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Penelitian ini mendeskripsikan efektivitas program pembelajaran menggunakan Model Pembelajaran ExPRession untuk meningkatkan keterampilan sistem berpikir siswa pada materi interferensi cahaya. Sampel penelitian terdiri dari siswa kelas XI MIPA 1 dan XI MIPA 2 dari salah satu SMA di Lampung Timur. Desain penelitian yang digunakan adalah Non-Equivalent Pretest-Posttest Control Group Design. Teknik pengumpulan data menggunakan teknik tes. Instrumen tes berupa 7 soal esai yang mengakomodasi tujuh indikator kemampuan berpikir sistem, yaitu: 1) mengeksplorasi berbagai sudut pandang; 2) mempertimbangkan permasalahan secara tepat; 3) menetapkan batasan; 4) unsur pembeda dan pengukur; 5) mengidentifikasi dan mengkarakterisasi hubungan; 6) mengidentifikasi dan mengkarakterisasi umpan balik; 7) menjelaskan dan memprediksi perilaku sistem. Teknik analisis data menggunakan data deskriptif dan parametrik dengan menggunakan Independent Sample T-Test dan N-gain. Hasil Uji Independent Sample T-Test menunjukkan bahwa Sig. (2-tailed) posttest 0,001 kurang dari 0,05 yang berarti terdapat perbedaan yang signifikan rata-rata skor keterampilan berpikir sistem siswa pada kelas eksperimen dan kontrol setelah diberikan perlakuan. Peningkatan keterampilan berpikir sistem siswa pada kelas eksperimen yang melaksanakan program pembelajaran dengan model Pembelajaran ExPRession sebesar 0,6 termasuk dalam kategori sedang, lebih signifikan dibandingkan dengan kelas kontrol dengan pembelajaran konvensional yaitu sebesar 0,2 dengan kategori rendah.

Kinship recognition that primarily focuses on exploiting facial micro expressions is an interesting and challenging problem that aims to determine whether multiple individuals belong to the same family. Previous approaches have been limited by model capacity and insufficient training data, resulting in low-level features and shallow model learning. These common manual features cannot capture information effectively, leading to suboptimal accuracy. In this paper, we propose a kinship recognition that exploits facial micro expressions using a hybrid Vision Transformer and Mamba (ViTMa) model with modified Deep Feature Fusion, which combines different backbone architectures and feature fusion strategies. The ViTMa model is pre-trained on a large dataset and adapted to Indonesian facial images. The Siamese architecture processes two input images, extracts features fused with feature fusion, and passes them to a classification network. Experiments on the FIW-Local Indonesia dataset demonstrate the effectiveness of this method, with the best model using B16 quadratic features and multiplicative fusion achieving an average accuracy of 85.18% across all kinship categories, outperforming previous approaches. We found that B16, despite being the smallest backbone, has the best performance compared to larger backbones such as L16 with an average accuracy of 67.99%, B32 with an average accuracy of 72.98%, and L32 with an average accuracy of 71.69%. Thus, the ViTMa model with our proposed B16 quadratic feature fusion and multiplicative fusion strategy achieves the best performance and achieves better accuracy outperforming previous studies.

Latar Belakang: Penggerakan kader posyandu merupakan bentuk implementasi pilar ke-3 penanganan stunting yaitu konvergensi, koordinasi dan konsolidasi program nasional, daerah dan masyarakat. Peran kader yang baik dapat mempengaruhi status gizi balita karena mampu menjadi motivator dan edukator. Melalui analisis SWOT penggerakan masyarakat melalui kader menjadi strategi tepat yang dapat digunakan dalam penurunan stunting meskipun dalam pelaksanaannya mengalami berbagai hambatan baik dari internal kader maupun eksternal kader. Tujuan: mengeksplorasi peran kader, faktor pendukung, faktor penghambat dan strategi yang dilakukan selama menjalankan tugas dalam program penurunan stunting di wilayah kerja Puskesmas Wori. Metode: jenis penelitian kualitatif dengan pendekatan studi kasus di wilayah kerja Puskesmas Wori. teknik pemilihan informan menggunakan purposive sampling yaitu sesuai dengan kriteria inklusi dan kriteria eksklusi yang ditetapkan peneliti dengan jumlah informan utama sebanyak 8 dan informan tambahan 7 orang. Teknik pengumpulan data menggunakan wawancara mendalam, Focus Group Disscussion (FGD) dan observasi. Instrumen penelitian menggunakan pedoman wawancara, alat dokumentasi dan alat perekam/ voice recorder. Penggolahan data mengguanak open code. Hasil: Hasil analisis menunjukkan bahwa peran kader untuk menjalankan tugasnya dalam penurunan stunting dipengaruhi beberapa faktor predisposing, penguat dan penghambat. Faktor predisposing terdiri motivasi dan keterampilan kader dalam membantu menurunkan angka stunting. Faktor penguat karena adanya dukungan dari puskesmas dan desa dengan pelibatan di pelatihan maupun kegiatan lain sehingga program penurunan stunting semakin maksimal. Dalam menjalankan tugas, kader juga mengalami hambatan dalam penerimaan di masyarakat. Ibu dengan balita stunting sulit menerima informasi dari kader karena faktor budaya dan ibu juga sulit untuk dilibatkan dalam kegiatan posyandu namun faktor tersebut dapat diatasi dengan keterampilan kader dalam berkomunikasi. Kesimpulan: Peran kader sangat penting dan strategis dalam mendukung program puskesmas untuk menurunkan stunting. Dukungan yang besar baik material maupun non material membuat kader terus bersemangat dalam menjalankan tugas. Pentingnya pelatihan secara rutin dan refreshing mampu meningkatkan kemampuan dan pengetahuan mereka dalam menghadapi masyarakat.

