More foreign-accented but more comprehensible: Attrition and amelioration of L1 speech in proficient L2 learners.

Background: There is an increasing interest in cross-linguistic influences of the second language (L2) on the first (L1), but its communicative impact remains to be elucidated. This study investigates how L2 learners' L1 pronunciation is perceived as foreign-accented and (in) comprehensible as a function of their L2 learning experience and proficiency levels. Methods: Read speech of 154 L1 Japanese learners of L2 English in the J-AESOP corpus was examined, where approximately one-third of them had lived in English-speaking countries and the rest had never lived outside of Japan. Their L1 speech was rated by another group of native Japanese listeners for accentedness and comprehensibility (from October 25, 2022 to August 20, 2023), while their L2 speech was previously rated by native American English listeners for nativelikeness or proficiency. The speakers' vowel acoustics were also examined. Results: More proficient L2 speakers were perceived as more foreign-accented in their L1, but only if they had lived overseas; their length of residence abroad predicted the degree of perceived accentedness. In contrast, more proficient L2 speakers were consistently perceived as more comprehensible in the L1, regardless of prior overseas experience. Acoustic analyses indicated that perceived accentedness is associated with a clockwise chain shift of all vowel categories in the vowel space. It was also revealed that the dispersion, rather than compactness, of vowel production contributed to perceived comprehensibility, although the degree of L1 vowel dispersion did not predict L2 proficiency. Conclusions: The overall results suggest two main conclusions. First, perceptible L1 foreign accent likely results from L1 disuse rather than L2 interference, thereby L1 pronunciation differs from native norms at a system-wide rather than category-specific level. Second, L2 learning has a positive influence on perceived L1 comprehensibility, rather than individuals with clearer and more comprehensible L1 speech being inherently better L2 learners.

Crystal structure of tick tyrosylprotein sulfotransferase reveals the activation mechanism of the tick anticoagulant protein madanin.

Ticks pose a substantial public health risk as they transmit various pathogens. This concern is related to the adept blood-sucking strategy of ticks, underscored by the action of the anticoagulant, madanin, which is known to exhibit an approximately 1000-fold increase in anticoagulant activity following sulfation of its two tyrosine residues, Tyr51 and Tyr54. Despite this knowledge, the molecular mechanism underlying sulfation by tick tyrosylprotein sulfotransferase (TPST) remains unclear. In this study, we successfully prepared tick TPST as a soluble recombinant enzyme. We clarified the method by which this enzyme proficiently sulfates tyrosine residues in madanin. Biochemical analysis using a substrate peptide based on madanin and tick TPST, along with the analysis of the crystal structure of the complex and docking simulations, revealed a sequential sulfation process. Initial sulfation at the Tyr51 site augments binding, thereby facilitating efficient sulfation at Tyr54. Beyond direct biochemical implications, these findings considerably improve our understanding of tick blood-sucking strategies. Furthermore, combined with the utility of modified tick TPST, our findings may lead to the development of novel anticoagulants, promising avenues for thrombotic disease intervention and advancements in the field of public health.

A Novel High-Throughput Screening Method for a Human Multicentric Osteosarcoma-Specific Antibody and Biomarker Using a Phage Display-Derived Monoclonal Antibody.

Osteosarcoma is a malignant tumor that produces neoplastic bone or osteoid osteoma. In human multicentric osteosarcoma (HMOS), a unique variant of human osteosarcoma (HOS), multiple bone lesions occur simultaneously or asynchronously before lung metastasis. HMOS is associated with an extremely poor prognosis, and effective treatment options are lacking. Using the proteins in our previously generated HMOS cell lines as antigens, we generated antibodies using a human antibody phage library. We obtained antibody clones recognizing 95 independent antigens and developed a fluorescence probe-based enzyme-linked immunosorbent assay (ELISA) technique capable of evaluating the reactivity of these antibodies by fluorescence intensity, allowing simple, rapid, and high-throughput selection of antibody clones. These results were highly correlated with those using flow cytometry. Subsequently, the HMOS cell lysate was incubated with the antibody, the antigen-antibody complex was recovered with magnetic beads, and the protein bands from electrophoresis were analyzed using liquid chromatography-mass spectrometry (LC/MS). CAVIN1/polymerase I transcript release factor was specifically detected in the HMOS cells. In conclusion, we found via a novel high-throughput screening method that CAVIN1/PTRF is an HMOS-specific cell membrane biomarker and an antigen capable of producing human antibodies. In the future, antibody-drug conjugate targeting of these specific proteins may be promising for clinical applications.

