virus in Colombia. Methods: Cross-sectional study of diagnostic tests based on a secondary analysis of a previous study in 14 health care institutions in Valle del Cauca department. Results of dengue rapid test, clinical diagnosis, and reference tests ELISA NS1, ELISA IgM, and RT-PCR were obtained for 632 participants. The sensitivity, specificity, predictive values and likelihood ratios of the use alone, serial and parallel combinations of NS1, IgM, NS1/IgM of the rapid test and clinical diagnosis were compared using Cochran´s Q and MacNemar tests for paired data. Results: The sensitivity of clinical diagnosis (61.4% 95%IC 56-66.7) was higher than the sensitivity of rapid tests (37% 95% IC 29.6-44.7) (P<0.001). The serial used of NS1/IgM rapid test when clinical diagnosis was negative increased the sensitivity to 79.5% and, the serial use when clinical diagnosis was positive increased the specificity (from 66.3% to 98.7%). However, the latter decreased the sensitivity to 32.2%. While all negative likelihood ratios (LR-) were close to 1, the serial use of rapid tests when clinical diagnosis was positive had LR+ higher than 10. Conclusion: The clinical diagnosis is more sensitive than rapid tests, but by itself does not confirm or rule out dengue. A positive result in rapid tests is useful to confirm dengue but a negative result does not rule it out.

Complex secretory products in the oviduct of the plethodontid salamander Bolitoglossa dofleini (Amphibia, Urodela).

The ultrastructure of the secretory granules in the cells of the subdivisions of the oviduct in the neotropical plethodontid salamander Bolitoglossa dofleini was studied by transmission electron microscopy. In addition, we applied the cationic dye Cuprolinic Blue (CB) at different electrolyte concentrations to demonstrate proteoglycans, and the pyrogallol red-copper (PR-C) method to stain proteins at the ultrastructural level. The entire oviduct is lined by a simple epithelium that contains ciliated and microvillous cells in the first subdivision, the aglandular pars recta; microvillous cells show a moderate secretory activity. The following pars convoluta is differentiated into five glandular subdivisions and the aglandular "uterine portion". Especially in the glandular parts, the epithelium is arranged in longitudinal folds. At their crests ciliated and microvillous cells similar to those in the pars recta occur. Gland cells are crowded with secretory granules that differ in their structural complexity (with and without electron-dense spheres or masses; elaborated, homogeneous or granular matrix; spherical; distorted) along the various subdivisions. Further, as suggested by the CB-technique, the cranial subdivisions contain large amounts of sulphated proteoglycans that decrease in the caudal direction. Carboxylated proteglycans appear to be present in all subdivisions examined. Electron-dense spheres of secretory granules are largely free of CB-precipitates, but stain more or less intensely with PR-C. The ultrastructure of the pars recta, and especially the "uterine portion" indicates transporting capability. The epithelial cells of the "uterus" have coated pits and a considerable amount of lysosome-like bodies.

Characterization of glycosaminoglycans during tooth development and mineralization in the axolotl, Ambystoma mexicanum.

Glycosaminoglycans (GAGs) involved in the formation of the teeth of Ambystoma mexicanum were located and characterized with the cuprolinic blue (CB) staining method and transmission electron microscopy (TEM). Glycosaminoglycan-cuprolinic blue precipitates (GAGCB) were found in different compartments of the mineralizing tissue. Various populations of elongated GAGCB could be discriminated both according to their size and their preferential distribution in the extracellular matrix (ECM). GAGCB populations that differ in their composition could be attributed not only to the compartments of the ECM but also to different zones and to different tooth types (early-larval and transformed). Larger precipitates were only observed within the dentine matrix of the shaft of the early-larval tooth. The composition of the populations differed significantly between the regions of the transformed tooth: pedicel, shaft and dividing zone. In later stages of tooth formation, small-sized GAGCBs were seen as intracellular deposits in the ameloblasts. It is concluded that the composition of GAGCB populations seems to play a role in the mineralization processes during tooth development in A. mexicanum and influence qualitative characteristics of the mineral in different tooth types and zones, and it is suggested that GAGs might be resorbed by the enamel epithelium during the late phase of enamel formation.

