bluuebunny/crossref_metadata_2025 huggingface dataset:
Kentucky Antimicrobial Stewardship Innovation Consortium (KASIC): A Program to Improve Antibiotic Use in the Commonwealth
Authors: Song Matthew , Wilde Ashley M. , Song Chelsea M. , Moore Sarah E. , Swingler Elena A. , Johnson Wes M. , Montes de Oca Jamison E. , Truong Clover N. , Carrico Ruth M. , Furmanek Stephen , Shipley Madeline , Chandler Thomas , Schulz Paul S. , Ramirez Julio A.
Published: July 2023

Antimicrobial resistance is a significant threat to modern healthcare and is driven by antimicrobial overuse. Hospital antimicrobial use in Kentucky exceeds national rates and long-term care facility antimicrobial stewardship implementation is low. Partnering with the Kentucky Department for Public Health, the Kentucky Antimicrobial Stewardship Innovation Consortium (KASIC) was created to facilitate antimicrobial stewardship across the Commonwealth of Kentucky. This manuscript describes KASIC objectives, defines the KASIC target audience, and describes KASIC methods
Isolamento social entre pessoas idosas em tempo de distanciamento social na pandemia de Covid-19
Authors: Almeida Maria Weila Coêlho , Moura Leides Barroso Azevedo , Squinca Flávia Aparecida
Published: July 2023

Analisar a situação de isolamento social de pessoas idosas em tempo de distanciamento social na pandemia de Covid-19. Métodos: Trata-se de revisão de literatura por meio de pesquisa bibliográfica, em literatura nacional e internacional. Os dados foram coletados nos meses de agosto a dezembro de 2021, a partir das Bases de Dados disponíveis na Biblioteca Central (BCE) da Universidade de Brasília (UnB) por meio do acesso remoto ao Portal de Periódicos da Capes, via Rede CAFÉ. A amostra final foi composta por 10 artigos. Resultados: A revisão de literatura revelou que os estudos apresentam lacunas na conceituação e descrição teórica de isolamento social ao tratar a temática no período pandêmico de covid-19. Apontaram a existência de um número reduzido de estudos específicos relacionados ao isolamento social em tempo de distanciamento social na pandemia de Covid-19; a não delimitação clara e objetiva acerca da diferença entre isolamento social e distanciamento social com o uso desses termos retratados como sinônimos; e que apesar de o distanciamento social ser uma medida fundamental para evitar a disseminação da Covid-19 e de o isolamento social representar um grave problema de saúde pública, a literatura revela que suas consequências podem ser mitigadas ou mesmo prevenidas por meio de ações do poder público, família e toda a sociedade a fim de superar suas múltiplas dimensões e impactos. Conclusão: Conclui-se que há necessidade da definição conceitual de isolamento social nos estudos e a distinção da medida de distanciamento social, a fim de evidenciar melhor os impactos do isolamento social devido à pandemia da Covid-19.
Aesthetic Rehabilitation of Anterior Malalinged Dentition with E-Max Laminate Veneers-A Case Report
Authors: Idnani Simran , Rishu Tushar , khan Shuja
Published: July 2023

Various different procedures have been advocated in literature to correct dental anomalies specially in the aesthetic region such as discolouration of tooth due to fluorosis or spacing in dentition due to change in shape of tooth like peg laterals. One of the most common treatment modalities to treat such cases are using VENEERS. This case report presents conservative and aesthetic procedure in the management of malalinged anterior teeth with E max lithium disilicate laminate veneers.
REVITALISASI PROSES SELEKSI MAHASISWA BARU: IMPLEMENTASI APLIKASI UJIAN SARINGAN MASUK BERBASIS ONLINE (STUDI KASUS: STMIK KAPUTAMA)
Authors: Restuono Langgeng
Published: July 2023

Aplikasi Ujian saringan masuk merupakan suatu aplikasi yang menjadi gerbang penyajian soal-soal yang bisa tambah, maupun di jawab secara online ataupun offline tergantung bagaimana aplikasi ini digunakankan. STMIK Kaputama selama beberapa tahun ini menjalankan proses penerimaan mahasiswa baru, dan ujian saringan masuk masih secara manual. Untuk memudahkan calon peserta dan juga panitia dalam rangka penerimaan mahasiswa baru, serta ujian saringan masuk, alangkah baiknya jika aplikasi yang mengelola itu semua agar menjadi mudah dan tepat dalam penerimaan serta ujian saringan masuk. Pada skripsi ini, penulis mecoba membuat aplikasi Ujian Saringan Masuk Secara Online untuk proses Penerimaan Mahasiswa Baru dan Ujian Saringan Masuk yang berbentuk aplikasi web dengan implementasi menggunakan PHP dan MySQL.
JARINGAN SENSOR NIRKABEL DAN IoT UNTUK KOTA PINTAR PAMEKASAN
Authors: Baidawi Asmat
Published: July 2023

Inovasi sekarang dalam kehidupan sehari – hari terutama dalam dunia komputer dan smartphone, sensor dan jaringan sensor terutama  yang berhubungan dengan jaringan seluler generasi berikutnya membuka peluang besar bagi para peneliti dan pengembang berbagai sistem dan aplikasi di beberapa Smart City. Beberapa aplikasi umum termasuk pemantauan sampah, sensor kelembaban, sensor pasokan air, sensor lampu jalan, sensor polusi udara dll. Memerlukan gateway pintar untuk menyediakan kecepatan data yang tinggi, konektivitas dari ujung ke ujung diperlukan bandwith dengan kualitas yang lebih tinggi. Jurnal ini ini mengusulkan model baru untuk konsep kota pintar pamekasan menggunakan IoT yang lebih penting dan bermanfaat.
PERANCANGAN APLIKASI DETEKSI ORISINALITAS CITRA DIGITAL MENERAPKAN METODE WHIRLPOOL
Authors: Adhar Deny
Published: July 2023