Kegiatan pengabdian kepada masyarakat (PkM) Pelatihan Metode Ilmiah dan Analisis Statistika untuk siswa SMKS Arjuna ini merupakan bagian dari upaya untuk menumbuhkan bakat, minat dan kemampuan meneliti siswa. Hal yang melatarbelakangi kegiatan ini adalah sedikitnya pemenang lomba karya ilmiah dari SMK di Sumatera Utara. SMK Swasta Arjuna merupakan salah satu SMK di sekitar Institut Teknologi Del yang membuka jurusan farmasi dan kesehatan, yang bekerja sama dengan Pemerintahan kabupaten Toba untuk pengembangan obat herbal. Pelaksanaan pelatihan ini telah berjalan lancar dengan baik. Meskipun siswa belum dapat menulis topik sesuai jurusan mereka, namun pelatihan ini telah mampu memberikan pemahaman dasar menulis karya ilmiah dan analisis statistika. Siswa telah mampu mengangkat topik di sekitar mereka dan menuliskannya ke dalam karya ilmiah serta mempresentasikan dan mempertanggungjawabkan hasil karyanya di depan juri. Berdasarkan hasil kuesioner, peserta menyampaikan bahwa pelatihan ini bermanfaat karena memberikan pemahaman serta motivasi untuk menulis karya ilmiah.

ABSTRAK Verifikasi kekerabatan berbasis citra wajah merupakan salah satu penerapan sistem Artificial Intelligence yang berguna dalam kehidupan, misalnya untuk penyelidikan kriminal, analisis silsilah, dan lainnya. Perancangan sistem pengenalan wajah pada verifikasi kekerabatan dapat dilakukan menggunakan salah satu algoritma deep learning yaitu metode Convolutional Neural Network. Penelitian ini dilakukan dengan tujuan untuk mengetahui kinerja dari 3D ResNet-18 pada verifikasi kekerabatan berdasarkan sistem pengenalan wajah dan mengetahui kinerja 3D ResNet-18 saat menggunakan embedded system secara real time. Hasil penelitian kinerja ResNet-18 tanpa embedded system memperoleh nilai akurasi training sebesar 0,9771 menggunakan optimizer RMSprop dengan epoch 30 dan batch size 25. Pada pengujian kinerja real time ResNet-18, optimizer SGD berhasil pada ukuran batch size 10, 15, dan 25. Namun untuk pengujian pada perangkat Jetson Nano, optimizer RMSprop gagal akibat ukuran model yang terlalu besar. Kata kunci: embedded sistem, CNN, 3D Resnet18, RMSprop, kekerabatan  ABSTRACT Face-based kinship verification is one of the applications of artificial intelligence systems that are useful in various aspects of life, such as criminal investigations, pedigree analysis, and more. The design of a face recognition system for kinship verification can be done using one of the deep learning algorithms, namely the convolutional neural network method. This research was conducted with the aim of determining the performance of 3D ResNet-18 in kinship verification based on face recognition systems and assessing the performance of 3D ResNet-18 when using an embedded system in real time. The results of the ResNet-18 performance research without an embedded system obtained a training accuracy of 0.9771 using the RMSprop optimizer with 30 epochs and a batch size of 25. In real-time performance testing of ResNet-18, the SGD optimizer succeeded with batch sizes of 10, 15, and 25. However, during testing on the Jetson Nano device, the RMSprop optimizer failed due to the size of the model being too large. Keywords: Embedded System, CNN, 3D Resnet-18, RMSprop, Kinship

This study aims to find out how the efforts of PAI teachers in improving students' social deviant behavior at SMP Negeri 22 Padang, the first is what are the forms of social deviant behavior of students at SMP Negeri 22 Padang, what are the causal factors in students' social deviant behavior at SMP Negeri 22 Padang , and how the efforts of PAI teachers in improving students' social deviant behavior at SMP Negeri 22 Padang. This research is a research using qualitative methods, with a case study approach. Sources of data from this study were three informants through interviews consisting of PAI teachers, BK teachers, and students at SMP Negeri 22 Padang. The data collection techniques of this research are observation, interviews, and documentation. The data analysis technique used is data reduction, data presentation and conclusion drawing. The data validation technique is by using source triangulation. The results of this study indicate that the efforts of PAI teachers in improving social deviant behavior at SMP Negeri 22 Padang are: First: related to the form of social deviant behavior of students at SMP Negeri 22 Padang, namely: a) being rude to teachers, b) saying rudely, c) carrying cellphones going to school, d) not collecting PAI assignments, e) skipping school. Second, there are three factors that cause students' social deviant behavior, namely: a) family factors, b) friend factors. c) community environmental factors. Third, regarding the efforts of PAI teachers in improving students' social deviant behavior, there are three, namely: a) preventive efforts, b) repressive efforts, and c) curative efforts.

Kegemukan merupakan masalah yang serius di dunia, karena terus meningkat di semua negara sejak tahun 1980 - 2014 yaitu 5% - 11% pada pria dan 8% - 15% pada wanita, serta menyebabkan 3,4 juta orang meninggal dunia setiap tahunnya. Sementara di Indonesia kegemukan pada anak usia sekolah tahun 2018 masih tinggi yaitu 11,2% gemuk dan 4,8% obesitas pada usia 13-15 tahun, serta 9,5% gemuk dan 4% obesitas pada usia 16-18 tahun. Tinjauan literatur sistematis ini bertujuan untuk mengkonfirmasi hasil penelitian sebelumnya dan menentukan tindakan apa yang perlu dilakukan khususnya mengenai kegemukan pada remaja. Metode kompilasi literatur menggunakan tinjauan sistematis, diperoleh dari database PubMed.gov, The BMJ, Cambridge.Core dan Google Scholar yang tersedia online dan diakses pada 25 November 2019 – 25 Januari 2020, yang berupa penelitian asli, diterbitkan sejak Desember 2014 – Desember 2019 tersedia teks secara lengkap, dengan kata kunci "Aktivitas Fisik dan Kegemukan Pada Remaja", intervensi dan/atau observasi, pada remaja laki-laki dan perempuan, rentang usia dalam kategori anak dan remaja. Setelah dilakukan pencarian dan skrining dengan kriteria inklusi dan eksklusi, diperoleh total 145 jurnal penelitian yang sesuai hingga pada akhir sejumlah 19 penelitian dianalisis secara sistematis, dengan tambahan 11 literatur dari Kemenkes RI, WHO dan beberapa penelitian terkait, sehingga total menjadi 30 literatur. Hasil tinjauan literatur ini menunjukkan bahwa anak-anak yang aktif secara fisik memiliki tingkat kebugaran jasmani dan status gizi yang lebih baik. Maka hendaknya anak-anak dan remaja diberikan lebih banyak kesempatan untuk melakukan aktivitas fisik, karena hal itu akan meningkatkan kebugaran jasmani dan menjaga status gizi anak di sekolah, sehingga secara sinergis dapat menekan tingginya angka kegemukan pada anak usia sekolah.