Plasma membrane proteomic changes of Arabidopsis DRP1E during cold acclimation in association with the enhancement of freezing tolerance.

The freezing tolerance of plants that live in cold regions increases after exposure to low temperature, a process termed cold acclimation (CA). During CA, restructuring of the plasma membrane (PM) is important to enhance freezing tolerance. We have previously shown that the function of DYNAMIN-RELATED PROTEIN 1 E (DRP1E), which regulates endocytosis by pinching vesicles from the PM, is associated with the enhancement of freezing tolerance during CA in Arabidopsis. DRP1E is predicted to play a role in reconstituting the PM composition during CA. In this study, to test the validity of this hypothesis, we studied the changes in PM proteome patterns induced by drp1e mutation. In a detailed physiological analysis, after 3 days of CA, only young leaves showed significantly less increase in freezing tolerance in the mutant than in the wild type (WT). Using nano-liquid chromatography-tandem mass spectrometry, 496 PM proteins were identified. Among these proteins, 81 or 71 proteins were specifically altered in the WT or the mutant, respectively, in response to CA. Principal component analysis showed that the proteomic pattern differed between the WT and the mutant upon cold acclimation (CA), suggesting that DRP1E contributes to reconstruction of the PM during CA. Cluster analysis revealed that proteins that were significantly increased in the mutant after CA were biased toward glycosylphosphatidylinositol-anchored proteins, such as fasciclin-like arabinogalactan proteins. Thus, a primary target of DRP1E-associated PM reconstruction during CA is considered to be glycosylphosphatidylinositol-anchored proteins, which may be removed from the PM by DRP1E in young leaves after 3 days of CA.

The pentameric hydrocoel lobes organize adult pentameral structures in a sea cucumber, Apostichopus japonicus.

Despite being one of the bilaterians, the body plan of echinoderms shifts from bilateral symmetry to five-fold radial, or pentaradial symmetry during embryogenesis or their metamorphosis. While the clarification of the developmental mechanism behind this transition will be a basis for understanding their unique body plan evolution, it is still poorly understood. With this regard, the hydrocoel, a mesodermal coelom formed on the left side of bilateral larva, would be a clue for understanding the mechanism as it is the first pentaradial structure that appears before metamorphosis and develops into the water vascular system of adults. By analyzing the development of a sea cucumber, Apostichopus japonicus, we found that the hydrocoel expresses genes related in muscle and neural formation such as myosin heavy chain, tropomyosin, soxC, and elav, implying that cells of the hydrocoel contributes to muscle and neural structures in the adult. Furthermore, ablation of one of the hydrocoel lobes led to incomplete development of adult pentameral structures. The ablation of primary hydrocoel lobes resulted in the reduction in tentacles and the ablation of secondary hydrocoel lobes resulted in the reduction in water vascular canals and nerve cords. Our findings suggest that the hydrocoel lobes may serve as a potential organizing center for establishing the pentaradial body plan in echinoderms.

Predicting and classifying Japanese singleton and geminate consonants using logarithmic duration.

To clarify the acoustic variables for predicting and classifying Japanese singleton and geminate consonants, raw and logarithmic durations of the consonants and their related segments were examined using 12 minimal pair words that were pronounced in a carrier sentence at various speaking rates by 20 native Japanese speakers. Regression and discriminant analyses revealed that the logarithmic durations were better at predicting and classifying Japanese singleton and geminate consonants than the raw durations used in many previous studies. Specifically, the best acoustic variables were the logarithmic duration of the consonant's closure or frication and the logarithmic average duration of the mora in the preceding carrier phrase. These results suggest that logarithmic durations are relational invariant acoustic variables that can cope with the durational variations of singleton and geminate consonants in a wide range of speaking rates.