Salah satu dampak buruk teknologi adalah pemalsuan data yang tidak terlihat oleh mata manusia. Dokumen tersebut terlihat seperti aslinya padahal sebenarnya data tersebut telah dipalsukan. Pemalsuan ini sering terjadi pada dokumen gambar. Gambar dibuat semirip mungkin bahkan tidak terlihat berbeda dengan aslinya. Sehingga diperlukan suatu program yang dapat mendeteksi keaslian data dan algoritma yang tepat untuk mendeteksi keaslian citra, sedangkan algoritma whirpool merupakan algoritma yang paling cocok digunakan dalam penelitian ini. Whirlpool adalah fungsi hash aman berbasis blok-cipher. Whirlpool menghasilkan kode hash 512 bit untuk pesan input dengan panjang maksimum kurang dari 2256 bit. Blockcipher yang mendasarinya, berdasarkan Advanced Encryption Standard (AES), menggunakan kunci 512-bit dan beroperasi pada blok teks biasa 512-bit. Whirlpool telah disahkan oleh Nessie (Skema Eropa Baru untuk Tanda Tangan, Integritas, dan Enkripsi).
Aplikasi Media Pembelajaran Diagnosa Kerusakan Pada Handphone Berbasis Android
Authors: Setiawan Nur Zulkhairi , Yusfrizal Yusfrizal
Published: July 2023

Dalam dunia globalisasi saat ini teknologi dan komunikasi sangat dibutuhkan oleh masyarakat, salah satu perangkat komunikasi saat ini yang sangat populer adalah handphone. Penggunanya pada saat ini bukan hanya kalangan pengusaha bahkan dari kalangan bawah sampai atas pun menggunakanya bahkan anak-anak pun sudah banyak menggunakan handphone sebagai media komunikasi. Seiring berjalannya waktu perkembangan handphone semakin meningkat dan fasilitas-fasilitasnya pun semakin banyak dan berkembang. Adapun kerusakan pada handphone ada dua macam yaitu kerusakan pada perangkat hardware dan software. Oleh karena itu, keadaan yang telah diuraikan di atas menjadi acuan untuk mengembangkan sebuah aplikasi yang dapat membantu memecahkan masalah seperti seorang ahli atau pakar di bidangnya, dari permasalahan tersebut penulis akan mencoba membuat sebuah aplikasi yang berhubungan dengan masalah kerusakan handphone, supaya permasalahan tersebut lebih mudah ditelusuri kerusakannya, tetapi kita cukup dengan menggunakan aplikasi ini maka semua permasalahan kerusakan handphone dapat diketahui. Dari penelitian yang dilakukan diperoleh aplikasi pembelajaran kerusakan baik hardware ataupun software yang dibuat menggunakan Eclipse dan bahasa pemrograman Java.
Kecerdasan Spiritual sebagai Dasar Terbentuknya Profesionalitas Guru Pendidikan Agama Kristen
Authors: Anjaya Carolina Etnasari , Arifianto Yonatan Alex , Fernando Andreas
Published: July 2023

This study aims to describe the role of spiritual intelligence in increasing the professionalism of Christian Religious Education teachers. The professionalism of Christian Religious Education teachers plays an important role in the success of giving birth to students with Christ's character. Spiritual intelligence is the result of an intimate and intense relationship with God. Through this research, the author conveys that through spiritual intelligence, God actually speaks and guides teachers in living their professional lives and responsibilities. The indicators of spiritual intelligence presented in this study help to understand how spiritual intelligence actually works. The research method used is qualitative. Collecting data using the method of observation and literature study. The results of the study concluded that when a teacher has spiritual intelligence, the ability to understand the nature of oneself is created, able to understand God's will so that the spirit to move his life is the Holy Spirit. Thus his existence both body and soul (all thoughts and feelings) is directed to become a teacher similar to the Lord Jesus Christ. AbstrakPenelitian ini bertujuan untuk mendiskripsikan peranan kecerdasan spiritual dalam peningkatan profesionalitas guru pendidikan agama Kristen.  Profesionalitas pada guru PAK memegang berperan penting dalam keberhasilan melahirkan anak didik yang berkarakter Kristus. Kecerdasan spiritual merupakan hasil dari hubungan yang intim dan intens dengan Tuhan. Melalui penelitian ini penulis menyampaikan bahwa melalui kecerdasan spiritual, sejatinya Allah berbicara dan menuntun para guru dalam menjalani hidup dan tanggung jawab profesinya. Indikator-indikator kecerdasan spiritual yang dikemukakan dalam penelitian ini membantu memahami bagaimana sesungguhnya kecerdasan spiritual bekerja. Metode penelitian yang digunakan adalah kualitatif. Pengumpulan data menggu-nakan metode observasi dan studi pustaka. Hasil penelitian menyimpulkan bahwa ketika seorang guru memiliki kecerdasan spiritual maka tercipta kemampuan memahami hakikat diri, mampu memahami kehendak Tuhan sehingga spirit untuk menggerakkan hidupnya adalah Roh Kudus. Dengan demikian keberadaan dirinya baik tubuh dan jiwa (segenap pikiran dan perasaan) terarah untuk menjadi guru yang serupa dengan Tuhan Yesus Kristus.
Palestinian EFL Students' Perceptions of Using Edmodo in Developing their Writing Skills
Authors: Mahmoud Itmeizeh  , Zeiadee Khalil  , Matt Smith
Published: July 2023

