Characterization of seven microsatellite marker loci in a genus of Malagasy lemurs (Propithecus)

The sifakas (genus Propithecus) are large, diurnal, lemurs endemic to the island of Madagascar. Throughout the island, lemur species are particularly vulnerable to extinction due to relatively small geographical ranges (Martin 2000). Thus, understanding the taxonomy of these highly endangered primates is essential for taking appropriate conservation action. For this study we collected tissue samples from 68 wild sifakas representing various Propithecus species. Genomic DNA was isolated and polymerase chain reaction (PCR) amplification was carried out. We describe seven microsatellites found to be polymorphic and useful for analysis of the population genetics of sifakas.     

Dietary Flexibility and Feeding Strategies of <Emphasis Type="Italic">Eulemur</Emphasis>: A Comparison with <Emphasis Type="Italic">Propithecus</Emphasis>

Despite the great variety of habitats in Madagascar, Eulemur has successfully populated most forested habitats on the island. Although the high dietary flexibility of Eulemur is often credited as one of the drivers of its evolutionary success, other behavioral evidence suggests a limited capacity for dietary switching. To shed light on the feeding strategies of Eulemur, we compared the dietary flexibility between populations of this genus with that of another widespread lemur taxon, Propithecus. We hypothesized that Eulemur would show greater dietary flexibility than Propithecus, which has a digestive system specialized for folivory, and that Eulemur living in dry forests would switch its diet from fruit to other food seasonally. To examine these hypotheses, we performed a phylogenetic least-squares analysis on 10 populations of Eulemur and 7 of Propithecus to assess the contribution of environmental variables and body mass on their dietary flexibility while controlling for phylogenetic relatedness. Eulemur relied heavily on fruit and did not show large variations in primary food over the year. Propithecus consumed leaves and fruits equally and exhibited considerable flexibility across seasons. Therefore, in contrast to our predictions, the anatomical specialization for fiber digestion heightens dietary flexibility in Propithecus. At the intrageneric level, we found similar ecogeographic variation; populations of both genera with heavier body mass consumed more fruit. As we predicted, Eulemur in drier habitats switched the diet from fruit to alternative food more frequently. To compensate for low dietary flexibility, Eulemur mostly adopts a power-feeding strategy by which it increases energy expenditure to acquire patchily distributed fruit resources.     

Morphometric analysis of Tremadocian (earliest Ordovician) kirkocystid mitrates (Echinodermata, Stylophora) from the Taebaeksan Basin, Korea

Abundant isolated remains of stylophoran echinoderms (cornutes and mitrates) are reported for the first time in the late Tremadocian (Asaphellus Zone) Tumugol Formation of Korea. Mitrate remains include numerous adorals of Kirkocystidae. Several new important anatomical features have been observed on these adorals, as an internal calcitic layer that is associated to s₂ and possibly also to the palmar complex. This observation suggests that the palmar complex would be present not only in mitrocystitid mitrates, but also in peltocystitids. For the first time, several morphometric analyses have been undertaken based on isolated kirkocystid adorals, so as to explore the morphological diversity displayed by Korean adorals, but also in order to compare their morphology with that of other Gondwanan kirkocystids. Morphometric analyses indicate the occurrence of two contrasted morphologies within Korean adorals (morphotypes A and B), and of three distinct morphologies within European and North African forms (“Anatifopsis”, “Balanocystites”, and “escandei” morphotypes). Comparison of Korean adorals with those from Europe and North Africa shows that: (1) morphotypes B and “Anatifopsis” are equivalent; consequently, the two Korean specimens referred to morphotype B are assigned to the genus Anatifopsis; (2) morphology of most Korean adorals, which belong to morphotype A, is clearly distinct from that of all other described kirkocystids from Europe and North Africa. The small size, juvenile morphology, and great morphological variability observed in the morphotype A of the Korean adorals are suggestive of possible heterochronic processes (peramorphosis).     

