OR15-5 Human Sex Determination at the Edge of Ambiguity: Impaired SRY Phosphorylation Attenuates Expression of the Male Program

A paradox is posed by metazoan gene-regulatory networks (GRNs) that are robust yet evolvable. Insight may be obtained through studies of bistable genetic circuits mediating developmental decisions. A model in organogenesis is provided by the sex-specific differentiation of the embryonic gonadal ridge to form a testis or ovary. Here, we investigated a Swyer mutation in human testis-determining factor SRY that impairs its phosphorylation in association with variable developmental outcomes: fertile male, intersex, or infertile female (46, XY pure gonadal dysgenesis). The mutation (R30I) abrogates serine phosphorylation within a putative target site for protein kinase A (PKA) N-terminal to the HMG box. Diverse processes can be regulated by protein phosphorylation, including DNA recognition by transcription factors (TFs). Phosphorylation of this site in human SRY (LRRSSSFLCT; italics) in vitro was previously shown to enhance specific DNA affinity. Biological consequences of the mutation were evaluated in SRY-responsive mammalian cell lines following transient transfection. The mutation attenuated in concert occupancy of a target enhancer (TESCO) and SOX9 transcriptional activation. These perturbations were mitigated by acidic substitution (LRIDDDFL) whereas Ala substitutions (RRAAAFL or RIAAAFL) attenuated activity to an extent similar to R30I alone. No differences were observed in nuclear localization. Mutagenesis suggested that the central Ser is most efficiently phosphorylated in accord with PKA targeting rules. Replacement of the native site by an optimized “Kemptide” PKA site (LRRASLGCT) enhanced both SRY phosphorylation and SOX9 transcriptional activation whereas a “swapped” protein-kinase C determinant (LRRSSFRRCT) blocked phosphorylation. Among SRY variants, extent of cellular phosphorylation mirrored relative in vitro efficiencies of synthetic SRY-derived peptides as PKA-specific substrates. Although several kinases are predicted in silico to target this tri-serine motif, cell-based studies implicate PKA as the relevant kinase in vivo. Our results provide evidence that primate Sry requires its phosphorylation for full gene-regulatory activity. A PKA site N-terminal to the SRY HMG box, unique to primates, exemplifies network “tinkering” through recruitment of a new regulatory linkage. Molecular characterization of the R30I inherited Swyer mutation in SRY thus demonstrates that impaired TF phosphorylation can attenuate a human developmental switch at the edge of ambiguity

Discrimination of human and dog faces and inversion responses in domestic dogs (Canis familiaris)

Although domestic dogs can respond to many facial cues displayed by other dogs and humans, it remains unclear whether they can differentiate individual dogs or humans based on facial cues alone and, if so, whether they would demonstrate the face inversion effect, a behavioural hallmark commonly used in primates to differentiate face processing from object processing. In this study we first established the applicability of the Visual Paired Comparison (VPC or preferential looking) procedure for dogs using a simple object discrimination task with 2D pictures. The animals demonstrated a clear looking preference for novel objects when simultaneously presented with prior-exposed familiar objects. We then adopted this VPC procedure to assess their face discrimination and inversion responses. Dogs showed a deviation from random behaviour, indicating discrimination capability when inspecting upright dog faces, human faces and object images; but the pattern of viewing preference was dependent upon image category. They directed longer viewing time at novel (vs. familiar) human faces and objects, but not at dog faces, instead, a longer viewing time at familiar (vs. novel) dog faces was observed. No significant looking preference was detected for inverted images regardless of image category. Our results indicate that domestic dogs can use facial cues alone to differentiate individual dogs and humans, and that they exhibit a non-specific inversion response. In addition, the discrimination response by dogs of human and dog faces appears to differ with the type of face involved

DCU search runs at MediaEval 2014 search and hyperlinking

We described Dublin City University (DCU)'s participation in the Search sub-task of the Search and Hyperlinking Task at MediaEval 2014. Exploratory experiments were carried out to investigate the utility of prosodic prominence features in the task of retrieving relevant video segments from a collection of BBC videos. Normalised acoustic correlates of loudness, pitch, and duration were incorporated in a standard TF-IDF weighting scheme to increase weights for terms that were prominent in speech. Prosodic models outperformed a text-based TF-IDF baseline on the training set but failed to surpass the baseline on the test set