The present study explored Palestinian EFL students’ perceptions towards the use of Edmodo in developing their writing skills at Palestine Ahliya University, Bethlehem. Sixteen sophomore English major students taking the Oral Communication II course in the Spring Semester 2019 - 2020 served as the participants of the study. The researchers adopted a mixed methodological approach. A questionnaire consisting of 27 items was adapted by the researchers for the purpose of eliciting students’ perceptions towards the use of Edmodo in developing their writing skills and the barriers they faced while using the platform. In addition, in-depth qualitative data was gathered from students' written responses to an end-of-course assignment posted on Edmodo. In general, the results of the study revealed that students had positive perceptions towards using Edmodo in terms of writing development and collaborative learning. However, it is worth noting that several participants reflected negative responses which resulted from the technological difficulties and their limited experiences with Edmodo.
Kegiatan Ekstrakurikuler Sebagai Wadah Pengembangan Bakat dan Minat Siswa
Authors: Hana Salsabila Putri , Syaflita Dina , Indriani Novi
Published: July 2023

Humans are born with innate abilities which are potentials that still need to be developed. Every human being has their own characteristics that can distinguish one human from another. With the existence of human abilities, it is demanded to be able to develop his potential so that he can become a quality and qualified human being. One way to hone these talents is through extracurricular activities with the aim that students' talents and creativity can be displayed to the fullest. The purpose of this study was to determine the effectiveness of these extracurricular activities as a forum for developing the talents and interests of the students of the Pekanbaru Bina Professional Vocational School. This research method is a qualitative method. The data collection technique used in this study is observation with data analysis techniques using interactive data analysis techniques. This extracurricular activity makes the students look so excited and enthusiastic. This is because students are given the freedom to be able to choose extracurricular activities that interest them. Extracurricular activities can be a success in exploring students' talents and interests.
Report of the Erasmus+ KA171 Agreement’s First-Period
Authors: OZBAY Betul
Published: July 2023

From the end of the 18th and the beginning of the 19th century, the classical monarchy systems of the old era gradually ended, and national states were established. The administrative order of the states of today’s world is mostly in the form of nation-states governed by representative democracy. However, especially since the second half of the 20th century, international partnerships and collaborative studies in different channels such as science, art, education, culture and trade have begun to be carried out with transnational agreements that have become increasingly important. Especially with education programs at different grades, individuals from a younger age can get to know various cultures by living in different societies. In this way, the opportunity to receive education at universities in other countries creates an important awareness for the individuals, the hosting institutions, and future generations. The increase in such cultural exchanges will contribute to the formation of more tolerant people and societies that understand each other more. In this context, one of the issues worth investigating is what characteristics were inherited by later generations from old monarchies as societies transformed. This question has been the source of motivation for our KA171 project proposal. Certainly, the most important legacy we inherit from our ancestors is the language we speak. We learn the language of the society we were born in from our parents and then pass it on to new generations. Although it is almost impossible to draw the boundaries of this heritage; it is possible to identify some areas that it has particularly affected.
Vecdî'nin Şiir Dünyası
Authors: UĞURLU İrem
Published: July 2023

Vecdî’nin Şiir Dünyası başlıklı çalışmanın amacı 17. asrın başlarında yaşamış ve döneminde belli bir etkiye ulaştığı günümüze ulaşan yazma nüshalarından ve kendisine yazılan nazirelerden anlaşılan Vecdî’nin eserlerindeki dünyaya nüfuz edebilmek ve çevreyi, yaşadığı kültürel dünyayı ve toplumu eserlerine ne derece yansıttığını tespit etmektir. 3 Mayıs 1661 tarihinde bir iftira sonucu 2 arkadaşıyla birlikte asılarak idam edilen Vecdî’nin asıl adının Abdülbaki, lakabının ise Boğukzâde olduğu kaynaklarda belirtilmektedir. Döneminin önemli siyasi figürlerinin dikkatini çekmiş ve bu dikkat yazın hayatının henüz baharında iken dünyadan ayrılmasına sebep olmuştur. Vecdî’nin bilinen tek eseri Dîvânçe-i Vecdî ismi ile anılmaktadır ve eserin önemli bir bölümünü gazeller oluşturmaktadır. Dîvânçe-i Vecdî dışında bir eseri var ise de henüz ele geçmiş değildir. Eserin çeşitli kütüphanelerde çok sayıda nüshasının olması döneminde ve sonrasında beğenilen şairlerden olduğuna işaret etse de hayatına dair yeterli bilgi mevcut değildir. Eserlerinde Fehim-i Kadîm gibi isimlerin etkisi gözlenebilen şairin Sebk-i Hindî üslubunda şiirler kaleme aldığı bilinmektedir. Şairin eserlerinde uzun tamlamalara ve söz sanatlarına yer vermesi, gazellerindeki beyit sayısını genellikle 7 ila 9 arasında tutması ve kaside hüviyeti gösteren gazellerinin bulunması bunların yanı sıra şiirlerindeki akıcılık eserin dikkat çekici özelliklerindendir.
Stance in CELiST: A vindication of text - reading
Authors: Moskowich Isabel , Crespo García Begoña
Published: July 2023

