Characterization of ethoxyresorufin O-de-ethylase in grey seal Halichoerus grypus

Ethoxyresorufin O-de-ethylase (EROD), a typical mixed function oxidase enzyme, was studied in liver and kidney of adult and pup grey seals, Halichoerus grypus. EROD activity was present in most liver samples but was only barely detectable in a few of the kidney samples. Liver EROD activity was greatest at pH 7.5-8.0; activity increased with temperature from 20 degrees C to a plateau starting at about 33 degrees C and extending to 37 degrees C, and fell off sharply above 37 degrees C. Km values ranged from 3 X 10(-7) to 3 X 10(-6)M; Vmax ranged from 5 X 10(-12) to 1 X 10(-10) moles product formed X min-1 X mg protein-1. Liver EROD activity was not detectable in pups less than 1 day old, and activity increased with age in pups up to 5-12 days old. Adults had greater EROD activity than did pups.     

Molecular Phylogenetics and Chronometrics of Tarsiidae Based on 12S mtDNA Haplotypes: Evidence for Miocene Origins of Crown Tarsiers and Numerous Species within the Sulawesian Clade

We report new mitochondrial DNA sequence data from tarsiers sampled from several populations, including the extreme northeast and southwest of the range of the Tarsius tarsier species complex, the most extensive sampling ever reported for this taxon. Our results provide the opportunity to produce the first ever molecular chronometric analysis of Tarsiidae. These results date the age of crown tarsiers, minimally, to the middle Miocene, and each of the 3 tarsier species groups, Tarsius bancanus, T. syrichta, and the T. tarsier complex, to the early or middle Miocene. Thus, each of these 3 species has evolved in isolation for a period of time that is consistent with that which would be expected for multiple speciation events. Our analysis of the Tarsius tarsier complex reveals 5 subclades, each of which is interpreted to represent a haplogroup at, or above, the species level, a result that is consistent with current hypotheses about numerous cryptic species within this species complex. The implications for conservation within the Sulawesi biogeographic region are that Sulawesi is subdivided into numerous subregions of endemism and that, by extrapolating the example of cryptic tarsier species to other taxa, biodiversity may be underestimated by an order of magnitude. The practical realties of conservation in Sulawesi are such that it is most reasonable to assume that anthropogenic extinctions are occurring, and that some species will go extinct before they have even been identified.     

Liver and kidney glucose-6-phosphatase levels in carbon tetrachloride- and DDT-administered mice.

Mouse liver and kidney glucose-6-phosphatase levels were found to be decreased 24 h after administration of various doses of carbon tetrachloride (CCl4) when compared to controls. Liver glucose-6-phosphatase levels were always decreased to a greater extent than the kidney enzyme in mice given the same amount of CCl4. Administration of p,p'-1,1,1,-trichloro-2,2-bis (p-chlorophenyl) ethane (p,p'-DDT) to mice did not significantly alter the glucose-6-phosphatase levels of liver or kidney.     

Primate evolution: a biology of holocene extinction and survival on the southeast Asian Sunda Shelf islands

What biological traits distinguish taxa susceptible to extinction from less susceptible taxa? Substantiated island biogeographic theory suggests that after insularization, small islands lose more species than do large islands. Thus, susceptible taxa are those now found on only large islands. The traits of susceptible taxa can thus be found by comparing the biology of species found only on large islands with those also found on small islands. The islands examined here are those of the Sunda Shelf, created as a result of the Holocene rise in sea levels of 120 m. We use four statistical comparisons: comparative analysis by (phylogenetically) independent contrasts (N = 8 contrasts at the subgeneric or deeper level), Spearman correlations, stepwise regression, and principle components analysis (N = 9 subgenera/genera). The genera and one subgenus considered are: Hylobates, Macaca, Nasalis, Nycticebus, Pongo, Presbytis, Symphalangus, Tarsius, and Trachypithecus. Traits of risk appear to be large body mass, low density, large annual home range, and low maximum latitude. Expected traits that did not correlate with susceptibility were low interbirth interval, high percent frugivory, high group mass, low altitudinal range, and small geographic range. The risky traits also apply to just the anthropoids (i.e., prosimians excluded). The risky traits are explained if susceptibility is induced by requirements for a large extent of habitat, a small population size, and specialization. These findings, which indicate that efficiency and plasticity of use of the environment separate susceptible from successful primate taxa, might be relevant to an understanding of hominoid evolution.     

