bluuebunny/crossref_metadata_2025 huggingface dataset:
PENGARUH LAMA WAKTU FERMENTASI TERHADAP YIELD DAN SIFAT ORGANOLEPTIK VIRGIN COCONUT OIL (VCO) YANG DIHASILKAN DARI KELAPA DAERAH BALI
Authors: Nurida Ulfa , Lusiani Cucuk Evi
Published: August 2021

Bali adalah salah satu daerah di Indonesia yang berpotensi menghasilkan kelapa dalam jumlah yang banyak.  Kelapa daerah Bali memiliki ukuran buah yang besar, bentuk buah bulat telur dan kadar minyak kopra  mencapai 69,28%. Buah kelapa tersebut dapat dijadikan berbagai macam produk olahan, salah satunya yaitu  minyak kelapa murni atau virgin coconut oil (VCO) yang dibuat dengan metode fermentasi. Proses fermentasi  dapat dilakukan dengan alat sederhana, waktu yang efisien, dan produk dengan tingkat ketengikan yang  rendah dan daya simpan yang lama. Tujuan dari penelitian ini adalah untuk mengetahui pengaruh lama waktu  fermentasi terhadap yield dan sifat organoleptik (aroma, rasa dan warna) produk VCO yang dihasilkan dari  kelapa daerah Bali. Proses fermentasi dilakukan menggunakan penambahan ragi tempe dengan konsentrasi 2% b/v selama 6, 12, 18, 24 dan 30 jam. Produk VCO yang dihasilkan pada penelitian ini dianalisis untuk  perhitungan nilai yield, sifat organoleptik, dan nilai pH. Nilai yield produk VCO mengalami peningkatan seiring  dengan lamanya waktu fermentasi dengan nilai tertinggi yaitu 4,30% v/v pada waktu fermentasi 30 jam. Semua  produk VCO yang dihasilkan dalam penelitian ini memiliki aroma khas kelapa dan tidak tengik, rasa khas minyak  kelapa, berwarna bening kekuningan dan nilai pH 5. Dengan demikian, produk VCO yang dihasilkan memenuhi  syarat standar mutu VCO, yaitu Standar Nasional Indonesia (SNI) 7381:2008.
STUDI LITERATUR PERBANDINGAN PRODUKSI CRUDE SELULASE DARI BAHAN BERLIGNOSELULOSA UNTUK PEMBUATAN BIOETANOL
Authors: Hasanah Nikmatul , Nalaway Ifan Nida Nusha , Rulianah Sri
Published: August 2021

Bahan berlignoselulosa yaitu biomassa dari tanaman yang memiliki komponen utama selulosa dan hemiselulosa.  Karena memiliki kandungan selulosa yang cukup tinggi, maka bahan berlignoselulosa dapat dimanfaatkan  sebagai bahan baku produksi crude selulase dengan bantuan kapang. Crude selulase dapat diaplikasikan dalam  pembuatan bioetanol. Tujuan studi literatur ini adalah untuk membandingkan produksi crude selulase dari bahan  berlignoselulosa dengan kapang. Selain itu, juga bertujuan untuk membandingkan pembuatan bioetanol dari  bahan berlignoselulosa menggunakan metode Simultaneous Saccharification and Fermentation (SSF) dengan  hidrolisis enzimatis. Berdasarkan studi literatur yang telah dilakukan, dapat disimpulkan bahwa adanya pengaruh  kandungan lignoselulosa bahan, waktu inkubasi, dan konsentrasi substrat terhadap aktivitas enzim yang  dihasilkan. Nilai aktivitas enzim paling tinggi yaitu ditunjukkan pada produksi crude selulase dari ampas tebu  menggunakan kapang Phanerochaete chrysosporium dengan kondisi terbaik yaitu waktu inkubasi 17 hari dan  konsentrasi substrat 7% sebesar 91,304 U/mL. Studi literatur mengenai pembuatan bioetanol dapat disimpulkan  bahwa penambahan crude selulase dan waktu inkubasi berpengaruh terhadap kadar etanol yang dihasilkan.  Kadar etanol tertinggi sebesar 11,04% dari bagasse menggunakan crude selulase Phanerochaete chrysosporium pada penambahan 50% dan waktu fermentasi 144 jam.
ANALISIS EKONOMI PRA-RANCANGAN PABRIK HAND SANITIZER DAUN SIRIH DENGAN KAPASITAS PRODUKSI 480 TON/TAHUN
Authors: Pradana Wulandari Novi , Suharti Profiyanti Hermien
Published: August 2021

Hand sanitizer adalah cairan antiseptik untuk membersihkan atau menghilangkan kuman pada tangan. Peran  antiseptik dalam hand sanitizer biasanya bergantung pada komposisi alkohol. Alkohol membuat tangan menjadi  kering serta menimbulkan dehidrasi pada kulit. Penggunaan alkohol sebagai antiseptik dapat diganti dengan  bahan-bahan alami. Bahan alami yang digunakan harus memiliki sifat antibakteri seperti daun sirih serta jeruk  nipis. Daun sirih (Piper betle Linn) sangat terkenal dalam kehidupan warga di Indonesia. Daun sirih bermanfaat  untuk kesehatan gigi serta kerap digunakan selaku obat kumur, menghilangkan bau tubuh serta mulut, mengobati  sariawan, mimisan, gatal- gatal, koreng serta keputihan pada perempuan. Daun sirih memiliki senyawa antibakteri  yang terdiri dari senyawa fenol serta turunannya. Daun sirih memiliki berbagai kandungan kimia yaitu minyak  atsiri. Proses pembuatan hand sanitizer ini diawali dengan pembuatan ekstrak daun sirih menggunakan metode  soxhlet. Hasil ekstrak daun sirih ditambahkan bahan lainnya yaitu etanol, na-cmc, natrium metabisulfit, dan  gliserin dalam rangka menghasilkan hand sanitizer berbentuk gel. Perhitungan analisis ekonomi pabrik ini  didasarkan pada rencana pendirian pabrik di Rembang pada tahun 2024 untuk hasil dari perhitungan didapatkan  nilai Return on Investment (ROI) saat sebelum pajak sebesar 90% serta Return on Investment (ROI) setelah pajak  sebesar 54%. Nilai Pay Out Time (POT) sehabis pajak sebesar 1,8 tahun dan sebelum pajak sebesar 1,1 tahun.  Break Even Poin (BEP) sebesar Rp. 99.064.027.413,39 serta Shut Down Point (SDP) sebesar Rp. 28.773.654.292,14.  Berdasarkan hasil analisis yang sudah dilakukan, pra-perancangan pabrik hand sanitizer menggunakan daun sirih  dengan formulasi na-cmc dan gliserin dengan kapasitas 480 ton/ tahun layak untuk didirikan.
EVALUASI FOULING FAKTOR TERHADAP KINERJA HEAT EXCHANGER PADA GAS COOLER UNIT CO2 LIQUID PLANT
Authors: Prasasti Tri Endang , Udjiana S. Sigit , Muharram Yuliman
Published: August 2021

