bluuebunny/crossref_metadata_2025 huggingface dataset:
EFL TEACHERS’ PERSPECTIVES ON MOBILE-ASSISTED LANGUAGE LEARNING (MALL) RESOURCES FOR VOCATIONAL HIGH SCHOOL STUDENTS
Authors: Sari Nara , Sulistyo Teguh
Published: April 2022

This study aimed to ascertain vocational EFL teachers’ perspectives on the development of Mobile-Assisted Language Learning (MALL) resources in vocational schools. It employed a mixed-method approach in which data were collected simultaneously using an open-ended questionnaire and semi-structured interviews. The study involved 95 English teachers from vocational schools in two cities of East Java, Indonesia. Among the participants, four were invited to participate in an online interview to discuss their perspectives on the characteristics, use and development of MALL material for vocational students. Data analysis was performed using descriptive statistics. The analysis results showed that more than half of the respondents agreed that MALL resources were more accessible (65.4%), easier to generate (65.4%) and more successful at improving learners’ autonomy (73.1%), compared to traditional materials. Additionally, 40% of teachers thought that MALL materials could be used to promote classroom interaction, and it was much easier for teachers to provide immediate feedback through mobile phone. In summary, 42% of teachers agreed that MALL resources could aid in foreign language acquisition. Due to students’ proximity to technology and mobile devices, teachers recognized that MALL materials can be applied inside and outside the classrooms. Therefore, vocational EFL teachers must be able to develop and implement MALL materials in their classrooms.
DISCOURSE ANALYSIS ON SELECTED TEXTS OF BOG BOG BALI CARTOON MAGAZINE
Authors: Hamdani Beny , Luthfiyati Dian , Husnulhotimah Husnulhotimah , Kartika Sania Dina
Published: April 2022

The recent article intends to describe the application of grammatical cohesive devices on the articles of Bog-Bog Bali Cartoon Magazine and to figure out the effect of grammatical cohesive devices on text to be coherent and unified. The article uses library research with collected from written texts. The data are qualitative data taken from Bog-Bog Bali Cartoon Magazine which were published in 2003. The articles are written texts. The Text is realized by 38 ranks (non-embedded) clauses and 6 ranks shifted (embedded) clauses. There is also cohesion between elements in each text of Bog-Bog Bali Magazine. The use of reference such as personal (e.g., we, us, their, them, it, etc), demonstrative reference (one, the, that, this), comparative reference which shows degree of comparison (e.g., same, finest, most, latest) is often used in the articles. In fact, text one consists of 37 elements of anaphoric reference, 7 elements of cataphoric reference and 7 elements of exophoric reference. . In summary, it reveals that grammatical cohesive  can make the text coherent and unified.
THE ANALYSIS OF GENERIC STRUCTURE OF DESCRIPTIVE TEXT USED BY STUDENTS OF ELEMENTARY SCHOOL 1 PENAMBANGAN PROBOLINGGO
Authors: Rosa Indah Chindy Hanggara
Published: April 2022

The difficulty found in writing is not only due to the need to generate and organize ideas using the appropriate choice of vocabulary, sentence, and paragraph organization. A descriptive text is one of the texts that can help students to improve their ideas into a text. The researcher used the qualitative method as the way to analyze the data in this research. It focused on analyzing the student’s skill of applying the generic structure of descriptive texts. The writer used the data from the students’ descriptive texts taken from the worksheet of Elementary school 1 Penambangan Probolinggo. The findings are as follows; they do not master the language features of descriptive text and lack vocabulary and got difficulties describing with effective word choice. They failed to show well-organized writing and did not use identification appropriately, the last they were still unable to transfer and develop their ideas into a written form well in composing a text and do not use the description process in writing appropriately. Finally, the researcher concluded that some students at  Elementary school 1 Penambangan Probolinggo still need more practice to choose a topic that they are about to discuss and to arrange the descriptions with proper connectives.
THE CHALLENGES OF USING TIK-TOK MEDIA IN ELT
Authors: Agting Ovani , Suhartatik Suhartatik , Pusparini Indrawati
Published: April 2022

Tik-Tok application as teaching media is fit in with the students’ need and interests. The goal is to develop Tik-Tok application as media for teaching Speaking at  the level of senior high school.  Research and Development focus on the steps how to develop Tik-Tok media in the teaching of Speaking. The finding show that Tik-Tok media is needed in learning speaking with the score of 85%. The result of compatibility is also absolutely agreed that Tik-Tok media is compatible with the students’ characteristic (85%). The expertise  judgment of Material and Media vaalidate Tik-Tok media with the  criteria  of  “very good” in the aspects of  content and  video application. It means that the development of Tik-Tok application is feasible to be used as media on Speaking class.
AN ANALYSIS OF STUDENTS’ ABILITY IN WRITING CAUSE AND EFFECT ESSAY AT SMAN 1 KEDIRI
Authors: Aini Nurul , Widayanti Deny Ega , Winarto Agus Edi
Published: April 2022

This study aims to find the ability, the problems as well as how they solve them in writing cause and effect essay. This research used descriptive qualitative. The data were obtained by analyzing the students’ essay writing based on Jacobs et al, assessing rubric instrument, calculating questionnaire from 32 respondents, and transcribing interview from the English teacher. The subject is the students of eleventh-grade SMAN 1 Kediri. The result showed that the ability of students in writing cause and effect classified as “Good” based on the means score of the students’ cause and effect essay. It was found twelve students had problem in language use, seven students had problem in mechanics, there were six students had problem in content, five students had problem in organization and two students had problem in vocabulary. Also, there were three kinds of solutions offered by the students such as using internet to access various source, using dictionary, and practice writing more.
PELATIHAN DAN SIMULASI DATA MINING PADA SMA NEGERI 3 SEMARANG
Authors: Zami Farrikh Al , Novianto Sendi , Rohmani Asih , Winarno Sri , Gamayanto Indra , Sukamto Titien Suhartini , Wibowo Sasono , Sani Ramadhan Rakhmat , Budi Setyo
Published: April 2022

