Metabolic alterations during the growth of tumour spheroids

Solid tumours undergo considerable alterations in their metabolism of nutrients in order to generate sufficient energy and biomass for sustained growth and proliferation. During growth, the tumour microenvironment exerts a number of influences (e.g. hypoxia and acidity) that affect cellular biology and the flux or utilisation of fuels including glucose. The tumour spheroid model was used to characterise the utilisation of glucose and describe alterations to the activity and expression of key glycolytic enzymes during the tissue growth curve. Glucose was avidly consumed and associated with the production of lactate and an acidified medium, confirming the reliance on glycolytic pathways and a diminution of oxidative phosphorylation. The expression levels and activities of hexokinase, phosphofructokinase-1, pyruvate kinase and lactate dehydrogenase in the glycolytic pathway were measured to assess glycolytic capacity. Similar measurements were made for glucose-6-phosphate dehydrogenase, the entry point and regulatory step of the pentose-phosphate pathway (PPP) and for cytosolic malate dehydrogenase, a key link to TCA cycle intermediates. The parameters for these key enzymes were shown to undergo considerable variation during the growth curve of tumour spheroids. In addition, they revealed that the dynamic alterations were influenced by both transcriptional and posttranslational mechanisms

Assessing the Participants of UNDANA Teacher Pro Profiles with Digital-Based Learning Model; Cat. 1, 2022

This study aims to profile teachers in the 1st category of Undana Teacher Professional Education in 2022 using a digital-based learning model. The expected results are the right model in evaluating digital-based learning models to be ready to become professional teachers, graduates from Category I Teacher Professional Education Participants in 2022 at Nusa Cendana University, Kupang, East Nusa Tenggara. Conditions in the field show that student learning strategies and activities play a very important role in digital-based learning, where students sometimes lack concentration and are less skilled in learning activities with digital-based models. The research uses qualitative methods and aims to evaluate the effectiveness of digital-based learning models in preparing teachers for professionalism. Research innovations at the theoretical level, for the development of studies on population mobility, who work as teachers, practical levels for regions of origin, the potential for remittances of migrants between regions in NTT, and their impact on destination areas. The design of this study was carried out in two stages within a span of 1 year of research procedures in the first semester: field observations which were exploratory studies of learning strategies for In-service PPG students in Kupang City and its surroundings. In various study programs where learning uses Digital-based. Followed by the preparation of a draft field observation report. Seminars or workshops around the effect of full IT/Digital use between learning strategies and Digital mastery which are still minimal/insufficient in the 2 (two) PPG Study Programs referred to above and with reporting. The second semester, Conducting Interactive Analysis of the Milles and Humburman model where IT media learning strategies with Online learning strategies affect the ability to learn to operate a Computer and continue with paying attention to Digital students high ability to operate Computers / IT in lectures at PPG Undana

Metabolic alterations during the growth of tumour spheroids

Solid tumours undergo considerable alterations in their metabolism of nutrients in order to generate sufficient energy and biomass for sustained growth and proliferation. During growth, the tumour microenvironment exerts a number of influences (e.g. hypoxia and acidity) that affect cellular biology and the flux or utilisation of fuels including glucose. The tumour spheroid model was used to characterise the utilisation of glucose and describe alterations to the activity and expression of key glycolytic enzymes during the tissue growth curve. Glucose was avidly consumed and associated with the production of lactate and an acidified medium, confirming the reliance on glycolytic pathways and a diminution of oxidative phosphorylation. The expression levels and activities of hexokinase, phosphofructokinase-1, pyruvate kinase and lactate dehydrogenase in the glycolytic pathway were measured to assess glycolytic capacity. Similar measurements were made for glucose-6-phosphate dehydrogenase, the entry point and regulatory step of the pentose-phosphate pathway (PPP) and for cytosolic malate dehydrogenase, a key link to TCA cycle intermediates. The parameters for these key enzymes were shown to undergo considerable variation during the growth curve of tumour spheroids. In addition, they revealed that the dynamic alterations were influenced by both transcriptional and posttranslational mechanisms

Pengembangan dan penerapan sistem keamanan berbasis Internet of Things (IoT) dan monitoring berbasis website bagi kinerja Atmospheric Water Generator (AWG) untuk sistem pertanian lahan kering di Desa Tesiayofanu, Kecamatan Kie, Kabupaten Timor Tengah Selatan

