RANK rewires energy homeostasis in lung cancer cells and drives primary lung cancer.

Lung cancer is the leading cause of cancer deaths. Besides smoking, epidemiological studies have linked female sex hormones to lung cancer in women; however, the underlying mechanisms remain unclear. Here we report that the receptor activator of nuclear factor-kB (RANK), the key regulator of osteoclastogenesis, is frequently expressed in primary lung tumors, an active RANK pathway correlates with decreased survival, and pharmacologic RANK inhibition reduces tumor growth in patient-derived lung cancer xenografts. Clonal genetic inactivation of KRasG12D in mouse lung epithelial cells markedly impairs the progression of KRasG12D -driven lung cancer, resulting in a significant survival advantage. Mechanistically, RANK rewires energy homeostasis in human and murine lung cancer cells and promotes expansion of lung cancer stem-like cells, which is blocked by inhibiting mitochondrial respiration. Our data also indicate survival differences in KRasG12D -driven lung cancer between male and female mice, and we show that female sex hormones can promote lung cancer progression via the RANK pathway. These data uncover a direct role for RANK in lung cancer and may explain why female sex hormones accelerate lung cancer development. Inhibition of RANK using the approved drug denosumab may be a therapeutic drug candidate for primary lung cancer.

Nuttalliellidae in Burmese amber: implications for tick evolution.

Ticks are composed of 3 extant families (Argasidae, Ixodidae and Nuttalliellidae) and 2 extinct families (Deinocrotonidae and Khimairidae). The Nuttalliellidae possess one extant species (Nuttalliella namaqua) limited to the Afrotropic region. A basal relationship to the hard and soft tick families and its limited distribution suggested an origin for ticks in the Afrotropics. The Deinocrotonidae has been found in Burmese amber from Myanmar and Iberian amber from Spain, suggesting a wider distribution of the lineage composed of Deinocrotonidae and Nuttalliellidae. The current study describes 8 fossils from mid-Cretaceous (ca. 100 Ma) Burmese amber: 2 Deinocroton species (Deinocroton bicornis sp. nov.; Deinocroton lacrimus sp. nov.), 5 Nuttalliella species (Nuttalliella gratae sp. nov., Nuttalliella tuberculata sp. nov., Nuttalliella placaventrala sp. nov., Nuttalliella odyssea sp. nov., Nuttalliella tropicasylvae sp. nov.) and a new genus and species (Legionaris nov. gen., Legionaris robustus sp. nov.). The argument is advanced that Deinocroton do not warrant its own family, but forms part of the Nuttalliellidae comprising 3 genera, Deinocroton, Legionaris nov. gen. and Nuttalliella). Affinities of Burmese tick fossils to the Australasian region, specifically related to rifting of the Burma terrane from northern Australia ~150 million years ago, suggest that Nuttalliella had a much wider distribution than its current limited distribution. The distribution of Nuttalliella likely stretched from Africa over Antarctica and much of Australia, suggesting that extant members of this family may still be found in Australia. Considerations for the geographic origins of ticks conclude that an Afrotropic origin can as yet not be discarded.

The visible skeleton 2.0: phenotyping of cartilage and bone in fixed vertebrate embryos and foetuses based on X-ray microCT.

For decades, clearing and staining with Alcian Blue and Alizarin Red has been the gold standard to image vertebrate skeletal development. Here, we present an alternate approach to visualise bone and cartilage based on X-ray microCT imaging, which allows the collection of genuine 3D data of the entire developing skeleton at micron resolution. Our novel protocol is based on ethanol fixation and staining with Ruthenium Red, and efficiently contrasts cartilage matrix, as demonstrated in whole E16.5 mouse foetuses and limbs of E14 chicken embryos. Bone mineral is well preserved during staining, thus the entire embryonic skeleton can be imaged at high contrast. Differences in X-ray attenuation of ruthenium and calcium enable the spectral separation of cartilage matrix and bone by dual energy microCT (microDECT). Clearing of specimens is not required. The protocol is simple and reproducible. We demonstrate that cartilage contrast in E16.5 mouse foetuses is adequate for fast visual phenotyping. Morphometric skeletal parameters are easily extracted. We consider the presented workflow to be a powerful and versatile extension to the toolkit currently available for qualitative and quantitative phenotyping of vertebrate skeletal development.

