RhoGAP18B Isoforms Act on Distinct Rho-Family GTPases and Regulate Behavioral Responses to Alcohol via Cofilin

Responses to the effects of ethanol are highly conserved across organisms, with reduced responses to the sedating effects of ethanol being predictive of increased risk for human alcohol dependence. Previously, we described that regulators of actin dynamics, such as the Rho-family GTPases Rac1, Rho1, and Cdc42, alter Drosophila’s sensitivity to ethanol-induced sedation. The GTPase activating protein RhoGAP18B also affects sensitivity to ethanol. To better understand how different RhoGAP18B isoforms affect ethanol sedation, we examined them for their effects on cell shape, GTP-loading of Rho-family GTPase, activation of the actin-severing cofilin, and actin filamentation. Our results suggest that the RhoGAP18B-PA isoform acts on Cdc42, while PC and PD act via Rac1 and Rho1 to activate cofilin. In vivo, a loss-of-function mutation in the cofilin-encoding gene twinstar leads to reduced ethanol-sensitivity and acts in concert with RhoGAP18B. Different RhoGAP18B isoforms, therefore, act on distinct subsets of Rho-family GTPases to modulate cofilin activity, actin dynamics, and ethanol-induced behaviors.     

Latency of auditory P300 response is related with cognitive deficits in Obstructive Sleep Apnea Syndrome

Sleep and Biological Rhythms - Obstructive Sleep Apnea Syndrome (OSAS) is characterized by desaturation in blood oxygen level and sleep fragmentation because of repeated upper airway obstruction....     

A molecular phylogeny of Basidiophora reveals several apparently host-specific lineages on Astereae

The genus Basidiophora has long been thought to contain only two species, Basidiophora entospora and Basidiophora kellermanii, the latter of which was transferred to a newly described monotypic genus, Benua, at the end of the twentieth century, leaving Basidiophora monotypic, despite its vast host range, including a member of the Eupatoriae and several genera in the subfamily Asteroideae of the Asteraceae. Using historic herbarium specimens, we demonstrate that while Benua kellermanii is genetically highly homogenous, at least seven distinct phylogenetic lineages exist within Basidiophora, which, based on sequence divergence, most likely constitute hitherto overlooked cryptic species. As the specimens from Symphyotrichum novae-angliae formed a well-supported clade with little variation, we consider Peronospora simplex described on this host as an independent species, which is transferred to the genus Basidiophora in this study. The phylogeny of the pathogens corresponds well to the phylogeny of the respective hosts, which is unusual in downy mildews and might hint at clade-limited colonisation and subsequent radiation to closely related hosts of Astereae or even suggest a co-evolution scenario. Our findings provide further evidence that species with assumed broad host ranges should be thoroughly evaluated with respect to their phylogenetic relationships, especially in biotrophic genera with only limited morphological diversity. In some cases, host specificity of genetically divergent lineages might be the only phenotypic trait remaining for species delimitation. Future detailed morphological comparisons are needed to reveal if the seemingly cryptic species of Basidiophora can be distinguished based on subtle morphological characteristics.     

An Analysis of the Proportion of the U.S. Population that Ingests Soil or Other Non-Food Substances

Reliable quantitative data are lacking that document the prevalence of ingestion of soil and other “non-food” substances among U.S. children and adults. This article explores the proportion of the U.S. population that ingests substances such as soil, clay, starch, paint, or plaster. We compiled data from the National Health and Nutrition Examination Survey (NHANES) collected from years 1971–1975 (NHANES I) and 1976–1980 (NHANES II) because these particular surveys asked participants specific questions about non-food ingestion practices. We examined the prevalence of the behavior across multiple demographic variables, such as age, gender, education, and income level. Approximately 1% (NHANES II) to 2.5% (NHANES I) of the U.S. population ingests some type of non-food substance. The most notable variation across the demographic subgroups studied was the difference in estimated prevalence among young children (1 to <3 years) compared to older children and adults. Estimated prevalence was also higher among blacks compared to whites and within lower compared to higher socioeconomic groups. This analysis helps fill data gaps on the relative pattern of non-food ingestion practices on a national scale. This information provides perspective for risk assessors when evaluating exposure variables and for risk managers when weighing risk management alternatives.     

mTOR controls ovarian follicle growth by regulating granulosa cell proliferation

We have shown that inhibition of mTOR in granulosa cells and ovarian follicles results in compromised granulosa proliferation and reduced follicle growth. Further analysis here using spontaneously immortalized rat granulosa cells has revealed that mTOR pathway activity is enhanced during M-phase of the cell cycle. mTOR specific phosphorylation of p70S6 kinase and 4E-BP, and expression of Raptor are all enhanced during M-phase. The predominant effect of mTOR inhibition by the specific inhibitor Rapamycin (RAP) was a dose-responsive arrest in the G1 cell cycle stage. The fraction of granulosa cells that continued to divide in the presence of RAP exhibited a dose-dependent increase in aberrant mitotic figures known as anaphase bridges. Strikingly, estradiol consistently decreased the incidence of aberrant mitotic figures. In mice treated with RAP, the mitotic index was reduced compared to controls, and a similar increase in aberrant mitotic events was noted. RAP injected during a superovulation regime resulted in a dose-dependent reduction in the numbers of eggs ovulated. Implications for the real-time regulation of follicle growth and dominance, including the consequences of increased numbers of aneuploid granulosa cells, are discussed.     