DCU at the NTCIR-12 SpokenQuery&Doc-2 task

We describe DCU’s participation in the NTCIR-12 SpokenQuery&Doc (SQD-2) task. In the context of the slide-group retrieval sub-task, we experiment with a passage retrieval method that re-scores each passage according to the relevance score of the document from which the passage is taken. This is performed by linearly interpolating their relevance scores which are calculated using the Okapi BM25 model of probabilistic retrieval for passages and documents independently. In conjunction with this, we assess the benefits of using pseudo-relevance feedback for expanding the textual representation of the spoken queries with terms found in the top-ranked documents and passages, and experiment with a general multidimensional optimisation method to jointly tune the BM25 and query expansion parameters with queries and relevance data from the NTCIR-11 SQD-1 task. Retrieval experiments performed over the SQD-1 and SQD-2 queries confirm previous findings which affirm that integrating document information when ranking passages can lead to improved passage retrieval effectiveness. Furthermore, results indicate that no significant gains in retrieval effectiveness can be obtained by using query expansion in combination with our retrieval models over these two query sets

DCU at the NTCIR-11 SpokenQuery&Doc task

We describe DCU's participation in the NTCIR-11 Spoken-Query&Document task. We participated in the spoken query spoken content retrieval (SQ-SCR) subtask by using the slide group segments as basic indexing and retrieval units. Our approach integrates normalised prosodic features into a standard BM25 weighting function to increase weights for terms that are prominent in speech. Text queries and relevance assessment data from the NTCIR-10 SpokenDoc-2 passage retrieval task were used to train the prosodic-based models. Evaluation results indicate that our prosodic-based retrieval models do not provide significant improvements over a text-based BM25 model, but suggest that they can be useful for certain queries

Expression of the gene encoding secretor type galactoside 2 ? fucosyltransferase (FUT2) and ABH antigens in patients with oral lesions

Objective: The aim of this work was to evaluate the expression of FUT2 gene in saliva and histo ABH antigens of patients with oral lesions. Study Design: In total 178 subjects were examined, half of whom suffered from oral pre-cancerous and cancerous lesions, while the other half were the healthy control group We analyzed the FUT 2 polymorphism by ASO-PCR (allele specific oligonucleotid - polymerase chain reaction) with specific primers for G428 allele and the wild type allele of FUT2 gene. To reveal A, B and H antigens in tissue sections of the patients (n= 89) we used a modified specific red cell adherence technique. Results: We found a high intensity of oral disease in the non-secretor group (OR = 2.43). A total of 58% of the patients with oral pre-cancerous and cancerous lesions was non secretors (se_/_), in contrast with the healthy population (21.5%). A strongly positive reaction was defined as a sheet of indicator erythrocytes adhered to the epithelial cells. In 31 of the 54 samples analyzed the test showed slightly positive results on atypical areas, and there was a complete antigen deletion in areas affected by neoplasia. Nineteen samples showed a total absence of ABH antigens in both histologically normal and pathological areas. Blood group antigens were expressed at a high level in benign and highly differentiated malignant tumors. In poorly differentiated malignant tumors, they were mostly absent. Conclusion: Considering these results we suggest the use of this method to monitor probable preneoplastic lesions in risk population, especially in those with no secretor status (absence of FUT2 gene). © Medicina Oral S. L