Rapid evolution of the primate larynx?

Tissue vibrations in the larynx produce most sounds that comprise vocal communication in mammals. Larynx morphology is thus predicted to be a key target for selection, particularly in species with highly developed vocal communication systems. Here, we present a novel database of digitally modeled scanned larynges from 55 different mammalian species, representing a wide range of body sizes in the primate and carnivoran orders. Using phylogenetic comparative methods, we demonstrate that the primate larynx has evolved more rapidly than the carnivoran larynx, resulting in a pattern of larger size and increased deviation from expected allometry with body size. These results imply fundamental differences between primates and carnivorans in the balance of selective forces that constrain larynx size and highlight an evolutionary flexibility in primates that may help explain why we have developed complex and diverse uses of the vocal organ for communication.

Hard ticks in Burmese amber with Australasian affinities.

Three examples of metastriate hard ticks (Ixodida: Ixodidae) with apparent affinities to modern Australasian genera are described from the mid-Cretaceous (ca. 100 Ma) Burmese amber of Myanmar. Two nymphs of Bothriocroton muelleri sp. nov. represent the oldest (and only) fossil record of this genus, living members of which are restricted to Australia and predominantly feed on monitor lizards, snakes and echidnas. A female of Archaeocroton kaufmani sp. nov. shares its basis capitulum shape with the tuatara tick Archaeocroton sphenodonti (Dumbleton, 1943), the only extant member of this genus and an endemic species for New Zealand. The presence of 2 Australasian genera in Burmese amber is consistent with a previous record of an Ixodes Latreille, 1795 tick from this deposit which resembles Australian members of this genus. They further support an emerging hypothesis that fauna of the amber forest, which may have been on an island at the time of deposition, was at least partly Gondwanan in origin. A revised evolutionary tree for Ixodida is presented compiling data from several new Burmese amber ticks described in the last few years.

Morphology and volume of Meibomian glands ex vivo pre and post partial tarsal plate excision, cryotherapy and laser therapy in the dog using microCT.

OBJECTIVE: To determine the morphology and volume of Meibomian glands (MG) of dogs with microCT before and after partial tarsal plate excision (PTPE), cryotherapy, and laser therapy. PROCEDURE: MicroCT scans were made of 12 upper lids (ULs) and lower lids (LLs) of 12 dogs. After undergoing PTPE, 10 ULs and LLs were scanned again, and one UL and one LL was scanned after laser therapy and one UL and one LL after cryotherapy. RESULTS: The length of the area containing MGs did not change pre- and post-PTPE, and cryo- or laser therapy. The mean number of MGs in the ULs and LLs was 30.50 and 29.42, respectively, and did not change during the procedures. The average length of one individual MG was 2.60 mm. The mean volume of MGs in the 10 ULs and LLs pre-PTPE was 21.45 and 17.2 mm3 , respectively, and 12.84 and 11.25 mm3 in the UL and LL after PTPE, respectively. The mean volume of MGs decreased from 29.78 mm3 precryotherapy to 28.91 mm3 post-treatment and in the lower eyelid from 22.87 to 22.4 mm3 after cryotherapy. The mean volume of MGs in the UL and LL before laser therapy was 8.95 and 6.78 mm3 , respectively, and after 9.25 and 6.38 mm3 , respectively. CONCLUSION: MicroCT is a valuable tool to determine the morphology and the volume of MGs and to demonstrate changes that occur after PTPE, laser-, and cryotherapy. There is no need for additional preparation, such as staining, of the specimen prior to scanning.

X-ray computerized microtomography and confocal Raman microscopy as complementary techniques to conventional imaging tools for the microstructural characterization of Cheddar cheese.