The complete mitochondrial sequence of Tarsius bancanus: evidence for an extensive nucleotide compositional plasticity of primate mitochondrial DNA

Inconsistencies between phylogenetic interpretations obtained from independent sources of molecular data occasionally hamper the recovery of the true evolutionary history of certain taxa. One prominent example concerns the primate infraordinal relationships. Phylogenetic analyses based on nuclear DNA sequences traditionally represent Tarsius as a sister group to anthropoids. In contrast, mitochondrial DNA (mtDNA) data only marginally support this affiliation or even exclude Tarsius from primates. Two possible scenarios might cause this conflict: a period of adaptive molecular evolution or a shift in the nucleotide composition of higher primate mtDNAs through directional mutation pressure. To test these options, the entire mt genome of Tarsius bancanus was sequenced and compared with mtDNA of representatives of all major primate groups and mammals. Phylogenetic reconstructions at both the amino acid (AA) and DNA level of the protein-coding genes led to faulty tree topologies depending on the algorithms used for reconstruction. We propose that these artifactual affiliations rather reflect the nucleotide compositional similarity than phylogenetic relatedness and favor the directional mutation pressure hypothesis because: (1) the overall nucleotide composition changes dramatically on the lineage leading to higher primates at both silent and nonsilent sites, and (2) a highly significant correlation exists between codon usage and the nucleotide composition at the third, silent codon position. Comparisons of mt genes with mt pseudogenes that presumably transferred to the nucleus before the directional mutation pressure took place indicate that the ancestral DNA composition is retained in the relatively fossilized mtDNA-like sequences, and that the directed acceleration of the substitution rate in higher primates is restricted to mtDNA.     

Front dentition of the omomyiformes (Primates)

Within the primates the front dentition plays a major role in the classification of the different taxa. The extant strepsirhines are clearly characterized by the possession of a tooth comb. Tarsius differs in this complex by a construction which remembers the beak of an owl (vertically implanted medial incisors, upper pair bigger than mandibular pair). The front teeth of the extinct Microchoeridae are described. The dental formula of the group is discussed and compared with the Omomyidae. The wear pattern presents evidence of a dental comb as early as the Late Eocene.     

Biosynthesis of glycosaminoglycan precursors: evidences for different tissue specific forms of UDP-glucose dehydrogenase

UDP-glucose dehydrogenase (UDPGDH) was extracted and partially purified from different rat tissues and the kinetic parameters and some properties of the enzyme were determined and compared. The pH optimum ranged between 8.6 and 9.4 for liver and kidney UDPGDH and between 8.4 and 8.6 for skin and lung UDPGDH. Liver and kidney enzymes showed a similar affinity for both UDPG and NAD. Lung and skin enzymes also showed similar affinity for both substrates, which differed however from that of liver and kidney UDPGDH. Both liver and kidney enzymes had a higher heat stability and a different electrophoretic mobility compared to skin and lung UDPGDH. These data suggest the existence of different tissue specific forms of the enzyme.     

Multiple forms of mutarotases from the kidney, liver, and small intestine of rats: purification, properties, subcellular localization and developmental changes

