Plasma membrane proteome of adhesion‐competent endometrial epithelial cells and its modulation by Rab11a

Human endometrial epithelium (EE) is composed of a multitude of proteins, amongst which those localized on the plasma membrane [plasma membrane proteins (PMPs)] are of critical relevance in the early stages of implantation. Evidence supports the key role of few PMPs in implantation. However, many remain unidentified, as efforts have not been made till date to generate the plasma membrane proteome of human EE cells, using a gel‐free approach. This study presents a protein catalog of the PMP enriched fraction of Ishikawa cell line; often used as an in vitro model for embryo‐adhesive EE. Liquid chromatography with tandem mass spectrometry identified 3,598 proteins. Of these, 1,963 proteins were annotated for their membrane localization. Of 1,963 proteins, 1,321 were found to have a transmembrane domain and 43 proteins had glycophosphatidylinositol (GPI) anchor. Extensive data mining revealed endometrial expression of 943 proteins reported in humans and/or rodents. Further, quantitative alterations were observed in the plasma membrane proteome on the perturbation of intracellular trafficking. Silencing of Rab11a (known for its role in plasma membrane organization) expression caused alteration in the abundance of 74 proteins. Caveolin‐1 and EpCAM levels were reduced whereas Rab4a abundance increased in the PMP extracts of Rab11a deficient cells, compared with control cells. Briefly, the study reports the identity of several novel plasma membrane‐localized proteins. A major spin‐off of the study is the identification of novel proteins trafficked by Rab11a to the plasma membrane. Targeted analysis of novel PMPs may reveal their specific roles in endometrial receptivity and implantation.     

Molecular and morphological boundaries of the predatory mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae)

Neoseiulus californicus (McGregor) is a natural enemy of pest mites used worldwide in many crops. Its correct identification is thus essential to ensure biological control success. The present study aimed to characterize molecular and morphological intraspecific variations for assisting in the diagnosis of the species and to build baseline information about expected variations within a commercially important phytoseiid species. Morphological and molecular [12S rRNA, cytochrome b mitochondrial (mt)DNA, and internal transcribed spacer] analyses were carried out on fourteen populations collected worldwide and on one mass‐reared strain. The genetic distances between the specimens of N. californicus and another related species were high and no overlap was observed, sustaining the reliability of such molecular methods for assisting a specific diagnosis. Furthermore, the genetic distances between populations of N. californicus were very low and overlap between intra‐ and interpopulations distances was observed, except for two populations collected in France (Marsillargues and Midi‐Pyrénées). The high mitochondrial differentiation between these two latter populations and the others questions their specific status: do they belong to the species N. californicus or to another cryptic species? However, using nuclear DNA marker analyses, no distinct differences were observed. Furthermore, even if significant morphological differences were observed between the populations, these differences were very small and the standard errors within each population were very low. We thus concluded that all the populations studied belong to the species N. californicus, despite unexpected high mitochondrial variations. The present study thus displays the importance of an integrative taxonomic approach for avoiding misidentifications. A discussion on morphological and mtDNA variations in relation to diagnostic reliability is developped. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 393–406.     

In-Vivo Efficacy of Compliant 3D Nano-Composite in Critical-Size Bone Defect Repair: a Six Month Preclinical Study in Rabbit

Bone defects above critical size do not heal completely by itself and thus represent major clinical challenge to reconstructive surgery. Numerous bone substitutes have already been used to promote bone regeneration, however their use, particularly for critical-sized bone defects along with their long term in vivo safety and efficacy remains a concern. The present study was designed to obtain a complete healing of critical-size defect made in the proximal tibia of New Zealand White rabbit, using nano-hydroxyapatite/gelatin and chemically carboxymethylated chitin (n-HA/gel/CMC) scaffold construct. The bone-implant interfaces and defect site healing was evaluated for a period up to 25 weeks using radiography, micro-computed tomography, fluorescence labeling, and histology and compared with respective SHAM (empty contra lateral control). The viscoelastic porous scaffold construct allows easy surgical insertion and post-operatively facilitate oxygenation and angiogenesis. Radiography of defect treated with scaffold construct suggested expedited healing at defect edges and within the defect site, unlike confined healing at edges of the SHAM sites. The architecture indices analyzed by micro-computed tomography showed a significant increase in percentage of bone volume fraction, resulted in reconciled cortico-trabecular bone formation at n-HA/gel/CMC constructs treated site (15.2% to 52.7%) when compared with respective SHAM (10.2% to 31.8%). Histological examination and fluorescence labeling revealed that the uniformly interconnected porous surface of scaffold construct enhanced osteoblasts’ activity and mineralization. These preclinical data suggest that, n-HA/gel/CMC construct exhibit stimulation of bone''s innate regenerative capacity, thus underscoring their use in guided bone regeneration.     

Hip extension, knee flexion paradox: a new mechanism for non-contact ACL injury

Considering that an athlete performs at-risk sports activities countless times throughout the course of his or her career prior to the instance of anterior cruciate ligament (ACL) injury, one may conclude that non-contact ACL injury is a rare event. Nevertheless, the overall number of non-contact ACL injuries, both in the US and worldwide, remains alarming due to the growing number of recreational and professional athletes participating in high-risk activities. To date, numerous non-contact ACL injury mechanisms have been proposed, but none provides a detailed picture of sequence of events leading to injury and the exact cause of this injury remains elusive. In this perspective article, we propose a new conception of non-contact ACL injury mechanism that comprehensively integrates risk factors inside and outside the knee joint. The proposed mechanism is robust in the sense that it is biomechanically justifiable and addresses a number of confounding issues related to ACL injury.     