This study explored the use of X-ray computerized microtomography (micro-CT) and confocal Raman microscopy to provide complementary information to well-established techniques, such as confocal laser scanning microscopy (CLSM), for the microstructural characterization of cheese. To evaluate the potential of these techniques, 5 commercial Cheddar cheese samples, 3 with different ripening times and 2 with different fat contents, were analyzed. Confocal laser scanning microscopy was particularly useful to describe differences in fat and protein distribution, especially between the 2 samples with different fat contents. The quantitative data obtained through image analysis correlated well with the nutritional information provided in the product labels. Conversely, micro-CT was more advantageous for studying the size and spatial distribution of microcrystals present within the cheese matrix. Two types of microcrystals were identified that differed in size, shape, and X-ray attenuation. The smallest, with a diameter of approximately 10 to 20 µm, were more abundant in the samples and presented a more uniform roundish shape and higher X-ray attenuation. Larger and more heterogeneous crystals with diameters reaching 50 µm were also observed in scarcer numbers and showed lower X-ray attenuation. Confocal Raman microscopy was useful not only for identifying the distribution of all these components but also allowed comparing the presence of micronutrients such as carotenoids in the cheeses and provided compositional information on the crystals detected. Small and large crystals were identified as calcium phosphate and calcium lactate, respectively. Overall, using micro-CT, confocal Raman microscopy, and CLSM in combination generated novel and complementary information for the microstructural and nutritional characterization of Cheddar cheese. These techniques can be used to provide valuable knowledge when studying the effect of milk composition, processing, and maturation on the cheese quality attributes.

Digital dissection of the head of the frogs Calyptocephalella gayi and Leptodactylus pentadactylus with emphasis on the feeding apparatus.

Micro-computed tomography (microCT) of small animals has led to a more detailed and more accurate three-dimensional (3D) view on different anatomical structures in the last years. Here, we present the cranial anatomy of two frog species providing descriptions of bone structures and soft tissues of the feeding apparatus with comments to possible relations to habitat and feeding ecology. Calyptocephalella gayi, known for its aquatic lifestyle, is not restricted to aquatic feeding but also feeds terrestrially using lingual prehension. This called for a detailed investigation of the morphology of its feeding apparatus and a comparison to a fully terrestrial species that is known to feed by lingual prehension such as Leptodactylus pentadactylus. These two frog species are of similar size, feed on similar diet but within different main habitats. MicroCT scans of both species were conducted in order to reconstruct the complete anatomical condition of the whole feeding apparatus for the first time. Differences in this regard are evident in the tongue musculature, which in L. pentadactylus is more massively built and with a broader interdigitating area of the two main muscles, the protractor musculus genioglossus and the retractor musculus hyoglossus. In contrast, the hyoid retractor (m. sternohyoideus) is more massive in the aquatic species C. gayi. Moreover, due to the different skull morphology, the origins of two of the five musculi adductores vary between the species. This study brings new insights into the relation of the anatomy of the feeding apparatus to the preferred feeding method via 3D imaging techniques. Contrary to the terrestrially feeding L. pentadactylus, the skeletal and muscular adaptations of the aquatic species C. gayi provide a clear picture of necessities prescribed by the habitat. Nevertheless, by keeping a certain amount of flexibility of the design of its feeding apparatus, C. gayi is able to employ various methods of feeding.

First description of freshwater mite Unionicola sauerensis sp. nov. infesting thick-shelled river mussel Unio crassus.

A sample of 30 thick-shelled river mussels Unio crassus Philipsson (Unionida: Unionidae) was collected from the River Sauer in Luxembourg to acquire data on parasitic infestations of the mussels. Among other parasites, different development stages of freshwater mites were collected from the gills and the mantle of the mussels and were documented with bright-field, stereo, and confocal laser scanning microscopy and microscopic X-ray computed tomography. The retrieved data allowed a morphological description of larvae and female adults of the mites and assigning them to the genus Unionicola Haldeman (Trombidiformes: Unionicolidae) and the subgenus Pentatax Thor. Additionally, adult stages and larvae were barcoded by sequencing a section of the mitochondrial COI and 18S rRNA genes. This resulted in 4 new, similar Unionicola lineages from the adult stages, which differ in at least 14.7% (uncorrected p distance) from those already published. Barcoding of larval DNA was not successful. The comparison with known European species of the genus Unionicola and analysis of the barcoding results allowed the proposal of a new species of the genus Unionicola. The species was named Unionicola sauerensis sp. nov. after the River Sauer in Luxembourg, where the infested mussels were collected.

Labial and buccal minor salivary glands of the dog - location, three-dimensional arrangement and histology.