Alat penukar kalor atau yang biasa disebut dengan heat exchanger adalah suatu peralatan yang dapat digunakan  untuk menukar kalor dari suatu fluida ke fluida lain yang punya perbedaan suhu, penukaran kalor tersebut dapat  terjadi dengan kontak langsung atau secara kontak tidak langsung. Dilihat dari fungsinya yang sangat penting,  maka alat ini digunakan secara luas dalam dunia perindustrian. Ada banyak macam heat exchanger seperti  contohnya Plate Heat Exchanger (PHE). Plate Heat Exchanger (PHE) merupakan salah satu jenis dari alat penukar  kalor (Heat Exchanger) yang terdiri dari pelat dan rangka, PHE mempunyai proses perpindahan kalor yang terjadi  diantara kedua fluida pada sisi-sisi pelat. Suatu alat penukar kalor akan sangat mempengaruhi keberhasilan  dalam keseluruhan rangkaian proses industri, karena jika ada kegagalan operasi, baik kegagalan mekanik maupun  operasional maka dapat berakibat berhentinya operasi unit dalam suatu industri. Maka dari itu sebuah alat  penukar kalor (Heat Exchanger) harus memiliki kinerja yang sangat baik sehingga mendapatkan hasil yang  maksimal dan dapat bekerja secara penuh terhadap suatu unit yang sedang beroperasi. Salah satu faktor baik  tidaknya suatu heat exchanger yaitu dengan melihat nilai fouling factor. Fouling adalah fenomena menempel  dan menumpuknya abu pada dinding penghantar panas yang dipasang tepat di lingkungan dimana suhu gas pada  bagian belakang furnace lebih rendah dibandingkan suhu untuk melunak abu. Hasil dari evaluasi ini adalah tetap mempertahankan nilai fouling 0,0003.
EFEK LAMA WAKTU FERMENTASI TERHADAP YIELD VIRGIN COCONUT OIL (VCO) DARI KELAPA DAERAH MALANG DENGAN KONSENTRASI RAGI 2% B/V
Authors: Jannah Ainiyah Fithriyatul , Lusiani Cucuk Evi
Published: August 2021

Virgin Coconut Oil (VCO) merupakan salah satu produk olahan dari daging buah kelapa tua yang diolah tanpa  melalui pemanasan sehingga tidak merusak komponen penting yang terkandung dalam VCO. Buah kelapa untuk  pembuatan VCO dapat diperoleh dari daerah Malang yang memiliki ketebalan daging buah ± 1 cm dengan  kandungan minyak sebesar 30-35%. Pembuatan VCO dapat dilakukan dengan beberapa metode salah satunya  yaitu metode fermentasi yang menghasilkan VCO dengan kemurnian tinggi sebesar 53,2% b/b. Penelitian ini  bertujuan untuk mengetahui efek lama waktu fermentasi terhadap yield, pH, warna, aroma, dan rasa dari produk  VCO. Metode fermentasi dalam penelitian ini dilakukan menggunakan ragi tempe selama 6, 12, 18, 24, 30 jam. Berdasarkan hasil analisis produk VCO, dapat diketahui bahwa semakin lama waktu fermentasi menghasilkan nilai  yield yang juga semakin tinggi. Nilai yield tertinggi diperoleh pada waktu fermentasi 30 jam sebesar 16,00% v/v.  Produk VCO yang dihasilkan pada waktu fermentasi 6 sampai 30 jam memiliki nilai pH yang sama yaitu 6 dengan  sifat fisik yang sesuai dengan syarat mutu Standar Nasional Indonesia (SNI) 7381:2008 yaitu memiliki warna  (transparan) dan jernih, aroma khas kelapa segar, dan rasa khas minyak kelapa.
PEMANFAATAN KOAGULAN ALAMI DARI CAMPURAN BIJI TREMBESI DAN KITOSAN PADA PENGOLAHAN LIMBAH PENYAMAKAN KULIT
Authors: Nurismasari Shilma Ananta , Hardjono Hardjono
Published: August 2021

Industri penyamakan kulit merupakan industri yang menghasilkan air limbah dengan kategori B3 yang akan  berbahaya jika dibuang langsung ke sungai. Karena pada limbah tersebut mengandung unsur Krom (Cr) yang  berasal dari proses tanning (penyamakan). Untuk mengurangi kandungan logam Cr pada limbah industri dapat  menggunakan koagulan alami yang merupakan salah satu alternatif dalam pengolahan limbah karena koagulan  alami lebih ramah lingkungan dan mudah didapatkan disekitar lingkungan kita. Penelitian ini bertujuan untuk  menurunkan kadar krom, TSS, TDS, Turbiditas dan pH pada limbah penyamakan kulit menggunakan koagulan  alami yaitu campuran dari biji trembesi dan kitosan. Metode yang digunakan yaitu koagulasi-flokulasi dengan  alat jar test dengan perbandingan variasi koagulan alami campuran biji trembesi dan kitosan sebesar 50%:50%,  25%:75%, 75%:25%, 100%:0, 0:100% dengan kecepatan pengadukan cepat 200 Rpm selama 2 menit dan  dilanjutkan pengadukan lambat dengan kecepatan 30, 60, dan 100 Rpm selama 30 menit. Berdasarkan  penelitian yang dilakukan didapatkan dosis koagulan alami campuran yang efektif dalam menurunkan kadar  krom yaitu koagulan trembesi 75% + kitosan 25% dengan penurunan kadar krom mencapai 1,479 mg/L atau  sebesar 82,445% dan memiliki penurunan nilai TSS sebesar 96,825%, TDS sebesar 9131 mg/L dan nilai turbiditas  mencapai 37,789%.
PERHITUNGAN NERACA MASSA PADA STRIPPER (DA-101) DI PABRIK UREA DEPARTEMEN PRODUKSI I A PT PETROKIMIA GRESIK
Authors: Prastika Dhevita Tara Dwi , Qurrota’Ayuni Dini , Chumaidi Achmad , Matmulyanto Matmulyanto
Published: August 2021