Craniodental Characters in the Taxonomy of Propithecus

The indriid genus Propithecus comprises the sifakas, medium-sized lemurs endemic to the forests of Madagascar. Traditionally, scientists divided the genus into only 2 or 3 species —Propithecus diadema, P. verreauxi, and, since 1988, P. tattersalli— with 4 or 5 subspecies in each of the first 2 taxa, but recent authors have suggested that many more distinct species should be recognized. We draw from quantitative and qualitative studies of craniodental traits to evaluate further the phenetic distinctiveness and taxonomic status of each named form of Propithecus. We recognize 9–10 species in the genus. The 4 or 5 species of the Propithecus diadema group —P. diadema, P. candidus, P. perrieri, P. edwardsi, and perhaps P. holomelas, if distinct— share several derived features, including large average body size and a mandible specialized for rotational chewing, and clearly comprise a closely related complex. The 5 species of the Propithecus verreauxi group —P. verreauxi, P. coquereli, P. deckenii, P. coronatus, P. tattersalli— are each highly distinctive morphologically and likely do not comprise a monophyletic group. In particular, we point out the highly distinctive cranial features of Propithecus coronatus, which researchers have traditionally largely overlooked.     

Effects of bleaching on the pharmacological and toxicological activities elicited by the aqueous extracts prepared from two “fire corals” collected in the Mexican Caribbean

Hydrocorals of the genus Millepora are abundant skeleton-forming inhabitants of coral reefs around the world. These species are popularly known as “fire corals” since contact with them causes severe pain, skin eruptions and blisters as a result of the release of unidentified toxins. Millepora species associate with photosynthetic dinoflagellates of the genus Symbiodinium (“zooxanthellae”), and up to now the role of these symbionts in the toxic effects induced by the “fire corals” is unknown. In this study, we compared the hemolytic, vasoconstrictor, and phospholipase A₂ (PLA₂) activities of the crude aqueous extracts prepared from normal and bleached specimens of two hydrocorals collected in the Mexican Caribbean, Milleporaalcicornis and Millepora complanata. Electrophoretic analysis revealed some differences between the protein profiles of the extracts prepared from normal and bleached specimens. Bleaching decreased, but not abolished, the hemolytic effect induced by the hydrocorals extracts and the phospholipase A₂ activity of M. complanata extract. Furthermore, it did not modify the enzymatic activity of M. alcicornis extract and vasoconstriction elicited by both extracts. Our results suggest that the presence of the symbionts does not importantly influence the pharmacological and toxic effects induced by Millepora ssp. extracts, and indicate that cnidarians are the main source of the bioactive compounds.     

Fish otoliths from the Priabonian (Late Eocene) of North Italy and South-East France - Their paleobiogeographical significance

The study of the Late Eocene (Priabonian) otolith associations from Possagno, North-East Italy, and from the Synclinal d’Allons in Haute Provence, South-East France, allows for the reconstruction of a teleost fauna of 55 taxa, which is the most diversified assemblage presently known from the Upper Eocene Paleo-Mediterranean basin. Thirty-six taxa are identified at the species level, and five of those are new: “genus Alepocephalidarum” astrictus, “genus Lophiiformorum” canovae, “genus Agonidarum” sudans, “genus Uranoscopidarum” cochlearis and Aseraggodes laganum. In the Synclinal d’Allons, the otolith associations reflect a tropical to subtropical neritic environment with a few mesopelagic fishes. At Possagno, the associations indicate an environment that changed from one that was deep and exposed to the pelagic realm and then evolved to a more shallow sea with a well-diversified benthic life and less mesopelagic fishes. A paleobiogeographical analysis of all known data on Priabonian otoliths, worldwide, shows clearly a western Atlantic (Louisiana) and an eastern Atlantic-Paleomediterranean association. In the eastern Atlantic-Paleomediterranean association, the Aquitaine association not only differs from the Possagno-Allons association in function of its more distant geographical position, but also by its stronger oceanic character in the southern part of the basin, and by the occurrence in the north, of a very shallow water facies (Saint-Estephe Formation) that contains some taxa which are known nowhere else in the Priabonian. The Ukraine fauna is characterized by a high number of species, which have an Oligocene record in other European sites. The northern geographic location of Ukraine, combined with the good connections to both the North Sea Basin and the Turgai street can provide the explanation. Many Oligocene species (or their close relatives) probably already existed at Eocene times in more northern regions, but could penetrate only in more southern European seas since the strong cooling at the beginning of the Oligocene.     