OBJECTIVE: Transplantation of minor salivary glands (MSGs) to the conjunctiva is a treatment option for patients suffering from dry eye disease. As there is not enough information about labial and buccal MSGs in dogs, the aim of this study was to provide evidence of the presence of these glands and to investigate their spatial arrangement and excretory ducts. METHODS: The oral mucosa of the lower lip of 4 dogs and the whole lower jaw of 1 dog were used for histological and microCT analysis. Presence, number, volumes and the tissue depth of MSGs were assessed. RESULTS: Histological analysis showed that compact tubulo-acinar glands were located in the submucosal connective tissue. MicroCT images revealed that 9 to 21 MSGs were arranged in a single row at the level of the dental alveolae. The volume of the MSGs increased from rostral to caudal and the total volume of glandular tissue per animal ranged from 35.01 mm3 to 549.43 mm3 . The mean tissue depth of MSGs ranged from 0.57 mm to 1.37 mm (upper surface of glands) and between 1.43 mm and 3.09 mm (lower surface of the glands). Excretory ducts left the dorsal part of the glands and ran in dorso-rostral direction. CONCLUSIONS: The location, number and volume of the labial and buccal MSGs in the dog could be detected and described using microCT scans and histology. The present results can provide valuable information for future transplantation of labial MSGs as therapeutic measure against keratoconjunctivitis sicca.

Hippocampal size did not differ between epileptic and non-epileptic dogs using volumetric and subjective methods.

Hippocampal changes in epilepsy may manifest as hippocampal atrophy/sclerosis. A recent human study suggests that the demonstration of hippocampal volume loss is more reliable using quantitative evaluation methods. The aim of the present study was to obtain volumetric data in both epileptic and healthy dogs, to compare hippocampal volumes in both groups, and to compare subjective and volumetric assessment. Volumetric measurements of the hippocampi, lateral ventricles and hemispheria were performed in 31 epileptic and 15 control dogs. There was a positive association between the body weight and the hemispheric volume, as well as between the hemispheric volume and the ipsilateral hippocampal volume. There was no significant correlation between age and the volume of any measured brain structures. There was no statistically significant difference between the hippocampal volumes of the control group and the epileptic group. A statistically significant difference between the two groups for hippocampus/hemispherium ratio or hippocampal asymmetric ratio was not identified. An extrapolated hippocampal volume based on body weight was not possible in this study population.

Tenocytes form a 3-D network and are connected via nanotubes.

Cells use different cell adhesion and communication structures to promote tissue development, maintenance of tissue integrity as well as repair and regenerative processes. Another recently discovered way of information exchange is long-distance thin cellular processes called nanotubes (NTs), mainly studied in vitro. Information on the existence and relevance of NTs in vivo is sparse. Building on two references which hint at the potential existence of longitudinally directed cell processes resembling NTs, we investigated tendons from young (3 weeks) and adult (9 weeks, 4 and 8 months) Fisher rats. Whole mounts of rat tail tendon fascicles (RTTfs) and sections of Achilles, flexor, extensor and patellar tendons were stained with Deep Red plasma membrane and DAPI nuclear stain and immunolabelled with Connexin43 (Cx43). In addition, 3-D reconstruction of serial semithin sections and TEM was used to verify the presence of NTs. We were able to demonstrate NTs as straight thin longitudinal processes (Ø 100-500 nm) reaching up to several 100 µm in length, mainly originating from lateral sheet-like cell processes or cell bodies in all tendon types investigated. NTs were observed to distend between tenocyte rows at the same level but also connect cells of different rows, thus leading to a complex 3-D cellular scaffold. Shorter NTs connected lateral cell sheets of tenocytes in the same row, omitting one or two cells. In addition, we detected links or potential branching of NTs. Cx43 immunostaining for the detection of gap junctions revealed Cx43-positive foci at the end-to-end contacts of tenocyte cell bodies as well as along their contacting sheet-like processes. Only rarely, we found clear Cx43 signals at their potential contact points between NTs and tendon cells as well as along the course of NTs, and most NTs appeared completely devoid of Cx43 signals. Therefore, we conclude that NTs in tendons could have a twofold function: long-distance communication as well as stabilization of a mechanically challenged tissue. From in vitro studies it is known that NTs allow intercellular transmission of various cell components, offering potential protective effects for the respective tissue. Further studies on functional properties of NTs in tendons under changing mechanical loading regimens are required in the future. The fact that NTs are present in tendons may necessitate the reconsideration of our traditional understanding of cell-to-cell communication.