PT Petrokimia Gresik merupakan salah satu perusahaan pupuk terbesar yang ada di Indonesia. PT Petrokimia  Gresik mengoperasikan 23 pabrik yang terdiri dari 16 pabrik yang memproduksi pupuk serta 7 pabrik yang  memproduksi produk non-pupuk. Pada proses produksi urea terdapat alat yang bernama stripper. Stripper  adalah alat yang digunakan untuk memisahkan satu komponen atau lebih dari campurannya menggunakan  prinsip perbedaan kelarutan. Departemen Produksi I A Urea memiliki kapasitas produksi sebesar 460.000  ton/tahun dan konsentrasi produk minimal 98,4%. Pada penelitian ini, dilakukan perhitungan neraca massa  untuk mengevaluasi perhitungan neraca massa stripper pabrik urea PT Petrokimia Gresik. Perhitungan neraca  massa yaitu berdasarkan banyaknya bahan yang masuk serta keluar dari sistem. Untuk menghitung neraca  massa harus diketahui terlebih dahulu data bahan baku yang masuk dalam bentuk laju mol dan %mol. Hasil  perhitungan neraca massa yang dilakukan pada alat stripper diperoleh total neraca massa input sama dengan  total neraca massa output (balance) sebesar 232403 kg/jam.
STUDI AKTIVASI LIMBAH CANGKANG KERANG SEBAGAI KATALIS RAMAH LINGKUNGAN DALAM PEMBUATAN BIODESEL
Authors: Rohmana Galuh Citra Cahya , Sudarminto Hadi Priya , Damayanti Sasmita Eka
Published: August 2021

Biodiesel merupakan salah satu energi alternatif pengganti bahan bakar fosil dari hasil reaksi transesterifikasi  minyak nabati dengan metanol. Dalam pembuatan biodiesel umumnya menggunakan katalis homogen  seperti NaOH atau KOH. Seperti yang dapat kita pahami, bahan tersebut menimbulkan produk samping  berupa gliserol dan rumitnya pemisahan produk biodiesel dengan katalis. Disamping itu potensi gangguan  kesehatan akibat kontak dengan katalis bisa membahayakan. Maka, mulai dikembangkan katalis heterogen.  Suatu proses yang dapat menjadi solusi dari masalah tersebut adalah diberlakukannya limbah cangkang  kerang sebagai katalis ramah lingkungan. Keberadaan cangkang kerang di sepanjang Pelabuhan Kota  Pasuruan yang saat ini menjadi masalah dapat diatasi dengan aktivasi limbah tersebut menjadi sebuah  katalis. Tujuan dari penelitian ini untuk menganalisis biodiesel hasil aktivasi limbah cangkang kerang,  membandingkan kualitas dan kuantitas biodiesel. Membandingkan perbedaan hasil biodiesel menggunakan  katalis homogen NaOH/KOH dan katalis heterogen cangkang kerang, Mengetahui pengaruh katalis cangkang  kerang terhadap % konversi yield dari produk biodiesel. Dari hasil penelitian ini, dihasilkan % konversi  biodiesel dengan katalis limbah cangkang kerang sebesar 0,403 gram biodiesel/gram mol dan yield dari  proses sebesar 0,596 gram biodiesel/gram minyak. % konversi biodiesel dengan katalis homogen KOH sebesar 0,424 gram biodiesel/gram mol dan yield dari proses sebesar 0,525 gram biodiesel/gram minyak dengan hasil lebih besar menggunakan KOH.
PENGARUH WAKTU PEMANASAN TERHADAP RENDEMEN MINYAK KELAPA PADA METODE BASAH
Authors: Firdana Khoridho Putra , Dewi Ernia Novika
Published: August 2021

Minyak yang diekstrak dari kelapa dapat menjadi bahan baku dalam pembuatan minyak goreng, margarin, selai,  mentega, biodisel dan kosmetik. Dari sisi ekonomi, minyak kelapa memiliki nilai ekonomi yang yang cukup tinggi  karena permintaan di pasaran terus meningkat. Metode pembuatan minyak kelapa yang umum digunakan  adalah metode basah dan metode kering. Metode basah lebih dipilih karena lebih ekonomis dalam hal  penggunaan alat. Disamping itu metode ini lebih singkat karena tanpa pengeringan kelapa menjadi kopra.  Tujuan dari penelitian ini adalah untuk mengetahui pengaruh waktu pemanasan terhadap rendemen minyak  kelapa yang dihasilkan pada metode basah. Pembuatan minyak kelapa dengan metode basah diawali dengan  pemisahan daging kelapa dari kulitnya, pencucian, pengecilan ukuran sampai didapatkan santan (coconut milk).  Santan kemudian dilakukan pemanasan pada suhu 90 C disertai pengadukan selama variabel waktu 100, 120,  140, 180 sampai dihasilkan minyak kelapa. Hasil penelitian menunjukkan bahwa rendemen terbesar yaitu 22,22 % diperoleh pada waktu pemanasan 120 menit.
Pengembangan Sistem Pakar Diagnosis Penyaki Kelinci dengan Menggunakan Metode Case Based Reasoning
Authors: Luqman Affandi  , Mamluatul Hani'ah  , Nita Komalasari
Published: August 2021