Stance in academic writing has been discussed extensively within the fields of discourse analysis andpragmatics (Alonso-Almeida, 2015; Hyland,2005; White, 2003). Thus, Hyland and Jiang (2016) identify certain linguistic elements that are said to be indexical of stance: hedges, boosters, attitude markers, and self-mentions. While that model aims at a macroscopic analysis, the present study is a microscopic one, and compares two scientific texts written by a male and a female author to detect possible differences in the way that these authors present themselves or give their opinions in their writings. To this end, we have sought to apply Hyland and Jiang’s (2016) three-aspects modelbutusing Cesiri’s inventory (2012) as a starting point. We have applied this adapted model to two samples from the Corpus of English Life Sciences Texts (CELiST), one of the subcorpus of the Coruña Corpus of English Scientific Writing. Both texts belong to the same genre, are dated at an almost identical time, and deal with similar subjects. The only difference is that one was written by a woman, Emily Gregory, and theother by a man, Alpheus Packard. Although these texts are part of an electronic corpus, on this occasion we will avoid the automatic analytical techniques of corpus linguistics asfaras possible. Rather, we will conduct a microscopic-level study by means of close reading, although some quantification of data will precede the qualitative analysis where this is useful. It is hoped that the qualitative focus presented in theanalysis might open up new paths in the study of stance.
Stance matrices licensing that-clauses and interpersonal meaning in nineteenth-century women's instructive writing in English
Authors: Alonso-Almeida Francisco
Published: July 2023

This paper examines stance matrices licensing that-clauses in a corpus of instructional texts authored by women during the nineteenth century, gathered under the label COWITE19. These matrices can reveal various aspects of the authors' assessment, involvement and understanding of the information they present. In other words, their use discloses the authors' evaluation of their text while conveying a wide range of interpersonal meanings without disregarding their organising potential as textual markers. The types of matrices explored in this article precede the that-clauses and generally contain information denoting authorial perspective and involvement. The data used to demonstrate their forms and functions derive from analysing all cases found in the Corpus of Women's Instructive Texts in English (1550–1900) (COWITE); for the current study, only the nineteenth-century sub-corpus, henceforth COWITE19, has been considered. This corpus exclusively comprises instructional texts penned by women during the nineteenth century. The findings reveal that although the corpus primarily encompasses recipes from a diverse range of registers, the authoritative voice of women is distinctly evident in the matrices analysed, conveying a series of interpersonal meanings that unequivocally highlight the experience of women writers and their adept command of the content and techniques being discussed.
« Vous me taxerez de “séparatiste” sans doute ! Je le prendrais comme un compliment »1 : les intellectuelles indépendantistes de la Révolution tranquille (1957-1968)2
Authors: Carlos Jean-Philippe
Published: July 2023

Notre étude porte sur les idées des principales figures intellectuelles féminines et la place qu’elles occupent dans le réseau des revues indépendantistes entre 1957 et 1968. En nous basant sur l’analyse des revues L’Action nationale, Liberté, L’Indépendance, Parti pris et La Nation, nous montrons que ces intellectuelles ont joué un rôle important dans la diffusion de l’idéal indépendantiste durant la décennie 1960. En suivant le parcours de ces femmes, nous pouvons ainsi mieux appréhender les transformations de la culture politique et intellectuelle du Québec durant la Révolution tranquille.
Assessing MSDs before Introduction of a Cobot: Psychosocial Aspects and Employee’s Subjective Experience
Authors: Cippelletti Emma , Azouaghe Soufian , Pellier Damien , Landry Aurélie
Published: July 2023

Musculoskeletal disorders (MSDs) are one of the main causes of work disability (EU-OSHA, 2019; WHO, 2019). Several solutions, including the cobotic system (EUROGIP, 2017), have been put forward to improve unhealthy working conditions and prevent MSDs. We sought to identify the MSD risk factors of workers on a screen-printed glass production line prior to introduction of a cobot. We used a mixed data collection technique: video observations and assessment of MSD risk factors by expert ergonomists, and then self-confrontation interviews with six production-line operators and subjective perception of risk factors. The two types of assessment (by experts and by operators) showed that the most demanding risk factors were physical (e.g., work posture) and psychosocial (e.g., mental workload). Certain risk factors were viewed differently by the experts and the operators. One question remains: How can a cobot make work more meaningful for operators?
Rate-distortion theory of neural coding and its implications for working memory
Authors: Jakob Anthony MV , Gershman Samuel J
Published: July 2023

Rate-distortion theory provides a powerful framework for understanding the nature of human memory by formalizing the relationship between information rate (the average number of bits per stimulus transmitted across the memory channel) and distortion (the cost of memory errors). Here, we show how this abstract computational-level framework can be realized by a model of neural population coding. The model reproduces key regularities of visual working memory, including some that were not previously explained by population coding models. We verify a novel prediction of the model by reanalyzing recordings of monkey prefrontal neurons during an oculomotor delayed response task.
Influenza virus transcription and progeny production are poorly correlated in single cells
Authors: Bacsik David J , Dadonaite Bernadeta , Butler Andrew , Greaney Allison J , Heaton Nicholas S , Bloom Jesse D
Published: July 2023