Comparative studies of mutarotase [aldose 1-epimerase, EC 5.1.3.3] from the kidney, liver and small intestine of rats were performed placing in the focus on the study of multiple forms. The findings obtained are as follows. Mutarotases from the kidney and liver of adult rats were both separated into four forms (types I-IV) by DEAE-cellulose column chromatography, whereas only two forms (types I and II) were detected in the small intestine. Liver mutarotase type I was further separated into types I1 and I2 by column chromatography on hydroxylapatite. Types I and II from the kidney and type II from the liver were purified to homogeneity as judged by isoelectric focusing on thin layer polyacrylamide gel. Of various physicochemical properties, only the Km for alpha-D-xylose and the isoelectric point were different among the multiple forms. Liver mutarotase was immunohistochemically localized in the nuclei of parenchymal cells and small intestine enzyme in the nuclei of mucosal cells, indicating similarity with the localization of kidney enzyme (in the nuclei of epithelial cells of renal tubules and glomeruli) which was reported in our previous paper [Experientia (1979) 35, 1094-1097]. The kidney mutarotase level increased gradually after birth and reached a maximum near adult level within 20 days. This developmental pattern was essentially the same as that in the liver but clearly different from that in the small intestine, in which the mutarotase activity of suckling rats was several times higher than that of adult rats. Distribution patterns of multiple forms (types I-IV) of the enzyme in the kidney and liver of 10-day-old rats were similar to those in respective tissues of adult rats. On the other hand, the small intestine of 10-day-old rats contained four forms (types I-IV), whereas there were only two forms (types I and II) in adult rats.     

Isolation and characterization of 12 microsatellite loci for population studies of Sulawesi tarsiers (Tarsius spp.)

This study reports the development and characterization of the first 12 microsatellite markers for tarsiers. Nine loci were isolated from Dian's tarsier, Tarsius dianae and three from the Philippine tarsier, Tarsius syrichta. The 12 markers were used to screen 40 individuals of Dian's tarsier and 40 individuals of the Lariang tarsier, Tarsius lariang for allelic diversity. This suite of highly polymorphic microsatellites provides the first chance to genetically study parentage patterns in tarsiers.     

A new Middle Miocene tarsier from Thailand and the reconstruction of its orbital morphology using a geometric-morphometric method

Tarsius is an extant genus of primates endemic to the islands of Southeast Asia that is characterized by enormously enlarged orbits reflecting its nocturnal activity pattern. Tarsiers play a pivotal role in reconstructing primate phylogeny, because they appear to comprise, along with Anthropoidea, one of only two extant haplorhine clades. Their fossils are extremely rare. Here, we describe a new species of Tarsius from the Middle Miocene of Thailand. We reconstructed aspects of its orbital morphology using a geometric-morphometric method. The result shows that the new species of Tarsius had a very large orbit (falling within the range of variation of modern Tarsius) with a high degree of frontation and a low degree of convergence. Its relatively divergent lower premolar roots suggest a longer mesial tooth row and therefore a longer muzzle than in extant species. The new species documents a previous unknown Miocene group of Tarsius, indicating greater taxonomic diversity and morphological complexity during tarsier evolution. The current restriction of tarsiers to offshore islands in Southeast Asia appears to be a relatively recent phenomenon.     

Geographical Variation in Duet Songs of Sulawesi Tarsiers: Evidence for New Cryptic Species in South and Southeast Sulawesi

Geographic populations of Sulawesi tarsiers differ conspicuously in the acoustic structure of the loud duet calls produced by mated pairs. There is evidence that the differences in call features relate to species differentiation. Duet calls can thus be used as a tool to identify cryptic species and for their monitoring in the field. We recorded duets from various geographically isolated populations of Tarsius tarsier in south and southeast Sulawesi to survey the species identity of the tarsiers in these regions. We compared their calls with the duet calls of the Manado tarsier (= Tarsius spectrum), Tarsius dianae, and the Togian Island tarsier. In addition, we recorded and analyzed the duet call of the tarsier from Peleng Island, Tarsius pelengensis. Qualitative characteristics of duet songs of both sexes, analysis of frequency and temporal parameters, and multivariate statistics show 4 distinct vocal forms that predict the existence of 4 new species. These are found on Selayar Island, on the south mainland (Bantimurung), on the southeast mainland (Puwato), and on the Islands of Kabaena and Buton. Our analysis of the call of the Peleng tarsier revealed striking similarities with the call of Tarsius dianae. Further morphological analyses are needed to support the proposed new species in south and southeast Sulawesi and to clarify the taxonomic affinity between the central Sulawesi tarsier species Tarsius pelengensis and T. dianae.     

