A new species of Pseudoloris (Omomyidae,Primates) from the middle Eocene of Sant Jaume de Frontanyà (Eastern Pyrenees,Spain)

In this article we describe a new species of Pseudoloris (Omomyidae, Primates) from the Robiacian (middle Eocene) locality of Sant Jaume de Frontanyà (Eastern Pyrenees, Spain). Pseudoloris pyrenaicus is characterized by its medium size, thickened paracristid, absence of a distinct paraconid, and well‐developed buccal cingulid in the lower molars, large hypoconulid in the M₃, paraconule and hypocone reduced in the M¹ and M² and absent in the M³. The material from Sant Jaume de Frontanyà constitutes the most abundant sample of the genus Pseudoloris found until now in the Iberian Peninsula. Almost all the dental elements have been recovered, including those teeth hardly known for other species of the genus, such as lower and upper incisors. The new species shows intermediate features between Pseudoloris isabenae from Capella and Pseudoloris parvulus, present in different Spanish and French sites. Therefore, we consider that Pseudoloris pyrenaicus is an intermediate form between these two species. Am J Phys Anthropol 143:92–99, 2010. © 2010 Wiley‐Liss, Inc.     

Locking Yourself Out: Diversity Among Dentally Zalambdodont Therian Mammals

We review the evolution of dental zalambdodonty across therian mammals. Among zalambdodonts, there is little or no occlusion between the protocone and talonid basin and one of the central cusps of the upper molars (metacone or paracone) and the talonid basin of the lower molars are lost or reduced. Over two dozen genera of therian mammals show zalambdodont molars, including tenrecids, chrysochlorids, Solenodon, the marsupial mole Notoryctes, the extinct placentals Apternodus, Oligoryctes, Parapternodus, and Koniaryctes, and the extinct marsupials Necrolestes and Yalkaparidon. The chiropteran Harpiocephalus is nearly zalambdodont. Transformation series provided by paleontological or ontogenetic data, plus occlusal relationships, can be used to determine the homology of molar cusps in zalambdodont taxa. Zalambdodont dental specialization does not appear to have led to ecological specialization. With the important exception of golden moles and tenrecs, dentally zalambdodont taxa are less speciose than their nonzalambdodont sister taxa.     

New remains of Chambius kasserinensis from the Eocene of Tunisia and evaluation of proposed affinities for Macroscelidea (Mammalia,Afrotheria)

Abstract

Chambius kasserinensis from the late Early or early Middle Eocene Chambi locality, central Tunisia, is undoubtedly the oldest known macroscelidid and possibly the basalmost representative of the order Macroscelidea. Hence, since its discovery in 1986, Chambius has played a key role in analyses focusing on afrotherian and eutherian phylogeny; for instance, as early as 1995, Butler’s review of fossil macroscelideans highlighted the central position of Chambius in the origin of the order. Despite this, Chambius remained poorly known until recently. Here based on new mandibular fragments, well-preserved upper molars and CT scan analysis of the holotype maxilla, Chambius is revised. Its dentition is first described in detail, providing a precise characterization of the genus. Chambius is notably defined by a submolariform P₄ with a three-cusped talonid, a reduced talonid on M₂, and a prominent metaconule on M^1?2. Interestingly, the two transverse lophs of the upper molars are basically formed by preconulecristae, evoking the recently defined peculiar bilophodonty of paenungulates. Comparisons with other Paleogene and modern macroscelidids, European Louisinidae, and North American Apheliscidae are also made, allowing the various hypotheses about the origin and early evolution of macroscelidids to be reviewed.     