Growth and cellular patterning during fetal human inner ear development studied by a correlative imaging approach.

BACKGROUND: Progressive transformation of the otic placode into the functional inner ear during gestational development in humans leads to the acquisition of hearing perception via the cochlea and balance and spatial orientation via the vestibular organ. RESULTS: Using a correlative approach involving micro-computerized tomography (micro-CT), transmission electron microscopy and histological techniques we were able to examine both the morphological and cellular changes associated with human inner ear development. Such an evaluation allowed for the examination of 3D geometry with high spatial and temporal resolution. In concert with gestational progression and growth of the cochlear duct, an increase in the distance between some of the Crista ampullaris is evident in all the specimens examined from GW12 to GW36. A parallel increase in the distances between the macular organs - fetal utricle and saccule - is also evident across the gestational stages examined. The distances between both the utricle and saccule to the three cristae ampullares also increased across the stages examined. A gradient in hair cell differentiation is apparent from apex to base of the fetal cochlea even at GW14. CONCLUSION: We present structural information on human inner ear development across multiple levels of biological organization, including gross-morphology of the inner ear, cellular and subcellular details of hearing and vestibular organs, as well as ultrastructural details in the developing sensory epithelia. This enabled the gathering of detailed information regarding morphometric changes as well in realizing the complex developmental patterns of the human inner ear. We were able to quantify the volumetric and linear aspects of selected gestational inner ear specimens enabling a better understanding of the cellular changes across the fetal gestational timeline. Moreover, these data could serve as a reference for better understanding disorders that arise during inner ear development.

Cranial kinesis in the miniaturised lizard Ablepharus kitaibelii (Squamata: Scincidae).

Cranial kinesis refers to intracranial movements in the vertebrate skull that do not concern the jaw joint, the middle ear or the hypobranchial skeleton. Different kinds of cranial kinesis have been reported for lizards, including mesokinesis, metakinesis, amphikinesis (simultaneous mesokinesis and metakinesis) and streptostyly. Streptostyly is considered relatively widespread within lizards, whereas mesokinesis has been documented only for geckos, varanids and anguids. The present study investigated cranial kinesis in the miniaturised scincid Ablepharus kitaibelii by integrating morphological and experimental data. Based on micro computed tomography, we provide a description of skull osteology. Cranial joints were studied with histology, which results in the first detailed description of cranial joint histology for a member of the Scincidae. Taken together, the morphological data indicate a high potential for amphikinesis and streptostyly, which was also corroborated by skull manipulations. High-speed cinematography demonstrated that mesokinesis occurs during food uptake, processing and intraoral transport cycles. Bite force measurements showed prolonged and reasonably hard biting even at large gape angles. Based on these data, we formulate a model of the amphikinetic A. kitaibelii skull mechanism, which provides an extension of Frazzetta's quadric-crank model by placing a special emphasis on metakinesis. According to this model, we hypothesise that metakinetic intracranial movements may provide a means for reducing strain in jaw adductor muscles. Presented hypotheses can be addressed and tested in future studies.

Surgical enlargement of the nasomaxillary aperture and transnasal conchotomy of the ventral conchal sinus: Two surgical techniques to improve sinus drainage in horses.