This study aimed to describe the efforts made by teachers in fulfilling the children’s rights and preventing of violence against children through child-friendly school policy in the Islamic elementary schools. This research was qualitative descriptive research, and the research subjects were of teachers and principals of 7 several Islamic elementary schools located in Lampung Province (Metro, East Lampung, Pringsewu, and Tanggamus). The sampling technique was purposive random sampling, while the data were collection through interviews, observation, documentation, and literature study. The results of this research showed that: 1) providing protection2) providing consultation rooms, counseling guidance to parents and students, 3) doing fun activities 4) providing facilities that ensure the safety of learners 5) socializing, opening communication 6) providing good example 7) supervision, and mentoring 8) create regulations that guarantee the rights and safety of learners both physical and psychological, 9) attach posters about child-friendly education, and 10) Give a admonition to teachers who commit acts of Violence toward students. The implementation of child-friendly learning in madrasas during the covid-19 pandemic has been carried out well online, offline, and a home visit.

Background: Population growth in Indonesia remains high and itconstitutes a population problem which requires the enactment of population policy, namely by reducing the growth rate as low as possible through the family planning programs. 3.8% of women of childbearing age (WCA) in Indonesia still choose to use the traditional family planning methods, including the use of natural ingredients or herbal medicine as the traditional contraception, the use of coitus interruptus method, and the use of calendar method which is lower in effectiveness than modern contraceptive methods. Purpose: The purpose of this study is to review determinants of traditional contraceptive use among women. Methods: Four databases were used to organize this scoping review, namely PubMed, JSTOR, Science Direct, and Google Scholar. These databases wereconsidered to be relevant on condition thatthe scoping review research questions could be answered through the search ofarticles using keywords. To obtain the relevant articles, the inclusion and exclusion criteria were set, then the articles were selected through the PRISMA flow chart. After that, a critical appraisal was conducted to assess the quality of each article, the articles were further condensed using data charting, an evidence-based analysis was compiled, and the results were reported. Results: This study resulted in thefinding of 9 theme-related articles whose research characteristics consisted of grade A for 6 articles, grade B for 1 article, which all employed cross sectional research method, and grade Good for 2 qualitative design articles.These articles were obtained from developing countries. The limitation of this study was that not all factors contained in the collection of articles were discussed by the researchersdue to the lack of comparison journals which matched the theme. Conclusion: The results of scoping review from the titledeterminants of traditional contraceptive use among womenrevealed that there were several related factors including age, education level, economic status, parity, husband’s support, culture and information systems. Based on the 9 articles in this scoping review, the research results on factors related to the use of traditional family planningmethods were supported. However, not all factors such as sexual behavior, smoking, religious prohibitions, health problems and service quality could be studied in depth due to the limited comparison journals. Therefore, the researchers expect that further research can cover all factors related to the use of traditional family planning methods in order to develop knowledge concerning the traditional contraceptive use, and it is expected that the results of this studycanserve the material for government policy making in future family planning programs.

Indonesia is an archipelagic country that has a very wide sea area. Thus, Indonesian sea has a huge potential of natural resources that can be utilized to grow the nation's economy. There are many occupations and efforts that can be done to increase the income from the sea and also to conserve it. Fishery is one of the most effective way to gain the sea resources; however, fishery is limited by the weather condition on the sea. This is also a problem that happened in Puger Beach. Puger Beach is located in the south Jember and it faces the Hindia Ocean, which means the weather condition is more dangerous for fishermen than other part of coastal. To ensure the safety of the fishermen, the weather condition on the sea must be evaluated and predicted before the fishery. This study designed a system to provide fishermen in Puger Beach an information about sea and beach weather condition which consist of wave height prediction, wind speed, temperature, humidity and weather prediction. The wind speed is obtained from self-designed anemometer system, the temperature is measured using LM35 sensor, and the humidity is assessed using DHT22. The wave height in the sea was predicted by calculating the wind speed value and effective average fetch value using neural network algorithm. The weather on the sea and on the beach were predicted by rain and light sensor. This weather prediction would be classified into three different results, namely raining, cloudy and bright. After some experiments, the result showed that the device can provide the information needed for fishermen and it has a high sensing accuracy. The humidity measurement had an average error of 1.1%, the temperature measurement had 1.42% average error, and 2.37% for the wind speed measurement. The wave height measurement system worked out and found the average wave height in Puger Beach 0.37 meters.

Grouper (Epinephelus sp.) is a reef fish commodity that are currently utilized in a large enough scale. Of the types of grouper, tiger grouper has advantages compared to other types. This fish hight economic value because it has a delicious meat, nutritious and contain unsaturated fatty acids. Hazard Analysis and Critical Control Point (HACCP) is a systematic approach to food safety management based on the principles there that aims to identify the hazards that tend to occur at every step of the food chain. One of the principles of HACCP is to determine the CCP using Decision Tree Diagram CCP (Decision Tree). Field Work Practice was conducted at PT. Alam Jaya, District Rungkut, Surabaya City, East Java Province on 18 January to 26 February , 2016. The working method that used in Field Work Practice is descriptive method with data collection included primary data and secondary data. Data is collected by observation, interview and active participation. CCP determination based on the presence of danger in a process that can not be removed in the next process. Determination of CCP based on the observation of every production process is known that the process of receiving raw materials and metal detectors are CCP.