SMA Negeri 3 Semarang adalah salah satu dari beberapa SMA terbaik di kota semarang yang Siswa/i serta Gurunya mempunyai keahlian tinggi. Unsur utama yang dapat menaikkan keahlian SMA tersebut adalah telah dilakukannya kaloborasi dengan perguruan tinggi dalam kegiatan pelatihan dan peningkatan ilmu pengetahuan dari berbagai bidang, agar supaya dapat bersaing diera globalisasi. Sekarang ini menguatkan keahlian Siswa/i dan Guru merupakan hal utama, karena nantinya akan dapat bersaing dengan negara maju lainnya. Untuk menguatkan keahlian tersebut maka dilakukan pelatihan dan simulasi data mining di SMA Negeri 3 Semarang. Diawali dengan survei, kemudian dianalisis dan akhirnya dilaksanakan kegiatan pelatihan data mining. Hasil dari pengabdian masyarakat data mining ini adalah Siswa dan Guru mampu memahami data mining, contoh dan penerapannya, sehingga Siswa dan Guru memiliki pengetahuan khusus data mining yang berguna untuk meningkatkan kompetensi dan persaingan global.
SOSIALISASI LITERASI DATA WEBINAR CREATIVE TALK SERIES, MENGENAL BIG DATA: JEMPOLMU, DATAMU
Authors: Nugraha Febrianta Surya , Rihastuti Siti , Rosyidi Afnan
Published: April 2022

Tren Big Data di kalangan akademisi yang mempelajari tentang teknologi informasi sudah beredar sejak lama, akan tetapi tren big data pada masyarakat umum muncul pada era berkembangnya penggunaan smartphone yang masif. Persepsi masyarakat akan big data yang masih banyak kurang paham yang mendasari diselenggarakannya kegiatan ini untuk mengenalkan tentang big data. Kegiatan ini dilakukan dengan model web seminar (Webinar) karena pada saat perlaksanaan kegiatan ini, terdapat pandemi COVID-19. Pendekatan untuk mendapatkan umpan balik dari peserta melalui 3 cara, yaitu pada sesi tanya jawab, pemberian waktu kepada peserta untuk menyampaikan pandangan dan kesimpulan setelah pemberian materi baik secara lisan maupun tulisan, dan kesan serta masukan pada saat absensi tentang kegiatan yang telah dilakukan. Kegiatan Pengabdian kepada Masyarakat Webinar Creative Talk Series, Mengenal Big Data: Jempolmu Datamu telah terlaksana dengan baik. Sebanyak 166 peserta yang terdaftar dalam absensi dan mengikuti acara ini hingga selesai. Berdasarkan umpan balik peserta, tingkat pemahaman tentang big data cukup baik dan peserta ingin mengetahui keterkaitan antara teknologi big data dengan cabang teknologi informasi yang lain. Meskipun tingkat pemahaman tentang teknologi big data yang meningkat, masih ada permasalahan tentang pemahaman ilmu dan teknologi yang menunjang teknologi big data.
Evaluation Of Pavement Me Locally Calibrated Distress Models For Jointed Plain Concrete Pavements In Pennsylvania
Authors: Ramirez Luis , Morian Dennis
Published: April 2022

The Pennsylvania Department of Transportation is currently in the process of implementing the Mechanistic Empirical Pavement Design Guide and its associated software AASHTOWare Pavement ME Design into its standard procedure for designing pavements. Among the key tasks of this implementation effort are the verification of the software's ``global'' pavement distress and smoothness prediction models and their local calibration to Pennsylvania conditions. These key tasks have been completed in a different study based on test sections located in Pennsylvania. This paper presents an evaluation of the locally calibrated pavement distress models for new Jointed Plain Concrete Pavements using three pavement sections outside the pool of test sections included in the verification and calibration tasks. Three approaches were used in the evaluation. The first two correspond to global and local models with field and laboratory measured input values, and the third one correspond to local coefficients with PennDOT default recommended input values. Additionally, 20-year designs were executed using Pavement ME and compared with designs completed using the 1993 AASHTO design guide. The results of this investigation indicated that for the sections evaluated the local calibration improved the prediction of joint faulting but had a negligible effect in the prediction of fatigue cracking. The global and local distress models under-predicted fatigue cracking and over-predicted faulting for the majority of cases evaluated. Additionally, it was found that the difference in slab thickness designs for Pavement ME and AASHTO93 tend to be larger when the input values deviate more from the local default recommended values.
Investigation of TiO2 Photocatalytic Properties of Concrete Samples with Dielectric Constant Applications
Authors: Joshaghani Alireza , Zollinger Dan
Published: April 2022

There is a going interest to reduce emission and improve air quality of the environment through the use of titanium dioxide (TiO2) topical treatments. TiO2 is regarded as one of the most efficient and environment-friendly ultraviolet light-sensitive materials for the photocatalytic degradation of various pollutants. The effort to reduce nitrogen dioxides in the ambient air is proposed to be implemented on the surfaces of streets and sidewalks. Two methods are used to quantify photocatalytic performance: direct and the indirect. The direct method measures the difference of NO concentrations before and after surface treatment, but this method may be highly inaccurate due to the influence of various parameters, time, and costs. The indirect method, on the other hand, quantifies the photocatalysis performance by using the measured surface dielectric to estimate the amount of oxidized NOx byproducts that have accumulated on the surface. This approach is entirely non-destructive and constitutes a new protocol to assess the field-effectiveness of photocatalytic topical treatments based on reproducible and logical monitoring procedures. Finally, mass balance computation was used to validate the effectiveness of this protocol and check the accuracy of the proposed method.
Industry Training for Concrete Paving Crews in Australia (2006-2020): The “Grey Card” Course
Authors: Hodgkinson John
Published: April 2022