Basic oxidative stress metabolites in eastern Pacific green turtles (Chelonia mydas agassizii)

Analysis of hematological and biochemical parameters, including oxidative stress indicators, is an invaluable tool in wildlife health assessment, particularly for threatened or endangered species. This study was aimed at obtaining baseline information of oxidative stress indicators in eastern Pacific green turtles (Chelonia mydas agassizii) from a relatively undisturbed habitat at Bahía Magdalena, Baja California Sur, Mexico. Tissues were analyzed for superoxide radical (O(2)(*-) production, lipid peroxidation (measured as thiobarbituric acid reactive substances, TBARS), and antioxidant enzyme activity (superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST)). Overall levels for all variables were within ranges reported for other reptile species. Results suggest differences in oxidative metabolism among tissues (p< or =0.05). Liver, lung and muscle had the highest levels of O(2)(*-) production. Liver revealed the highest TBARS levels. Liver and muscle showed the highest SOD activity, while liver and kidney had the highest CAT and GST activities. These data provide baseline values of the oxidative stress indicators in tissues from eastern Pacific green turtles. Development of a biomarker system to assess the health of wildlife species, especially one that could detect early exposure to environmental pollutants or emerging diseases, would provide a useful tool in the long-term conservation of the species.     

Specificity and multiple forms of beta-galactosidase in the rat.

1. Ten rat tissues and organs have been assayed for beta-galactosidase with phenyl beta-d-galactoside, p-nitrophenyl beta-d-galactoside, p-aminophenyl beta-d-galactoside and 4-methylumbelliferyl beta-d-galactoside as substrates. 2. The relative activities of these tissues are independent of the mode of assay, and maximum rates of hydrolysis are not greatly affected by the nature of the substrate. 3. Inhibition studies suggest the liver enzyme has no associated beta-glucosidase activity. 4. There is no cellular localization of preferential activity towards any of the four substrates in liver, kidney or spleen. 5. Evidence suggesting the non-destructive penetration of liver lysosomal membranes by p-nitrophenyl beta-d-galactoside is presented. 6. Liver lysosomal beta-galactosidase exists in multiple forms that can be separated on DEAE-cellulose, and the enzyme components that are bound to the membrane appear to be similar to those of the lysosome sap. 7. The chromatographic pattern of enzyme excreted in the urine is compared with those from the kidney, intestine, spleen and liver.     

Osteological evidence for the evolution of activity pattern and visual acuity in primates

Examination of orbit size and optic foramen size in living primates reveals two adaptive phenomena. First, as noted by many authors, orbit size is strongly correlated with activity pattern. Comparisons of large samples of extant primates consistently reveal that nocturnal species exhibit proportionately larger orbits than diurnal species. Furthermore, nocturnal haplorhines (Tarsius and Aotus) have considerably larger orbits than similar-sized nocturnal strepsirrhines. Orbital hypertrophy in Tarsius and Aotus accommodates the enormously enlarged eyes of these taxa. This extreme ocular hypertrophy seen in extant nocturnal haplorhines is an adaptation for both enhanced visual acuity and sensitivity in conditions of low light intensity. Second, the relative size of the optic foramen is highly correlated with the degree of retinal summation and inferred visual acuity. Diurnal haplorhines exhibit proportionately larger optic foramina, less central retinal summation, and much higher visual acuity than do all other primates. Diurnal strepsirrhines exhibit a more subtle but significant parallel enlargement of the optic foramen and a decrease in retinal summation relative to the condition seen in nocturnal primates. These twin osteological variables of orbit size and optic foramen size may be used to draw inferences regarding the activity pattern, retinal anatomy, and visual acuity of fossil primates. Our measurements demonstrate that the omomyiforms Microchoerus, Necrolemur, Shoshonius, and Tetonius, adapiform Pronycticebus, and the possible lorisiform Plesiopithecus were likely nocturnal on the basis of orbit diameter. The adapiforms Leptadapis, Adapis, and Notharctus, the phylogenetically enigmatic Rooneyia, the early anthropoids Proteopithecus, Catopithecus, and Aegyptopithecus, and early platyrrhine Dolichocebus were likely diurnal. The activity pattern of the platyrrhine Tremacebus is obscure. Plesiopithecus, Pronycticebus, Microchoerus, and Necrolemur probably had eyes that were very similar to those of extant nocturnal primates, with a high degree of retinal summation and rod-dominated retinae. Leptadapis and Rooneyia likely had eyes similar to those of extant diurnal strepsirrhines, with moderate degrees of retinal summation, a larger cone:rod ratio than in nocturnal primates, and, more speculatively, well-developed areae centrales similar to those of diurnal strepsirrhines. Adapis exhibited uncharacteristically high degrees of retinal summation for a small-eyed (likely diurnal) primate. None of the adapiform or omomyiform taxa for which we were able to obtain optic foramen dimensions exhibited the extremely high visual acuity characteristic of extant diurnal haplorhines.     