AbstrakProgram Pengabdian kepada Masyarakat melalui kemitraan masyarakat dilaksanakan  dalam rangka mendukung program nasional dalam peningkatan sumberdaya manusia melalui pendidikan kejuruan. Mitra merupakan Desa yang berada di Kabupaten TTS, Provinsi Nusa Tenggara Timur. Kegiatan PkM berlangsung selama 10 (enam) bulan dengan keterlibatan 3 (tiga) dosen dan 5 (lima) mahasiswa. Melalui kegiatan ini diharapkan masyarakat memiliki ketrampilan untuk dapat menggunakan sistem kontrol keamana AWG dan sistem monitoring berasis IoT. Kegiatan ini disampaing meningkatkan ketrampilan dan pengetahuan dalam berbagai sistem kontrol di industri. Metode yang akan dilaksanakan pada PKM ini meliputi tahap persiapan, pelaksanaan kegiatan, monitoring dan evaluasi, serta penyusunan laporan. Persiapan yang akan dilakukan meliputi  observasi,  survey  lokasi,  dan  penyampaian  masalah  serta  solusi  yang  akan ditempuh dalam pelaksanaan kegiatan. Pelaksanaan berupa perancangan alat, ujicoba alat, peningkatan kapasitas mitra melalui pelatihan dan pelatihan penggunaan alat, pendampingan dalam pemeliharaan alat, tahap monitoring oleh tim pelaksana dan evaluasi pelaksanaan program.   Kata kunci: sistem keamanan; IoT; AWG, Website. Abstract The Community Service Program through community partnerships is carried out in order to support the national program in improving human resources through vocational education. Mitra is a village located in TTS Regency, East Nusa Tenggara Province. PkM activities lasted for 10 (six) months with the involvement of 3 (three) lecturers and 5 (five) students. Through this activity, it is hoped that the community will have the skills to be able to use the AWG safety control system and the IoT-based monitoring system. This activity is aimed at improving skills and knowledge in various control systems in the industry. The methods that will be implemented in this PKM include the preparation stage, the implementation of activities, monitoring and evaluation, and the preparation of reports. The preparations that will be carried out include observation, location surveys, and submission of problems and solutions that will be taken in the implementation of activities. The implementation is in the form of tool design, tool testing, partner capacity building through training and training in the use of tools, assistance in tool maintenance, monitoring stages by the implementation team and evaluation of program implementation. The outputs of this PKM activity include the achievement of program target outputs, and output achievement targets in the form of publication of activity results in national journals and scientific meetings, and publications in newspapers and social media. Keywords: security system; IoT; AWG; Website

Perancangan dan pelatihan pemanfaatan kit PLC berbasis microcontroller bagi guru dan siswa program keahlian teknik instalasi tenaga listrik di SMK Negeri 1 Kupang Barat

AbstrakProgram Pengabdian kepada Masyarakat melalui kemitraan masyarakat dilaksanakan  dalam rangka mendukung program nasional dalam peningkatan sumberdaya manusia melalui pendidikan kejuruan. Mitra merupakan lembaga pendidikan dibawah pemerintah daerah Kabupaten Kupang, Provinsi Nusa Tenggara Timur. Kegiatan PkM berlangsung selama 6 (enam) bulan dengan keterlibatan 5 (lima) dosen dan 3 (tiga) mahasiswa. Melalui kegiatan ini diharapkan masyarakat (siswa) memiliki ketrampilan untuk dapat menggunakan tranier sistem kontrol PLC berbasis mikrokontroler.  Melalui kegiatan ini diharapkan  Metode yang akan dilaksanakan pada PKM ini meliputi tahap persiapan, pelaksanaan kegiatan, monitoring dan evaluasi. Persiapan yang akan dilakukan meliputi  observasi,  survey  lokasi,  dan  penyampaian  masalah  serta  solusi  yang  akan ditempuh dalam pelaksanaan kegiatan. Pelaksanaan berupa perancangan alat, ujicoba alat, peningkatan kapasitas mitra melalui pelatihan dan pelatihan penggunaan alat, pendampingan dalam pemeliharaan alat, tahap monitoring oleh tim pelaksana dan evaluasi pelaksanaan program menunjukan kit telah digunakan sebagai media pembelajaran dan memudahkan guru dan siswa dalam memepelajari konsep dan praktek PLC.. Luaran kegiatan PKM yaitu mitra (siswa) memiliki ketrampilan untuk dapat menggunakan tranier sistem kontrol PLC berbasis mikrokontroler dan mengasai konsep PLC dengan mudah. Kegiatan ini dapat meningkatkan ketrampilan dan pengetahuan dalam berbagai sistem kontrol di industri. pencapaian  output  sasaran  program,  dan  target  capaian  luaran  berupa  publikasi  hasil kegiatan pada jurnal nasional maupun pada pertemuan ilmiah, dan publikasi pada koran dan media sosial. Kata kunci: trainer; PLC; microcontroler; SMK AbstractThe Community Service Program through community partnerships is carried out in order to support national programs in improving human resources through vocational education. Mitra is an educational institution under the local government of Kupang Regency, East Nusa Tenggara Province. PkM activities lasted for 6 (six) months with the involvement of 5 (five) lecturers and 3 (three) students. Through this activity, it is hoped that the community (students) will have the skills to be able to use a microcontroller-based PLC control system handrail. Through this activity, it is hoped that the methods that will be implemented in this PKM include the preparation stage, implementation of activities, monitoring and evaluation. The preparations that will be carried out include observation, location surveys, and the submission of problems and solutions that will be taken in the implementation of activities. The implementation in the form of tool design, tool trials, partner capacity building through training and training in the use of tools, assistance in equipment maintenance, monitoring stages by the implementation team and evaluation of program implementation show that kits have been used as learning media and make it easier for teachers and students to learn PLC concepts and practices. The output of PKM activities is that partners (students) have the skills to be able to use microcontroller-based PLC control system and understand the PLC concept easily. This activity can improve skills and knowledge in various control systems in the industry. achievement of program target output, and output achievement targets in the form of publication of the results of activities in national journals and scientific meetings, and publications in newspapers and social media. Keywords: trainer; PLC; microcontroller; vocational high schoo