Tooth Wear Difference Between the Yuanmou Hominoid and <Emphasis Type="Italic">Lufengpithecus</Emphasis>

The Late Miocene hominoids recovered from Lufeng (Lufengpithecus) and Yuanmou of Yunnan Province, China, are among the most numerous hominoid fossils in Eurasia. They have yielded critical evidence for the evolutionary history, biogeography and paleobiology of Miocene hominoids. We examined and compared the wear pattern and differences of 804 molars of the Yuanmou hominoid and Lufengpithecus. Our results indicate that both the upper and lower molars of the Yuanmou hominoids were more heavily worn than those of Lufengpithecus. The wear patterns of the individual molars between the Yuanmou hominoid and Lufengpithecus also are different. The heaviest wear of lower molars of the Yuanmou hominoid occur in M₂, followed by M₁ and M₃. In Lufengpithecus, M₁ and M₃ were more heavily worn than M₂. There are differences in wear between the upper and lower molars for the two hominoids. Among the various factors related to tooth wear, we suggest that the main reason for the tooth wear differences between the Yuanmou hominoid and Lufengpithecus may be that they had different diets. More soft dietary items like leaves and berries were probably consumed by Lufengpithecus, and the Yuanmou hominoid may mainly have feed on harder or frugivorous diets. This result complements findings from previous studies of tooth size proportion, and the development of lower molar shearing crests in the 2 samples. Enamel thickness, living environment, behavior patterns, and population structure also might account for dental wear differences between the Yuanmou hominoid and Lufengpithecus.     

Tooth Replacement in Late Jurassic Dryolestidae (Eupantotheria, Mammalia)

The discovery of juvenile dentitions of late Jurassic (Kimmeridgian) Dryolestidae (Eupantotheria, Mammalia) from Guimarota, Portugal, yields for the first time information on the mode of tooth replacement in therian mammals prior to the dichotomy of placentals and marsupials. As in extant placentals, tooth replacement occurs at all antemolar positions [incisors (I1–I4), canine (C), premolars (P1–P4)]. P1 and P2 have premolariform milk predecessors, whereas the large premolariform third (P3) and fourth premolars (P4) are preceded by molariform deciduous premolars (dP3, dP4). Tooth replacement takes place in two waves, at least in the lower jaw, with I₂, I₄, P₁, and P₃ in the first series and I₁, I₃, C, P₂, and P₄ in the second. P₄ is the last premolar to erupt, and it is present when the sixth molar (M₆) starts to break through. The reduced tooth replacement pattern of marsupials (with only dP3 being replaced postnatally) evolved secondarily from the primitive or plesiomorphic mammalian condition, which was retained in Dryolestidae and Eutheria.     

Fossil remains of Macaca sylvanus florentina (Cocchi, 1872) (Primates, Cercopithecidae) from the Early Pleistocene of Quibas (Murcia, Spain)

The macaque material from the Early Pleistocene site of Quibas (Albanilla, Murcia, Spain), including dentognathic remains, isolated teeth and some postcranial bone fragments, is described. Both metrically and morphologically, this sample must be attributed to Macaca sylvanus (the Barbary macaque). This species is currently distributed through North Africa and Gibraltar, but was much more widely distributed during the Plio-Pleistocene, being represented by several European fossil subspecies. Metrical comparisons of dental size and proportions between extant M. s. sylvanus and fossil Macaca sylvanus florentina from the type locality and other Italian sites are undertaken, in order to classify the remains from Quibas at the subspecies level. The results show that the Quibas sample not only fits the range of variation of M. s. florentina from the type locality, but also differs from the extant Barbary macaque condition in several regards. This permits us to formally attribute the material from Quibas to M. s. florentina. The material described in this paper therefore significantly improves the knowledge of this fossil taxon, particularly regarding the upper dentition, and further confirms the taxonomic distinctiveness of this extinct taxon at the subspecies rank. Taken as a whole, M. s. florentina largely overlaps in dental dimensions with M. s. sylvanus, but differs from the latter by displaying (on average): (1) absolutely longer upper molars (especially M¹ and M³); (2) relatively wider upper molars (especially M¹ and M²); (3) longer M³ as compared with the M²; (4) absolutely longer M₁ and M₃; and (5) relatively narrower M₃.     