Le genre Nucleolites (Echinoidea, Cassiduloidea) du Bajocien à l'Oxfordien dans le Bassin de Paris : apport des données architecturales à la systématique et à la phylogénie

The classification of the Paris basin Bajocian-Oxfordian species of the genus Nucleolites is revised. A classic point of view, until now used by authors and mainly based on the general morphology of the test, is first developed. Then, this last is modified with new architectural data, which refer to the extraxial-axial theory concerning the structure of the apical system as well as the presence of supplementary and catenal plates. These architectural data are first used facing the general morphology of the test. Such approach leads to a new point of view for the classification of the species of the genus Nucleolites. The systematics is particularly significantly simplified. The deduced and proposed phylogenetic hypothesis shows that the genus Nucleolites is composed of two parallel lineages as soon as the beginning of the genus in the Bajocian: a group with a so-called “primitive” architecture, and a group with an “advanced” one. On and after the Late Callovian, the species with a primitive organisation give birth to advanced species, which continue in younger time. In this way, the genus Nucleolites may be paraphyletic since the Late Callovian onward.     

The first complete skull of Simopithecus darti from Makapansgat,South Africa,and its systematic position

The skull of a subadult female of Simopithecus darti from the “Upper Phase I” of the Makapansgat Limeworks, South Africa, is described. It is the first complete skull of Simopithecus which is recorded from the lower Pleistocene “Australopithecine cave deposits” of southern Africa. The present skull shows an interesting mixture of primitive and advanced features. In some respects, it is reminiscent of Parapapio, in others it seems to have affinities with the extant Theropithecus gelada rather than with Simopithecus oswaldi. The systematics of the fossil genus Simopithecus and of the tribe Theropithecini Jolly (1966) are discussed.     

Two reaction pathways for transformation of high potential cytochrome b559 of PS II into the intermediate potential form

This study describes an analysis of different treatments that influence the relative content and the midpoint potential of HP Cyt b559 in PS II membrane fragments from higher plants. Two basically different types of irreversible modification effects are distinguished: the HP form of Cyt b559 is either predominantly affected when the heme group is oxidized (“O-type” effects) or when it is reduced (“R-type” effects). Transformation of HP Cyt b559 to lower potential redox forms (IP and LP forms) by the “O-type” mechanism is induced by high pH and detergent treatments. In this case the effects consist of a gradual decrease in the relative content of HP Cyt b559 while its midpoint potential remains unaffected. Transformation of HP Cyt b559 via an “R-type” mechanism is caused by a number of exogenous compounds denoted L: herbicides, ADRY reagents and tetraphenylboron. These compounds are postulated to bind to the PS II complex at a quinone binding site designated as Q_C which interacts with Cyt b559 and is clearly not the Q_B site. Binding of compounds L to the Q_C site when HP Cyt b559 is oxidized gives rise to a gradual decrease in the E_m of HP Cyt b559 with increasing concentration of L (up to 10 K_ox(L) values) while the relative content of HP Cyt b559 is unaffected. Higher concentrations of compounds L required for their binding to Q_C site when HP Cyt b559 is reduced (described by K_red(L)) induce a conversion of HP Cyt b559 to lower potential redox forms (“R-type” transformation). Two reaction pathways for transitions of Cyt b559 between the different protein conformations that are responsible for the HP and IP/LP redox forms are proposed and new insights into the functional regulation of Cyt b559 via the Q_C site are discussed.     