In addition to advances in materials and construction equipment there is a continuing need to ensure adequate skills are available within paving crews. Since 2006 concrete pavement crews and their supervisors in Australia have been required to undertake a mandatory one-day course. Known in industry as the ``Grey Card'' course it was developed jointly by NSW Roads and Maritime Services (RMS) and industry applicable to highway and similar classification roads. It demonstrates a commitment from both sectors to relevant training. Over 180 courses with 3,800 participants have been completed in four States and the Australian Capital Territory. Successful participants are issued with a card that is recognised throughout industry irrespective of changes in an individual's employment. The course is presented by instructors accredited by RMS and drawn from industry professionals who have demonstrated considerable construction experience. There is no other course of this type in Australia. The course has the primary objective of consistently high quality construction. Based on agency Specifications the course sets out the reasons for various construction requirements and site practices necessary to achieve them at paving crew level. Sessions include the basics of making good concrete, setting forms reinforcement and dowels, placing paving and compaction, surface finishing and texturing, curing and protection. This paper sets out the development and presentations of the courses.
SYNTHESIS, SPECTRAL CHARACTERIZATION AND STABILITY OF THE NEW ND (III) AND CO (II) HETEROMETALLIC COMPLEXES WITH AMINOPOLYCARBOXYLIC ACIDS
Authors: Trunova Elena , Mishchenko Artem , Makotryk Tamara
Published: April 2022

New heterometallic complexes of Nd(III) and Co(II) based on ethylenediaminetetraacetic and ethylenediaminedisuccinic acids have been synthesized. The complexes studied by electron absorption spectroscopy, IR-spectroscopy, and thermogravimetric analysis. The influence of a number of factors on the properties and structure of heteronuclear complexes is discussed. The spectral characteristics of homonuclear and heteronuclear complexes are compared. A theoretical analysis of the geometry of Co(II) and Nd(III) complexes with ethylenediaminesuccinic acid was carried out by the semi-empirical PM7 method. It has been shown that the f-d-complexes are of the folded type, in which the ligand-complexone realizes the maximum denticity to Nd(III), and the Co(II) coordination sphere is formed by bridging edta/edds carboxyl groups and intrasphere water molecules. The Co(II) coordination polyhedron corresponds to a distorted octahedron, and the Nd(III) coordination polyhedron corresponds to a one-capped square antiprism (C4v) with the coordination number Nd(III)=8. During study fixed that heterometallic complexes have the same structure in solutions and in the solid state.
HYDROGEN’S EVOLUTION ON TITANIUM ELECTRODE FROM SULFURIC ACID SOLUTION
Authors: Kozin Valentin , Bliznyuk Antonina
Published: April 2022

Thanks to the unique combination of physicochemical properties, metal hydrides (MH) are widely used in various fields of science and technology. High thermal, chemical and radiation resistance MH allows it to be used in nuclear industry as a material for nuclear reactors, chemical engineering, metallurgy for the production and refining of metals, for the production of devices operating at elevated temperatures and in adverse conditions. Unique properties MH are widely used in hydrogen energy as hydrogen storage, in rechargeable batteries. Analysis of literature data has shown that most studies are devoted to the practical use of the unique MH ability of reversibly absorbs a large amount of hydrogen. However, the electrochemical characteristics of the electrode processes of metal hydride systems were practically not investigated.

The aim of this work is to study the processes that take place on the titanium electrode in sulfuric acid solutions, and the composition of titanium compounds that are formed. The paper presents the results of studies of the hydride formation reactions occurring on a polarized titanium cathode in aqueous solution 2 N sulfuric acid. The study was carried out using the following methods: the cyclic voltammetry and the method of x-ray phase analysis of the composition of the surface. It has been found that the reduction of hydrogen at the titanium cathode from a solution of sulfuric acid is accompanied by the formation of hydrides, which causes an increase in the overvoltage of hydrogen evolution and high values of the angular coefficients of the Tafel dependence. The kinetic parameters of this process have been calculated: the transfer coefficients α, theoretical and experimental angular coefficients btheor  and bexp , exchange currents ie, the dependence of the hydrogen overvoltage from temperature. It has been found that exchange current density of hydrogen evolution reaction in this system is close by value to the current exchange of hydrogen evolution at the noble metals Pd, Pt, Rh, Ir and are (2,70∙÷0,8)∙10-3А·sm-2 in the temperature range 298 ÷ 343 K. The value of activation energy of 19,83 kJ mol-1 indicates the diffusion control of this process.
KAJIAN PERUBAHAN STATUS KELURAHAN MENJADI DESA DI KABUPATEN TAKALAR STUDY OF CHANGE STATUS URBAN VILLAGE TO VILLAGE IN TAKALAR REGENCY
Authors: Nain Umar
Published: April 2022