Kelinci merupakan salah satu binatang yang tidak lepas dari ancaman serangan penyakit yang apabila dibiarkan akan berdampak buruk bagi kesehatannya. Faktor yang dapat membuat kelinci sering kali terkena penyakit yaitu kondisi kebersihan kandangnya dan dari makanannya. Beberapa penyakit yang dapat menyerang kelinci diantaranya diare, scabies, dan hairball. Selain itu, jarang peternak dan pemelihara kelinci yang kurang memiliki waktu untuk melakukan pemeriksaan langsung ke dokter hewan. Kelinci yang sakit namun tidak segera diberikan penangan, akan membuat penyakitnya menjadi lebih parah bahkan bisa sampai menimbulkan kematian. Dengan adanya masalah tersebut maka dibuatlah pengembangan sistem pakar diagnosis penyakit kelinci dengan metode CBR. Sedangkan metode Case Based Reasoning atau CBR merupakan metode pemecahan masalah yang dalam mencari solusi dari suatu kasus yang baru, sistem akan melakukan pencarian terhadap solusi dari kasus lama. Dalam metode ini terdapat 4 proses yaitu retrieve, reuse, revise dan retain. Penelitian pengembangan sistem pakar diagnosis penyakit kelinci menggunakan metode Case Based Reasoning bertujuan agar dapat membantu memberikan penanganan pertama pada penyakit yang diderita oleh kelinci berdasarkan kesamaan gejala yang ada dan mengetahui penanganan pertama dalam menanggulangi penyakit pada kelinci dengan cara menginputkan gejala-gejala penyakit agar diketahui penyakitnya dan penanganannya. Hasil dari sistem ini berupa diagnosis penyakit dan solusi utama yang diperlukan sesuai dari inputan gejala yang dipilih user. Dalam penelitian ini menghasilkan akurasi kesesuain antara hasil dari pakar dan aplikasi sebesar 83%.
PENERAPAN ALGORITME FP-GROWTH UNTUK MENENTUKAN PELETAKAN BARANG PEDAGANG SAYUR
Authors: Wahyu Alfafisabil  , Dermawan Budi Arif , Padilah Tesa Nur
Published: August 2021

Sayuran merupakan sumber vitamin dan protein. Setiap ibu rumah tangga membutuhkan sayuran untuk memasak dalam kehidupan sehari-hari. Sayuran didapatkan di pasar sehingga akan mempersulit ibu rumah tangga yang rumahnya jauh dari pasar. Pedagang sayur keliling merupakan pedagang yang menjual berbagai macam sayuran yang dibawa ke rumah-rumah untuk memenuhi kebutuhan ibu rumah tangga. Pedagang sayur keliling bertujuan untuk mencari keuntungan, sehingga untuk memaksimalkan tingkat penjualan diperlukan strategi penjualan. Association rules adalah metode untuk mencari hubungan antar item pada suatu dataset. Data mining dapat disebut salah satu langkah dari proses KDD. FP-Growth merupakan algoritme untuk mencari himpunan data yang paling sering muncul. Penelitian ini menganalisis data transaksi untuk memprediksi peletakan barang dipedagang sayur dengan tujuan memaksimalkan tingkat penjualan menggunakan algoritme FP-Growth dan bahasa pemrograman python. Pada proses data mining dengan menggunakan algoritme FP-Growth peneliti menjelaskan langkah-langkah FP-Growth dengan perhitungan manual. Evaluasi peneliti melakukan pencocokan hasil perhitungan manual dengan program. Setelah perhitungan sesuai, peneliti menggunakan data tota transaksi untuk mengetahui rules-nya dengan syarat minimum support 0.01 atau 1% dan minimum confidence 0.9 atau 90%. Pada hasil terdapat 44 rules yang memenuhi syarat.
Implementasi GraphQL Untuk Mengatasi Under-Fetching pada Pengembangan Sistem Informasi Pelacakan Alumni Politeknik Negeri
Authors: Firdausi Annisa Taufika , Hormansyah Dhebys Suryani , Fany Ervansyah
Published: August 2021

Pendistribusian data pada sistem informasi memiliki berbagai macam cara yang digunakan. Diantaranya adalah dengan menggunakan metode REST API. Namun, terdapat beberapa kekurangan yang menjadi masalah pada REST API. Salahsatunya adalah masalah under-fetching, yaitu masalah dimana bagian frontend harus melakukan lebih dari 1 kali request untuk memenuhi kebutuhan data yang diperlukan. Masalah ini dapat diselesaikan dengan menerapkan GraphQL sebagai metode pendistribusian data. Tujuan dari penelitian kali ini adalah untuk membandingkan performa antara sistem informasi dengan REST API dan sistem informasi dengan GraphQL. Studi kasus pada penelitian ini adalah pada pengembangan sistem informasi pelacakan alumni Politeknik Negeri Malang. Hasil penelitian ini berupa perbandingan performa antara sistem informasi dengan metode pendistribusian data REST API dan sistem informasi dengan metode pendistribusian data GraphQL. GraphQL menunjukkan performa yang baik pada jumlah data yang besar dan kompleks, serta ketika terdapat banyak pengguna yang mengakses data dalam waktu yang bersamaan. Sedangkan untuk data yang sederhana dan sistem informasi yang tidak memiliki banyak pengguna yang akan mengakses data secara bersamaan, maka REST API masih lebih unggul.
A New Pipeline to Automatically Segment and Semi-Automatically Measure Bone Length on 3D Models Obtained by Computed Tomography
Authors: Beltran Diaz Santiago , H’ng Chee Ho , Qu Xinli , Doube Michael , Nguyen John Tan , de Veer Michael , Panagiotopoulou Olga , Rosello-Diez Alberto
Published: August 2021

The characterization of developmental phenotypes often relies on the accurate linear measurement of structures that are small and require laborious preparation. This is tedious and prone to errors, especially when repeated for the multiple replicates that are required for statistical analysis, or when multiple distinct structures have to be analyzed. To address this issue, we have developed a pipeline for characterization of long-bone length using X-ray microtomography (XMT) scans. The pipeline involves semi-automated algorithms for automatic thresholding and fast interactive isolation and 3D-model generation of the main limb bones, using either the open-source ImageJ plugin BoneJ or the commercial Mimics Innovation Suite package. The tests showed the appropriate combination of scanning conditions and analysis parameters yields fast and comparable length results, highly correlated with the measurements obtained via ex vivo skeletal preparations. Moreover, since XMT is not destructive, the samples can be used afterward for histology or other applications. Our new pipelines will help developmental biologists and evolutionary researchers to achieve fast, reproducible and non-destructive length measurement of bone samples from multiple animal species.
Real-Time Induction of Macrophage Apoptosis, Pyroptosis, and Necroptosis by Enterococcus faecalis OG1RF and Two Root Canal Isolated Strains
Authors: Chi Danlu , Lin Xinwei , Meng Qingzhen , Tan Jiali , Gong Qimei , Tong Zhongchun
Published: August 2021