Dimensions of the heads of the fast and slow sedimenting forms of bacteriophage T2L

Bacteriophage T2L undergoes a reversible conversion between two conformations having S_{20. w} values of about 1000 s and 700 s. These are known, respectively, as the “fast” and “slow” sedimenting forms. Preparations of both forms of the bacteriophage were fixed with formaldehyde, negatively stained with uranyl acetate, and observed in the electron microscope. The tail fibers of the fast form generally extended upward, in proximity to the tail and terminating on the head. The tail fibers of the slow form extended outward, away from the tail.     

Crystal Structure of Glyceraldehyde-3-Phosphate Dehydrogenase 1 from Methicillin-Resistant Staphylococcus aureus MRSA252 Provides Novel Insights into Substrate Binding and Catalytic Mechanism

The dreaded pathogen Staphylococcus aureus is one of the causes of morbidity and mortality worldwide. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), one of the key glycolytic enzymes, is irreversibly oxidized under oxidative stress and is responsible for sustenance of the pathogen inside the host. With an aim to elucidate the catalytic mechanism and identification of intermediates involved, we describe in this study different crystal structures of GAPDH1 from methicillin-resistant S. aureus MRSA252 (SaGAPDH1) in apo and holo forms of wild type, thioacyl intermediate, and ternary complexes of active-site mutants with physiological substrate d-glyceraldehyde-3-phosphate (G3P) and coenzyme NAD⁺. A new phosphate recognition site, “new P_i” site, similar to that observed in GAPDH from Thermotoga maritima, is reported here, which is 3.40 Å away from the “classical P_i” site. Ternary complexes discussed are representatives of noncovalent Michaelis complexes in the ground state. d-G3P is bound to all the four subunits of C151S.NAD and C151G.NAD in more reactive hydrate (gem-di-ol) form. However, in C151S + H178N.NAD, the substrate is bound to two chains in aldehyde form and in gem-di-ol form to the other two. This work reports binding of d-G3P to the C151G mutant in an inverted manner for the very first time. The structure of the thiaocyl complex presented here is formed after the hydride transfer. The C3 phosphate of d-G3P is positioned at the “P_s” site in the ternary complexes but at the “new P_i” site in the thioacyl complex and C1-O1 bond points opposite to His178 disrupting the alignment between itself and NE2 of His178. A new conformation (Conformation I) of the 209-215 loop has also been identified, where the interaction between phosphate ion at the “new P_i” site and conserved Gly212 is lost. Altogether, inferences drawn from the kinetic analyses and crystal structures suggest the “flip-flop” model proposed for the enzyme mechanism.     

Juvenile ornithopod (Dinosauria: Rhabdodontidae) remains from the Upper Cretaceous (Lower Campanian, Gosau Group) of Muthmannsdorf (Lower Austria)

The fragmentary remains of a juvenile rhabdodontid ornithopod from the Coal-bearing Complex of the Gosau Group (Lower Campanian, Grünbach syncline) at Muthmannsdorf near Wiener Neustadt, Lower Austria are revised. The material, probably belonging to a single individual, includes a right dentary (lectotype of Iguanodon suessi Bunzel, 1871, designated herein), teeth, a fragmentary parietal, fragments of scapula, ?radius, femur, tibia, two vertebrae (lost) and a manual ungual.The lectotype dentary does not provide clear autapomorphies or sufficient diagnostic features to determine its position within the Rhabdodontidae at generic level. By this “Iguanodon suessi” Bunzel, 1871 and the genus “Mochlodon” Seeley, 1881, to which it was latter referred as type species, cannot be characterized sufficiently by differential diagnosis and these are best considered nomina dubia. Based upon combined character comparisons (mainly postcranial features) the Muthmannsdorf ornithopod is referred herein to Zalmoxes Weishampel, Jianu, Csiki and Norman, 2003, a genus so far known from the late Maastrichtian of Romania. It probably but not evidently represents a yet unnamed species, most closely related to Zalmoxes shqiperorum Weishampel, Jianu, Csiki and Norman, 2003. At the present state of knowledge the Austrian material is not further diagnostic at the species level and kept in open nomenclature as Zalmoxes sp.     