Regional autonomy gives authority to the regions to regulate their household affairs in accordance with the potential of the region. The purpose of this study was to determine the problems of the urban village based on the condition of the underdeveloped community into urban areas; analyze changes in urban village into Villages based on various regulations on villages; analyze the stages of the process and various requirements 60 60  that must be met in making changes to the status of the urban village to village. The approach used in this research is through literature studies and regulatory studies. To strengthen both studies are supported by field data conducted through observation and interviews. The informants in this study are the head of urban village and his apparatus, community leaders, religious leaders, youth leaders and social groups and members of the Community Empowerment Institution in the urban village. Data analysis through 3 stages, namely data reduction, data presentation, and verification and conclusions. The results showed that the Rajaya urban village, Parangluara urban village, Takalar urban village and Bontolebang urban village were sufficiently fulfilling the requirements, rational and worthy of changing the status from urban village to village
ANALISIS PEMILIHAN SERENTAK KETUA RW DAN KETUA RT KOTA MAKASSAR DITINJAU DARI ASPEK ETIKA POLITIK
Authors: Patarai Idris , Burhanuddin Burhanuddin , Saleh Akhir
Published: April 2022

This study aims to analyze the election of RW and RT Chairpersons in terms of political ethics in Makassar City. The research method used isqualitative. Informants are voter or non-voters who live in six sub-districts. Based on geographical considerations, six districts were selected, namelyUjung Tanah, Makassar, Tamalate, Tamalanrea, Manggala and Rappocini. Data was extracted by researchers as human instruments, throughinterviews with informants. In addition, observation and documentation were also carried out. Data analysis proceeded by induction-interpretationconceptualization. The results showed that accountability for the legal aspects was largely ignored by the organizing agency such as therequirements of the 5% candidate, the completeness of the voting booths, and the completeness of the TPS administration. What is encouraging isthe enthusiasm of the community as a means of political education, it turns out that in general it can be well received, there does not appear to be anymeaningful friction, prolonged conflict and administrative demands that are compensatory and organizational. The legal legitimacy at the democraticparty is not relevant to the application of the rule of law based on values in the sociological principle, that in the existence of a statutory regulation,it should be interpreted as a need or an interest that can regulate society and felt as a demand that must be obeyed together. The regulation wasmade unilaterally by the city government without regard to the rules and legal values that live in the community
KARAKTER DAN NILAI-NILAI KEPEMIMPINAN PAMONGPRAJA UNTUK SINERGITAS ORGANISASI DAN PELAYANAN PUBLIK DI KOTA MAKASSAR
Authors: Saleh Akhir
Published: April 2022

ABSTRACTThe research aims to analyze the character and values of civil service leadership for organizational synergy and public service. method used is qualitative. The data source in this study is the general public, service users, sub-district head and a portion of the village head and one section, namely the Government Section as an element of the Makassar City Secretariat. Likewise with the regional government partners, among others were several leaders of the ORW and other community organization leaders. The main instrument in data collection is the researchers themselves as human instruments with interview and observation techniques. Data analysis proceeded by inductioninterpretation- conceptualization. The results showed that the Camats had taken the initiative to make new breakthroughs taking initiatives that have access to the reach of far-reaching thinking with visionary and innovative insight, able to build cooperation, and create regional conduciveness through coordination and peace making, although on the other hand is still weak in law enforcement. The results of this study also show that leadership in Makassar City has been able to overcome and make decisions when facing very emergency conditions.

Keywords: character, leadership value, civil service.
Dampak Pandemi Covid-19 Terhadap Kinerja Perusahaan Farmasi yang terdaftar di BEI
Authors: Dewi Iriana Kusuma , Kencana Putri Nilam
Published: April 2022

The purpose of the study was to see differences in financial performance before and when the covid-19 virus was first announced in Indonesia, in pharmaceutical companies listed on the IDX. The analytical method used is the Paired Sample T-Test Difference. The population in this study were all pharmaceutical companies listed on the Indonesia Stock Exchange as many as 12 companies. The sample used in this study were 11 companies in the 2019-2021 period consisting of Darya-Varia Laboratoria Tbk (DVLA), Indofarma Tbk (INAF), PT Kimia Farma Tbk (KAEF), Kalbe Farma Tbk (KLBF), Merck Tbk (BRAND), PT Phapros Tbk (PEHA), Pyridam Farma Tbk (PYFA), PT Organon Pharma Indonesia Tbk (SCPI), PT Sido Muncul Herbal and Pharmaceutical Industry Tbk (SIDO), Tempo Scan Pacific Tbk (TSPC), and Millennium Pharmacon International Tbk (SDPC). The results of the study found that there were no differences in financial performance before and during the covid-19 virus in pharmaceutical companies listed on the IDX in terms of liquidity ratios, activity ratios, and profitability ratios. Based on the results of the study, it can be concluded that there is no difference in financial performance before and during the covid-19 pandemic.

Abstrak

Tujuan Penelitian untuk melihat perbedaan kinerja keuangan sebelum dan saat virus covid-19 pertama kali di umumkan di Indonesia, pada perusahaan Farmasi yang terdaftar di BEI. Metode analisis yang digunakan adalah Uji Beda Paired Sample T-Test. Populasi dalam penelitian ini adalah seluruh perusahaan Farmasi yang terdaftar di Bursa Efek Indonesia yakni sebanyak 12 Perusahaan. Sampel yang digunakan dalam penelitian ini adalah 11 perusahaan pada periode 2019-2021 yang terdiri dari Darya-Varia Laboratoria Tbk (DVLA), Indofarma Tbk (INAF), PT Kimia Farma Tbk (KAEF), Kalbe Farma Tbk (KLBF), Merck Tbk (MERK), PT Phapros Tbk (PEHA), Pyridam Farma Tbk (PYFA), PT Organon Pharma Indonesia Tbk (SCPI), PT Industri Jamu Dan Farmasi Sido Muncul Tbk (SIDO), Tempo Scan Pacific Tbk (TSPC), dan Millennium Pharmacon International Tbk (SDPC). Hasil Penelitian menemukan bahwa tidak terdapat perbedaan kinerja keuangan sebelum dan saat virus covid-19 pada perusahaan farmasi yang terdaftar di BEI ditinjau dari rasio likuiditas, rasio aktivitas, dan rasio profitabilitas. Berdasarkan hasil penelitian maka dapat disimpulkan tidak terdapat perbedaan kinerja keuangan sebelum dan saat pandemi virus covid-19.