Abstract Background Barriers to receiving optimal healthcare exist for Indigenous populations globally for a range of reasons. To overcome such barriers and enable greater access to basic and specialist care, developments in information and communication technologies are being applied. The focus of this scoping review is on web-based therapeutic interventions (WBTI) that aim to provide guidance, support and treatment for health problems. Objectives This review identifies and describes international scientific evidence on WBTI used by Indigenous peoples in Australia, New Zealand, Canada and USA for managing and treating a broad range of health conditions. Eligibility criteria Studies assessing WBTI designed for Indigenous peoples in Australia, Canada, USA and New Zealand, that were published in English, in peer-reviewed literature, from 2006 to 2018 (inclusive), were considered for inclusion in the review. Studies were considered if more than 50% of participants were Indigenous, or if results were reported separately for Indigenous participants. Sources of evidence Following a four-step search strategy in consultation with a research librarian, 12 databases were searched with a view to finding both published and unpublished studies. Charting methods Data was extracted, synthesised and reported under four main conceptual categories: (1) types of WBTI used, (2) community uptake of WBTI, (3) factors that impact on uptake and (4) conclusions and recommendations for practice. Results A total of 31 studies met the inclusion criteria. The WBTI used were interactive websites, screening and assessment tools, management and monitoring tools, gamified avatar-based psychological therapy and decision support tools. Other sources reported the use of mobile apps, multimedia messaging or a mixture of intervention tools. Most sources reported moderate uptake and improved health outcomes for Indigenous people. Suggestions to improve uptake included as follows: tailoring content and presentation formats to be culturally relevant and appropriate, customisable and easy to use. Conclusions Culturally appropriate, evidence-based WBTI have the potential to improve health, overcome treatment barriers and reduce inequalities for Indigenous communities. Access to WBTI, alongside appropriate training, allows health care workers to better support their Indigenous clients. Developing WBTI in partnership with Indigenous communities ensures that these interventions are accepted and promoted by the communities.

Tujuan penelitian ini yakni untuk melihat upaya menumbuhkan solidaritas kebangsaan anakusia dini melalui permainan bakiak. Metode penelitian ini menggunakan Jenis penelitian penelitian deskriptif kualitatif. Dengan subjek penelitian anak-anak yang tinggal di desa Lembah Kecamatan Dolopo Kabupaten Madiun. Data penelitian ini terdiri dari data primer dan data skunder. Data primer merupakan data yang dihasilkan dari wawancara, observasi. Sedangkan data skunder merupakan data yang dihasilkan dari dokumen. Instrumen yang digunakan yakni merupakan peneliti sendiri dan memakai tahapan obsrvasi dan lembar pedoman wawancara yang dimana pada lembar wawancara tersebut merupakan pedoman peneliti untuk mengetahui pendapat dari orang tua anak usia dini yang dilaksanakan di desa lembah kecamatan dolopo kabupaten madiun Berdasarkan hasil penelitian maka dapat disimpulkan permainan Bakiak pada Anak Usia Dini di desa lembah kecematan dolopo kabupaten madiun, dapat menumbuhkan rasa solidaritas kebangsaan pada diri Anak Usia Dini yang tercermin dari kemampuan anak usia dini dalam membangun kebersamaan, rasa simpati, rasa empati, kekompakan dan mewujudkan tujuan atau kepentingan bersama.

This study aims to examine the prediction of rainfall and river water debit using the Back Propagation Neural Network (BP-NN) method. Prediction results are classified using the Support Vector Machine (SVM) method to predict flooding. The parameters used to predict rainfall with BP-NN are minimum, maximum and average temperature, average relative humidity, sunshine duration, and average wind speed. The debit of Ular Pulau Tagor river is predicted by BP-NN. BPNN and SVM modeling using software R. Daily climate data from 2015-2017 were taken from three stations, namely Sampali climatology station, Kualanamu meteorological station, and Tuntung geophysics station. Prediction of river water debit is for 6 days and 30 days in the future. The best dataset is a 6 day prediction with a combination of 60% training and 40% testing. Flood prediction accuracy with SVM was 100% in predicting flood events for the next 6 days.

Background: Infectious diseases such as diarrhea and Acute Respiratory Infection (ARI) lead to loss of appetite in children and stunting growth. This study analyzes the relationship between the incidence of stunting and the frequency/duration of diarrhea and ARI in children under five years. Design and Methods: The stratified random sampling method was used to obtain data from 152 children in 4 villages in Surabaya, East Java, Indonesia. Results: The results showed that children under the age of five, experience higher stunting duration and longer frequency of diarrhea. The Rho Spearmen Test showed differences in the incidence of stunting with the frequency of diarrhea P = 0.005 (P

Quality of Service merupakan metode pengukuran tentang seberapa baik jaringan yang terpasang dan juga merupakan suatu usaha untuk mendefinisikan karakteristik dan sifat dari satu layanan. Dengan dibuatnya sistem pembayaran online yang terdapat di PT. PLN (Persero) Jember, layanan internet yang digunakan hendaknya harus memenuhi standar TIPHON (Telecommunications and Internet Protocol Harmonization Over Networks). Maka diperlukan optimasi kinerja QoS sebagai salah satu cara untuk mengetahui seberapa besar kualitas layanan data yang harus dipenuhi. Parameter QoS yang digunakan untuk analisis layanan komunikasi data adalah jitter, packet loss, throughtput, dan delay. Dari hasil analisis data menunjukan bahwa pada jam sibuk (09.00-11.00 WIB) dan non sibuk (11.00-13.00 WIB) mendapatkan hasil rata – rata indeks QoS sebesar 2,125 dalam kategori “kurang memuaskan”. Dengan kapasitas bandwidth yang disediakan sebesar 3 Mbps. Kemudian dari hasil perhitungan optimasi bandwidth yang diperlukan sebesar 7,154 Mbps dan disimulasikan mendapatkan rata–rata indeks QoS yang sebesar 3,5 dalam kategori “sangat memuaskan”.