New eutherian mammals from the Late Cretaceous of Aix‐en‐Provence Basin,south‐eastern France

In Europe, the fossil record of Late Cretaceous eutherian mammals is very poor, being limited to only three genera (Labes, Lainodon, and Valentinella). Labes and Lainodon are well‐supported members of Zhelestidae, a stem eutherian clade, whereas Valentinella is more problematic, being recently considered as a nomen dubium. Based on X‐ray computed microtomography scan analysis of the holotype and thanks to the discovery of new specimens from the type locality (Vitrolles‐La Plaine, south‐eastern France, late Maastrichtian), we reassessed Valentinella. This genus is unique by the association of an enlarged and rounded jaw angle with an assumed relatively elevated angular process, a bulbous protoconid and an unbasined heel on p4, a p5 with a wide molariform talonid and a hypoflexid, a robust molar morphology with a potential specialized crushing‐grinding function (bulbously constructed cusps, large talonid, and horizontal apical wear facet of the hypocone), a somewhat reduced m3 relative to m2, a premolariform ?P3 or ?P4 lacking a metacone, and a relatively large hypocone on upper molars. These characters reinstate Valentinella as a valid genus. We also describe Mistralestes arcensis gen. et sp. nov. from a newly discovered locality (La Cairanne‐Highway, south‐eastern France, late Campanian). Mistralestes is defined by a robust premolariform p5 with no cingulid, paraconid, or metaconid; molars with a transverse protocristid, a gradual compression of the trigonid from m1 to m3, and paracristid and protocristid probably confluent on m3. Based on comparisons and phylogenetic analyses, Valentinella and Mistralestes may belong to Zhelestidae but this systematic attribution remains poorly supported. © 2013 The Linnean Society of London     

Examples of short- and long-term changes of dental pattern in Scottish voles (Rodentia; Microtinae)

An isolated population of the bank vole, Clethrionomys glareolus, at Loch Tay, Perthshire, which was found to have abnormally complex third upper molars in 1955-57, was re-examined in 1972 and found to have reverted to normal. Subfossil teeth of the field vole, Microtus agrestis, from the island of Jura, Inner Hebrides, dating back to at least 2000 BP have been compared with recent specimens from the island. They demonstrate a slight but progressive reduction in the size of the postero-medial ridge of M¹ that is characteristic of certain marginal     

Localized and internal effect of nitrate on symbiotic dinitrogen fixation

Alfalfa (Medicago sativa L.) is a deeply rooted perennial legume which, under field conditions, may be exposed to varying NO₃^? concentrations with depth. Our objective was to characterize the effect of localized (deep vs shallow) exposure of alfalfa root systems to NO₃^? on symbiotic N₂ fixation and NO₃^?-N uptake. Cuttings of a single alfalfa plant were grown in vertical split root systems in a controlled environment chamber. The split root system was a rigid acrylic tube (5 cm diam. by 60 cm long) filled with silica sand and divided into upper and lower sections at the 30-cm depth by a 5-mm-thick wax layer. Roots penetrated the wax layer, but mixing of nutrient solutions between the sections was prevented. Nodulation was restricted to the upper section. The plants were subjected for 10 days to the following treatments: both sections of the split root system received nutrient solution containing either 0.5, 5.25, or 10 mM NO₃^?; the upper section received 0.5 mM NO₃^? while the lower section received 10 mM NO₃^?; or the upper section received 10 mM NO₃^? while the lower section received 0.5 mM NO₃^?. Increasing supply of NO₃^? in the nutrient solution to both sections resulted in higher NO₃^?-N uptake, lower nodule mass and lower specific nitrogenase activity. Although NO₃^?-N uptake did not differ, plants exposed to 10 mM NO₃^? for 10 days in the upper, nodulated section of the root system had a 20% lower nodule mass than plants exposed to the same NO₃^? concentration in the lower, non-nodulated section of the root system. Specific nitrogenase activity was not different between these two treatments. Therefore, we conclude that: (1) nodule mass was dependent on two factors, the amount of NO₃^?-N taken up and the concentration of NO₃^? within the nodulated root zone; and (2) specific nitrogenase activity was little affected by the concentration of NO₃^? surrounding the nodules, but was strongly inhibited by NO₃^?-N taken up.     