Kata Kunci: Likuiditas, Aktivitas, Profitabilitas, Kinerja Keuangan
Relationship Between Plantar Tissue Hardness and Plantar Pressure Distributions in People With Diabetic Peripheral Neuropathy
Authors: Duan Yijie , Ren Weiyan , Liu Wei , Li Jianchao , Pu Fang , Jan Yih-Kuen
Published: April 2022

Objective: People with diabetic peripheral neuropathy (DPN) are usually accompanied with increased plantar pressure. Such high plantar loading during daily activities may cause changes in the biomechanical properties of plantar soft tissue, whose viability is critical to the development of foot ulcers. This study aimed to investigate the relationship between plantar tissue hardness and plantar pressure in people with and without DPN, and preliminarily explore the influence of plantar loading patterns on the plantar pressure and tissue hardness.

Methods: The study was conducted on 14 people with DPN and 14 diabetic people without DPN. The Shore durometer and MatScan System were used to measure the plantar tissue hardness and plantar pressure, respectively. The plantar loading level was evaluated by the duration of daily weight-bearing activity and was used to group diabetic participants with and without DPN into two subgroups (lower loading group and higher loading group).

Results: The plantar tissue hardness was significantly correlated with static peak plantar pressure (PPP, p < 0.05) and dynamic pressure-time integral (PTI, p < 0.05) in the forefoot region in people with DPN. Results of variance analysis showed a significant interaction effect between peripheral neuropathy and plantar loading on tissue hardness (p < 0.05), but not plantar pressure. For people with DPN, significant differences in tissue hardness between the higher loading group and lower loading group were observed in the forefoot, midfoot and hindfoot regions. In the higher loading group, people with DPN had significantly greater tissue hardness than that in people without DPN in the toes, forefoot, midfoot and hindfoot regions (p < 0.05).

Conclusions: There is a significant correlation between tissue hardness and PPP, and between tissue hardness and PTI in people with DPN. Plantar loading associated with daily activities plays a significant role on the plantar tissue hardness in people with DPN. The findings of this study contribute to further understand the relationship between increased plantar tissue hardness and high plantar pressure in people with diabetic peripheral neuropathy.
Droplet-Microfluidic-Based Promoter Engineering and Expression Fine-Tuning for Improved Erythromycin Production in Saccharopolyspora erythraea NRRL 23338
Authors: Yun Kaiyue , Zhang Yue , Li Shixin , Wang Yan , Tu Ran , Liu Hao , Wang Meng
Published: April 2022

Erythromycin is a clinically important drug produced by the rare actinomycete Saccharopolyspora erythraea. In the wide-type erythromycin producer S. erythraea NRRL 23338, there is a lack of systematical method for promoter engineering as well as a well-characterized promoter panel for comprehensive metabolic engineering. Here we demonstrated a systematical promoter acquiring process including promoter characterization, engineering and high-throughput screening by the droplet-microfluidic based platform in S. erythraea NRRL 23338, and rapidly obtained a panel of promoters with 21.5-fold strength variation for expression fine-tuning in the native host. By comparative qRT-PCR of S. erythraea NRRL 23338 and a high-producing strain S0, potential limiting enzymes were identified and overexpressed individually using two screened synthetic promoters. As a result, erythromycin production in the native host was improved by as high as 137.24 folds by combinational gene overexpression. This work enriches the accessible regulatory elements in the important erythromycin-producing strain S. erythraea NRRL 23338, and also provides a rapid and systematic research paradigm of promoter engineering and expression fine-tuning in the similar filamentous actinomycete hosts.
Breast Cancer Patients With Positive Apical or Infraclavicular/Ipsilateral Supraclavicular Lymph Nodes Should Be Excluded in the Application of the Lymph Node Ratio System
Authors: Wang Zhe , Chong Wei , Zhang Huikun , Liu Xiaoli , Zhao Yawen , Guo Zhifang , Fu Li , Ma Yongjie , Gu Feng
Published: April 2022

Aim: Increasing studies have demonstrated lymph node ratio (LNR) to be an accurate prognostic indicator in breast cancer and an alternative to pN staging; however, the AJCC-TNM staging system classified apical or infraclavicular/ipsilateral supraclavicular lymph node-positive (APN(+)) patients with a worse prognosis as the pN3 stage. Until now, different reports on LNR in breast cancer have ignored this possibility. Consequently, it is necessary to discuss the role of APN(+) patients in the LNR system to obtain a precise LNR that predicts the prognosis accurately.

Materials and Methods: We collected data on 10,120 breast cancer patients, including 3,936 lymph node-positive patients (3,283 APN(−) and 653 APN(+) patients), who visited our hospital from 2007 to 2012. Then we applied X-tile analysis to calculate cut-off values and conduct survival analysis and multivariate analysis to evaluate patients’ prognosis.

Results: We confirmed that some APN(+) patients were mis-subgrouped according to previously reported LNR, indicating that APN(+) patients should be excluded in the application of LNR to predict prognosis. Then we applied X-tile analysis to calculate two cut-off values (0.15 and 0.34) for LNR-APN(−) patients and conducted survival analysis and found that LNR-APN(−) staging was superior to pN staging in predicting the prognosis of APN(−) breast cancer patients.