Development of larval and transformed teeth in Ambystoma mexicanum (Urodela, Amphibia): an ultrastructural study.

Odontogenesis of early larval non-pedicellate teeth, late larval teeth with a more or less distinct dividing zone and fully transformed pedicellate teeth in Ambystoma mexicanum (Urodela) was studied to obtain insights into the development of differently structured teeth in lower vertebrates. Using transmission electron microscopy we investigated five developmental stages: (1) papilla; (2) bell stage (secretion of the matrix begins); (3) primordium (mineralization and activity of ameloblasts starts); (4) replacement tooth (young, old); and (5) established, functional tooth. Development of the differently structured teeth is largely identical in the first three stages. Mineralization takes place in apico-basal direction up to the (prospective) pedicel (early and some late larvae) or up to the zone that divides the late larval and transformed tooth in pedicel and dentine shaft (pedicellate condition). Mineralization starts directly at the collagen and by means of matrix vesicles. First odontoblasts develop small processes that extend to the basal lamina of the inner epithelial layer of the enamel organ. The processes are small and lack organelles in early larval teeth, but become larger, arborescent, and contain some organelles in late larval and transformed teeth. The processes are surrounded by unmineralized matrix (predentine). Odontoblasts at the basis of the teeth, at the pedicel, and in the zone of division do not develop significant cytoplasmic processes that extend into the matrix. Cells of the inner enamel epithelium differentiate to ameloblasts that secrete the enamel. In the early larval tooth they show an extensive basal labyrinth that becomes regressive when the enamel layer is completed. In late larval and transformed teeth, however, a large cavity arises between the basal ruffled border of ameloblasts and their basal lamina. This cavity appears to mediate amelogenesis. A small apical zone in early, but not in late larval teeth directly below the thin enamel layer consists of enameloid and is free of dentine channels.

The secondary tongue of Salamandra salamandra: histochemical and ultrastructural aspects of the developing lingual epithelium.

The development of the lingual epithelium of Salamandra salamandra was investigated with emphasis on histochemical and ultrastructural aspects. The temporal and spatial occurrence and the typical appearance of various cell types; i.e. pavement cells, replacement pavement cells, basal cells, mitochondria rich cells, goblet cells and glandular cells have been analysed and documented in detail from the young larval stage up to the metamorphosed animal (2 months after metamorphosis). It is shown that anatomical re- and de novo-constructions related to the formation of the secretory tongue led to distinct changes in the cellular equipment of the epithelium of the tongue, including various histochemical properties. Finally, functional aspects of the morphological characteristics are discussed in detail and compared with respective findings in other species.

The development of the tooth pattern and dentigerous bones in Polypterus senegalus (Cladistia, Actinopterygii).

The formation sequence of the tooth-bearing bones and the tooth pattern in early ontogeny of Polypterus senegalus is investigated using transparent preparation, histological sections, and SEM. During the attachment step of the yolk-sac larva the first dermal bones and teeth are formed. Teeth appear simultaneously in the areas of the maxillary, dentary, dermopalatine, prearticular, and coronoid 1 along with the first separate anlagen of these bones. A monostichous arrangement of primary teeth is established on the maxillary, dentary, and dermopalatine. Polystichous tooth arrangements do not occur before the early pterolarval phase, and then only in connection with bones of the palate and inner dental arcades. Especially pronounced is the influence of tooth formation on the structure of the parasphenoid that becomes much thickened by accretion of denticulate platelets, but we found neither evidence for a distinct vomeral contribution to the parasphenoid, nor a composite origin of the ectopterygoid in ontogeny. First replacement teeth are found in association with the maxillary and dentary as early as the late apterolarval phase. Primary teeth are of a single general type, whereas from the pterolarval phase onward three tooth types can be distinguished that are restricted to certain tooth bearing bones. Relatively late in ontogeny, dermo-metapterygoid and entopterygoid become formed and colonised by teeth, whereas first branchial teeth and tooth plates appear earlier during the first phase of extrinsic larval feeding. Characteristics of development of the dentition are discussed in comparison with character states of other better known fossil and recent taxa among Actinopterygii and Sarcopterygii. Compared to the assumed basic pattern of actinopterygian fishes, Polypteriformes show a derived condition with respect to structure, arrangement, replacement, and differentiation of teeth, which arises in sequence during larval development. This also corresponds to observed changes of feeding behaviour and functional demands during larval life.

