A new species of Pliopithecus Gervais, 1849 (Primates: Pliopithecidae) from the Middle Miocene (MN8) of Abocador de Can Mata (els Hostalets de Pierola,Catalonia, Spain)

Pliopithecus (Pliopithecus) canmatensis sp. nov. is described from several Late Aragonian localities from Abocador de Can Mata (ACM) in els Hostalets de Pierola (Vallès‐Penedès Basin, Catalonia, Spain), spanning from ～11.7 to 11.6 Ma (C5r.3r subchron), and being correlated to the MN8 (reference locality La Grive L3). The ACM remains display a pliopithecine dental morphology with well‐developed pliopithecine triangles on M/2 and M/3. This, together with other occlusal details, negates an attribution to the subgenus Epipliopithecus. Although slightly smaller, the ACM remains are most similar in size to comparable elements of P. piveteaui and P. antiquus. Several occlusal details (such as the greater development of the buccal cingulid in lower molars) and dental proportions (M/3 much longer than M/2), however, indicate greater similarities with P. antiquus from Sansan and La Grive. The ACM remains, however, differ from P. antiquus in dental proportions as well as occlusal morphology of the lower molars (including the less peripheral position of the protoconid and more medial position of the hypoconulid, the more mesial position of the buccal cuspids as compared to the lingual ones, the narrower but distinct mesial fovea, the higher trigonid, and the more extensive buccal cingulid, among others). These differences justify a taxonomic distinction at the species level of the ACM pliopithecid remains with respect to P. antiquus. Previous pliopithecid findings from the Vallès‐Penedès Basin, previously attributed to P. antiquus, are neither attributable to the latter species nor to the newly erected one. Am J Phys Anthropol, 2010. © 2009 Wiley‐Liss, Inc.     

Kretzoiarctos gen. nov., the Oldest Member of the Giant Panda Clade

The phylogenetic position of the giant panda, Ailuropoda melanoleuca (Carnivora: Ursidae: Ailuropodinae), has been one of the most hotly debated topics by mammalian biologists and paleontologists during the last century. Based on molecular data, it is currently recognized as a true ursid, sister-taxon of the remaining extant bears, from which it would have diverged by the Early Miocene. However, from a paleobiogeographic and chronological perspective, the origin of the giant panda lineage has remained elusive due to the scarcity of the available Miocene fossil record. Until recently, the genus Ailurarctos from the Late Miocene of China (ca. 8–7 mya) was recognized as the oldest undoubted member of the Ailuropodinae, suggesting that the panda lineage might have originated from an Ursavus ancestor. The role of the purported ailuropodine Agriarctos, from the Miocene of Europe, in the origins of this clade has been generally dismissed due to the paucity of the available material. Here, we describe a new ailuropodine genus, Kretzoiarctos gen. nov., based on remains from two Middle Miocene (ca. 12–11 Ma) Spanish localities. A cladistic analysis of fossil and extant members of the Ursoidea confirms the inclusion of the new genus into the Ailuropodinae. Moreover, Kretzoiarctos precedes in time the previously-known, Late Miocene members of the giant panda clade from Eurasia (Agriarctos and Ailurarctos). The former can be therefore considered the oldest recorded member of the giant panda lineage, which has significant implications for understanding the origins of this clade from a paleobiogeographic viewpoint.     

The effect of organic nitrogen and glucose on the production of recombinant human insulin-like growth factor in high cell densityEscherichia coli fermentations

Summary Two kinds of fed batch fermentation processes were compared at a 10-liter scale to examine their effect on recombinant human insulin-like growth factor (IGF-1) gene expression inEscherichia coli. The difference between the two processes was the feed medium composition and whether the process used a single or dual feed during the course of the fermentation. In the dual feed system, organic nitrogen was delivered at a higher rate (50 g/h) than in the single feed system (5 g/h). The dual feed process resulted in a significant increase in IGF-1 yield. 30 mg IGF-1/g dry cell weight was synthesized in the dual feed system compared to 3 mg IGF-1/g dry cell weight obtained in the single feed system. The presence of high levels of organic nitrogen during the induction period may enhance IGF-1 synthesis by protecting the IGF-1 from proteolytic degradation. The IGF-1 yield decreased to 17 mg/g dry cell weight when the glucose supply was decreased from 17 g/h to 8 g/h during the induction period; however, an increase in glucose supply from 17 g/h to 50 g/h during the induction period did not enhance the IGF-1 synthesis. Thus, the enhancement of IGF-1 gene expression in the dual feed process was mainly dependent on a high level of organic nitrogen and an appropriate level of glucose in the medium during the induction period.     