The ultimate success of a viral infection at the cellular level is determined by the number of progeny virions produced. However, most single-cell studies of infection quantify the expression of viral transcripts and proteins, rather than the amount of progeny virions released from infected cells. Here, we overcome this limitation by simultaneously measuring transcription and progeny production from single influenza virus-infected cells by embedding nucleotide barcodes in the viral genome. We find that viral transcription and progeny production are poorly correlated in single cells. The cells that transcribe the most viral mRNA do not produce the most viral progeny and often represent aberrant infections that fail to express the influenza NS gene. However, only some of the discrepancy between transcription and progeny production can be explained by viral gene absence or mutations: there is also a wide range of progeny production among cells infected by complete unmutated virions. Overall, our results show that viral transcription is a relatively poor predictor of an infected cell’s contribution to the progeny population.
Influenza virus transcription and progeny production are poorly correlated in single cells
Authors: Bacsik David J. , Dadonaite Bernadeta , Butler Andrew , Greaney Allison J. , Heaton Nicholas S. , Bloom Jesse D.
Published: July 2023

The ultimate success of a viral infection at the cellular level is determined by the number of progeny virions produced. However, most single-cell studies of infection quantify the expression of viral transcripts and proteins, rather than the amount of progeny virions released from infected cells. Here we overcome this limitation by simultaneously measuring transcription and progeny production from single influenza-virus-infected cells by embedding nucleotide barcodes in the viral genome. We find that viral transcription and progeny production are poorly correlated in single cells. The cells that transcribe the most viral mRNA do not produce the most viral progeny, and often represent aberrant infections that fail to express the influenza NS gene. However, only some of the discrepancy between transcription and progeny production can be explained by viral gene absence or mutations: there is also a wide range of progeny production among cells infected by complete unmutated virions. Overall, our results show that viral transcription is a relatively poor predictor of an infected cell’s contribution to the progeny population.
Statistical analysis supports pervasive RNA subcellular localization and alternative 3' UTR regulation
Authors: Bierman Rob , Dave Jui M , Greif Daniel M , Salzman Julia
Published: July 2023

Targeted low-throughput studies have previously identified subcellular RNA localization as necessary for cellular functions including polarization, and translocation. Furthermore, these studies link localization to RNA isoform expression, especially 3’ Untranslated Region (UTR) regulation. The recent introduction of genome-wide spatial transcriptomics techniques enables the potential to test if subcellular localization is regulated in situ pervasively. In order to do this, robust statistical measures of subcellular localization and alternative poly-adenylation (APA) at single-cell resolution are needed. Developing a new statistical framework called SPRAWL, we detect extensive cell-type specific subcellular RNA localization regulation in the mouse brain and to a lesser extent mouse liver. We integrated SPRAWL with a new approach to measure cell-type specific regulation of alternative 3’ UTR processing and detected examples of significant correlations between 3’ UTR length and subcellular localization. Included examples, Timp3, Slc32a1, Cxcl14, and Nxph1 have subcellular localization in the mouse brain highly correlated with regulated 3’ UTR processing that includes the use of unannotated, but highly conserved, 3’ ends. Together, SPRAWL provides a statistical framework to integrate multi-omic single-cell resolved measurements of gene-isoform pairs to prioritize an otherwise impossibly large list of candidate functional 3’ UTRs for functional prediction and study. In these studies of data from mice, SPRAWL predicts that 3’ UTR regulation of subcellular localization may be more pervasive than currently known.
Evolution of a functionally intact but antigenically distinct DENV fusion loop
Authors: Meganck Rita M , Zhu Deanna , Dong Stephanie , Snoderly-Foster Lisa J , Dalben Yago R , Thiono Devina , White Laura J , DeSilva Arivianda M , Baric Ralph S , Tse Longping V
Published: July 2023

A hallmark of dengue virus (DENV) pathogenesis is the potential for antibody-dependent enhancement, which is associated with deadly DENV secondary infection, complicates the identification of correlates of protection, and negatively impacts the safety and efficacy of DENV vaccines. Antibody-dependent enhancement is linked to antibodies targeting the fusion loop (FL) motif of the envelope protein, which is completely conserved in mosquito-borne flaviviruses and required for viral entry and fusion. In the current study, we utilized saturation mutagenesis and directed evolution to engineer a functional variant with a mutated FL (D2-FL), which is not neutralized by FL-targeting monoclonal antibodies. The FL mutations were combined with our previously evolved prM cleavage site to create a mature version of D2-FL (D2-FLM), which evades both prM- and FL-Abs but retains sensitivity to other type-specific and quaternary cross-reactive (CR) Abs. CR serum from heterotypic (DENV4)-infected non-human primates (NHP) showed lower neutralization titers against D2-FL and D2-FLM than isogenic wildtype DENV2 while similar neutralization titers were observed in serum from homotypic (DENV2)-infected NHP. We propose D2-FL and D2-FLM as valuable tools to delineate CR Ab subtypes in serum as well as an exciting platform for safer live-attenuated DENV vaccines suitable for naïve individuals and children.
Delayed postglacial colonization of Betula in Iceland and the circum North Atlantic
Authors: Harning David J , Sacco Samuel , Anamthawat-Jónsson Kesara , Ardenghi Nicolò , Thordarson Thor , Raberg Jonathan H , Sepúlveda Julio , Geirsdóttir Áslaug , Shapiro Beth , Miller Gifford H
Published: July 2023