The structure and development of the skull of Costa Rican plethodontid salamanders (Amphibia: Urodela).

We describe the structure of the skulls of the Costa Rican plethodontid salamanders Bolitoglossa subpalmata, Oedipina uniformis and Nototriton abscondens, and the characteristic sequences of development and ossification of the bony elements resulting from direct development using mainly cleared and stained specimens. Significant differences between the species studied are observed. N. abscondens possesses the broadest premaxillary pars dentalis and O. uniformis the narrowest one. The premaxillary dorsal processes are fused over their rostral third only in B. subpalmata; over half their extention in N. abscondens and almost completely in O. uniformis. A prefrontal is always present in N. abscondens; it is hidden underneath the nasal or missing in B. subpalmata, and it is always absent in O. uniformis. The skull bones, with the exception of the orbitosphenoid, develop and ossify sequentially from caudal to rostral in these directly developing species. A more massive pars dentalis of a generally narrower premaxillary are found as typical characters in males.

Teeth and their sex-dependent dimorphic shape in three species of Costa Rican plethodontid salamanders (Amphibia: Urodela).

The shape of the teeth and their sex-dependent dimorphic expression in three species of Costa Rican plethodontids (Bolitoglossa subpalmata, Oedipina uniformis and Nototriton abscondens) were studied using light microscopy and scanning electron microscopy. The teeth of the vomerine tooth patches are about one third larger than the teeth of the jaws in B. subpalmata and O. uniformis, whereas all teeth of N. abscondens are of about uniform size. The occurrence of bicuspid tooth germs in the fetus proves that primary teeth are bicuspid in these directly developing plethodontids. Females possess only bicuspid teeth consisting of a pedicel and a crown, as is considered characteristic for urodeles after metamorphosis. Adult males possess conical monocuspid teeth on the premaxillary. These teeth--which are similar to the typical late larval tooth of salamanders presenting a larval stage--are about twice as big as the neighbouring bicuspid maxillary teeth. N. abscondens males possess some monocuspid teeth and teeth of aberrant shapes on the premaxillary and the maxillaries. A tendency to build more monocuspid teeth in the premaxillary region than in the maxillary region can be observed in this species. We suppose that different degrees of sensitivity to androgens in each section of the dental lamina of the upper jaw cause the secondary occurrence of conical monocuspid teeth predominantly on the premaxillary section.

The odontoclasts of Ambystoma mexicanum.

The resorption of teeth in Ambystoma mexicanum during postembryonal ontogenesis and induced metamorphosis occurs by means of light-microscopic detectable giant-cells. These have morphological and functional characters similar to those of odontoclasts of other vertebrates. The multinucleated odontoclasts resorb not only the pedicel (base), but the stalk of the tooth, too. When active, the cells form a ruffled border and a sealing zone. In this way they are able to demineralize the hard tissues of teeth (dentin and mineral of the pedicel) and to dissolve the extracellular matrix. Resorption of enamel has not been observed. Marks of resorption resemble the Howship's lacunae of other tetrapods. TRAP as a typical enzyme of odontoclasts could not be detected histochemically. Dependence of PTH, which is supposed to be necessary for the formation and activation of odontoclasts as well as of thyroxine can be excluded, although the resorbing cells are functionally and cytologically identical with those of other vertebrates. This demands some other mechanism for the formation and regulation of the odontoclasts in A. mexicanum.

Development of the pattern of dentition and dental laminae of Costa Rican plethodontid salamanders (Amphibia: Urodela).