Partial purification and isoelectric focusing patterns of the buffalo (Bubalus bubalis) and the Kedah-Kelantan cattle (Bos indicus) glutathione transferases.

1. Glutathione transferases from the liver, lung and kidney tissues of the buffalo (Bubalus bubalis) and the Kedah-Kelantan cattle (Bos indicus) were partially purified by ammonium sulphate precipitation and Sephadex G-75 gel filtration. 2. Liver tissue contains the highest enzyme activity when compared to the lung and kidney tissues. 3. The activity in cattle is higher than that in the buffalo. 4. Isoelectric focusing separates the activities into the acidic, near neutral and basic fractions. 5. The focused patterns are different for each of the tissues and in each of the species investigated.     

Cranial morphology of Aegyptopithecus and Tarsius and the question of the tarsier-anthropoidean clade

New crania of the Oligocene anthropoidean Aegyptopithecus provide a test of the hypothesized tarsier-anthropoidean clade. Three cranial characters shared by Tarsius and some modern anthropoideans (apical interorbital septum, postorbital septum, "perbullar" carotid pathway) were examined. 1) An apical interorbital septum is absent in Aegyptopithecus. A septum does occur in Galago senegalensis (Lorisidae) and Microcebus murinus (Cheirogaleidae), so the presence of a septum is not strong evidence favoring a tarsiiform-anthropoidean clade. 2) In Aegyptopithecus and other anthropoideans, the postorbital septum is formed mainly by a periorbital flange of the zygomatic that extends medially from the lateral orbital margin onto or near the braincase. The postorbital plate of Tarsius is formed by frontal and alisphenoid flanges that extend laterally from the braincase to the zygomatic's frontal process, which is not broader than the postorbital bars of other prosimians. Periorbital flanges evolved in Tarsius for support or protection of the enormous eyes, as suggested by the occurrence of maxillary and frontal flanges that cup portions of the eye but do not separate it from temporal muscles. 3) The internal carotid artery of Aegyptopithecus enters the bulla posteriorly and crosses the anteroventral part of the promontorium. The tympanic cavity was probably separated from the anteromedial cavity by a septum stretching from the carotid channel to the ventrolateral bullar wall. In Tarsius, the carotid pathway is prepromontorial, and a septum stretches from the carotid channel to the posteromedial bullar wall. Quantitative analyses indicate that anterior carotid position has evolved because of erect head posture. The cranium of Oligocene anthropoideans thus provides no support for the hypothesized tarsier-anthropoidean clade.     