To investigate the effects of two Enterococcus faecalis root canal isolated strains (CA1 and CA2) and of the OG1RF strain on apoptosis, pyroptosis, and necroptosis in macrophages. The virulence factors of E. faecalis CA1 and CA2 pathogenic strains were annotated in the Virulence Factors Database (VFDB). E. faecalis CA1, CA2, and OG1RF strains were used to infect RAW264.7 macrophages (MOI, 100:1). We assessed the viability of intracellular and extracellular bacteria and of macrophages at 2, 6, and 12 h post-infection. We used a live cell imaging analysis system to obtain a dynamic curve of cell death after infection by each of the three E. faecalis strains. At 6 and 12 h post-infection, we quantified the mRNA expression levels of PANoptosis-related genes and proteins by RT-qPCR and western blot, respectively. We identified ultrastructural changes in RAW264.7 cells infected with E. faecalis OG1RF using transmission electron microscopy. We found 145 and 160 virulence factors in the CA1 and CA2 strains, respectively. The extracellular CA1 strains grew faster than the CA2 and OG1RF strains, and the amount of intracellular viable bacteria in the OG1RF group was highest at 6 and 12 h post-infection. The macrophages in the CA1 infection group were the first to reach the maximum PI-positivity in the cell death time point curve. We found the expressions of mRNA expression of caspase-1, GSDMD, caspase-3, MLKL, RIPK3, NLRP3, IL-1β and IL-18 and of proteins cleaved caspase-1, GSDMD, cleaved caspase-3 and pMIKL in the macrophages of the three infection groups to be upregulated (P<0.05). We detected ultrastructural changes of apoptosis, pyroptosis, and necroptosis in macrophages infected with E. faecalis. The three E. faecalis strains induced varying degrees of apoptosis, pyroptosis, and necroptosis that were probably associated with PANoptosis in macrophages. The E. faecalis CA1 strain exhibited faster growth and a higher real-time MOI, and it induced higher expression levels of some PANoptosis-related genes and proteins in the infected macrophages than the other strains tested.
A Predictive Maintenance Model for Flexible Manufacturing in the Context of Industry 4.0
Authors: Sang Go Muan , Xu Lai , de Vrieze Paul
Published: August 2021

The Industry 4.0 paradigm is the focus of modern manufacturing system design. The integration of cutting-edge technologies such as the Internet of things, cyber–physical systems, big data analytics, and cloud computing requires a flexible platform supporting the effective optimization of manufacturing-related processes, e.g., predictive maintenance. Existing predictive maintenance studies generally focus on either a predictive model without considering the maintenance decisions or maintenance optimizations based on the degradation models of the known system. To address this, we propose PMMI 4.0, a Predictive Maintenance Model for Industry 4.0, which utilizes a newly proposed solution PMS4MMC for supporting an optimized maintenance schedule plan for multiple machine components driven by a data-driven LSTM model for RUL (remaining useful life) estimation. The effectiveness of the proposed solution is demonstrated using a real-world industrial case with related data. The results showed the validity and applicability of this work.
Vitamin D3 Attenuates Viral-Induced Inflammation and Fibrotic Responses in Bronchial Smooth Muscle Cells
Authors: Plesa Maria , Gaudet Mellissa , Mogas Andrea , Jalaleddine Nour , Halayko Andrew , Al Heialy Saba , Hamid Qutayba
Published: August 2021

Toll-like receptor 3 (TLR3) activation by viral infections plays a key role in promoting inflammatory immune responses that contribute to pulmonary fibrosis in chronic inflammatory respiratory diseases. Vitamin D3 has been shown to be beneficial to patients with asthma and chronic obstructive pulmonary disease (COPD) through its anti-inflammatory and anti-fibrotic properties. Smooth muscle cells are one of the major contributors to airway remodeling in asthma and COPD. We therefore aimed to investigate the effect of vitamin D3 treatment on viral-induced TLR3 responses in Bronchial Smooth Muscle Cells (BSMCs) as a mechanism contributing to pulmonary fibrosis in asthma and COPD. Primary BSMCs from patients with asthma (n=4), COPD (n=4), and healthy control subjects (n=6) were treated with polyinosinic: polycytidylic acid (polyI:C), TLR3 agonist in the presence or absence of vitamin D3 (1,25D3). Here we report the mRNA expression and protein levels of pro-inflammatory and pro-fibrotic markers (IL-6, IFN-β1, CCL2/MCP-1, fibronectin 1 and type I collagen) among BSMCs groups: asthma, COPD, and healthy controls. We show that at the baseline, prior to polyI:C stimulation, asthma and COPD BSMCs presented increased pro-inflammatory and pro-fibrotic state compared to healthy control subjects, as measured by quantitative PCR and immunoassays (ELISA/Flow Cytometry. Ligation of TLR3 by polyI:C in BSMCs was associated with increased TLR3 mRNA expression, and 1,25D3 treatment significantly reduced its expression. In addition, 1,25D3 decreased the expression of IL-6, IFN-β1, CCL2, FN1 and COL1A1 induced by polyI:C in BSMCs. The regulatory effect of 1,25D3 treatment on polyI:C-stimulated BSMCs was further confirmed at protein levels. Our findings suggest that vitamin D3 attenuates TLR3 agonist-induced inflammatory and fibrotic responses in BSMCs and support the clinical relevance of vitamin D3 supplementation in patients with viral infections having chronic respiratory diseases, such as asthma and COPD.
Antigen Specific Regulatory T Cells in Kidney Transplantation and Other Tolerance Settings
Authors: Hu Min , Rogers Natasha M. , Li Jennifer , Zhang Geoff Y. , Wang Yuan Min , Shaw Karli , O’Connell Philip J. , Alexander Stephen I.
Published: August 2021

Kidney transplantation is the most common solid organ transplant and the best current therapy for end-stage kidney failure. However, with standard immunosuppression, most transplants develop chronic dysfunction or fail, much of which is due to chronic immune injury. Tregs are a subset of T cells involved in limiting immune activation and preventing autoimmune disease. These cells offer the potential to provide tolerance or to allow reduction in immunosuppression in kidney transplants. The importance of Tregs in kidney transplantation has been shown in a number of seminal mouse and animal studies, including those with T cell receptors (TCRs) transgenic Tregs (TCR-Tregs) or Chimeric Antigen Receptor (CAR) Tregs (CAR-Tregs) showing that specificity increases the potency of Treg function. Here we outline the animal and human studies and clinical trials directed at using Tregs in kidney transplantation and other tolerance settings and the various modifications to enhance allo-specific Treg functionin vivoandin vitro.
COVID-19 Immunobiology: Lessons Learned, New Questions Arise
Authors: Kaklamanos Aimilios , Belogiannis Konstantinos , Skendros Panagiotis , Gorgoulis Vassilis G. , Vlachoyiannopoulos Panayiotis G. , Tzioufas Athanasios G.
Published: August 2021