Penelitian ini bertujuan untuk: (1) mengetahui karakteristik modul fisika berbasis masalah pada materi listrik dinamis, (2) mengetahui peningkatan kemampuan berpikir kreatif siswa setelah mengikuti pembelajaran menggunakan modul fisika berbasis masalah pada materi listrik dinamis. Jenis penelitian ini adalah research and development yang menggunakan model 4-D meliputi: define, design, develop, dan disseminate, untuk mengembangkan modul fisika berbasis masalah. Teknik pengumpulan data dimulai dengan tahap pra penelitian; analisis siswa, kurikulum, dan materi; tujuan pengembangan modul; pemilihan format berdasarkan kriteria modul; desain awal modul, validasi draf I, uji coba kecil, dan uji coba di kelas. Instrumen yang digunakan adalah angket untuk validasi modul, silabus, RPP, kisi-kisi soal kemampuan berpikir kreatif, serta angket respon siswa dan guru. Pengembangan modul dinilai berdasarkan kelayakan isi, penyajian, dan bahasa oleh 2 dosen, 2 guru, dan 3 peer review. Hasil penelitian ini adalah (1) kualitas modul fisika berbasis masalah pada materi listrik dinamis yang dikembangkan berkategori baik, dan (2) kemampuan berpikir kreatif siswa setelah mengikuti proses pembelajaran menggunakan modul fisika berbasis masalah pada materi listrik dinamis mengalami peningkatan.

Rodentibacter spp. are opportunistic pathogens that are often isolated from the upper respiratory tracts of laboratory rodents. In particular, R. pneumotropicus and R. heylii require considerable caution in rodent colonies, as they cause lethal pneumonia in rodents. A new species, R. haemolyticus, has recently been classified in the genus, and a very closely related strain, Rodentibacter sp. strain JRC, has been isolated in Japan. This study focused on strain JRC by performing genomic and pathogenic analyses. Draft genome sequencing of strain JRC identified several genes coding for putative virulent proteins, including hemolysin and adhesin. Furthermore, we found a new RTX (repeats-in-structural toxin) toxin gene in the genome, which was predicted to produce a critical virulence factor (RTXIA) similar to Enterobacteriaceae. The concentrated culture supernatant containing RTX toxin (RTXIA) showed cytotoxicity toward RAW264.7 cells. Pre-incubation with anti-CD11a attenuated the cytolysis, suggesting that the concentrated culture supernatant containing RTXIA is cell surface LFA-1 mediated cytolysin. Experimental infection of strain JRC intranasally with 5 female BALB/c-Rag2 -/- mice showed 60% lethality and was not significantly different from those of R. pneumotropicus ATCC 35149 T using the log-rank test. Combined with our finding that RTXIA has an almost identical amino acid sequence (98% identity) to that of R. haemolyticus 1625/19 T , these results strongly suggest that RTXIA-producing strain JRC (and related R. haemolyticus) is pathogenic to immunodeficient rodents, and both agents should be excluded in laboratory rodent colonies., (Copyright © 2022. Published by Elsevier B.V.)

The RIKEN BioResource Research Center (BRC) was established in 2001 as a comprehensive biological resource center in Japan. The Experimental Animal Division, one of the BRC infrastructure divisions, has been designated as the core facility for mouse resources within the National BioResource Project (NBRP) by the Japanese government since FY2002. Our activities regarding the collection, preservation, quality control, and distribution of mouse resources have been supported by the research community, including evaluations and guidance on advancing social and research needs, as well as the operations and future direction of the BRC. Expenditure for collection, preservation, and quality-control operations of the BRC, as a national core facility, has been funded by the government, while distribution has been separately funded by users' reimbursement fees. We have collected over 9000 strains created mainly by Japanese scientists including Nobel laureates and researchers in cutting-edge fields and distributed mice to 7000 scientists with 1500 organizations in Japan and globally. Our users have published 1000 outstanding papers and a few dozen patents. The collected mouse resources are accessible via the RIKEN BRC website, with a revised version of the searchable online catalog. In addition, to enhance the visibility of useful strains, we have launched web corners designated as the "Mouse of the Month" and "Today's Tool and Model." Only high-demand strains are maintained in live colonies, while other strains are cryopreserved as embryos or sperm to achieve cost-effective management. Since 2007, the RIKEN BRC has built up a back-up facility in the RIKEN Harima branch to protect the deposited strains from disasters. Our mice have been distributed with high quality through the application of strict microbial and genetic quality control programs that cover a globally accepted pathogens list and mutated alleles generated by various methods. Added value features, such as information about users' publications, standardized phenotyping data, and genome sequences of the collected strains, are important to facilitate the use of our resources. We have added and disseminated such information in collaboration with the NBRP Information Center and the NBRP Genome Information Upgrading Program. The RIKEN BRC has participated in international mouse resource networks such as the International Mouse Strain Resource, International Mouse Phenotyping Consortium, and Asian Mouse Mutagenesis and Resource Association to facilitate the worldwide use of high-quality mouse resources, and as a consequence it contributes to reproducible life science studies and innovation around the globe., (© 2021. The Author(s).)

Background: Cilia-associated respiratory bacillus (CARB; now known as Filobacterium rodentium gen. nov., sp. nov.) is a primary pathogen of rodents. A CARB-like organism was reported in post-mortem lung samples of cats using light and electron microscopy. Here we explore by molecular procedures if a Filobacterium sp. is a part of the normal feline lower respiratory microbiome and whether it could in some cats contribute to the development of chronic bronchial disease., Methodology: A Filobacterium sp. was identified in three Czech cats clinically diagnosed as having chronic neutrophilic bronchitis. Bronchoalveolar lavage fluid (BALF) specimens obtained from these cats were subjected to panbacterial 16S rDNA PCR followed by Sanger sequencing of the V5 to V8 region. After these cats were treated with specific antimicrobials, their clinical signs resolved promptly, without recurrence. Next, BALF specimens from 13 Australian and 11 Italian cats with lower respiratory disease and an additional 16 lung samples of Italian cats who died of various causes were examined using next generation sequencing (NGS). Subsequently, a Filobacterium-specific qPCR assay was developed and used to re-test BALF specimens from the 11 Italian cats and lung tissue homogenates from the additional 16 deceased cats., Principal Findings: An amplicon of 548 bp with 91.24% sequence agreement with Filobacterium rodentium was obtained from all three patients, suggesting the novel Filobacterium sp. was the cause of their lower respiratory disease. The novel Filobacterium sp., which we propose to call F. felis, was detected in 3/3 Czech cats with chronic neutrophilic bronchitis, 13/13 Australian cats and 6/11 Italian cats with chronic lower respiratory disease, and 14/16 necropsy lung specimens from Italian cats. NGS and qPCR results all showed identical sequences. The Filobacterium sp. was sometimes the preponderant bacterial species in BALF specimens from cats with lower airway disease. There was an association between the presence of large numbers (greater than 105 organisms/mL) of Filobacterium and the presence of neutrophilic and/or histiocytic inflammation, although only a subset of inflammatory BALF specimens had F. felis as the preponderant organism., Conclusion: The novel Filobacterium sp. comprises a finite part of the normal feline lower respiratory microbiome. Under certain circumstances it can increase in absolute and relative abundance and give rise to neutrophilic and/or histiocytic bronchitis, bronchiolitis and bronchopneumonia. These findings strongly suggest that F. felis could be an underdiagnosed cause of feline bronchial disease., Competing Interests: The authors have declared that no competing interests exist.