Conclusion: From this study, we conclude that excluding APN(+) patients is the most necessary condition for effective implementation of the LNR system. LNR-APN(−) staging could be a more comprehensive approach in predicting prognosis and guiding clinicians to provide accurate and appropriate treatment.
Salt-Sensitive Hypertension Induces Osteoclastogenesis and Bone Resorption via Upregulation of Angiotensin II Type 1 Receptor Expression in Osteoblasts
Authors: Pramusita Adya , Kitaura Hideki , Ohori Fumitoshi , Noguchi Takahiro , Marahleh Aseel , Nara Yasuhiko , Kinjo Ria , Ma Jinghan , Kanou Kayoko , Tanaka Yukinori , Mizoguchi Itaru
Published: April 2022

Hypertension is a chronic-low grade inflammatory disease, which is known to be associated with increased bone loss. Excessive activity of the local renin–angiotensin system (RAS) in bone leads to increased bone resorption. As inflammatory cytokines may activate RAS components, we hypothesized that the elevated proinflammatory cytokine levels in hypertension activate bone RAS and thus lead to increased bone resorption. To investigate whether salt-sensitive hypertension (SSHTN) induces osteoclastogenesis and bone resorption, we generated a model of SSHTN in C57BL/6J mice by post-Nω-nitro-l-arginine methyl ester hydrochloride (l-NAME) high-salt challenge. SSHTN led to the reduction of distal femur trabecular number and bone volume fraction, while trabecular separation of femoral bone showed a significant increase, with no change in cortical thickness. Histomorphometric examination showed a significant reduction in trabecular bone volume fraction with an increased number of multinucleated tartrate-resistant acid phosphatase (TRAP)-positive cells and increased osteoclast surface fraction in the trabecular distal femur of hypertensive mice. Furthermore, analysis of gene expression in bone tissue revealed that TRAP and RANKL/OPG mRNA were highly expressed in hypertensive mice. TNF-α and angiotensin II type 1 receptor (AGTR1) mRNA and protein expression were also upregulated in SSHTN mice. These observations suggested that TNF-α may have an effect on AGTR1 expression leading to osteoclast activation. However, TNF-α stimulation did not promote AGTR1 mRNA expression in osteoclast precursors in culture, while TNF-α increased AGTR1 mRNA expression in osteoblast culture by activation of downstream p38. Angiotensin II was also shown to increase TNF-α-induced RANKL/OPG mRNA expression in primary osteoblast culture and osteoclastogenesis in a TNF-α-primed osteoblast and osteoclast precursor co-culture system. In addition, local injection of lipopolysaccharide into the supracalvariae of SSHTN mice markedly promoted osteoclast and bone resorption. In conclusion, mice with SSHTN show increased osteoclastogenesis and bone resorption due mainly to increased TNF-α and partly to the upregulation of AGTR1 in osteoblasts.
A Putative Plasmodium RNA-Binding Protein Plays a Critical Role in Female Gamete Fertility and Parasite Transmission to the Mosquito Vector
Authors: Kumar Sudhir , Abatiyow Biley A. , Haile Meseret T. , Oualim Kenza M. Z. , Leeb Amanda S. , Vaughan Ashley M. , Kappe Stefan H.I.
Published: April 2022

Plasmodium falciparum sexual stage gametocytes are critical for parasite transmission from the human host to the mosquito vector. Mature gametocytes generate fertile male (micro-) or female (macro-) gametes upon activation inside the mosquito midgut. While a number of parasite genes have been described that are critical for P. falciparum gametogenesis and fertility, no parasite gene has been shown to have a unique function in macrogametes. The genome of P. falciparum encodes numerous RNA-binding proteins. We identified a novel protein containing a putative RNA-binding domain, which we named Macrogamete-Contributed Factor Essential for Transmission (MaCFET). This protein is expressed in the asexual and sexual stages. Parasites that carry a deletion of MaCFET (Pfmacfet¯), developed normally as asexual stages, indicating that its function is not essential for the asexual proliferation of the parasite in vitro. Furthermore, Pfmacfet¯ male and female gametocytes developed normally and underwent activation to form microgametes and macrogametes. However, by utilizing genetic crosses, we demonstrate that Pfmacfet¯ parasites suffer a complete female-specific defect in successful fertilization. Therefore, PfMaCFET is a critical female-contributed factor for parasite transmission to the mosquito. Based on its putative RNA-binding properties, PfMaCFET might be in involved in the regulation of mRNAs that encode female-specific functions for fertilization or female-contributed factors needed post fertilization.
Development and Application of CRISPR-Cas Based Tools
Authors: Hu Yanping , Li Wei
Published: April 2022

Abundant CRISPR-Cas systems in nature provide us with unlimited valuable resources to develop a variety of versatile tools, which are powerful weapons in biological discovery and disease treatment. Here, we systematically review the development of CRISPR-Cas based tools from DNA nuclease to RNA nuclease, from nuclease dependent-tools to nucleic acid recognition dependent-tools. Also, considering the limitations and challenges of current CRISPR-Cas based tools, we discuss the potential directions for development of novel CRISPR toolkits in the future.
miR-513c-5p Suppression Aggravates Pyroptosis of Endothelial Cell in Deep Venous Thrombosis by Promoting Caspase-1
Authors: Chu Chu , Wang Bin , Zhang Zhen , Liu Wen , Sun Shangwen , Liang Gang , Zhang Xiaoshan , An Hongqiang , Wei Ran , Zhu Xiaoxiao , Guo Qiang , Zhao Lin , Fu Xiaoxiao , Xu Ke , Li Xia
Published: April 2022