The Phosphodiesterase-5 Inhibitor Vardenafil Is a Potent Inhibitor of ABCB1/P-Glycoprotein Transporter

One of the major causes of chemotherapy failure in cancer treatment is multidrug resistance (MDR) which is mediated by the ABCB1/P-glycoprotein. Previously, through the use of an extensive screening process, we found that vardenafil, a phosphodiesterase 5 (PDE-5) inhibitor significantly reverses MDR in ABCB1 overexpressing cancer cells, and its efficacy was greater than that of tadalafil, another PDE-5 inhibitor. The present study was designed to determine the reversal mechanisms of vardenafil and tadalafil on ABC transporters-mediated MDR. Vardenafil or tadalafil alone, at concentrations up to 20 µM, had no significant toxic effects on any of the cell lines used in this study, regardless of their membrane transporter status. However, vardenafil when used in combination with anticancer substrates of ABCB1, significantly potentiated their cytotoxicity in ABCB1 overexpressing cells in a concentration-dependent manner, and this effect was greater than that of tadalafil. The sensitivity of the parenteral cell lines to cytotoxic anticancer drugs was not significantly altered by vardenafil. The differential effects of vardenafil and tadalafil appear to be specific for the ABCB1 transporter as both vardenafil and tadalafil had no significant effect on the reversal of drug resistance conferred by ABCC1 (MRP1) and ABCG2 (BCRP) transporters. Vardenafil significantly increased the intracellular accumulation of [3H]-paclitaxel in the ABCB1 overexpressing KB-C2 cells. In addition, vardenafil significantly stimulated the ATPase activity of ABCB1 and inhibited the photolabeling of ABCB1 with [125I]-IAAP. Furthermore, Western blot analysis indicated the incubation of cells with either vardenafil or tadalafil for 72 h did not alter ABCB1 protein expression. Overall, our results suggest that vardenafil reverses ABCB1-mediated MDR by directly blocking the drug efflux function of ABCB1

Fitting the Elementary Rate Constants of the P-gp Transporter Network in the hMDR1-MDCK Confluent Cell Monolayer Using a Particle Swarm Algorithm

P-glycoprotein, a human multidrug resistance transporter, has been extensively studied due to its importance to human health and disease. In order to understand transport kinetics via P-gp, confluent cell monolayers overexpressing P-gp are widely used. The purpose of this study is to obtain the mass action elementary rate constants for P-gp's transport and to functionally characterize members of P-gp's network, i.e., other transporters that transport P-gp substrates in hMDR1-MDCKII confluent cell monolayers and are essential to the net substrate flux. Transport of a range of concentrations of amprenavir, loperamide, quinidine and digoxin across the confluent monolayer of cells was measured in both directions, apical to basolateral and basolateral to apical. We developed a global optimization algorithm using the Particle Swarm method that can simultaneously fit all datasets to yield accurate and exhaustive fits of these elementary rate constants. The statistical sensitivity of the fitted values was determined by using 24 identical replicate fits, yielding simple averages and standard deviations for all of the kinetic parameters, including the efflux active P-gp surface density. Digoxin required additional basolateral and apical transporters, while loperamide required just a basolateral tranporter. The data were better fit by assuming bidirectional transporters, rather than active importers, suggesting that they are not MRP or active OATP transporters. The P-gp efflux rate constants for quinidine and digoxin were about 3-fold smaller than reported ATP hydrolysis rate constants from P-gp proteoliposomes. This suggests a roughly 3∶1 stoichiometry between ATP hydrolysis and P-gp transport for these two drugs. The fitted values of the elementary rate constants for these P-gp substrates support the hypotheses that the selective pressures on P-gp are to maintain a broad substrate range and to keep xenobiotics out of the cytosol, but not out of the apical membrane