Development of the orangutan permanent dentition: Assessing patterns and variation in tooth development

This study examines dental formation and alveolar emergence in a large cross-sectional sample composed primarily of wild-reared orangutans (N = 89) in order to provide information on the development of the permanent dentition in this hominoid and to address questions of variation in individual tooth formation, between teeth and between individuals. All specimens have been radiographed in lateral aspect and stages of crown and root formation recorded for all teeth. The ranges of crown and root formation of I¹₁, C¹₁, P⁴₄, and M³₃ have been calculated relative to the stage of M₁¹ development within a specific tooth quadrant. Then, for each specimen, BMDP scatterplot and nonparametric statistics have been used to graph changes in stages of these teeth relative to M¹₁ stages and to examine relationships between pairs of upper and lower dental counterparts and between teeth of each jaw. Results indicate 1) high correlations between upper and lower tooth pairs and between many of the permanent teeth within individuals, 2) a relatively large range of variability in individual tooth development (multistage ranges relative to M¹₁), 3) greater variation in root development at emergence than earlier reports, and 4) evidence of variability within the sequence emergence pattern of the orangutan. © 1996 Wiley-Liss, Inc.     

Synthesis,DNA Binding,and Photonuclease Activity of New Tetraaza Macrocyclic Constrained Isoxazole Rings as Subunit in Metal Complexes

The DNA-binding and photonuclease activity of newly synthesized tetra-azamacrocyclic ligand L (C₃₂H₃₂N₈O₄) and its complexes of type [MLCl₂] and [ML]Cl₂ (where M = Co(II), Fe(II) and Cu(II); L = N,N′-[3-(4-{5-[(2-amino-ethylamino)-methyl]-isoxazol-3yl}-phenyl)-isoxazol-5-yl methyl-ethane-1,2-diamine] are specified. An octahedral geometry has been proposed for Fe(II) and Co(II) complexes, while the Cu(II) complex has a square planar environment. The absorption spectral results indicate that the complexes bind with the base pairs of DNA, with an intrinsic binding constant K_b of Fe(II), Co(II), and Cu(II) complexes found to be 3.2 × 10⁴ M^?1, 5.3 × 10⁴ M^?1, and 4.2 × 10⁴ M^?1, respectively, in 5 mM Tris-HCl/50 mM NaCl buffer at pH 7.2. The large enhancement in the relative viscosity of DNA on binding to the complexes supports the proposed DNA binding modes. The viscosity and thermal denaturation studies sustain the effective intercalation with DNA. The DNA photocleavage studies demonstrated that compounds exhibit significant photonuclease activity by a concentration dependent on singlet oxygen mediated mechanism.     

Variation and evolutionary trends in the dentition of late pleistocene Mcrotus pennsylvanicus from three levels in bell cave,Alabama

Dental remains of the meadow vole, Microtus pennsylvanicus, were recovered from three superposed day units (zones) and four excavations (pits) in Bell Cave, Alabama. Bone collagen ¹⁴C dates of 26,500 + 480/ ‐ 500 and 11,800 + 480/‐ 500 yr BP. are associated with the bottom and top units, respectively. Length, width, number of closed triangles (five or six), and a measurement to determine the amount of closure of the sixth triangle were taken on the first lower molar (M₁) from the three zones in Pits 1 and 4. Length and width of the third upper molar (M₃) were also determined for the three zones in Pit 4. Analysis of Variance revealed no effect of pit origin or number of triangles on M₁ length or width. A trend from large to small size in M³ is documented, but is not statistically significant The same trend is evidenced by length and width of M₁. Size changes in length and width of M₁ from Pit 4 are statistically significant, but those from Pit 1 are not Although there is no difference in the lengths or widths of five versus six triangle M¹s from any pit or zone, there may be a trend towards increase in isolation of the sixth triangle from the remainder of the anteroconid complex. This trend is statistically significant for zone when ANOVA is performed on the variable, called closure, including both pit and zone origin, but is not significant when teeth from each pit are tested separately. An independent test of the length of M₁ from additional zones in Pits 2 and 3 confirms the low probability that directional size change is due to accidental sorting of fossil materials. These data demonstrate statistically significant intraspecific differences during a 15,000 year period of the late Pleistocene, and also provide evidence of variation that can be expected due to the combined effects of depositional reorganization, sampling and either stochastic processes or selection. The slight dwarfing recorded, while not particularly supportive of any evolutionary mode, may be correlated with climatic modification during the Wisconsinan glacial period. The evolutionary rate of dwarfing, as measured by mean reduction in length of the first lower molar, is 4.47 darwins. Although faster than many rates for fossil vertebrates and invertebrates calculated over longer periods of time, it is probably not unusual for Quaternary mammals during the time interval represented.     