Deep vein thrombosis (DVT) is a common peripheral vascular disease. Secondary pulmonary embolism (PE) caused by DVT leads to substantial patient death. Inflammation has been suggested as a key factor in the pathophysiology of DVT, however, involvement of pyroptosis-related inflammatory factors in DVT formation remains unclear. Here, we proposed that post-transcriptional modification of caspase-1 might be a crucial trigger for enhanced pyroptosis in vascular endothelial cells (VECs), and consequently contributed to severer symptoms in DVT patients. In order to explore the involvement of pyroptosis in DVT, peripheral blood mononuclear cells were collected from 30 DVT patients, and compared with the healthy controls, we found caspase-1 was increased both in mRNA and protein levels. miRNA microarray analysis demonstrated that down-regulated miR-513c-5p was significantly negatively correlated with the expression of caspase-1. In vitro assays suggested that miR-513c-5p overexpression could ameliorate the expression of caspase-1, and thus decreased the production of cleaved gasdermin D (GSDMD) and interleukin (IL)-1β and IL-18 in VECs. The dual-luciferase reporter assay identified direct binding between miR-513c-5p and the 3′ untranslated region of caspase-1 encoding gene. The administration of miR-513c-5p mimics through tail vein injection or caspase-1 inhibitor (vx-765) by intraperitoneal injection remarkably decreased the volume of blood clots in vivo, whereas miR-513c-5p inhibitor aggravated thrombosis formation and this effect was dramatically weakened when treated in combination with vx-765. Collectively, these results revealed that the pyroptosis of VECs induced by decreased miR-513c-5p was involved in DVT progression and indicated a potential therapeutic strategy of targeting the miR-513c-5p/caspase-1/GSDMD signal axis for DVT management.
Aβ Assemblies Promote Amyloidogenic Processing of APP and Intracellular Accumulation of Aβ42 Through Go/Gβγ Signaling
Authors: Antonino Magdalena , Marmo Paula , Freites Carlos Leandro , Quassollo Gonzalo Emiliano , Sánchez Maria Florencia , Lorenzo Alfredo , Bignante Elena Anahi
Published: April 2022

Alzheimer’s disease (AD) is characterized by the deposition of aggregated species of amyloid beta (Aβ) in the brain, which leads to progressive cognitive deficits and dementia. Aβ is generated by the successive cleavage of the amyloid precursor protein (APP), first by β-site APP cleaving enzyme 1 (BACE1) and subsequently by the γ-secretase complex. Those conditions which enhace or reduce its clearance predispose to Aβ aggregation and the development of AD. In vitro studies have demonstrated that Aβ assemblies spark a feed-forward loop heightening Aβ production. However, the underlying mechanism remains unknown. Here, we show that oligomers and fibrils of Aβ enhance colocalization and physical interaction of APP and BACE1 in recycling endosomes of human neurons derived from induced pluripotent stem cells and other cell types, which leads to exacerbated amyloidogenic processing of APP and intracellular accumulation of Aβ42. In cells that are overexpressing the mutant forms of APP which are unable to bind Aβ or to activate Go protein, we have found that treatment with aggregated Aβ fails to increase colocalization of APP with BACE1 indicating that Aβ-APP/Go signaling is involved in this process. Moreover, inhibition of Gβγ subunit signaling with βARKct or gallein prevents Aβ-dependent interaction of APP and BACE1 in endosomes, β-processing of APP, and intracellular accumulation of Aβ42. Collectively, our findings uncover a signaling mechanism leading to a feed-forward loop of amyloidogenesis that might contribute to Aβ pathology in the early stages of AD and suggest that gallein could have therapeutic potential.
Experimental and Natural Induction of de novo Centriole Formation
Authors: Takumi Kasuga , Kitagawa Daiju
Published: April 2022

In cycling cells, new centrioles are assembled in the vicinity of pre-existing centrioles. Although this canonical centriole duplication is a tightly regulated process in animal cells, centrioles can also form in the absence of pre-existing centrioles; this process is termed de novo centriole formation. De novo centriole formation is triggered by the removal of all pre-existing centrioles in the cell in various manners. Moreover, overexpression of polo-like kinase 4 (Plk4), a master regulatory kinase for centriole biogenesis, can induce de novo centriole formation in some cell types. Under these conditions, structurally and functionally normal centrioles can be formed de novo. While de novo centriole formation is normally suppressed in cells with intact centrioles, depletion of certain suppressor proteins leads to the ectopic formation of centriole-related protein aggregates in the cytoplasm. It has been shown that de novo centriole formation also occurs naturally in some species. For instance, during the multiciliogenesis of vertebrate epithelial cells, massive de novo centriole amplification occurs to form numerous motile cilia. In this review, we summarize the previous findings on de novo centriole formation, particularly under experimental conditions, and discuss its regulatory mechanisms.
Synthetic Circular gRNA Mediated Biological Function of CRISPR-(d)Cas9 System
Authors: Wang Mingxia , Xu Jinming , Meng Jialin , Huang Xinbo
Published: April 2022