The Pragian-Emsian conodont successions of the Barrandian area: search of an alternative to the GSSP polygnathid-based correlation concept

Six Pragian-Emsian boundary sections in the Barrandian area, western of Prague, provided evidence of well detectable entries of Latericriodus fauna probably at the earliest Emsian beds (particularly Latericriodus bilatericrescens gracilis Bultynck). The chance to find icriodontid conodonts increases with latest part of Praha Fm., which is apparently of Emsian age, whereas polygnathids are sparsely preserved to absent. The high icriodontid/polygnathid ratio links together all these Barrandian sections, although their open-sea depositional environments range widely from deep troughs with rapid calciturbidite accumulation (Pod Barrandovem section) to relatively starving slope environments on elevations (Na Po?árech sections). The reports on polygnathid occurrences around the Pragian-Emsian boundary beds of the Barrandian area are much biased by poor reproducibility of the results (the conodonts cannot be found again) as well as by different levels where they were randomly found and/or by major taxonomic problems with the “kitabicus” and “dehiscens” definitions and their stratigraphic use. Apart from the GSSP in the Zinzilban Gorge (Uzbekistan) and its “kitabicus” boundary, the newly introduced “gracilis” biostratigraphic-marker concept preserves the major volume of the Pragian and respects also approximately the base of the traditional Emsian. These “gracilis” entries are clustered around the dark-colored “graptolite-bearing interval” beds, which largely form a prominent lithological marker within the latest, light gray-colored Dvorce-Prokop Limestone of the Barrandian area. This “gracilis” biostratigraphic marker has a promising correlation potential relative to Spanish and Moroccan sections.     

An activation gating switch in Kv1.2 is localized to a threonine residue in the S2-S3 linker

The activation properties of Kv1.2 channels are highly variable, with reported half-activation (V_1/2) values ranging from ∼−40 mV to ∼+30 mV. Here we show that this arises because Kv1.2 channels occupy two distinct gating modes (“fast” and “slow”). “Slow” gating (τ_act = 90 ± 6 ms at +35 mV) was associated with a V_1/2 of activation of +16.6 ± 1.1 mV, whereas “fast” gating (τ_act = 4.5 ± 1.7 ms at +35 mV) was associated with a V_1/2 of activation of −18.8 ± 2.3 mV. It was possible to switch between gating modes by applying a prepulse, which suggested that channels activate to a single open state along separate “fast” and “slow” activation pathways. Using chimeras and point mutants between Kv1.2 and Kv1.5 channels, we determined that introduction of a positive charge at or around threonine 252 in the S2-S3 linker of Kv1.2 abolished “slow” activation gating. Furthermore, dialysis of the cytoplasm or excision of cell-attached patches from cells expressing Kv1.2 channels switched gating from “slow” to “fast”, suggesting involvement of cytoplasmic regulators. Collectively, these results demonstrate two modes of activation gating in Kv1.2 and specific residues in the S2-S3 linker that act as a switch between these modes.     