The pattern of development of teeth and dental laminae of three Costa Rican plethodontids (Amphibia, Urodela, Plethodontidae) was investigated using transparent preparations, light microscopy and scanning electron microscopy. The teeth of the jaws are monostichously positioned, those of the posterior vomeral parts are polystichously arranged. The anterior vomeral parts carry monostichously positioned teeth at the caudal margin; yet, the adult Bolitoglossa subpalmata possesses two lines. As a sex dimorphism adult males display long monocuspid premaxillary teeth which protrude to the outside of the mouth cavity. All species studied possess paired dental laminae in the lower jaw. Nototriton abscondens possesses an unpaired dental lamina in the upper jaw, which is constricted between the unpaired premaxillary and the maxillaries. In contrast, the dental laminae in the upper jaw of B. subpalmata and Oedipina uniformis are segmented into a premaxillary and two maxillary laminae. All species possess a pair of anterior vomeral and a pair of posterior vomeral dental laminae in the adults, whereas the vomeral dental laminae of the subadults are unsegmented. The pattern of dentition is compared with that of Gyrinophilus and Eurycea.

The epithelium of the tongue of Ambystoma mexicanum. Ultrastructural and histochemical aspects.

The distribution pattern of taste buds and goblet cells and histochemical and ultrastructural aspects of the tongue epithelium of Ambystoma mexicanum are here described. This study is also concerned with the developmental stages and origins of the epithelial cells. Pavement cells and goblet cells of the stratum superficiale are replaced by basal cells of the stratum germinativum in larvae and neotenous adults. The pavement cells of the larvae are characterized by a marginal layer of mucin grana. Decompaction of the mucins occurs immediately before extrusion in the adult. The larval goblet cell type (type I), which is also present in the adult, contains unfused grana of irregular shape. At the tip of the tongue, a further type (type II) of goblet cells is found. In the type II cells the intracellular secretory grana fuse to a single homogeneous mass. Leydig cells of the tongue epithelium are discerned by light microscopy first in the semi-adult, apparently correlated with partial metamorphosis. In the course of ontogenesis and induced metamorphosis the secretion changes to neutral glycoconjugates. The mucins of the pavement cells change first followed by those of the goblet cells. The glands of the secondary tongue show a dorso-ventral pattern of varying secretory qualities. Taste buds are found at the anterior margin of the tongue and along the base of the gill clasps in the early larva. They are already distributed all over the tongue at the end of the early larval phase.

The cycle duration of the seminiferous epithelium remains unaltered during GnRH antagonist-induced testicular involution in rats and monkeys.

Although the gonadotropic control of the spermatogenic process is well established, the endocrine regulation of the timing and kinetics of germ cell development has received little attention. We found previously that the administration of a GnRH antagonist (ANT) over a period of 25 days could retard spermatid development and slightly prolong cycle length in intact adult cynomolgus monkeys (Macaca fascicularis). The aim of the present study was to investigate the effects of extended exposure to ANT on the duration of the cycle of the seminiferous epithelium in the monkey. Additionally, the duration of spermatogenesis was studied in the ANT-exposed rat model. In experiment 1, monkeys were given either saline or ANT (n=6/group) and on day 30 all animals received a single injection of 5-bromodeoxyuridine (BrdU) to label S-phase germ cells. Testicular biopsies were taken on days 39, 43, 47 and 51 (end of treatment) for BrdU localization and flow cytometric analysis. ANT treatment suppressed hormone levels, reduced testis size by >70% and severely impaired germ cell production. Despite these alterations, cycle duration remained unchanged at all time-points compared with controls (10.12+/-0.15 days vs 10.16+/- 0.44 days). In experiment 2, adult male Sprague-Dawley rats (n=15/group) received either vehicle (VEH) or ANT for 14 days and received BrdU injection on day 2. Cycle duration was found to be shorter in the ANT-treated group (12.45+/-0.09 days) than in the control group (12.75+/-0.08, P<0.05). As spermatogenic cycle length in this control group was longer than that of our historical controls (range: 12.37-12.53 days), experiment 2 was repeated (n=10/group). In experiment 3, cycle duration was 12.51+/-0.02 for VEH and 12.46+/-0.05 for the ANT-treated group (P>0.05) in both species. We concluded that the duration of the cycle of the seminiferous epithelium in monkeys and rats is independent of gonadotropins but is rather regulated by the spermatogenic tissue itself.