There is strong evidence that COVID-19 pathophysiology is mainly driven by a spatiotemporal immune deregulation. Both its phenotypic heterogeneity, spanning from asymptomatic to severe disease/death, and its associated mortality, are dictated by and linked to maladaptive innate and adaptive immune responses against SARS-CoV-2, the etiologic factor of the disease. Deregulated interferon and cytokine responses, with the contribution of immune and cellular stress-response mediators (like cellular senescence or uncontrolled inflammatory cell death), result in innate and adaptive immune system malfunction, endothelial activation and inflammation (endothelitis), as well as immunothrombosis (with enhanced platelet activation, NET production/release and complement hyper-activation). All these factors play key roles in the development of severe COVID-19. Interestingly, another consequence of this immune deregulation, is the production of autoantibodies and the subsequent development of autoimmune phenomena observed in some COVID-19 patients with severe disease. These new aspects of the disease that are now emerging (like autoimmunity and cellular senescence), could offer us new opportunities in the field of disease prevention and treatment. Simultaneously, lessons already learned from the immunobiology of COVID-19 could offer new insights, not only for this disease, but also for a variety of chronic inflammatory responses observed in autoimmune and (auto)inflammatory diseases.
Scoring System Based on RNA Modification Writer-Related Genes to Predict Overall Survival and Therapeutic Response in Bladder Cancer
Authors: Zhang Pu , Liu Zijian , Wang Decai , Li Yunxue , Xing Yifei , Xiao Yajun
Published: August 2021

Introduction

It’s widely reported the “writer” enzymes mediated RNA adenosine modifications which is known as a crucial mechanism of epigenetic regulation in development of tumor and the immunologic response in many kinds of cancers. However, the potential roles of these writer genes in the progression of bladder cancer (BLCA) remain unclear.

Materials and Methods

We comprehensively described the alterations of 26 RNA modification writer genes in BLCA from the genetic and transcriptional fields and identified writer-related genes from four independent datasets. Utilizing least absolute shrinkage and selection operator (LASSO) regression and multivariate Cox regression, we constructed a ten writer-related gene signature. After that, we confirmed the predictive and prognostic value of this signature on another six independent datasets and established a nomogram to forecast the overall survival (OS) and mortality odds of BLCA patients clinically.

Results

The writer-related genes signature showed good performance in predicting the OS for BLCA patients. Moreover, the writer-related gene signature was related to EMT-related pathways and immune characteristics. Furthermore, the immune cell infiltration levels of CD8 T cells, cytotoxic cells, M1/2 macrophage cells and tumor mutation burden might be able to predict which patients will benefit from immunotherapy. This could also be reflected by the writer-related gene signature.

Conclusions

This signature might play an important role in precision individualized immunotherapy. The present work highlights the crucial clinical implications of RNA modifications and may help developing individualized therapeutic strategies for patients with BLCA.
Brd4 Regulates the Homeostasis of CD8+ T-Lymphocytes and Their Proliferation in Response to Antigen Stimulation
Authors: Peng Zhilin , Zhang Yiwen , Ma Xiancai , Zhou Mo , Wu Shiyu , Song Zheng , Yuan Yaochang , Chen Yingshi , Li Yuzhuang , Wang Guanwen , Huang Feng , Qiao Yidan , Xia Baijing , Liu Weiwei , Liu Jun , Zhang Xu , He Xin , Pan Ting , Xu Hanshi , Zhang Hui
Published: August 2021

CD8+ T cells are major components of adaptive immunity and confer robust protective cellular immunity, which requires adequate T-cell numbers, targeted migration, and efficient T-cell proliferation. Altered CD8+ T-cell homeostasis and impaired proliferation result in dysfunctional immune response to infection or tumorigenesis. However, intrinsic factors controlling CD8+ T-cell homeostasis and immunity remain largely elusive. Here, we demonstrate the prominent role of Brd4 on CD8+ T cell homeostasis and immune response. By upregulating Myc and GLUT1 expression, Brd4 facilitates glucose uptake and energy production in mitochondria, subsequently supporting naïve CD8+ T-cell survival. Besides, Brd4 promotes the trafficking of naïve CD8+ T cells partially through maintaining the expression of homing receptors (CD62L and LFA-1). Furthermore, Brd4 is required for CD8+ T cell response to antigen stimulation, as Brd4 deficiency leads to a severe defect in clonal expansion and terminal differentiation by decreasing glycolysis. Importantly, as JQ1, a pan-BRD inhibitor, severely dampens CD8+ T-cell immune response, its usage as an anti-tumor agent or latency-reversing agent for human immunodeficiency virus type I (HIV-1) should be more cautious. Collectively, our study identifies a previously-unexpected role of Brd4 in the metabolic regulation of CD8+ T cell-mediated immune surveillance and also provides a potential immunomodulation target.
The Dual Targeting of FcRn and FcγRs via Monomeric Fc Fragments Results in Strong Inhibition of IgG-Dependent Autoimmune Pathologies
Authors: Monnet Céline , Jacque Emilie , de Romeuf Christophe , Fontayne Alexandre , Abache Toufik , Fournier Nathalie , Dupont Gilles , Derache Delphine , Engrand Anais , Bauduin Aurélie , Terrier Aurélie , Seifert Alexander , Beghin Cécile , Longue Alain , Masiello Nicholas , Danino Laetitia , Nogre Michel , Raia Anais , Dhainaut Frederic , Fauconnier Louis , Togbe Dieudonnée , Reitinger Carmen , Nimmerjahn Falk , Stevens Wil , Chtourou Sami , Mondon Philippe
Published: August 2021