Strain SMR-CT, which was originally isolated from rats as the SMR strain, had been named 'cilia-associated respiratory bacillus' ('CAR bacillus'). 'CAR bacillus' was a Gram-stain-negative, filamentous argentophilic bacterium without flagella. SMR-CT grew at 37 °C under microaerobic conditions, showed gliding activity, hydrolysed urea and induced chronic respiratory diseases in rodents. The dominant cellular fatty acids detected were iso-C15 : 0 and anteiso-C15 : 0. The DNA G+C content was 47.7 mol%. 16S rRNA gene sequence analysis revealed SMR-CT and other strains of 'CAR bacillus' isolated from rodents all belonged to the phylum Bacteroidetes. The nearest known type strain, with 86 % 16S rRNA gene sequence similarity, was Chitinophaga pinensis DSM 2588T in the family Chitinophagaceae. Strain SMR-CT and closely related strains of 'CAR bacillus' rodent-isolates formed a novel family-level clade in the phylum Bacteroidetes with high bootstrap support (98-100 %). Based on these results, we propose a novel family, Filobacteriaceae fam. nov., in the order Sphingobacteriales as well as a novel genus and species, Filobacterium rodentium gen. nov., sp. nov., for strain SMR-CT. The type strain is SMR-CT ( = JCM 19453T = DSM 100392T).

Bordetella bronchiseptica is a pathogenic bacterium causing respiratory infections in a broad range of mammals. Recently, we determined the whole genome sequence of B. bronchiseptica S798 strain isolated from a pig infected with atrophic rhinitis and found four single-nucleotide polymorphisms (SNPs) at positions -129, -72, +22, and +38 in the region upstream of dnt encoding dermonecrotic toxin (DNT), when compared with a rabbit isolate, RB50. DNT is known to be involved in turbinate atrophy observed in atrophic rhinitis. Immunoblotting, quantitative real-time PCR, and β-galactosidase reporter assay revealed that these SNPs resulted in the increased promoter activity of dnt and conferred the increased ability to produce DNT on the bacteria. Similar or identical SNPs were also found in other pig isolates kept in our laboratory, all of which produce a larger amount of DNT than RB50. Our analysis revealed that substitution of at least two of the four bases, at positions -72 and +22, influenced the promoter activity for dnt. These results imply that these SNPs are involved in the pathogenicity of bordetellae specific to pig diseases.

Murine norovirus (MNV) has considerable genetical and biological diversity and is recognized worldwide as the most common contaminant in laboratory mouse colonies. This study developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method with the potential to detect a broad range of MNV. RT-LAMP, using a set of five primers containing mixed bases, obtained results under isothermal conditions at 62°C for 90min. Sensitivity of RT-LAMP was 50-fold less than that of two-step TaqMan real-time reverse transcription-polymerase chain reaction (TaqMan RT-PCR). Diagnostic performance of RT-LAMP on RNA extracted from mouse fecal specimens was compared with TaqMan RT-PCR and nested RT-PCR. MNV was detected in 54 of 120 mouse fecal specimens by RT-LAMP, and RT-LAMP had an estimated sensitivity and specificity of 96.4% and 100% compared with TaqMan RT-PCR, and 94.7% and 100% compared with nested RT-PCR. RT-LAMP, which does not require expensive instruments, might be useful for the screening of mice actively or persistently infected with MNV., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A one-step SYBR Green I real-time RT-PCR assay was developed for the detection and quantification of a broad range of murine noroviruses (MNVs). The primer design was based on the multiple sequence alignments of 101 sequences of the open reading frame (ORF)1-ORF2 junction of MNV. The broad reactivity and quantitative capacity of the assay were validated using 7 MNV plasmids. The assay was completed within 1 h, and the reliable detection limit was 10 copies of MNV plasmid or 0.063 median tissue culture infective doses per milliliter of RAW264 cell culture-propagated viruses. The diagnostic performance of the assay was evaluated using 158 mouse fecal samples, 91 of which were confirmed to be positive. The melting curve analysis demonstrated the diversity of MNV in the samples. This is the first report of a broadly reactive one-step SYBR Green I real-time RT-PCR assay for detecting of MNVs. The rapid and sensitive performance of this assay makes it a powerful tool for diagnostic applications.

Mouse hepatitis virus (MHV) is one of the most prevalent viruses detected in laboratory mouse colonies. Enterotropic strains predominate in natural infections, and molecular techniques for the detection of MHV shedding in feces are powerful enough to diagnose active infections. A reverse transcription-loop-mediated isothermal amplification (RT-LAMP) technique was developed for the detection of rodent coronaviruses within 90 min. The specificity of this technique was confirmed by its ability to detect all 17 different strains of MHV and 6 strains of rat coronaviruses as well as its failure to detect human, bovine, and porcine coronaviruses nonspecifically. The sensitivity of RT-LAMP was 3.2-fold higher than that of reverse transcription-polymerase chain reaction (RT-PCR) and 31.6-fold lower than that of nested RT-PCR. An evaluation of the diagnostic performance of RT-LAMP performed in duplicate using mouse fecal specimens showed that the sensitivity and specificity with respect to nested RT-PCR were 85.7% and 100%, respectively. RT-LAMP assays would be suitable for monitoring active MHV infection in mouse colonies., (Copyright © 2012 Elsevier B.V. All rights reserved.)