Ultrastructural aspects of cartilage formation, mineralization, and degeneration during primary antler growth in fallow deer (Dama dama).

Due to their rapid growth, regular replacement and easy accessibility, deer antlers are considered a useful model for the study of cartilage and bone differentiation and mineralization in mammals. The present study describes, for the first time, the cellular and extracellular matrix changes associated with cartilage formation, mineralization and degeneration in primary antlers on the ultrastructural level. Growing primary antlers of 3 to 4 cm length were obtained from six fallow bucks, aged about 10 months. It was shown that the chondroblasts were derived from progenitor cells of the antler perichondrium and differentiated into mature chondrocytes that subsequently underwent hypertrophic changes. Concomitant with cell hypertrophy, formation of a lacunar and a perilacunar extracellular matrix was observed, the latter containing numerous collagenous fibers. Mineralization of the extracellular matrix occurred via matrix vesicles and the formation of apatite crystals at distinct sites of the collagenous fibers. The hypertrophic chondrocytes of the mineralized cartilage then degenerated, a process that was also occasionally observed in more distally located cells surrounded by still unmineralized matrix. No morphological indications of a transdifferentiation of hypertrophic chondrocytes into bone forming cells, i.e., co-occurrence of a degenerating chondrocyte and a viable osteogenic cell in intact lacunae, were found. The cellular and extracellular matrix changes seen in primary antlers resemble those described for secondary antlers. Our results further indicate that the hypertrophic chondrocytes of primary antlers eventually undergo apoptosis, thereby providing further evidence that metaplastic conversion of cartilage into bone does not play a role in antler growth.

The lingual epithelium of Salamandra salamandra: metamorphotic changes during its ontogenesis.

The development of the epithelia of the secondary tongue of Salamandra salamandra is described on the basis of light microscopic and scanning electron microscopic studies of defined developmental stages. A glandular area with radial ridges and furrows is formed anterior to the primary tongue during the larval phase. Epithelial cones--each a compact anlage of a gland lying in the furrows--displace the lamina propria. The glandular area grows upward and latero-caudad during metamorphosis and forms the secondary tongue by fusing with the primary tongue. Lumina within the gland anlage appear at the beginning of metamorphosis. They open as glandular tubules towards the oral cavity at the climax of metamorphosis. The epithelial lining becomes single layered and differentiates into gland cells. The glands are increasingly surrounded by fibers of the musculus genioglossus. At the orifice of the gland, the gland cells mingle with the multilayered epithelium of the surface of the tongue. This contains two types of goblet cells in addition to the villus-shaped covering cells which leave gaps for the taste buds. The goblet cells are formed before (type I) and during (type II) metamorphosis and replace the typical larval goblet cells. The new mushroom-shaped part of the secondary tongue is characterized by aborally running septae of connective tissue, visible after digestion with pankreatin. The tip of the primary tongue which originally covers the glandular part becomes completely integrated. It is characterized by crypts which become shallower caudally.

Dentition and dentigerous bones in juveniles and adults of Polypterus senegalus (Cladistia, Actinopterygii).