Angiotensin carboxypeptidase activity in urine from normal subjects and patients with kidney damage

Angiotensin carboxypeptidase (ACP) activity has been detected in urine samples from normal subjects and patients with hypertension and diabetes by determining the enzyme's ability to convert angiotensin I to des-Leu angiotensin I. Gel filtration chromatography of a concentrated urine sample indicated that about equal amounts of the enzyme exist as 100 kDa and 500 kDa molecular weight forms, respectively. This ACP activity co-eluted with activity that cleaved histidine from des-Leu angiotensin I to form angiotensin II and activity that cleaved tyrosine from benzyloxycarbonyl-glutamyl-tyrosine (ZGT). These results suggest that the urinary ACP activity is due to cathepsin A as we have reported previously for the porcine kidney enzyme. Analysis of sequential urine samples from a single individual over a 6-day period revealed as much as a 6-fold fluctuation in creatinine-normalized ACP activity. Of five male healthy adult subjects, the creatinine-normalized urinary ACP activity ranged from 1.7 to 3.7 mU/mL with a mean of 2.8 mU/mL. However, five male patients with renovascular hypertension had elevated levels of ACP activity with a mean of 11.6 mU/mL. Of five male patients with diabetic nephropathy, all had elevated ACP activity levels with a mean of 21.0 mU/mL. It is concluded that ACP activity in the urine is due to cathepsin A probably derived from kidney tissue, and that the release is increased in patients with kidney damage. We suggest that urinary ACP activity should be evaluated further for a possible relationship to renal hypertension and as a potentially early marker for diabetic nephropathy.     

Occurrence and comparison of sulfite oxidase activity in mammalian tissues

Tissue extracts from six mammalian species have been assayed for sulfite oxidase (sulfite: ferricytochrome c oxidoreductase, EC 1.8.3.1) activity with cytochrome c as electron acceptor. Our results show a large distribution of sulfite oxidase activity in mammalian tissues. Liver, kidney, and heart tissues exhibit high activities whereas brain, spleen, and testis show very low activities. No significant species dependence was observed for the activity of this enzyme.     

An evolutionary switch in tissue-specific gene expression. Abundant expression of alpha 1-antitrypsin in the kidney of a wild mouse species

alpha 1-Antitrypsin (AT), one of the major proteinase inhibitors in mammalian serum, is generally considered to be synthesized exclusively in the liver. We have found that a wild-derived Mus species, Mus caroli, expresses AT mRNA in kidney at levels approaching that in liver; no other mouse, inbred or wild-derived, exhibits this striking property. Liver and kidney mRNAs from M. caroli encode very similar AT polypeptides that are distinct from that encoded by Mus musculus liver mRNA. In vivo, liver AT is secreted into the bloodstream, while kidney AT, which is processed differently from the liver protein, is excreted into the urine. Analysis of RNA from a hybrid between M. musculus and M. caroli indicates that a cis-acting genetic element may be responsible for the difference in AT expression. Restriction enzyme digestion patterns of AT genomic sequences in M. caroli DNA are considerably different from those in M. musculus; in addition, these sequences are undermethylated in liver DNA from M. musculus and in liver and kidney DNA from M. caroli, reflecting the respective patterns of expression. Further studies of the altered tissue specificity of AT expression that is apparent in these two related species should lead to new insights into the nature and evolution of genetic determinants of tissue-specific phenotypes.     

Locomotion and habitat utilization in free-rangingTarsius bancanus: A preliminary report

Four adultTarsius bancanus were followed for a total of over 120 hrs in Sepilok Forest Reserve, Sabah, using radiotracking techniques. Seven hundred and twenty-two records of locomotor and postural behaviour were gathered.Tarsius bancanus travels a mean of 1,800 m per night, over large (4.5 to 11.25 ha) ranges. They move at a mean height of 0.89 m, utilizing primarily vertical sapling trunks of diametre 2 to 4 cm. Lateral movement is carried out largely by leaping.Tarsius were found on the ground in 5.3% of cases, but movement on the ground is very largely restricted to short investigative movements by walking. Climbing is mostly related to foraging behaviour and characteristically occurs on relatively small supports.Tarsius bancanus' locomotion is similar to, but more specialized than that ofT. spectrum, Galago alleni, andGalago senegalensis, all of which have rather similar patterns of habitat utilization. We suggest that the energetic constraints of small body size together with the need to patrol large home ranges may have lead both toTarsius bancanus extreme degree of locomotor specialization and to its exclusive animalivory.