To evaluate murine norovirus (MNV) infection in laboratory mice, we attempted to develop an enzyme-linked immunosorbent assay (ELISA) system and an indirect immunofluorescent antibody (IFA) assay for detecting the anti-MNV-S7 antibody in mice. MNV-S7, which was isolated in Japan, was used in both assays. The antigen for ELISA was prepared by ultracentrifugation of culture supernatants of RAW 264 cells infected with MNV-S7. Positive sera were obtained from 6-week-old, female C57BL/6JJcl mice inoculated orally with MNV-S7. IFA against infected RAW 264 cells was able to discriminate positive sera from negative sera. Indirect ELISA was performed using 96-well ELISA plates coated with formalin-treated MNV-S7 antigen. In this ELISA system, mouse sera obtained 2 weeks after infection or later showed significantly high OD values and were judged positive. An equal level of anti-MNV-S7 antibody response was observed in BALB/cAJcl, C57BL/6JJcl, DBA/2JJcl, and Jcl:ICR mice; whereas, C3H/HeJJcl mice demonstrated slightly lower antibody production 4 weeks after infection. We also used this ELISA system to evaluate 77 murine serum samples obtained from 15 conventional mouse rooms in research facilities in Japan and found that approximately half of the serum samples contained antibody to MNV-S7. We found that some serum samples were negative for antibodies to mouse hepatitis virus and Mycoplasma pulmonis but positive for antibody to MNV-S7. The results suggest that the MNV infection is more prevalent than other infections such as mouse hepatitis virus and Mycoplasma pulmonis in conventional mouse colonies in Japan, as is the case in other areas of the world.

Mice are one of the most important model organisms for studying biological phenomena and diseases processes in life sciences. The biomedical research community has succeeded in launching large scale strategic knockout mouse projects around the world. RIKEN BRC, a comprehensive government funded biological resource center was established in 2001. RIKEN BRC has been acting as the core facility for the mouse resources of the National BioResource Project (NBRP) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan since 2002. RIKEN BRC is a founding member of the Federation of International Mouse Resources (FIMRe) together with the Jackson Laboratory, the European Mouse Mutant Archive, and other centers, and has participated in the International Mouse Strain Resource (IMSR) to distribute mouse strains worldwide. With the support of the scientific community, RIKEN BRC has collected over 3,800 strains including inbred, transgenic, knockout, wild-derived, and ENU-induced mutant strains. Excellent mouse models for human diseases and gene functions from academic organizations and private companies are distributed through RIKEN BRC. To meet research and social needs, our mice will be rederived to a specific pathogen-free state, strictly monitored for their health, and accurately tested for their genetic modifications and backgrounds. Users can easily access our mouse resources through the internet and obtain the mouse strains for a minimal fee. Cryopreservation of embryos and sperm is used for efficient preservation of the increasing number of mouse resources. RIKEN BRC collaborates with FIMRe members to support Japanese scientists in the use of valuable mouse resources from around the world.

Embryonic germ-line cells are unipotent cells that give rise to either sperm or oocytes. However, pluripotent stem cells can be derived from primordial germ cells (PGCs) or spermatogonia, suggesting that germ-line cells retain a capacity for pluripotency. Here, we made genome-wide comparisons of the gene expression profiles of freshly isolated PGCs, in vitro-formed PGCs (iPGCs), and other stem cell lines, including embryonic stem cells (ESCs), embryonic germ cells (EGCs) and germ-line stem (GS) cells. Comparing PGC with ESC, 382 genes/transcripts were significantly up-regulated in ESC, while 188 were elevated in PGC. This suggests that PGCs possess transcription program distinct from that of ESC, although both share expression of many pluripotency-associated genes. Our micro-array analysis showed that the analyzed samples could be classified into two groups: one consisting of all the ESCs and most of EGCs, and the other containing PGC samples, iPGC, one type of female EGC and GS cells. We then identified "signature" genes for the two groups, and used them to characterize GS cells, EGC, and iPGCs, and revealed developmental status of each cell type. The relationships between PGCs and stem cells derived from embryos or germ cells are discussed in light of these findings.

An enzyme-linked immunosorbent assay (ELISA) was developed to detect the antibody against lymphocytic choriomeningitis virus (LCMV) in sera of laboratory animals. In this ELISA system, LCMV-nucleoprotein (NP) expressed by recombinant baculovirus and purified with high molar urea was used as the antigen. Sera from laboratory animals experimentally infected with the Armstrong strain or the newly isolated M1 strain of LCMV were examined to detect anti-LCMV antibody by the ELISA system, and the reactivity was compared with that of IFA test. Regardless of LCMV strain, all the sera of adult mice infected with LCMV were positive with very high optical density (OD). Also, the sera from mice neonatally infected with LCMV M1 strain were positive with slightly lower OD than adult mice. In contrast, all the sera of uninfected mice were negative to LCMV-NP antigen. Similarly, anti-LCMV antibodies were detected in all the sera of hamsters, mastomyses, and gerbils infected with the LCMV Armstrong strain. The results of the ELISA were in complete agreement with those of IFA, and indicate the high sensitivity and specificity of the ELISA system in the detection of anti-LCMV antibody. Because this ELISA system does not require handling infectious LCMV in the course of the antigen preparation and serological assay, there is no risk of contamination in the laboratory or nearby animal facility. In addition, by using negative control antigen in parallel with positive antigen in ELISA, we can exactly check the LCMV contamination in laboratory animals.