Many-body energies during proton transfer in an aqueous system

The energetics of the mechanism of proton transfer from a hydronium ion to one of the water molecules in its first solvation shell are studied using density functional theory and the Møller–Plesset perturbation (MP2) method. The potential energy surface of the proton transfer mechanism is obtained at the B3LYP and MP2 levels with the 6-311++G** basis set. Many-body analysis is applied to the proton transfer mechanism to obtain the change in relaxation energy, two-body, three-body and four-body energies when proton transfer occurs from the hydronium ion to one of the water molecules in its first solvation shell. It is observed that the binding energy (BE) of the complex decreases during the proton transfer process at both levels of theory. During the proton transfer process, the % contribution of the total two-body energy to the binding energy of the complex increases from 62.9 to 68.09% (39.9 to 45.95%), and that of the total three-body increases from 25.9 to 27.09% (24.16 to 26.17%) at the B3LYP/6-311++G** (MP2/ 6-311++G**) level. There is almost no change in the water–water–water three-body interaction energy during the proton transfer process at both levels of theory. The contribution of the relaxation energy and the total four-body energy to the binding energy of the complex is greater at the MP2 level than at the B3LYP level. Significant differences are found between the relaxation energies, the hydronium–water interaction energies and the four-body interaction energies at the B3LYP and MP2 levels.     

Adenylyl cyclase expression and modulation of cAMP in rat taste cells

The present study determined Ca²⁺ handling in the hearts of rats subjected to chronic hypoxia (CH). Spectrofluorometry was used to measure intracellular Ca²⁺ concentration ([Ca²⁺]_i) and its responses to electrical stimulation, caffeine, and isoproterenol in myocytes from the right ventricle of rats breathing 10% oxygen for 1, 3, 7, 14, 21, 28, and 56 days and age-matched controls. The protein expression of sarco(endo)plasmic reticulum Ca²⁺-ATPase (SERCA) and its ryanodine receptor (RyR) were measured. The uptake of ⁴⁵Ca²⁺ by SERCA, release by RyR, and extrusion by Na⁺/Ca²⁺ exchange (NCX) were determined. It was found that Ca²⁺ homeostasis and Ca²⁺ responses to -adrenoceptor stimulation reached a new equilibrium after 4 wk of CH. Ca²⁺ content in the sarcoplasmic reticulum (SR) was reduced, but cytosolic Ca²⁺ remained unchanged after CH. Expression of SERCA and its Ca²⁺ uptake, Ca²⁺ release via RyR, and NCX activity were suppressed by CH. The results indicate impaired Ca²⁺ handling, which may be responsible for the attenuated Ca²⁺ responses to -adrenoceptor stimulation in CH. intracellular calcium ion concentration; calcium-adenosinetriphosphatase; ryanodine receptor; sodium/calcium exchange; sarcoplasmic reticulum; -adrenoceptor; chronic hypoxia     

Molecular phylogeny in endemic weevils: revision of the genera of Macaronesian Cryptorhynchinae (Coleoptera: Curculionidae)

A molecular phylogeny and lineage age estimates are presented for the Macaronesian representatives of the weevil subfamily Cryptorhynchinae, using two mitochondrial genes (cytochrome c oxidase subunit 1 and 16S). The Bayesian reconstruction is supplemented by observations on morphology, ecology, and reproductive biology. The present study often corroborates the groups previously outlined in higher‐level informal taxonomies. These and further groups are now assigned new taxonomic status. The following genera and subgenera are described (formerly Acalles): Aeoniacalles gen. nov. , Canariacalles gen. nov. , Ficusacalles gen. nov. , Madeiracalles gen. nov. , Silvacalles gen. nov. (with Tolpiacalles subgen. nov. , Tagasastacalles subgen. nov. ), Sonchiacalles gen. nov. , Echiumacalles gen. nov. (monotypic), Lauriacalles gen. nov. (monotypic), and Pseudodichromacalles gen. nov. (monotypic; formerly Dichromacalles). For the western Palaearctic genus Acalles Schoenherr, 1825 the first subgenus Origoacalles subgen. nov. is described and for the genus Onyxacalles Stüben, 1999 the first subgenus Araneacalles subgen. nov. ; Paratorneuma Roudier 1956 resyn. Except for one species of Acalles (Origoacalles), all of these new higher taxa are endemic to the Macaronesian Islands. All new taxa are presented, together with their host plants and further data, in a synoptic tabular overview. Based on the results of our phylogenetic analysis, we advocate the hypothesis that the evolution of the species in the new genera (of which most group into a ‘Macaronesian clade’) began in the comparatively arid succulent bush zone and that the shady and humid laurel forest of the thermo‐Canarian and thermo‐Madeiran zone was entered much later. Our reconstruction implies that the Canarian and Madeiran archipelagos were colonized by Cryptorhynchinae at least seven times from the continent but saw only one considerable adaptive radiation. It also becomes apparent that it is the ancestor species of the genus Canariacalles– and not Pseudodichromacalles– that features a close connection to the south‐western European and north‐western African species of Dichromacalles s.s. Finally, a key is presented for all genera and subgenera of the Macaronesian Cryptorhynchinae. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 160 , 40–87.     

The Onychophora of Trinidad, Tobago and the Lesser Antilles

This paper brings together the scattered literature on the Onychophora from Trinidad, Tobago and the Lesser Antilles. The 11 known species from this area are described, incorporating new material collected by the author. Scanning electron micrographs of the dorsal integument are presented for all but one species. Parthenogenesis is reported in a population of Epiperipatus imthurni from Trinidad.