Tooth types, their arrangement and the mode of tooth replacement were studied in juvenile and adult specimens of Polypterus senegalus by means of scanning electron microscopy of cleared and stained specimens as well as sections. All the dermal bones of the oropharynx are almost completely covered with teeth except for the angulare. The same is true for the branchial apparatus where only the hyoid skeleton is toothless. The teeth are uniformly monocuspid and conical, but can be classified according to shape and size into three types. These types and the mode of tooth replacement are characteristic for each dermal bone. In some of the jaw bones each tooth possesses a lingually situated replacement tooth. This is true for the teeth of the premaxillary, the maxillary, and the dentary which are arranged in a single line, and those of the dermopalatine, the coronoids, and the vomer which are in several lines and graded in size. Replacement teeth of all the other dentigerous elements develop on top of existing pulpal openings, forming an anastomosing common pulpal complex only after resorption of the previous tooth. The tooth plates of the dermal bones of the branchial apparatus are connected by syndesmosis only to the perichondrally ossified and to the cartilaginous or connective tissue material of the elements of the gill-arches. The dentition and its association with the bones of the head in Polypterus senegalus bear resemblances to advanced actinopterygians on the one hand (e.g. differentiation of tooth-types, arrangement), but also some similarities to living Amphibia (anchoring material and mode of replacement) on the other. The accentuation of a single marginal line of large teeth in both, the outer and the inner dental arcade of the jaws is a peculiarity of Polypterus that in a way parallels the derived state of similar monolinear tooth arrangements in Actinopterygii and Tetrapoda.

Changes of the lingual epithelium in Ambystoma mexicanum.

Changes in the lingual epithelium during ontogenesis and after induced metamorphosis in Ambystoma mexicanum are described as observed by light microscopy and scanning electron microscopy. The epithelium of the tongue is always multilayered in the larva as well as in the adult. It consists of a stratum germinativum with little differentiated basal cells and a stratum superficiale (superficial layer) with specialized superficial cells and goblet cells. Usually, there are more than two layers because of a stratum intermedium consisting of replacement cells. The apical cell membrane of the superficial cells is perforated by fine pores. Its most typical feature are microridges. Maturing superficial cells possess microvilli. Goblet cells occur in early larvae primarily in the centre of the tongue. They spread throughout the dorsal face of the tongue as their numbers increase during ontogenesis. The small apices of the goblet cells are intercalated in the wedges between the superficial cells. Leydig cells are not found on the larval tongue but on that of adults. Due to metamorphosis, the epithelium of the tongue changes. It is furrowed in its anterior part. The furrows house the openings of the lingual glands. The surface is further modulated by ridges which are densely coated by microvilli and which bear the taste buds. The villi of the tongue which lack extrusion pores show cilia and microvilli but lack microridges. The Leydig cells disappear during metamorphosis. In addition to the two types of goblet cells found in different regions of the glandular tubules, goblet cells occur in the caudal part. They secrete directly into the cavity of the mouth. The posterior part is characterised by a dense coat of cilia.

Estimation of the duration of the cycle of the seminiferous epithelium in the non-human primate Macaca mulatta using the 5-bromodeoxyuridine technique.

A comparatively low yield of germ cells has been reported for the spermatogenic process in primates. Kinetic studies of spermatogenesis and the spermatogenic cycle are needed to investigate this phenomenon but require the application of radioactively labeled compounds or irradiation. We have therefore investigated the suitability of a non-radioactive approach, viz., administration of 5-bromodeoxyuridine, for the determination of the kinetics of the spermatogenic cycle in a non-human primate, the rhesus monkey (Macaca mulatta). Four adult in-season animals received a bolus of 33 mg/kg 5-bromodeoxyuridine, one testis from each monkey was removed 3 h later and the other testis after 10 days and 11 h. Tissue was fixed in Bouin's solution and embedded in Paraplast. 5-Bromodeoxyuridine was localized by immunogold-silver staining with a monoclonal antibody. PAS-hematoxylin counterstaining was used for spermatogenic stage identification. At 3 h, the leptotene and zygotene spermatocytes in stages VII-IX were the most advanced 5-bromodeoxyuridine-positive cells. At 10 days 11 h, the label had advanced and pachytene spermatocytes in stages VI-IX contained 5-bromodeoxyuridine. The duration of the spermatogenic cycle was 10.42+/-0.07 days (range: 10.25-10.62 days). Peritubular cells and interstitial cells were rarely 5-bromodeoxyuridine-positive, and Sertoli cells were consistently negative for 5-bromodeoxyuridine. Importantly, our kinetic data closely resemble those obtained by means of the application of irradiation for this macaque species. We conclude that administration of 5-bromodeoxyuridine represents a non-radioactive reliable approach for studying kinetic aspects of the spermatogenic process in primates.