Human acute myeloblastic leukemia HL-60 cells become resistant to differentiation during long-term cultivation. After 150 passages, double minute chromosomes (dmins) found in early-passaged cells are replaced by large extrachromosomal elements (LEEs). In a DNA library derived from a purified fraction of LEEs, 12.6% (23/183) of clones were assigned to 8q24 and 9.2% (17/183) were assigned to 14q11 in the human genome. Fluorescence in situ hybridization (FISH) revealed a small aberrant chromosome, which had not been found in early-passaged cells, in addition to the purified LEEs. We determined that each LEE consisted of six discontinuous segments in a region that extended for 4.4Mb over the 8q24 locus. Five genes, namely, Myc (a proto-oncogene), NSMCE2 (for a SUMO ligase), CCDC26 (for a retinoic acid-dependent modulator of myeloid differentiation), TRIB1 (for a regulator of MAPK kinase) and LOC389637 (for a protein of unknown function), were encoded by the amplicon. Breaks in the chromosomal DNA within the amplicon were found in the NSMCE2 and CCDC26 genes. The discontinuous structure of the amplicon unit of the LEEs was identical with that of dmins in HL-60 early-passaged cells. The difference between them seemed, predominantly, to be the number (10-15 copies per LEE versus 2 or 3 copies per dmin) of constituent units. Expression of the Myc, NSMCE2, CCDC26 and LOC389637 and TRIB1 genes was constitutive in all lines of HL-60 cells and that of the first four genes was repressed during the terminal differentiation of early-passaged HL-60 cells. We also detected abnormal transcripts of CCDC26. Our results suggest that these genes were selected during the development of amplicons. They might be amplified and, sometimes, truncated to contribute to the maintenance of HL-60 cells in an undifferentiated state.

Persistent LCMV infection in wild-derived MAI/Pas mice housed under conventional conditions remained undetected for a decade, despite periodic health monitoring using dirty-bedding sentinels. When MAI/Pas mice were rederived by embryo transfer, recipient mothers produced antiLCMV antibodies, which first revealed the presence of the virus in the colony. Before this information was obtained, MAI/Pas mice had been shipped to another facility, undergone cesarean rederivation there, and been introduced into the recipient barrier. The foster mothers of rederived pups were LCMV-negative according to enzyme-linked immunosorbent assay, but sera of both cesarean-rederived MAI/Pas mice and their foster mothers were positive for LCMV infection by immunofluorescent assay (IFA). LCMV was isolated from the MAI/Pas mice, and its genomic RNA was sequenced. Examination of animal technicians in contact with LCMV-infected mice and of other mouse samples by IFA or a reverse transcriptase-polymerase chain reaction test (or both) revealed that neither the workers nor other animals had been infected with LCMV. Experimental data showed that LCMV transmission from persistently infected mice to naïve ones occurred only after direct contact of animals housed in the same cage. This experience demonstrates the importance of careful viral monitoring in the transfer of laboratory rodents between institutions, the limitation of dirty-bedding sentinels for detection of LCMV infection, and the inadequacy of cesarean rederivation for elimination of enzootic LCMV infection. 111

Mice without the gene for tenascin-C, a multifunctional extracellular matrix protein expressed in many important biological events, including wound healing, did not show any phenotype. However, it is now obvious that the phenotype of deletion of one gene frequently depends on the genetic background. Therefore, we have newly generated tenascin-C knockout mice (KO) by backcrossing original KO into three congenic lines: C57BL/6N, BALB/cA, and GRS/A (GR). And we investigated the disease course of reversible kidney injury, Habu-snake venom-induced proliferative glomerulonephritis. In all strains, the disease was more severe in KO, but the severity varied with the strain. The KO-GR showed irreversibility; all treated KO-GR died by the 4th month due to renal failure. The diseased KO-GR showed abnormal regenerative reactions, including reduced proliferation of mesangial cells, key players in glomerulonephritis, and reduced production of some kinds of cytokines and matrices, leading to poor formation of granulation tissue. In culture, the mesangial cells from the KO-GR had the same potential for proliferation and response to cytokines as controls, but interestingly, to achieve this potential, they required contact with tenascin-C. These reactions were blocked by an anti-tenascin monoclonal antibody. The results of the present study, the first report showing the most dramatic phenotype so far discovered, have strongly suggested the importance of tenascin-C in the resolution of the renal inflammation and that of the genetic background on which the KO was developed.

CD4 and CD8 double-positive (DP) T cells were transiently found in the Peyer's patch of mice 2 to 3 weeks after birth and preceded the appearance of DP intestinal intraepithelial lymphocytes (iIEL). DP iIEL were detected in euthymic-specific pathogen-free mice, but not in athymic and germ-free mice. The pattern of V beta genes expressed on these DP iIEL was similar to that of CD4 single-positive peripheral T cells, which reflected the negative selection by internal superantigens, M1s, and I-E. CD8 SP T cells did not receive such a strict negative selection with these antigens. DP iIEL expressed a high density of T cell receptor alpha beta and CD3 and responded to immobilized anti-CD3 to undergo blastogenesis and de novo IL2 receptor expression. These results indicate that DP iIEL are mature thymus-dependent T cells immigrated to intraepithelial space via the Peyer's patch in response to intestinal antigenic stimulation.

Rabbit mammary gland PRL receptors in the microsome fraction were solubilized with the zwitterionic detergent Chaps, and were separated into two fractions (Fr. A and B) by ion-exchange chromatography. The number of receptors in Fr. B was about 2.2 times greater than in Fr. A. In sucrose gradient centrifugation analysis, PRL receptors in Fr. A and Fr. B sedimented at different positions. After binding 125I-PRL, the apparent molecular weight (mol wt) of the PRL receptor in Fr. A changed from 42,400 to 65,500 and that in Fr. B changed from 89,400 to 108,000, suggesting that each binding subunit interacts with one PRL molecule. Cross-linking 125I-PRL to receptors revealed little change following SDS-PAGE, in the autoradiogram patterns of the microsome PRL receptors, either in the presence or absence of dithiothreitol. Both the microsome and the Chaps extract contained two major binding subunits (mol wt, 83,200 and 36,800) and one minor subunit (mol wt, 20,800). The mol wt of the dominant PRL receptors in Fr. A and Fr. B were 36,800 and 83,200, respectively. The latter form did not dissociate into a 36,800 mol wt form, suggesting that the rabbit mammary gland contains two independent binding subunits with mol wt of 36,800 and 83,200. Data showed that PRL receptors in the rabbit mammary gland are mostly the high Kd type receptor with a mol wt of 83,200.