CANDLES,an assay for monitoring GPCR induced cAMP generation in cell cultures

Background

G protein-coupled receptors (GPCRs) represent a physiologically and pharmacologically important family of receptors that upon coupling to Gα_S stimulate cAMP production catalyzed by adenylyl cyclase. Thus, developing assays to monitor cAMP production is crucial to screen for ligands in studies of GPCR signaling. Primary cell cultures represent a more robust model than cell lines to study GPCR signaling since they physiologically resemble the parent tissue. Current cAMP assays have two fundamental limitations: 1) absence of cAMP kinetics as competition-based assays require cell lysis and measure only a single time-point, and 2) high variation with separate samples needed to measure consecutive time points. The utility of real-time cAMP biosensors is also limited in primary cell cultures due to their poor transfection efficiency, variable expression levels and inability to select stable clones. We therefore, decided to develop an assay that can measure cAMP not only at a single time-point but the entire cAMP kinetics after GPCR activation in untransfected primary cells.

Results

CANDLES (C yclic A MP iN direct D etection by L ight E mission from S ensor cells) assay for monitoring cAMP kinetics in cell cultures, particularly in primary cultures was developed. The assay requires co-culturing of primary cells with sensor cells that stably express a luminescent cAMP sensor. Upon GPCR activation in primary cells, cAMP is transferred to sensor cells via gap junction channels, thereby evoking a luminescent read-out. GPCR activation using primary cultures of rat cortical neurons and mouse granulosa cells was measured. Kinetic responses of different agonists to adrenergic receptors were also compared using rat cortical neurons. The assay optimization was done by varying sensor-test cell ratio, using phosphodiesterase inhibitors and testing cell-cell contact requirement.

Conclusions

Here we present CANDLES assay based on co-culturing test cells with cAMP-detecting sensor cells. This co-culture setup allows kinetic measurements, eliminates primary cell transfections and reduces variability. A variety of cell types (rat cortical neurons, mouse granulosa cells and established cell lines) and receptors (adrenergic, follicle stimulating hormone and luteinizing hormone/chorionic gonadotropin receptors) were tested for use with CANDLES. The assay is best applied while comparing cAMP generation curves upon different drug treatments to untransfected primary cells.

     

A new species of Anchitherium (Equidae: Anchitheriinae) from the Middle Miocene of Abocador de Can Mata (Vallès-Penedès Basin, NE Iberian Peninsula)

Anchitherium nievei sp. nov. is erected on the basis of dentognathic remains from the late Aragonian (MN7+8, ca. 12.3–12.1 to 11.7–11.6 Ma, Middle Miocene) of three localities from the Abocador de Can Mata local stratigraphic series (els Hostalets de Pierola, Vallès-Penedès Basin, Catalonia, Spain). This taxon differs from endemic species of Anchitherium from the inner Iberian basins, most closely resembling A. aurelianense, A. hippoides and A. steinheimense by its marked brachyodonty and small dental size, although differing by dental proportions and several details of occlusal morphology. From a paleoenvironmental viewpoint, the presence of this taxon fits well with previous inferences that indicated a relatively closed and humid forested habitat, more similar to that of similarly aged French localities.     

Middle Miocene tragulid remains from Abocador de Can Mata: The earliest record of Dorcatherium naui from Western Europe

In the past, the tragulid species Dorcatherium crassum has been considered a synonym of Dorcatherium naui, although they are currently considered distinct species. The former has an exclusively Aragonian (MN4 to MN6) stratigraphic range, whereas D. naui is considered a typically Vallesian species (MN9 and MN10) that survives until the Turolian (MN11). There are only a few previous citations of Dorcatherium from the European latest Aragonian (MN7+8), and only that of Przeworno (Poland) can be securely attributed to D. naui. Here we describe dentognathic and postcranial remains of Dorcatherium from several localities from the stratigraphic series of the Abocador de Can Mata (ACM), in els Hostalets de Pierola (Vallès-Penedès Basin, Catalonia, Spain). These Dorcatherium remains, from both the lower (ca. 11.9-12.5 Ma, MN7) and upper (ca. 11.6 Ma, MN8) portions of the ACM series, constitute the earliest record of D. naui from the Iberian Peninsula and Western Europe. They most closely resemble D. naui from the Vallesian, but still share several features with D. crassum. On the basis of the unique combination of features displayed by the ACM material, a new subspecies, D. naui meini is erected. Its distinct status from the nominotypical subspecies, D. naui naui, is reinforced by the chronological differences between the two taxa. The morphological similarities between Dorcatherium and the extant water chevrotain (Hyemoschus aquaticus) tentatively suggest the presence of forested and humid paleoenvironments in this area of the Vallès-Penedès Basin during the latest Aragonian. Nevertheless, Dorcatherium is very rare in ACM and displays a very patchy chronological distribution. Together with the similarly spotty distribution of both primates and castorids along the ACM series, this suggests that very humid conditions might have been only intermittently present in this area throughout the latest Aragonian.