As the Arctic continues to warm, woody shrubs are expected to expand northward. This process, known as ‘shrubification,’ has important implications for regional biodiversity, food web structure, and high-latitude temperature amplification. While the future rate of shrubification remains poorly constrained, past records of plant immigration to newly deglaciated landscapes in the Arctic may serve as useful analogs. We provide one new postglacial Holocene sedimentary ancient DNA (sedaDNA) record of vascular plants from Iceland and place a second Iceland postglacial sedaDNA record on an improved geochronology; both show Salicaceae present shortly after deglaciation, whereas Betulaceae first appears more than 1000 y later. We find a similar pattern of delayed Betulaceae colonization in eight previously published postglacial sedaDNA records from across the glaciated circum North Atlantic. In nearly all cases, we find that Salicaceae colonizes earlier than Betulaceae and that Betulaceae colonization is increasingly delayed for locations farther from glacial-age woody plant refugia. These trends in Salicaceae and Betulaceae colonization are consistent with the plant families’ environmental tolerances, species diversity, reproductive strategies, seed sizes, and soil preferences. As these reconstructions capture the efficiency of postglacial vascular plant migration during a past period of high-latitude warming, a similarly slow response of some woody shrubs to current warming in glaciated regions, and possibly non-glaciated tundra, may delay Arctic shrubification and future changes in the structure of tundra ecosystems and temperature amplification.
FSHβ links photoperiodic signaling to seasonal reproduction in Japanese quail
Authors: Majumdar Gaurav , Liddle Timothy A , Stewart Calum , Marshall Christopher J , Bain Maureen , Stevenson Tyler
Published: July 2023

Annual cycles in daylength provide an initial predictive environmental cue that plants and animals use to time seasonal biology. Seasonal changes in photoperiodic information acts to entrain endogenous programs in physiology to optimize an animal’s fitness. Attempts to identify the neural and molecular substrates of photoperiodic time measurement in birds have, to date, focused on blunt changes in light exposure during a restricted period of photoinducibility. The objectives of these studies were first to characterize a molecular seasonal clock in Japanese quail and second, to identify the key transcripts involved in endogenously generated interval timing that underlies photosensitivity in birds. We hypothesized that the mediobasal hypothalamus (MBH) provides the neuroendocrine control of photoperiod-induced changes in reproductive physiology, and that the pars distalis of the pituitary gland contains an endogenous internal timer for the short photoperiod-dependent development of reproductive photosensitivity. Here, we report distinct seasonal waveforms of transcript expression in the MBH, and pituitary gland and discovered the patterns were not synchronized across tissues. Follicle-stimulating hormone-β (FSHβ) expression increased during the simulated spring equinox, prior to photoinduced increases in prolactin, thyrotropin-stimulating hormone-β, and testicular growth. Diurnal analyses of transcript expression showed sustained elevated levels of FSHβ under conditions of the spring equinox, compared to autumnal equinox, short (<12L) and long (>12L) photoperiods. FSHβ expression increased in quail held in non-stimulatory short photoperiod, indicative of the initiation of an endogenously programmed interval timer. These data identify that FSHβ establishes a state of photosensitivity for the external coincidence timing of seasonal physiology. The independent regulation of FSHβ expression provides an alternative pathway through which other supplementary environmental cues, such as temperature, can fine tune seasonal reproductive maturation and involution.
FSHβ links photoperiodic signalling to seasonal reproduction in Japanese quail
Authors: Majumdar Gaurav , Liddle Timothy A , Stewart Calum , Marshall Christopher J , Bain Maureen , Stevenson Tyler John
Published: July 2023

Endogenous programs in avian reproductive seasonal life history transitions have been known for over 30 years. Attempts to identify the neural and molecular substrates of photoperiodic time measurement have, to date, focussed on blunt changes in light exposure during a restricted period of photoinducibility. Here we report mediobasal hypothalamus (MBH) and pituitary gland specific transcript expression cascades associated with photoperiod-induced seasonal life history transitions. We identified multiple waves of transcript expression that were not synchronized across brain regions. Follicle-stimulating hormone-β (FSHβ) expression increased during the simulated vernal equinox, prior to photoinduced increases in prolactin, thyrotropin-stimulating hormone-β and testicular growth. Analyses showed sustained elevated levels of daily FSHβ expression in the pituitary under conditions of the vernal, compared to autumnal equinox, short (<12L) and long (>12L) photoperiods. FSHβ expression increased in Quail held in non-stimulatory photoperiod, indicative of an endogenous programmed change in transcription. These data identify that FSHβ provides an endogenous program for the photoperiod-dependent external coincidence timing for seasonal transitions in reproduction. FSHβ may prime the pituitary for subsequent stimulation by triiodothyronine-mediated changes in tanycytes during the photoinducible phase. Overall, the data indicate a multi-cellular, multi-neural interval timing mechanism resides in the brain to control seasonal life-history transitions in birds.
DYRK1A interacts with the tuberous sclerosis complex and promotes mTORC1 activity
Authors: Wang Pinhua , Sarkar Sunayana , Zhang Menghuan , Xiao Tingting , Kong Fenhua , Zhang Zhe , Balasubramanian Deepa , Jayaram Nandan , Datta Sayantan , He Ruyu , Wu Ping , Chao Peng , Zhang Ying , Washburn Michael , Florens Laurence A , Nagarkar-Jaiswal Sonal , Jaiswal Manish , Mohan Man
Published: July 2023