Ever since the gene editing function was discovered in the CRISPR-Cas9 system, numerous applications and utilities were investigated in order to apply this technique to medical use. However, the clinical practice was limited by unsatisfactory efficiency and unacceptable off-target editing. Modifications from different aspects of the Cas9 protein and gRNAs were published that aimed to improve its function in one way or another. Under the inspiration of Jacob L. Litke and Samie R. Jaffrey, we propose a novel gRNA design that could achieve rapid circular gRNA assembly inside the cells. This circular design consists of the gRNA of interested flanked by Twister ribozymes. The function of this circular gRNA was proved in vitro in both CRISPR-dCas9 and CRISPR-Cas9 systems. It presented a remarkable reduction in the off-target rate in accompany with reduced efficiency. With future improvement in its efficiency, this tool broadens our understanding and possibility of the CRISPR application.
Human Cytomegalovirus vMIA Inhibits MAVS Oligomerization at Peroxisomes in an MFF-Dependent Manner
Authors: Ferreira Ana Rita , Gouveia Ana , Magalhães Ana Cristina , Valença Isabel , Marques Mariana , Kagan Jonathan C. , Ribeiro Daniela
Published: April 2022

Upon intracellular recognition of viral RNA, RIG-I-like proteins interact with MAVS at peroxisomes and mitochondria, inducing its oligomerization and the downstream production of direct antiviral effectors. The human cytomegalovirus (HCMV) is able to specifically evade this antiviral response, via its antiapoptotic protein vMIA. Besides suppressing the programmed cell death of infected cells, vMIA inhibits the antiviral signalling at mitochondria by inducing the organelle’s fragmentation, consequently hindering the interaction between MAVS and the endoplasmic reticulum protein STING. Here we demonstrate that vMIA interferes with the peroxisomal antiviral signalling via a distinct mechanism that is independent of the organelle’s morphology and does not affect STING. vMIA interacts with MAVS at peroxisomes and inhibits its oligomerization, restraining downstream signalling, in an MFF-dependent manner. This study also demonstrates that vMIA is totally dependent on the organelle’s fission machinery to induce peroxisomal fragmentation, while this dependency is not observed at mitochondria. Furthermore, although we demonstrate that vMIA is also able to inhibit MAVS oligomerization at mitochondria, our results indicate that this process, such as the whole vMIA-mediated inhibition of the mitochondrial antiviral response, is independent of MFF. These observed differences in the mechanisms of action of vMIA towards both organelles, likely reflect their intrinsic differences and roles throughout the viral infection. This study uncovers specific molecular mechanisms that may be further explored as targets for antiviral therapy and highlights the relevance of peroxisomes as platforms for antiviral signalling against HCMV.
Development and Validation of a GC-EI-MS/MS Method for Ethyl Glucuronide Quantification in Human Hair
Authors: Mattia Alessandro , Moschella Clementina , David Maria Chiara , Fiore Marco , Gariglio Sara , Salomone Alberto , Vincenti Marco
Published: April 2022

Ethyl glucuronide (EtG) is a minor, non-oxidative ethanol metabolite detectable in several matrices for specific periods of time. In recent years, quantification of EtG in hair has been established as the most reliable biomarker for long-term alcohol consumption, with the Society of Hair Testing offering cut-off values for assessment of both abstinence and heavy drinking. Instrumental constrains and wide inter- and intra-laboratory variability represent the ultimate barriers to widespread acceptance of hair EtG determination in the forensic context. In this study, a new analytical method for hair EtG based on gas chromatographic (GC) separation, electron impact (EI) ionization, and tandem mass spectrometry (MS/MS) detection was developed and validated. At the same time, several parameters for sample pretreatment and instrumental analysis were optimized using real hair samples obtained from different drinking subjects. A full-factorial design-of-experiment approach included procedures for hair washing, pulverization, and extraction. Rigorous multi-step washing proved not to reduce the EtG content extracted in the subsequent sample incubation. Hair pulverization with a ball mill significantly improved the EtG extraction from the keratin matrix and allowed us to reduce the time needed for the subsequent extraction step, without affecting the extraction recovery. The hair extract was derivatized with N-methyl-N-(trimethylsilyl)-trifluoroacetamide. Upon electron impact ionization of the EtG-TMS derivative, triple quadrupole mass analyzers were operated in the selected reaction monitoring (SRM) mode using the fragment m/z 405 as the precursor ion (m/z 410 for the EtG-D5 internal standard), the transitions m/z 405 → 359 and m/z 410 → 359 for quantitation, and m/z 405 → 331 and m/z 405 → 287 for qualification/confirmation, all at 10 V collision energy. The final method was fully validated and then applied to 25 real hair samples. The calibration curve proved linear between 6 and 60 pg/mg. The limit of detection (LOD) was 4 pg/mg. Intra- and inter-assay precision and accuracy tests showed a variability and bias close to 15% or lower over the entire calibration range. The new method is routinely applied in the Italian State Police’s toxicology laboratory for hair analyses addressed to exclude excessive alcohol drinking and verify the psycho-physical requirements of the personnel.
A Copper-Based Biosensor for Dual-Mode Glucose Detection
Authors: Li Kai , Xu Xiaoyu , Liu Wanshan , Yang Shouzhi , Huang Lin , Tang Shuai , Zhang Ziyue , Wang Yuning , Chen Fangmin , Qian Kun
Published: April 2022

Glucose is a source of energy for daily activities of the human body and is regarded as a clinical biomarker, due to the abnormal glucose level in the blood leading to many endocrine metabolic diseases. Thus, it is indispensable to develop simple, accurate, and sensitive methods for glucose detection. However, the current methods mainly depend on natural enzymes, which are unstable, hard to prepare, and expensive, limiting the extensive applications in clinics. Herein, we propose a dual-mode Cu2O nanoparticles (NPs) based biosensor for glucose analysis based on colorimetric assay and laser desorption/ionization mass spectrometry (LDI MS). Cu2O NPs exhibited excellent peroxidase-like activity and served as a matrix for LDI MS analysis, achieving visual and accurate quantitative analysis of glucose in serum. Our proposed method possesses promising application values in clinical disease diagnostics and monitoring.