OBJECTIVE: To evaluate 2 surgical techniques for establishing and/or improving paranasal sinus drainage in cadaver heads and horses with sinusitis and evaluate the feasibility of postoperative transnasal sinus endoscopy. STUDY DESIGN: Ex vivo study (equine cadaver heads) and case series. SAMPLE POPULATION: Nine adult equine cadaver heads and 8 horses with recurrent sinusitis. METHODS: For the ex vivo study, the following procedures were performed on 9 cadaver heads: preoperative and postoperative computed tomography (heads 1-6), endoscopy-guided transnasal conchotomy of the ventral conchal sinus (TCVCS) and surgical enlargement of the nasomaxillary aperture (SENMAP) on opposite sides (heads 1-3), combined TCVCS and SENMAP on both sides (heads 4-9), evaluation of sinus drainage before and after surgery (heads 7-9), and postoperative transnasal endoscopy (heads 4-9). For the case series, 8 horses with secondary sinusitis were treated in standing position with SENMAP and/or TCVCS and postoperative transnasal endoscopy. RESULTS: Sinonasal communications were successfully created in all cadavers and affected live horses. Transnasal endoscopy of all sinuses except the middle conchal sinus was possible in heads 4-9 and in all clinical cases. Sinus drainage was improved (P = .028) by combining techniques. Blood loss in live horses ranged from 0.5-5.5 L (1.95 ± 1.5) per horse. Sinusitis resolved in all affected horses during follow-up of 3.2-25.5 months (13.5 ± 8.5). CONCLUSION: Transnasal conchotomy of the ventral conchal sinus and SENMAP consistently created large sinonasal communications, facilitating sinus endoscopy and improving sinus drainage. CLINICAL SIGNIFICANCE: Transnasal conchotomy of the ventral conchal sinus and SENMAP are viable options to treat horses with sinusitis and anatomical obstructions of the sinonasal communications.

Morphology of the bryozoan Cinctipora elegans (Cyclostomata, Cinctiporidae) with first data on its sexual reproduction and the cyclostome neuro-muscular system.

BACKGROUND: Cyclostome bryozoans are an ancient group of marine colonial suspension-feeders comprising approximately 700 extant species. Previous morphological studies are mainly restricted to skeletal characters whereas data on soft tissues obtained by state-of-the-art methods are still lacking. In order to contribute to issues related to cyclostome ground pattern reconstruction, we analyzed the morphology of the neuromuscular system Cinctipora elegans by means of immunocytochemical staining, confocal laser scanning microscopy, histological sections and microCT imaging. RESULTS: Polypides of C. elegans are located in elongated tubular skeletal cystids. Distally, the orifice leads into a prominent vestibulum which is lined by an epithelium that joins an almost complete perimetrical attachment organ, both containing radially arranged neurite bundles and muscles. Centrally, the prominent atrial sphincter separates the vestibulum from the atrium. The latter is enclosed by the tentacle sheath which contains few longitudinal muscle fibers and two principal neurite bundles. These emerge from the cerebral ganglion, which is located at the lophophoral base. Lateral ganglia are located next to the cerebral ganglion from which the visceral neurite bundles emerge that extend proximally towards the foregut. There are four tentacle neurite bundles that emerge from the ganglia and the circum-oral nerve ring, which encompasses the pharynx. The tentacles possess two striated longitudinal muscles. Short buccal dilatators are situated at the lophophoral base and short muscular sets are present at the abfrontal and frontal side of the tentacle base. The pharynx is myoepithelial and triradiate in cross-section. Oocytes are found inside the pharyngeal myoepithelium. The digestive tract contains dense circular musculature and few longitudinal muscles. The membranous sac contains regular, thin, circular and diagonal muscles and neurites in its epithelial lining. CONCLUSIONS: The general structure of the neuro-muscular system is more reminiscent of the condition found in Gymnolaemata rather than Phylactolaemata, which supports a close relationship between Cyclostomata and Gymnolaemata. Several characters of C. elegans such as the lateral ganglia or loss of the cardia are probably apomorphic for this species. For the first time, oocytes that surprisingly develop in the pharyngeal wall are reported for this species.

Application of a Parallelizable Perfusion Bioreactor for Physiologic 3D Cell Culture.

It is crucial but challenging to keep physiologic conditions during the cultivation of 3D cell scaffold constructs for the optimization of 3D cell culture processes. Therefore, we demonstrate the benefits of a recently developed miniaturized perfusion bioreactor together with a specialized incubator system that allows for the cultivation of multiple samples while screening different conditions. Hence, a decellularized bone matrix was tested towards its suitability for 3D osteogenic differentiation under flow perfusion conditions. Subsequently, physiologic shear stress and hydrostatic pressure (HP) conditions were optimized for osteogenic differentiation of human mesenchymal stem cells (MSCs). X-ray computed microtomography and scanning electron microscopy (SEM) revealed a closed cell layer covering the entire matrix. Osteogenic differentiation assessed by alkaline phosphatase activity and SEM was found to be increased in all dynamic conditions. Furthermore, screening of different fluid shear stress (FSS) conditions revealed 1.5 mL/min (equivalent to ∼10 mPa shear stress) to be optimal. However, no distinct effect of HP compared to flow perfusion without HP on osteogenic differentiation was observed. Notably, throughout all experiments, cells cultivated under FSS or HP conditions displayed increased osteogenic differentiation, which underlines the importance of physiologic conditions. In conclusion, the bioreactor system was used for biomaterial testing and to develop and optimize a 3D cell culture process for the osteogenic differentiation of MSCs. Due to its versatility and higher throughput efficiency, we hypothesize that this bioreactor/incubator system will advance the development and optimization of a variety of 3D cell culture processes.