The development of the dentigerous bones and teeth in the hemiramphid fish Dermogenys pusillus (Atheriniformes, Teleostei).

The development of the dentition and dentigerous bones was studied in the hemiraphid Dermogenys pusillus using histological sections, scanning electron microscopy, and cleared and stained specimens. Five days after birth, the toothless tip of the lower jaw begins to grow longer than the tip of the upper jaw. The growth originates from small cartilaginous triangles connected with Meckel's cartilage. The peri- and enchondrial ossification of the growing cartilaginous bars advances rostrad. The pharyngeal tooth plates are formed by fusion of the slat-like dentigerous dermal bones with the bony fractions of the gill branches. Hence, the tooth plates are composite bones. The ventral tooth plate ist formed by the two ceratobranchials V and the basibranchial IV together with the respective dermal bones. The paired pharyngobranchials III and IV are fused with the dorsal tooth plate, and the pharyngobranchials II is fused with the two respective lateral tooth plates. Mineralization starts after birth in elements of the pharyngeal tooth plates and their teeth. There are no indications that the pedicel on which the tooth is established is formed by the enamel organ, which is covered by pulpal cells. The enamel organ originates from the stratum basale of the oropharyngeal epithelium and moves within the jaw from labial toward lingual, the site of the establishment of the tooth. The anlage of the tooth on the tooth plates of the pharynx lies at the level of the tooth base.

Dentigerous bones and dentition in the hemiramphid fish Dermogenys pusillus (Atheriniformes, Teleostei).

Structure and arrangement of the teeth were studied in the hemiramphid Dermogenys pusillus, using scanning electron microscopy as well as cleared and stained specimens. The teeth of the jaws are small, monocuspid, and tilted towards the esophagus. They are arranged along the lateral edges of the premaxillas and dentaries. Each premaxilla bears additional teeth on an osseous bar extending from rostro-lateral to medio-lingual. The dentition of both dentaries curves slightly within the cavity of the mouth and gently tapers off laterorostrad just beyond the tip of the upper jaw. The part of the lower jaw, which typically protrudes beyond the upper jaw, is without teeth. One dorsal and two lateral tooth-bearing bony plates (tooth plates) are found in the pharyngeal region. Their teeth are largely irregular in arrangement. The teeth on the two lateral plates are small and monocuspid, whereas the dorsal and the ventral tooth plate possess additional strong bi- and tricuspid teeth. The teeth of the jaws and of the pharyngeal region obviously have a bony pedicel ("attachment bone") which is asymmetric in all teeth. An elastic suture connects the cone of dentine with the bony pedicel. The special construction of the teeth and their arrangement on the various dentigerous bones will be discussed with respect to their function in catching prey.

Hard tissue of teeth and their calcium and phosphate content in Ambystoma mexicanum (Urodela: Ambystomatidae).

The wall of the pulp cavity, fracture faces and the demineralized surfaces of teeth from larvae and adults of Ambystoma mexicanum were investigated by scanning electron microscopy (SEM). Calcium and phosphate contents were determined by microanalysis. The apical part of the tooth (crown, tooth apex) contains dentin canals. In the larva, these do not reach the enamel-dentin border but end below this border in front of a denser hard substance, possibly enameloid. The pedicel in the adult and the basal portion of the tooth in the larva (base) are without dentin canals. These parts of the teeth are characterized by longitudinally arranged collagen fibres as visualized on the demineralized surfaces. These observations indicate a congruency in early-larval and adult teeth between base and pedicel as well as apex and crown. This partition is also confirmed by the calcium and phosphate values which were identical in larvae and adults. Highest values are found in enamel and lowest values in the tooth-bearing bone. Calcium and phosphate content show a clear difference between dentin and the basal part of the tooth (pedicel and base). The ring-like dividing zone in the adult tooth is less well mineralized.