An adapid primate,sinoadapis, from lufeng,latest miocene,Yunnan, China

The paper deals with a new form ofSinoadapis from the Lufeng hominoid Locality, assigned toSinoadapis shihuibaensis sp. nov. Holotype PA 882 A fragment of right mandible with C,−M₃. Other materials. PA 903 A fragment of left maxilla with P³−M³. PA 959 Left lower tooth row with I₁−P², P₄. PA 902 Right upper tooth row with C′−P³. PA 964 An isolated left I¹. PA 907 An isolated right I³. PA 972 An isolated right M³.     

ATPase activity in plasmalemma-rich vesicles isolated by aqueous two-phase partitioning from Vicia faba mesophyll and epidermis: Characterization and influence of abscisic acid and fusicoccin

Plasmalemma-rich microsomal vesicles were prepared from whole leaf and acid-washed epidermal tissue of Vicia faba L. cv. Osnabrücker Markt by aqueous two-phase partitioning in dextran T-500 and polyethylenglycol 1350 aqueous phases. These vesicles were tightly sealed and predominantly right-side out, and contained a K⁺ -stimulated, mg²⁺-dependent and vanadate-sensitive ATPase. The enzyme from both tissues exhibited nearly identical properties: pH optimum 6.4, K_m for ATP 0.60 mM(whole leaf) and 0.67 mM (epidermis). V_max -480 nmol (mg protein)¹ min¹ (whole leaf) and 510 nmol (mg protein)¹ min¹ (epidermis), I₅₀ (Na₃,VO₄) 7.5 μM (whole leaf) and 15 μM (epidermis). The enzyme was not inhibited by NO₃(50 mM)or sodium azide (I mM). DCCD (20 μM) reduced enzyme activity to 50% (whole leaf) and 58% (epidermis), gramicidin S (20 μM) to 36% (whole leaf) and 41%(epidermis). Ca²⁺ inhibited the ATPase [I₅₀, C²⁺: 0.5 mM(whole leaf) and 0.8 mM(epidermis)]. Ca²⁺ inhibited the ATPase [I₅₀, C²⁺ 0.5 mM(whole leaf) und 0.8 (epidermis)]. The vanadate-sensitive ATPase from whole leaf and epidermal tissue was slightly but significantly stimulated by fusicoccin (FC) at a concentration (0.13 μM) promoting stomatal opening. The stimulation was not seen in the solubilized ATPase. Stomata of the cultivar used here were insensitive lo (±)ABA up to 2 μM level which is effective in most other cultivars and species. Likewise, at this concentration no effect of ABA on the activity of the epidermal ATPase was observed. The data are discussed with respect to the interaction of FC and ABA with the ATPase.     

Additional materials of Myanmarpithecus yarshensis (Amphipithecidae, Primates) from the middle Eocene Pondaung Formation

Myanmarpithecus yarshensis is an amphipithecid primate from the middle Eocene Pondaung Formation in Myanmar. It was previously known based on maxillary fragments with P⁴–M³ and mandibular fragments with C–P₃ and M_2–3. This study reports new materials for the genus, including a humeral head fragment, a lingual fragment of the right M², a lingual fragment of the right M³, and a left I¹. These new materials were collected from approximately the same point, and likely belonged to the same individual. The upper molar morphology and size of the new materials show similarity to those of the type specimen, indicating that the new materials can be assigned to M. yarshensis. The humeral head is the first postcranial element that is associated with dental materials for amphipithecids. The morphological similarity between the previously reported larger humerus and this specimen confirms the assignment of the former specimen to Amphipithecidae and suggests common locomotor adaptations in the family. The upper central incisor is large relative to the molar fragments, but is within the variation among extant platyrrhines. The tooth is spatulate-shaped and high crowned, and lacks the mesial process, indicating similarity to I¹ of haplorhines and clear differences from that of adapoids. It has been suggested that amphipithecids, including Myanmarpithecus, have affinities with notharctine adapoids, but the morphology of I¹ does not support the notharctine hypothesis of the Amphipithecidae.     