Novel molecules that directly target the neonatal Fc receptor (FcRn) and/or Fc gamma receptors (FcγRs) are emerging as promising treatments for immunoglobulin G (IgG)-dependent autoimmune pathologies. Mutated Fc regions and monoclonal antibodies that target FcRn are currently in clinical development and hold promise for reducing the levels of circulating IgG. Additionally, engineered structures containing multimeric Fc regions allow the dual targeting of FcRn and FcγRs; however, their tolerance needs to first be validated in phase I clinical studies. Here, for the first time, we have developed a modified monomeric recombinant Fc optimized for binding to all FcRns and FcγRs without the drawback of possible tolerance associated with FcγR cross-linking. A rational approach using Fc engineering allowed the selection of LFBD192, an Fc with a combination of six mutations that exhibits improved binding to human FcRn and FcγR as well as mouse FcRn and FcγRIV. The potency of LFBD192 was compared with that of intravenous immunoglobulin (IVIg), an FcRn blocker (Fc-MST-HN), and a trimeric Fc that blocks FcRn and/or immune complex-mediated cell activation through FcγR without triggering an immune reaction in several in vitro tests and validated in three mouse models of autoimmune disease.
Polycentricity and Regional Ocean Governance: Implications for the Emerging UN Agreement on Marine Biodiversity Beyond National Jurisdiction
Authors: Gjerde Kristina M. , Yadav Siddharth Shekhar
Published: August 2021

The governance of the two-thirds of the world’s ocean in areas beyond national jurisdiction (the high seas and deep seabed beyond national jurisdiction or ABNJ) is currently fragmented into diverse institutions addressing specific activities, issues or regions. This has hampered the international community’s ability to redress critical issues including biodiversity loss, pollution, climate change, ecosystem degradation and declining fisheries in an integrated and ecosystem-based manner. Our analysis of polycentricity theory and associated enabling conditions shows that the current polycentric approach to marine biodiversity in ABNJ is not yet fully functional: it is missing the two key attributes of: (1) generally applicable rules and norms structuring actions and behaviors, and (2) processes to enhance cooperation, coordination, and conflict resolution. Based on the enabling conditions conducive for achieving “a functional polycentric governance system” identified inCarlisle and Gruby (2019), combined with a prior analysis applying resilience principles for socio-ecological systems to ABNJ (Yadav and Gjerde, 2020), this article suggests seven ways the emerging United Nations agreement on the conservation and sustainable use of marine biodiversity of ABNJ (BBNJ Agreement) could stimulate coordinated and integrated action at both global and regional levels in ABNJ. These include: (1) overarching rules, goals and objectives; (2) formal and informal conflict resolution mechanisms; (3) robust global institutional arrangements; (4) strengthened global, regional and sectoral bodies with shared and overlapping responsibility for biodiversity conservation; (5) strengthened cooperation through integrated ecosystem assessments and strategic action programmes at ecologically meaningful scales that could include areas within and beyond national jurisdiction; (6) learning exchange mechanisms within and across regions; and (7) strengthened regional and national capacities for ecosystem-based management in ABNJ. Taken together, these tools could enhance the resilience of ocean institutions, ecosystems and biodiversity to cope with growing pressures, uncertainty and rapid change in ABNJ.
Potential Role of NEU1 in Hepatocellular Carcinoma: A Study Based on Comprehensive Bioinformatical Analysis
Authors: Wu Zhulin , He Li , Yang Lina , Fang Xuehong , Peng Lisheng
Published: August 2021

Background: Aberrant expression of NEU1 has been identified in many malignancies. Nevertheless, the clinical significance of NEU1 in hepatocellular carcinoma (HCC) has not been fully elucidated.

Methods: In our study, multiple databases, including ONCOMINE, The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), International Cancer Genome Consortium (ICGC), Cancer Cell Line Encyclopedia (CCLE), Human Protein Atlas (HPA), Kaplan-Meier (KM) plotter, MethSurv, Gene Expression Profiling Interactive Analysis (GEPIA), and Metascape, etc., were utilized to investigate the expression, prognostic value, and function of NEU1 in HCC.

Results: ONCOMINE, GEO, and TCGA analyses revealed that NEU1 was more highly expressed in HCC compared to normal tissues. Additionally, the mRNA and protein expression levels of NEU1 were increased in liver cancer cell lines and HCC tissues, respectively. Moreover, a trend toward increased NEU1 expression with advanced stage or grade was found. Furthermore, higher mRNA expression of NEU1 was found to be remarkably correlated with worse survival in HCC patients, and multivariate Cox analysis indicated that high mRNA expression of NEU1 was an independent prognostic factor for poor prognosis of HCC patients. Also, 21 methylated CpGs were found to be significantly related to HCC prognosis. Besides, functional enrichment analyses indicated that high NEU1 expression group had lower levels of B cells, CD8+ T cells, NK cells, and T helper cells, etc. than the low NEU1 expression group, and NEU1 may regulate a variety of tumor-related proteins and pathways, including lysosome, spliceosome, mTOR signaling pathway and so on.

Conclusion: High expression level of NEU1 was positively correlated with unfavorable prognosis of HCC patients, which may be related to the regulation of cancer-associated pathways and the inhibition of immune function by NEU1. Thus, NEU1 could be used as a potential prognostic biomarker and target for HCC.
Prediction of Cognitive Decline Using Heart Rate Fragmentation Analysis: The Multi-Ethnic Study of Atherosclerosis
Authors: Costa Madalena D. , Redline Susan , Hughes Timothy M. , Heckbert Susan R. , Goldberger Ary L.
Published: August 2021

Background: Heart rate fragmentation (HRF), a new non-invasive metric quantifying cardiac neuroautonomic function, is associated with increasing age and cardiovascular disease. Since these are risk factors for cognitive decline and dementia, in the Multi-Ethnic Study of Atherosclerosis (MESA), we investigated whether disrupted cardiac neuroautonomic function, evidenced by increased HRF, would be associated with worse cognitive function assessed concurrently and at a later examination, and with greater cognitive decline.

Methods: HRF was derived from the ECG channel of the polysomnographic recordings obtained in an ancillary study (n = 1,897) conducted in conjunction with MESA exam 5 (2010–2012). Cognitive function was assessed at exam 5 and 6.4 ± 0.5 years later at exam 6 (2016–2018) with tests of global cognitive performance (the Cognitive Abilities Screening Instrument, CASI), processing speed (Digit Symbol Coding, DSC) and working memory (Digit Span). Multivariable regression models were used to quantify the associations between HRF indices and cognitive scores.