Musculoskeletal architecture of the prey capture apparatus in salamandrid newts with multiphasic lifestyle: does anatomy change during the seasonal habitat switches?

Some newt species change seasonally between an aquatic and a terrestrial life as adults, and are therefore repeatedly faced with different physical circumstances that affect a wide range of functions of the organism. For example, it has been observed that seasonally habitat-changing newts display notable changes in skin texture and tail fin anatomy, allowing one to distinguish an aquatic and a terrestrial morphotype. One of the main functional challenges is the switch between efficient aquatic and terrestrial prey capture modes. Recent studies have shown that newts adapt quickly by showing a high degree of behavioral flexibility, using suction feeding in their aquatic stage and tongue prehension in their terrestrial stage. As suction feeding and tongue prehension place different functional demands on the prey capture apparatus, this behavioral flexibility may clearly benefit from an associated morphological plasticity. In this study, we provide a detailed morphological analysis of the musculoskeletal system of the prey capture apparatus in the two multiphasic newt species Ichthyosaura alpestris and Lissotriton vulgaris by using histological sections and micro-computed tomography. We then test for quantitative changes of the hyobranchial musculoskeletal system between aquatic and terrestrial morphotypes, The descriptive morphology of the cranio-cervical musculoskeletal system provides new insights on form and function of the prey capture apparatus in newts, and the quantitative approach shows hypertrophy of the hyolingual musculoskeletal system in the terrestrial morphotype of L. vulgaris but hypertrophy in the aquatic morphotype of I. alpestris. It was therefore concluded that the seasonal habitat shifts are accompanied by a species-dependent muscular plasticity of which the potential effect on multiphasic feeding performance in newts remains unclear.

Past climate change on Sky Islands drives novelty in a core developmental gene network and its phenotype.

BACKGROUND: A fundamental and enduring problem in evolutionary biology is to understand how populations differentiate in the wild, yet little is known about what role organismal development plays in this process. Organismal development integrates environmental inputs with the action of gene regulatory networks to generate the phenotype. Core developmental gene networks have been highly conserved for millions of years across all animals, and therefore, organismal development may bias variation available for selection to work on. Biased variation may facilitate repeatable phenotypic responses when exposed to similar environmental inputs and ecological changes. To gain a more complete understanding of population differentiation in the wild, we integrated evolutionary developmental biology with population genetics, morphology, paleoecology and ecology. This integration was made possible by studying how populations of the ant species Monomorium emersoni respond to climatic and ecological changes across five 'Sky Islands' in Arizona, which are mountain ranges separated by vast 'seas' of desert. Sky Islands represent a replicated natural experiment allowing us to determine how repeatable is the response of M. emersoni populations to climate and ecological changes at the phenotypic, developmental, and gene network levels. RESULTS: We show that a core developmental gene network and its phenotype has kept pace with ecological and climate change on each Sky Island over the last ~90,000 years before present (BP). This response has produced two types of evolutionary change within an ant species: one type is unpredictable and contingent on the pattern of isolation of Sky lsland populations by climate warming, resulting in slight changes in gene expression, organ growth, and morphology. The other type is predictable and deterministic, resulting in the repeated evolution of a novel wingless queen phenotype and its underlying gene network in response to habitat changes induced by climate warming. CONCLUSION: Our findings reveal dynamics of developmental gene network evolution in wild populations. This holds important implications: (1) for understanding how phenotypic novelty is generated in the wild; (2) for providing a possible bridge between micro- and macroevolution; and (3) for understanding how development mediates the response of organisms to past, and potentially, future climate change.