Immobilization of glucose isomerase to ion-exchange materials

Glucose isomerase (D -xylose ketol-isomerase EC 5.3.1.5) from Bacillus Coagulans was partially purified and immobilized by adsorption to anion exchangers. The highest activities were obtained when the enzyme was adsorbed to DEAE-cellulose. On immobilization to DEAE-cellulose the measured optimum pH value for enzyme activity shifted from 7.2 to 6.8. There was no appreciable difference between the heat stabilities of soluble and immobilized enzyme. The K_{m app} values for the immobilized enzyme were found to be 0.25M in the presence of 0.01M Mg²⁺ and 0.19M with 0.005M Mg²⁺, while those enzyme were 0.11 and 0.17M, re spectively. Under conditions of contimuous of D -glucose, a decrease of activity with time was observed, but this decrease was less at a low Mg²⁺ concentration and was affected by column geometry. There were no appreciable diffusional limitation effects in packed-bed columns.     

Enamel thickness and the helicoidal occlusal plane

In the present study 38 unworn maxillary molars (M¹ = 16, M²= 12, M³ = 10) of modern humans from a Slavic necropolis were sectioned through the mesial cusps in a plane perpendicular to the cervical margin of the crown. Five slightly worn M¹s and one slightly worn M³ were also used thus increasing the total sample to 44, but measurements made on the worn areas were coded as missing values. Seven measurements of enamel thickness as well as the heights of the protocone and the paracone dentine horns were recorded in order to analyze whether changes in these dimensions in anteroposterior direction can be related to the helicoidal occlusal plane. Uni- and multivariate analyses revealed that the distribution of enamel thickness within and between maxillary molars corresponds to a helicoidal occlusal wear pattern. Enamel thickness along the occlusal basin increases from anterior to posterior, which may lead to rapid development of a reverse curve of Monson in first molars when compared to posterior teeth. However, although these overall differences together with the serial, especially delayed eruption pattern of human molars, contribute to the marked expression of the helicoidal occlusal plane in Homo, differences in enamel patterning between molars indicate that a helicoidal plane is a structural feature of the orofacial skeleton. In contrast to first upper molars, second and third molars show absolutely and relatively thicker enamel under the Phase I wear facet of the paracone, i. e., the lingual slope of the paracone, than under the Phase II facet of the protocone, i. e., the buccal slope of that cusp. These proportional differences are most pronounced in M³, as evidenced by uni- and multivariate statistics. It thus appears that the pattern of enamel thickness distribution from M¹ to M³ follows a trend towards providing additional tooth material in areas that are under greater functional demands, that is, corresponding to a lingual slope of wear anteriorly and to a flat or even buccal one posteriorly. In addition, the heights of the dentine horns in anteroposterior direction change in a way that lends support to the hypothesis that the axial inclination of teeth could be one of the most important factors for the development of the helicoidal occlusal plane. Finally, the changes in morphology and enamel thickness distribution from first to third upper molars found in this study suggest that molars could be “specialized” in their function, i. e., from performing proportionally more shearing anteriorly to increased crushing and grinding activities posteriorly. © 1994 Wiley-Liss, Inc.     

Measurement of the extent of electron transfer to the bacteriopheophytin in the M-subunit in reaction centers of Rhodopseudomonas viridis

We have measured the extent of flash-induced electron transfer from the bacteriochlorophyll dimer, P, to the bacteriopheophytin in the M-subunit, H_M, in reaction centers of Rhodopseudomonas viridis. This has been done by measuring the transient states produced by excitation of reaction centers trapped in the PH_L ^–H_M state at 90 K. Under these conditions the normal forward electron transfer to the bacteriopheophytin in the L-subunit, H_L, is blocked and the yield of transient P⁺H_M ^– can be estimated with respect to the lifetime of P^*. Under these conditions flash induced absorbance decreases of the bacteriochlorophyll dimer 990 nm band suggest that a transient P⁺ state is formed with a quantum yield of 0.09±0.06 compared to that formed during normal photochemistry. These transient measurements provide an upper limited on the yield of a transient P⁺ H_M ^– state. An estimate of 0.09 as the yield of the P⁺ H_M ^– state is consistent with all current observations. This estimate and the lifetime of P^* suggest that the electron transfer rate from P^* to H_M, k_M, is about 5 × 10⁹ sec^–1 (_M = 200ps). These measurements suggest that the a branching ratio k_L/k_M is on the order of 200. The large value of the branching ratio is remarkable in view of the structural symmetry of the reaction center. This measurement should be useful for electron transfer calculations based upon the reaction center structure.     