Results: The participants’ mean age was 68 ± 9 years (54% female). Higher HRF at baseline was independently associated with lower cognitive scores at both exams 5 and 6. Specifically, in cross-sectional analyses, a one-standard deviation (SD) (13.7%) increase in HRF was associated with a 0.51 (95% CI: 0.17–0.86) points reduction in CASI and a 1.12 (0.34–1.90) points reduction in DSC. Quantitatively similar effects were obtained in longitudinal analyses. A one-SD increase in HRF was associated with a 0.44 (0.03–0.86) and a 1.04 (0.28–1.81) points reduction in CASI and DSC from exams 5 to 6, respectively. HRF added predictive value to the Cardiovascular Risk Factors, Aging, and Incidence of Dementia (CAIDE-APOE-ε4) risk score and to models adjusted for serum concentration of NT-proBNP, an analyte associated with cognitive impairment and dementia.

Conclusion: Increased HRF assessed during sleep was independently associated with diminished cognitive performance (concurrent and future) and with greater cognitive decline. These findings lend support to the links between cardiac neuroautonomic regulation and cognitive function. As a non-invasive, repeatable and inexpensive probe, HRF technology may be useful in monitoring cognitive status, predicting risk of dementia and assessing therapeutic interventions.
Reduced Hippocampal GABA+ Is Associated With Poorer Episodic Memory in Healthy Older Women: A Pilot Study
Authors: Jiménez-Balado Joan , Ycaza Herrera Alexandra , Igwe Kay , Klem Lynda , Buyukturkoglu Korhan , Irimia Andrei , Chen Liu , Guo Jia , Brickman Adam M. , Eich Teal S.
Published: August 2021

Background: The current pilot study was designed to examine the association between hippocampal γ-aminobutyric acid (GABA) concentration and episodic memory in older individuals, as well as the impact of two major risk factors for Alzheimer’s disease (AD)—female sex and Apolipoprotein ε4 (ApoE ε4) genotype—on this relationship.

Methods: Twenty healthy, community-dwelling individuals aged 50–71 (11 women) took part in the study. Episodic memory was evaluated using a Directed Forgetting task, and GABA+ was measured in the right hippocampus using a Mescher-Garwood point-resolved magnetic resonance spectroscopy (MRS) sequence. Multiple linear regression models were used to quantify the relationship between episodic memory, GABA+, ApoE ɛ4, and sex, controlling for age and education.

Results: While GABA+ did not interact with ApoE ɛ4 carrier status to influence episodic memory (p = 0.757), the relationship between GABA+ and episodic memory was moderated by sex: lower GABA+ predicted worse memory in women such that, for each standard deviation decrease in GABA+ concentration, memory scores were reduced by 11% (p = 0.001).

Conclusions: This pilot study suggests that sex, but not ApoE ɛ4 genotype, moderates the relationship between hippocampal GABA+ and episodic memory, such that women with lower GABA+ concentration show worse memory performance. These findings, which must be interpreted with caution given the small sample size, may serve as a starting point for larger studies using multimodal neuroimaging to understand the contributions of GABA metabolism to age-related memory decline.
Plasma Membrane Ca2+ Pump PMCA4z Is More Active Than Splicing Variant PMCA4x
Authors: Corradi Gerardo R. , Mazzitelli Luciana R. , Petrovich Guido D. , de Tezanos Pinto Felicitas , Rochi Lucia , Adamo Hugo P.
Published: August 2021

The plasma membrane Ca2+ pumps (PMCA) are P-ATPases that control Ca2+ signaling and homeostasis by transporting Ca2+ out of the eukaryotic cell. Humans have four genes that code for PMCA isoforms (PMCA1-4). A large diversity of PMCA isoforms is generated by alternative mRNA splicing at sites A and C. The different PMCA isoforms are expressed in a cell-type and developmental-specific manner and exhibit differential sensitivity to a great number of regulatory mechanisms. PMCA4 has two A splice variants, the forms “x” and “z”. While PMCA4x is ubiquitously expressed and relatively well-studied, PMCA4z is less characterized and its expression is restricted to some tissues such as the brain and heart muscle. PMCA4z lacks a stretch of 12 amino acids in the so-called A-M3 linker, a conformation-sensitive region of the molecule connecting the actuator domain (A) with the third transmembrane segment (M3). We expressed in yeast PMCA4 variants “x” and “z”, maintaining constant the most frequent splice variant “b” at the C-terminal end, and obtained purified preparations of both proteins. In the basal autoinhibited state, PMCA4zb showed a higher ATPase activity and a higher apparent Ca2+ affinity than PMCA4xb. Both isoforms were stimulated by calmodulin but PMCA4zb was more strongly activated by acidic lipids than PMCA4xb. The results indicate that a PMCA4 intrinsically more active and more responsive to acidic lipids is produced by the variant “z” of the splicing site A.
Neuronal and Astrocytic Regulations in Schizophrenia: A Computational Modelling Study
Authors: Fritschi Lea , Lindmar Johanna Hedlund , Scheidl Florian , Lenk Kerstin
Published: August 2021

According to the tripartite synapse model, astrocytes have a modulatory effect on neuronal signal transmission. More recently, astrocyte malfunction has been associated with psychiatric diseases such as schizophrenia. Several hypotheses have been proposed on the pathological mechanisms of astrocytes in schizophrenia. For example, post-mortem examinations have revealed a reduced astrocytic density in patients with schizophrenia. Another hypothesis suggests that disease symptoms are linked to an abnormality of glutamate transmission, which is also regulated by astrocytes (glutamate hypothesis of schizophrenia). Electrophysiological findings indicate a dispute over whether the disorder causes an increase or a decrease in neuronal and astrocytic activity. Moreover, there is no consensus as to which molecular pathways and network mechanisms are altered in schizophrenia. Computational models can aid the process in finding the underlying pathological malfunctions. The effect of astrocytes on the activity of neuron-astrocyte networks has been analysed with computational models. These can reproduce experimentally observed phenomena, such as astrocytic modulation of spike and burst signalling in neuron-astrocyte networks. Using an established computational neuron-astrocyte network model, we simulate experimental data of healthy and pathological networks by using different neuronal and astrocytic parameter configurations. In our simulations, the reduction of neuronal or astrocytic cell densities yields decreased glutamate levels and a statistically significant reduction in the network activity. Amplifications of the astrocytic ATP release toward postsynaptic terminals also reduced the network activity and resulted in temporarily increased glutamate levels. In contrast, reducing either the glutamate release or re-uptake in astrocytes resulted in higher network activities. Similarly, an increase in synaptic weights of excitatory or inhibitory neurons raises the excitability of individual cells and elevates the activation level of the network. To conclude, our simulations suggest that the impairment of both neurons and astrocytes disturbs the neuronal network activity in schizophrenia.