DYRK1A, a ubiquitously expressed kinase, is linked to the dominant intellectual developmental disorder, microcephaly, and Down syndrome in humans. It regulates numerous cellular processes such as cell cycle, vesicle trafficking, and microtubule assembly. DYRK1A is a critical regulator of organ growth; however, how it regulates organ growth is not fully understood. Here, we show that the knockdown of DYRK1A in mammalian cells results in reduced cell size, which depends on mTORC1. Using proteomic approaches, we found that DYRK1A interacts with the tuberous sclerosis complex (TSC) proteins, namely TSC1 and TSC2, which negatively regulate mTORC1 activation. Furthermore, we show that DYRK1A phosphorylates TSC2 at T1462, a modification known to inhibit TSC activity and promote mTORC1 activity. We also found that the reduced cell growth upon knockdown of DYRK1A can be rescued by overexpression of RHEB, an activator of mTORC1. Our findings suggest that DYRK1A inhibits TSC complex activity through inhibitory phosphorylation on TSC2, thereby promoting mTORC1 activity. Furthermore, using the Drosophila neuromuscular junction as a model, we show that the mnb, the fly homologs of DYRK1A, is rescued by RHEB overexpression, suggesting a conserved role of DYRK1A in TORC1 regulation.
Recursive self-embedded vocal motifs in wild orangutans
Authors: Lameira Adriano R. , Hardus Madeleine E. , Ravignani Andrea , Raimondi Teresa , Gamba Marco
Published: July 2023

Recursive procedures that allow placing a vocal signal inside another of similar kind provide a neuro-computational blueprint for syntax and phonology in spoken language and human song. There are, however, no known vocal patterns among nonhuman primates arranged in self-embedded combinations that evince vocal recursion or potential insipient forms and neuro-procedures thereof, suggesting a neuro-cognitive transformation exclusive to humans. Here, we uncover that wild flanged male orangutan long calls show two hierarchical strata, wherein rhythmically isochronous call sequences are nested within self-similar isochronous call sequences. Remarkably, three unrelated recursive motifs occurred simultaneously in long calls, refuting that motifs resulted from three parallel linear procedures or that motifs were simple anatomical artifacts of bodily constrains. Findings represent a case of recursive hominid vocal production in the absence syntax, semantics, phonology or music. Second-order combinatorics, ‘sequences within sequences’, involving hierarchically organized and cyclically structured vocal sounds in ancient hominids may have preluded the evolution of recursion in modern language-able humans.
C-type natriuretic peptide improves maternally aged oocytes quality by inhibiting excessive PINK1/Parkin-mediated mitophagy
Authors: Zhang Hui , Li Chan , Liu Qingyang , Li Jingmei , Wu Hao , Xu Rui , Sun Yidan , Cheng Ming , Zhao Xiaoe , Pan Menghao , Wei Qiang , Ma Baohua
Published: July 2023

The overall oocyte quality declines with ageing, and this effect is strongly associated with a higher reactive oxygen species (ROS) level and the resultant oxidative damage. C-type natriuretic peptide (CNP) is a well-characterized physiological meiotic inhibitor that has been successfully used to improve immature oocyte quality during in vitro maturation (IVM). However, the underlying roles of CNP in maternally aged oocytes have not been reported. Here, we found that the age-related reduction in the serum CNP concentration was highly correlated with decreased oocyte quality. Treatment with exogenous CNP promoted follicle growth and ovulation in aged mice and enhanced meiotic competency and fertilization ability. Interestingly, the cytoplasmic maturation of aged oocytes was thoroughly improved by CNP treatment, as assessed by spindle/chromosome morphology and redistribution of organelles (mitochondria, the endoplasmic reticulum [ER], cortical granules [CGs], and the Golgi apparatus). CNP treatment also ameliorated DNA damage and apoptosis caused by ROS accumulation in aged oocytes. Importantly, oocyte RNA-seq revealed that the beneficial effect of CNP on aged oocytes was mediated by restoration of mitochondrial oxidative phosphorylation, eliminating excessive mitophagy. CNP reversed the defective phenotypes in aged oocytes by alleviating oxidative damage and suppressing excessive PINK1/Parkin-mediated mitophagy. Mechanistically, CNP functioned as a cAMP/PKA pathway modulator to decrease PINK1 stability and inhibit Parkin recruitment. In summary, our results demonstrated that CNP supplementation constitutes an alternative therapeutic approach for advanced maternal age-related oocyte deterioration and may improve the overall success rates of clinically assisted reproduction in older women.
Ryanodine receptor 2 inhibition improves cardiac repolarization reserve and contractile function and prevents sudden arrhythmic death in failing hearts
Authors: Joshi Pooja , Estes Shanea , DeMazumder Deeptankar , Knollmann Bjorn C. , Dey Swati
Published: July 2023

Sudden cardiac death (SCD) from ventricular tachycardia/fibrillation (VT/VF) are a leading cause of death, but current therapies are limited. Despite extensive research on drugs targeting sarcolemmal ion channels, none have proven sufficiently effective for preventing SCD. Sarcoplasmic ryanodine receptor 2 (RyR2) Ca
2+
release channels, the downstream effectors of sarcolemmal ion channels, are underexplored in this context. Recent evidence implicates reactive oxygen species (ROS)-mediated oxidation and hyperactivity of RyR2s in the pathophysiology of SCD.To test the hypothesis that RyR2 inhibition of failing arrhythmogenic hearts reduces sarcoplasmic Ca
2+
leak and repolarization lability, mitigates VT/VF/SCD and improves contractile function.We used a guinea pig model that replicates key clinical aspects of human nonischemic HF, such as a prolonged QT interval, a high prevalence of spontaneous arrhythmic SCD, and profound Ca
2+
leak via a hyperactive RyR2. HF animals were randomized to receive dantrolene (DS) or placebo in early or chronic HF. We assessed the incidence of VT/VF and SCD (primary outcome), ECG heart rate and QT variability, echocardiographic left ventricular (LV) structure and function, immunohistochemical LV fibrosis, and sarcoplasmic RyR2 oxidation.DS treatment prevented VT/VF and SCD by decreasing dispersion of repolarization and ventricular arrhythmias. Compared to placebo, DS lowered resting heart rate, preserved chronotropic competency during transient β-adrenergic challenge, and improved heart rate variability and cardiac function.Inhibition of RyR2 hyperactivity with dantrolene mitigates the vicious cycle of sarcoplasmic Ca
2+
leak-induced increases in diastolic Ca
2+
and ROS-mediated RyR2 oxidation, thereby increasing repolarization reserve and protecting against VT/VF/SCD. Moreover, the consequent increase in sarcoplasmic Ca
2+
load improves contractile function. These potentially life-saving effects of RyR2 inhibition warrant further investigation, such as clinical studies of repurposing dantrolene as a potential new therapy for heart failure and/or SCD.
Epistasis facilitates functional evolution in an ancient transcription factor
Authors: Metzger Brian P.H. , Park Yeonwoo , Starr Tyler N. , Thornton Joseph W.
Published: July 2023