Isolation of plasma membrane and binding of the Ca2+ antagonist nimodipine in Chlamydomonas reinhardtii

Chlorophyll-free plasma membranes of the unicellular green alga Chlamydomonas reinhardtii Dangeard were purified from a microsomal fraction using an aqueous polymer two-phase system of 6.5% (w/w) dextran T500, 6·5% (w/w) polyethylene glycol 3350, 60 mM NaCI, 0 33 M sucrose and 5 mM potassium phosphate (pH 7·8). The plasma membrane fraction contained only 2·4% of the microsomal membrane protein. Specific activity of the plasma membrane marker enzyme, K*, Mg²⁺-ATPase (EC 3.6.1.3). was enriched 9-fold over the microsomal fraction, and 22% of total activity was recovered in the upper, polyethylene glycol-rich phase. Contamination from intracellular membranes was minimal. K*, Mg²⁺-ATPase showed a pH optimum at about 6·5, and addition of 0·05% (w/v) Triton X-100 stimulated the activity 3-fold. [³H]-Nimodipinc was employed to characterize 1,4-dihydropyridine-specific membrane receptors. Two apparent binding sites with different affinities to nimodipine were found in the crude microsomal fraction. The separation of plasma membranes from intracellular membranes revealed that one binding site with higher affinity (K_D= 9 nM) was located on the plasma membrane and a second binding site with lower affinity (K_D= 36 nM) on an intracellular membrane The apparent dissociation constants determined from the association and dissociation rate constants in kinetic experiments were comparable to those determined by equilibrium experiments. The maximum number of binding sites of the plasma membrane fraction and the intracellular membrane fraction was B_max= 440 and 470 fmol (mg protein)^-1, respectively. [³H]-Nimodipinc binding was inhibited by (±) verapamil and stimulated by D-cis-diltiazem in both fractions. Moreover, ethyle-neglycol-bis(2-aminoethylcther)-N, N'-tetraacctic acid (EGTA) inhibited [³H]-nimo-dipinc binding in the plasma membrane fraction but not in the intracellular membrane fraction This effect was cancelled by the addition of CaCl₂.     

Geometric morphometric analysis as a proxy to evaluate age‐related change in molar shape variation of low‐crowned Notoungulata (Mammalia)

Shape and age variation in dentition of Paleogene extinct native South American ungulates (Notoungulata) has been traditionally described using qualitative and quantitative approaches, and has played a controversial role in the systematics of several groups. Such is the case of the Notopithecidae, a group of notoungulates with low‐crowned teeth, known from the middle Eocene of Patagonia (Argentina). In this group, as well as in other contemporary families, extreme morphological changes associated to increasing dental wear were originally assumed to represent taxonomic differences; thus, dozens of species were erected, clearly reflecting the difficulty of defining discrete characters. In this contribution, a total of 89 upper molars and 91 lower molars were analyzed distributed in two factors, wear and species; three species of notopithecids were considered as study case, Notopithecus adapinus, Antepithecus brachystephanus, and Transpithecus obtentus, based on the large and well‐identified sample of upper and lower molars for each species. We have coupled geometric morphometric analyses with traditional comparative methods to get a better understanding and interpretation of both the changes in tooth shape contour and the link between shape and ontogeny. In addition, we evaluate the utility of this approach to identify which changes are strictly wear‐related and also test the qualitative characteristics used for diagnosing and differentiating notopithecid species. Our study yielded consistent results when applying independent geometric morphometric analyses on complex structures such as brachydont molar teeth. The landmark data is highly congruent with alternative sources of evidence, such as morphological studies using discrete characters. In notopithecid species, wear is the main factor affecting molar shape, followed by species (in lower molars) and allometry; in addition, lower teeth morphology is more definitive in separating species than upper molars, a fact that entails a key point for systematic studies of Paleogene brachydont notoungulates.