A 2MASS Analysis of the Stability of Southern Bok Globules

We used near-infrared 2MASS data to construct visual extinction maps of a sample of Southern Bok globules utilizing the NICE method. We derived radial extinction profiles of dense cores identified in the globules and analyzed their stability against gravitational collapse with isothermal Bonnor-Ebert spheres. The frequency distribution of the stability parameter xi_max of these cores shows that a large number of them are located in stable states, followed by an abrupt decrease of cores in unstable states. This decrease is steeper for globules with associated IRAS point sources than for starless globules. Moreover, globules in stable states have a Bonnor-Ebert temperature of T = 15 +- 6 K, while the group of critical plus unstable globules has a different temperature of T = 10 +- 3 K. Distances were estimated to all the globules studied in this work and the spectral class of the IRAS sources was calculated. No variations were found in the stability parameters of the cores and the spectral class of their associated IRAS sources. On the basis of 13CO J = 1-0 molecular line observations, we identified and modeled a blue-assymetric line profile toward a globule of the sample, obtaining an upper limit infall speed of 0.25 km/s.Comment: 53 pages, 15 figures, accepted for publication in Ap

Monitoring and impact mitigation during a 4D seismic survey near a population of gray whales off Sakhalin Island, Russia

A 4D seismic survey was conducted in 2010 near the feeding grounds of gray whales off Sakhalin Island, Russia. To minimize disruptions to the whales’ feeding activity and enhance understanding of the potential impacts of seismic surveys on gray whales Eschrichtius robustus, an extensive monitoring and mitigation plan (MMP) was developed. Typically, mitigation plans involve observers on seismic vessels to monitor for the presence of marine mammals in an exclusion zone so as to prevent physical injury to the animals. Due to the protected status of western gray whales, an additional protection zone based on a behavioural disturbance threshold of exposure of 156 dB re μPa2-s per pulse was applied for whales within their feeding habitat defined by the estimated 95% abundance contour. Real-time radio-transmitting acoustic recorders were deployed along this contour to verify modelled acoustic footprints within the feeding grounds. Shore- and vessel-based observation teams monitored for the presence and activity of whales. A real-time GIS workflow tracking procedure was developed that integrated acoustic and whale positioning data to determine if sound levels at a whales’ position within the feeding area exceeded the behavioural threshold, in which case a shut-down of the seismic source was implemented. Additionally, behaviour and distribution surveys were conducted before, during and after the seismic survey to evaluate the effectiveness of the MMP. No large changes in whale movement, respiration, or distribution patterns were observed during the seismic survey. This could be interpreted to mean that the MMP was effective in reducing the sound exposure and behavioural responses of gray whales to seismic sounds. KEY WORDS: Seismic · Mitigation · Western gray whale · Eschrichtius robustus · Sakhalin · Piltun · Sea of Okhotsk · Anthropogenic disturbanc

NMDA receptor content of synapses in stratum radiatum of the hippocampal CA1 area

Glutamate receptors activated by NMDA (NMDARs) or AMPA (AMPARs) are clustered on dendritic spines of pyramidal cells. Both the AMPAR-mediated postsynaptic responses and the synaptic AMPAR immunoreactivity show a large intersynapse variability. Postsynaptic responses mediated by NMDARs show less variability. To assess the variability in NMDAR content and the extent of their coexistence with AMPARs in Schaffer collateral-commissural synapses of adult rat CA1 pyramidal cells, electron microscopic immunogold localization of receptors has been used. Immunoreactivity of NMDARs was detected in virtually all synapses on spines, but AMPARs were undetectable, on average, in 12% of synapses. A proportion of synapses had a very high AMPAR content relative to the mean content, resulting in a distribution more skewed toward larger values than that of NMDARs. The variability of synaptic NMDAR content [coefficient of variation (CV), 0.64-0.70] was much lower than that of the AMPAR content (CV, 1.17-1.45). Unlike the AMPAR content, the NMDAR content showed only a weak correlation with synapse size. As reported previously for AMPARs, the immunoreactivity of NMDARs was also associated with the spine apparatus within spines. The results demonstrate that the majority of the synapses made by CA3 pyramidal cells onto spines of CA1 pyramids express both NMDARs and AMPARs, but with variable ratios. A less-variable NMDAR content is accompanied by a wide variability of AMPAR content, indicating that the regulation of expression of the two receptors is not closely linked. These findings support reports that fast excitatory transmission at some of these synapses is mediated by activation mainly of NMDARs