A protein’s genetic architecture – the set of causal rules by which its sequence determines its specific functions – also determines the functional impacts of mutations and the protein’s evolutionary potential. Prior research has proposed that proteins’ genetic architecture is very complex, with pervasive epistatic interactions that constrain evolution and make function difficult to predict from sequence. Most of this work has considered only the amino acid states present in two sequences of interest and the direct paths between them, but real proteins evolve in a multidimensional space of 20 possible amino acids per site. Moreover, almost all prior work has assayed the effect of sequence variation on a single protein function, leaving unaddressed the genetic architecture of functional specificity and its impacts on the evolution of new functions. Here we develop a new logistic regression-based method to directly characterize the global causal rules of the genetic architecture of multiple protein functions from 20-state combinatorial deep mutational scanning (DMS) experiments. We apply it to dissect the genetic architecture and evolution of a transcription factor’s specificity for DNA, using data from a combinatorial DMS of an ancient steroid hormone receptor’s capacity to activate transcription from two biologically relevant DNA elements. We show that the genetic architecture of DNA recognition and specificity consists of a dense set of main and pairwise effects that involve virtually every possible amino acid state in the protein-DNA interface, but higher-order epistasis plays only a tiny role. Pairwise interactions enlarge the set of functional sequences and are the primary determinants of specificity for different DNA elements. Epistasis also massively expands the number of opportunities for single-residue mutations to switch specificity from one DNA target to another. By bringing variants with different functions close together in sequence space, pairwise epistasis therefore facilitates rather than constrains the evolution of new functions.
The Toxoplasma monocarboxylate transporters are involved in the metabolism within the apicoplast and are linked to parasite survival
Authors: Dong Hui , Yang Jiong , He Kai , Zheng Wen-Bin , Lai De-Hua , Liu Jing , Ding Hui-Yong , Wu Rui-Bin , Brown Kevin M , Hide Geoff , Lun Zhao-Rong , Zhu Xing-Quan , Long Shaojun
Published: July 2023

The apicoplast is a four-membrane plastid found in the apicomplexans, which harbors biosynthesis and organelle housekeeping activities in the matrix. However, the mechanism driving the flux of metabolites, in and out, remains unknown. Here, we used TurboID and genome engineering to identify apicoplast transporters in Toxoplasma gondii. Among the many novel transporters, we show that one pair of apicomplexan monocarboxylate transporters (AMTs) appears to have evolved from a putative host cell that engulfed a red alga. Protein depletion showed that AMT1 and AMT2 are critical for parasite growth. Metabolite analyses supported the notion that AMT1 and AMT2 are associated with biosynthesis of isoprenoids and fatty acids. However, stronger phenotypic defects were observed for AMT2, including in the inability to establish T. gondii parasite virulence in mice. This study clarifies, significantly, the mystery of apicoplast transporter composition and reveals the importance of the pair of AMTs in maintaining the apicoplast activity in apicomplexans.
Functional ultrasound imaging of stroke in awake rats
Authors: Brunner Clément , Montaldo Gabriel , Urban Alan
Published: July 2023

Anesthesia is a major confounding factor in preclinical stroke research as stroke rarely occurs in sedated patients. Moreover, anesthesia affects both brain functions and the stroke outcome acting as neurotoxic or protective agent. So far, no approaches were well suited to induce stroke while imaging hemodynamics along with simultaneous large-scale recording of brain functions in awake animals. For this reason, the first critical hours following the stroke insult and associated functional alteration remain poorly understood. Here, we present a strategy to investigate both stroke hemodynamics and stroke-induced functional alterations without the confounding effect of anesthesia, i.e., under awake condition. Functional ultrasound (fUS) imaging was used to continuously monitor variations in cerebral blood volume (CBV) in +65 brain regions/hemisphere for up to 3hrs after stroke onset. The focal cortical ischemia was induced using a chemo-thrombotic agent suited for permanent middle cerebral artery occlusion in awake rats, and followed by ipsi- and contralesional whiskers stimulation to investigate on the dynamic of the thalamo-cortical functions. Early (0-3hrs) and delayed (day 5) fUS recording enabled to characterize the features of the ischemia (location, CBV loss), spreading depolarizations (occurrence, amplitude) and functional alteration of the somatosensory thalamo-cortical circuits. Post-stroke thalamo-cortical functions were affected not only early after the stroke onset but were also altered secondarly and remotely from the initial insult. Overall, our procedure enables early, continuous, and chronic evaluations of hemodynamics and brain functions which, combined to stroke or other pathologies, aims to better understand physiopathologies toward the development of clinically relevant therapeutic strategies.