Prostaglandin E2 enhances human cord blood stem cell xenotransplants and shows long-term safety in preclinical nonhuman primate transplant models

Hematopoietic stem cells (HSCs) are used in transplantation therapy to reconstitute the hematopoietic system. Human cord blood (hCB) transplantation has emerged as an attractive alternative treatment option when traditional HSC sources are unavailable; however, the absolute number of hCB HSCs transplanted is significantly lower than bone marrow or mobilized peripheral blood stem cells (MPBSCs). We previously demonstrated that dimethyl-prostaglandin E2 (dmPGE2) increased HSCs in vertebrate models. Here, we describe preclinical analyses of the therapeutic potential of dmPGE2 treatment by using human and nonhuman primate HSCs. dmPGE2 significantly increased total human hematopoietic colony formation in?vitro and enhanced engraftment of unfractionated and CD34(+) hCB after xenotransplantation. In nonhuman primate autologous transplantation, dmPGE2-treated CD34(+) MPBSCs showed stable multilineage engraftment over 1 year postinfusion. Together, our analyses indicated that dmPGE2 mediates conserved responses in HSCs from human and nonhuman primates and provided sufficient preclinical information to support proceeding to an FDA-approved phase 1 clinical trial.     

A novel sorting sequence in the beta2-adrenergic receptor switches recycling from default to the Hrs-dependent mechanism

Plasma membrane recycling of G protein-coupled receptors can occur by at least two distinct mechanisms as follows: a "default" mechanism that occurs nonselectively, and a specifically sorted mechanism that requires the endosome-associated protein Hrs. In this study we have defined a sequence in the beta2-adrenergic receptor cytoplasmic tail that confers Hrs dependence on receptor recycling. This sequence resembles acidic dileucine class motifs found in other membrane proteins but is structurally and functionally distinct from previously identified sorting sequences. Mutation of the novel sorting sequence rendered plasma membrane recycling independent of Hrs and independent of a distal PDZ ligand required for Hrs-dependent recycling. We propose that the novel sorting sequence functions to "switch" endocytic trafficking between mechanistically distinct recycling modes, thereby explaining failure of the wild type beta2-adrenergic receptor to recycle efficiently by default.     

Spatially Restricted G Protein-coupled Receptor Activity via Divergent Endocytic Compartments

Postendocytic sorting of G protein-coupled receptors (GPCRs) is driven by their interactions between highly diverse receptor sequence motifs with their interacting proteins, such as postsynaptic density protein (PSD95), Drosophila disc large tumor suppressor (Dlg1), zonula occludens-1 protein (zo-1) (PDZ) domain proteins. However, whether these diverse interactions provide an underlying functional specificity, in addition to driving sorting, is unknown. Here we identify GPCRs that recycle via distinct PDZ ligand/PDZ protein pairs that exploit their recycling machinery primarily for targeted endosomal localization and signaling specificity. The luteinizing hormone receptor (LHR) and β2-adrenergic receptor (B2AR), two GPCRs sorted to the regulated recycling pathway, underwent divergent trafficking to distinct endosomal compartments. Unlike B2AR, which traffics to early endosomes (EE), LHR internalizes to distinct pre-early endosomes (pre-EEs) for its recycling. Pre-EE localization required interactions of the LHR C-terminal tail with the PDZ protein GAIP-interacting protein C terminus, inhibiting its traffic to EEs. Rerouting the LHR to EEs, or EE-localized GPCRs to pre-EEs, spatially reprograms MAPK signaling. Furthermore, LHR-mediated activation of MAPK signaling requires internalization and is maintained upon loss of the EE compartment. We propose that combinatorial specificity between GPCR sorting sequences and interacting proteins dictates an unprecedented spatiotemporal control in GPCR signal activity.     

Toxic Metals in Breast Milk Samples from Ankara, Turkey: Assessment of Lead, Cadmium, Nickel, and Arsenic Levels

Toxic metals are one of the significant groups of chemical contaminants that humans are exposed to by oral, inhalation, and dermal routes. Exposure to these chemicals begins with intrauterine life and continues during lactation period at the first years of life. Breastfeeding has a much more special place than other nutrition options for infants. However, when possibility of contaminant transfer by breast milk is considered, its safety and quality is essential. Regarding infant and mother health and limited number of information on this field in Turkey, measuring contamination levels in breast milk is important. Therefore, in the present study, lead (Pb), cadmium (Cd), nickel (Ni), and arsenic (As) levels were measured by atomic absorption spectrometry in 64 breast milk samples obtained from mothers from Ankara, Turkey. Pb and Ni levels in breast milk samples were found to be 391.45?±?269.01?μg/l and 43.94?±?33.82?μg/l (mean ± SD), respectively. Cd was found only in one of 64 samples, and the level was 4.62?μg/l. As level was below the limit of quantification (LOQ, 7.6?μg/l) in all samples. These findings will accurately direct strategies and solutions of protection against contaminants in order to reduce their levels in biological fluids.