Point mutations in firefly luciferase C-domain demonstrate its significance in green color of bioluminescence

Firefly luciferase is a two-domain enzyme that catalyzes the bioluminescent reaction of firefly luciferin oxidation. Color of the emitted light depends on the structure of the enzyme, yet the exact color-tuning mechanism remains unknown by now, and the role of the C-domain in it is rarely discussed, because a very few color-shifting mutations in the C-domain were described. Recently we reported a strong red-shifting mutation E457K in the C-domain; the bioluminescence spectra of this enzyme were independent of temperature or pH. In the present study we investigated the role of the residue E457 in the enzyme using the Luciola mingrelica luciferase with a thermostabilized N-domain as a parent enzyme for site-directed mutagenesis. We obtained a set of mutants and studied their catalytic properties, thermal stability and bioluminescence spectra. Experimental spectra were represented as a sum of two components (bioluminescence spectra of putative “red” and “green” emitters); λ_max of these components were constant for all the mutants, but the ratio of these emitters was defined by temperature and mutations in the C-domain. We suggest that each emitter is stabilized by a specific conformation of the active site; thus, enzymes with two forms of the active site coexist in the reactive media. The rigid structure of the C-domain is crucial for maintaining the conformation corresponding to the “green” emitter. We presume that the emitters are the keto- and enol forms of oxyluciferin.     

Cultivation of the first mesophilic representative ("mesotoga") within the order Thermotogales

Cultivated members of the order Thermotogales comprise only thermophilic to hyperthermophilic anaerobic microorganisms. However, based on molecular studies, the existence of mesophilic members (“mesotoga”) within this order has been postulated but has not been demonstrated by cultural approaches so far. A “mesotoga” (strain PhosAc3) that belonged to an uncultivated lineage distantly related to the thermophilic Kosmotoga genus has now been cultivated in axenic culture. It grew between 30 °C and 50 °C (optimum 40 °C) and oxidized lactate using elemental sulphur as a terminal electron acceptor. Further genomic and physiological characterization of strain PhosAc3 will be important not only for understanding bacterial adaptation to high and moderate temperatures at small evolutionary scales, but also because “mesotoga” might play a crucial ecological role in ecosystems polluted by aromatic compounds.     

Cristatopecten nov. gen. benoisti (Pectinidae, Bivalvia) du Chattien d'Aquitaine (France). Révision systématique et réflexions sur l'origine et l'évolution des Cristatopecten

The detailed study of a sample of “Pecten (Amussiopecten) benoisti” Cossmann and Peyrot (1914) from the Chattian outcrops of the Adour Basin, allowed to improve our knowledge of this species poorly known till now and to show its important ornamental variability. It follows that some nomenclatural points have been clarified at the generic level: we refute the attribution of this species to the genus Amussiopecten Sacco (1897) as used by authors—genus which was besides emended several times subsequently— and we propose its allocation to Cristatopecten nov. gen. Furthermore, this study allowed to remove the ambiguity existing originally as for the attribution of the species benoisti and burdigalensis to a same genus: our comparison of the latter two taxa shows that if benoisti has to be henceforth included in the new genus Cristatopecten (as its type species), burdigalensis (which is the type species of Amussiopecten) must be incorporated in the genus Flabellipecten Sacco (1897). The two genera Amussiopecten and Flabellipecten are therefore synonymous, as Depéret and Roman had already stated in 1910. The Cristatopecten from the Old World seem to be derived from a group of “Aequipecten” notably known in the Italian Rupelian, the group of “Aequipecten deletus” (Michelotti, 1861), of which we study here a sample from the Aquitaine Chattian. In the Aquitaine Basin, the Cristatopecten are represented, in the Chattian, by Cristatopecten benoisti (Cossmann and Peyrot, 1914); in the Lower Aquitanian, by C. praehaueri (Bongrain, Cahuzac and Freneix, 1994); in the Burdigalian-Langhian of the Saubrigues paleocanyon, by C. haueri (Michelotti, 1847) and C. miogallicus (Cossmann and Peyrot, 1914). During the whole Neogene, they spread and evolved in the Tethys and the Paratethys areas, and seem to have become extinct in the Pliocene with the disappearance of C. cristatus (Bronn, 1827), a species that we consider not as an Amusium s.s., but as the last representative of the Cristatopecten phylum.