Phosphorylation of rat spermatidal protein TP2 by sperm-specific protein kinase A and modulation of its transport into the haploid nucleus

Transition protein 2 (TP2), which is expressed during stages 12-15 of mammalian spermiogenesis, has been shown to undergo phosphorylation immediately after its synthesis. We reported earlier that TP2 is phosphorylated in vitro at threonine 101 and serine 109 by the salt extract of sonication-resistant (elongating and elongated) spermatid nuclei and the protein kinase phosphorylating TP2 was identified to be protein kinase A (PKA). We now report that the cytosol from haploid spermatids but not from premeiotic germ cells is able to phosphorylate recombinant TP2 in vitro at threonine 101 and serine 109. The kinase present in the haploid spermatid cytosol that phosphorylates TP2 has been identified to be the sperm-specific isoform of protein kinase A (Cs-PKA). Reverse transcription-PCR analysis indicated that Cs-PKA was present in the haploid spermatids and absent from premeiotic germ cells. The rat Cs-PKA transcript was amplified and sequenced using the isoform-specific primers. The sequence of rat Cs-PKA at the N terminus differs from mouse and human by one amino acid. Western blot analysis using specific anti-Cα1 antibodies revealed that Cα1-PKA is absent in haploid spermatid cytosol. We have also established an in vitro nuclear transport assay for the haploid round spermatids. Using this assay, we have found that the cytoplasmic factors and ATP are absolutely essential for translocation of TP2 into the nucleus. Phosphorylation was found to positively modulate the NLS dependent import of TP2 into the nucleus

Rapid Detection of Staphylococcus aureus Using Graphene Oxide Coated Screen-Printed Sensor Strips

340–343Staphylococcus aureus (S. aureus) is the most common reason behind bacterial infections in humans. They cause a wide array of infections on skin, bone and joint medical implants and blood stream. In order to mitigate these infections, detecting the presence of this bacterium is of prime importance. This paper explores a new rapid detection technique to identify S aureus bacteria. A screen-printed sensor coated with graphene oxide was used as an identification platform. Graphene oxide was synthesized using modified Hummer’s method and was characterized using X-ray Diffractometer and Scanning Electron Microscope. The developed sensors showed significant decrease in the bulk resistance with the increase in bacterial concentrations. The developed sensors were subjected to selectivity, repeatability and reusability tests and showed good results

Interleaved lexical and audiovisual information can retune phoneme boundaries

To adapt to situations in which speech perception is difficult, listeners can adjust boundaries between phoneme categories using perceptual learning. Such adjustments can draw on lexical information in surrounding speech, or on visual cues via speech-reading. In the present study, listeners proved they were able to flexibly adjust the boundary between two plosive/stop consonants, /p/-/t/, using both lexical and speech-reading information and given the same experimental design for both cue types. Videos of a speaker pronouncing pseudo-words and audio recordings of Dutch words were presented in alternating blocks of either stimulus type. Listeners were able to switch between cues to adjust phoneme boundaries, and resulting effects were comparable to results from listeners receiving only a single source of information. Overall, audiovisual cues (i.e., the videos) produced the stronger effects, commensurate with their applicability for adapting to noisy environments. Lexical cues were able to induce effects with fewer exposure stimuli and a changing phoneme bias, in a design unlike most prior studies of lexical retuning. While lexical retuning effects were relatively weaker compared to audiovisual recalibration, this discrepancy could reflect how lexical retuning may be more suitable for adapting to speakers than to environments. Nonetheless, the presence of the lexical retuning effects suggests that it may be invoked at a faster rate than previously seen. In general, this technique has further illuminated the robustness of adaptability in speech perception, and offers the potential to enable further comparisons across differing forms of perceptual learning

Identification of two novel zinc finger modules and nuclear localization signal in rat spermatidal protein TP2 by site-directed mutagenesis

Spermatidal protein TP2, which appears transiently during stages 12-16 of mammalian spermiogenesis, is a DNA condensing zinc metalloprotein with a preference to GC-rich DNA. We have carried out a detailed site-directed mutagenesis analysis of rat spermatidal protein TP2 to delineate the amino acid residues involved in coordination with two atoms of zinc. Two zinc fingers modules have been identified involving 4 histidine and 4 cysteine residues, respectively. The modular structure of the two zinc fingers identified in TP2 define a new class of zinc finger proteins that do not fall into any of the known classes of zinc fingers. Transfection experiments with COS-7 cells using wild type and the two zinc finger pocket mutants have shown that TP2 preferentially localizes to nucleolus. The nuclear localization signal in TP2 was identified to be 87GKVSKRKAV95 present in the C-terminal third of TP2 as a part of an extended NoLS sequence

Neural correlates of phonetic adaptation as induced by lexical and audiovisual context

When speech perception is difficult, one way listeners adjust is by reconfiguring phoneme category boundaries, drawing on contextual information. Both lexical knowledge and lipreading cues are used in this way, but it remains unknown whether these two differing forms of perceptual learning are similar at a neural level. This study compared phoneme boundary adjustments driven by lexical or audiovisual cues, using ultra-high-field 7-T fMRI. During imaging, participants heard exposure stimuli and test stimuli. Exposure stimuli for lexical retuning were audio recordings of words, and those for audiovisual recalibration were audio–video recordings of lip movements during utterances of pseudowords. Test stimuli were ambiguous phonetic strings presented without context, and listeners reported what phoneme they heard. Reports reflected phoneme biases in preceding exposure blocks (e.g., more reported /p/ after /p/-biased exposure). Analysis of corresponding brain responses indicated that both forms of cue use were associated with a network of activity across the temporal cortex, plus parietal, insula, and motor areas. Audiovisual recalibration also elicited significant occipital cortex activity despite the lack of visual stimuli. Activity levels in several ROIs also covaried with strength of audiovisual recalibration, with greater activity accompanying larger recalibration shifts. Similar activation patterns appeared for lexical retuning, but here, no significant ROIs were identified. Audiovisual and lexical forms of perceptual learning thus induce largely similar brain response patterns. However, audiovisual recalibration involves additional visual cortex contributions, suggesting that previously acquired visual information (on lip movements) is retrieved and deployed to disambiguate auditory perception

Residual Energy-Aware Clustering Transformation for LEACH Protocol, Journal of Telecommunications and Information Technology, 2021, nr 2

Energy is one of the crucial performance parameters in wireless sensor networks (WSNs). Enhancement of network lifetime is an important consideration as well. Low energy-aware clustering hierarchy (LEACH) is one of the protocols proposed for WSNs. In this paper, a residual energy-aware clustering transformation protocol for LEACH (REACT-LEACH), enhancing performance of LEACH by introducing a clustering mechanism, is proposed. The proposed cluster head (CH) rotation and cluster reformation processes are more effective in REACT-LEACH, as residual energy is considered to be one of the metrics. Performance of REACT-LEACH is validated based on simulation

Rapid Detection of Staphylococcus aureus Using Graphene Oxide Coated Screen-Printed Sensor Strips

Staphylococcus aureus (S. aureus) is the most common reason behind bacterial infections in humans. They cause a wide array of infections on skin, bone and joint medical implants and blood stream. In order to mitigate these infections, detecting the presence of this bacterium is of prime importance. This paper explores a new rapid detection technique to identify S aureus bacteria. A screen-printed sensor coated with graphene oxide was used as an identification platform. Graphene oxide was synthesized using modified Hummer’s method and was characterized using X-ray Diffractometer and Scanning Electron Microscope. The developed sensors showed significant decrease in the bulk resistance with the increase in bacterial concentrations. The developed sensors were subjected to selectivity, repeatability and reusability tests and showed good results