Effect of cultivar and explants type on tissue culture regeneration of three Nigerian cultivars of tomatoes (Solanum lycopersicon)

&lt;p&gt;In order to assess the suitable explant(s) for &lt;em&gt;in-vitro&lt;/em&gt; regeneration of three local cultivars of Nigerian tomatoes, Ibadan local (IbL), Ife and JM94/46, cotyledon, hypocotyls and radicle explants were cultured in shoot regeneration medium consisting of&lt;strong&gt; &lt;/strong&gt;MS containing 30 g L&lt;sup&gt;-1&lt;/sup&gt; sucrose and 8 g L&lt;sup&gt;-1 &lt;/sup&gt;agar with no exogenous plant growth hormones. Forty-five of each explant type was cultured on the medium in triplicate experiments and results showed varied percentage survival and shooting for the various explants. Hypocotyl explants had the highest percentage of shooting explants at 13.3% for IbL; 6.67% for Ife and 20% in JM94/46. IbL cotyledon explants had 4.44% of shooting explants with no shoots recorded in Ife and JM94/46 cotyledon explants. IbL radicle explants had 2.22% shooting explants and no shoots recorded in Ife and JM94/46. Student Neuman Keuls (SNK) statistical analysis of cultivar-media interaction showed there was no significant difference (P &amp;gt; 0.05) among the three cultivars in number of calli and shooting calli. There was however significant difference among the cultivars in the number of shoots recorded. SNK values for explants-media interaction showed that cotyledon and radicle explants were significantly different (P &amp;lt; 0.05) from hypocotyl explants in the number of shoots produced.&lt;/p&gt;</jats:p

Progettazione e prova di servizi di orientamento e riallineamento on-line nel progetto UniSOFIA

In questo paper mostreremo il percorso svolto nell’ambito del sottoprogetto Orientamento e Riallineamento di UniSOFIA, illustrando la ricerca per la progettazione e la prova dei servizi realizzati con la finalità generale di costruire un ponte tra la scuola secondaria e l’università. La ricerca si è sviluppata prevalentemente on-line con l’individuazione e la comparazione di ‘casi’ e modelli disponibili, a partire dai quali sono state progettate le soluzioni adottate, compatibili con le condizioni di sistema disponibili. L’attenzione è rivolta in particolare alla presentazione di un questionario di auto-orientamento on-line con feedback automatico originale e del contesto funzionale di servizi di orientamento e riallineamento in cui è organicamente inserito. La prova è stata svolta monitorando l’utilizzo dei servizi disponibili on-line e con analisi in profondità resi possibili grazie alla collaborazione diretta degli utenti del corso di laurea in Scienze della comunicazione dell’Università di Cagliari che aderisce al progetto UniSOFIA. L’attività ha permesso di offrire servizi on-line di orientamento e riallineamento e di vagliarne l’utilizzabilità

Perbandingan Pengukuran Kadar Air Tanah Lempung Menggunakan Metode Gravimetry Dan Metode Gypsum Block Berdasarkan Variasi Kedalaman

Soil is one of the most important things in every civil engineering construction. But the problem is how to define physical and mechanic characteristic of the soil. Physical and mechanic characteristic that have influence on bearing capacity of the soil is water content. Size of soil also has effects on the changing of water content. To define water content, engineer is often using Gravimetry Method, but this method can only be doing in laboratory, so it can't specifically describe real condition of water content in the fields. Then, an in-site test such as Electrical Sensory Method is needed, in order To get correct result of water content in fields. In Electrical Sensory Method there is one method called Gypsum Block Method, this method is an in-situ water content test with individual calibration on laboratory.The results from Oebelo Village, Kupang District soils sample, the soils is CH soil, where CH is organic clay with high plasticity, Fat Clay. By using gypsum block method, water content results is in range of 19,35 % - 41,05 %, while with gravimetry method the water content is in range of 18,34 % - 34, 67 %. Water content test using gypsum block method can only be doing at 0,50 m dan 1,00 m depth range, at 1,50 m dan 2,00 m depth range cannot be using this method because is too difficult to install gypsum block and the gap results between gravimetry method and gypsum block method is in range of 4,19 % - 7,69 %

Kajian Spasial Faktor Risiko Terjadinya Kejadian Luar Biasa Campak dengan Geographical Information System

Cakupan imunisasi yang tinggi bukan jaminan tidak terjadi Kejadian Luar Biasa (KLB) campak. Penelitian ini bertujuan mengetahui faktor risiko penyebab terjadinya KLB campak di wilayah kerja Puskesmas KualinKabupaten TTS. Metode penelitian ini menggunakan pendekatan kualitatif dan kuantitatif. Data primer dari hasil investigasi KLB campak dan wawancara dengan responden. Jumlah sampel sebanyak 204 responden yang terdiri dari 102 kasus yang diambil dengan metode total sampling dan 102 kontrol yang diambil dengan metode purposive sampling. Desain penelitian yang digunakan adalah cross sectional study dan data analisis secaraspacial menggunakan GIS. Data dianalisis menggunakan uji chi-square. Penelitian ini menunjukkan sebanyak 94,1% berumur <15 tahun, dengan persentase laki-laki sebesar 54,9%, tidak diimunisasi 93,1%, dan sebesar 67,5% penderita tidak memperoleh vitamin A. Hasil uji chi-square diperoleh umur, vitamin A, status imunisasi, status gizi, pendapatan keluarga, kepadatan hunian, penyakit infeksi dan riwayat kontak sebagai faktor risiko terjadinya KLB Campak. Faktor risiko yang bersifat sebagai protektor adalah status imunisasi, pemberian vitamin A, status gizi, tingkat kepadatan hunian dan penyakit infeksi

Peptoid and Antibody-based GFP Sensors

In this work, we have made and characterized a pair of immunobiosensors for detecting the green fluorescent protein (GFP) in an aqueous matrix. An anti-GFP antibody-based biosensor was assembled to detect GFP, while a novel peptoid (N-substituted oligomers of glycine designated as IOS-1) biosensor was also assembled for GFP detection. A quartz crystal microbalance (QCM) gold sensor was used as the supporting substrate for self-assembly of the immunobiosensors. Gravimetric measurements of the QCM gold sensor during immunobiosensor construction and operation were available in real-time using a QCM instrument. X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and Fluorescence microscopy were used to characterize the immunobiosensors. Dose-dependent calibration curves were developed to contrast the performance of the peptoid immunobiosensor and the antibody-based immunobiosensor. The sensitivity of the biosensors shows that the peptoid could detect GFP at 8 nM, unlike the antibody immunobiosensor, which starts to measurably detect GFP at 40 nM. IOS-1 peptoid immunobiosensor had more adsorption capacity for GFP than the antibody-based immunobiosensor and could be reused through multiple adsorption/ desorption cycles. The peptoid immunobiosensor had a binding constant of 2.197 x 10(7) M(-1) with GFP

Optimizing Virus Prefiltration for Biopharmaceutical Manufacturing

Virus filters are single-use devices that use a size-based separation process. In virus filters, contaminating virus particles are retained while the therapeutic molecules pass through the membrane pores. Virus filters are an essential component of the overall virus clearance strategy. Sections 1 and 2 of this dissertation provide an introduction and extensive review of monoclonal antibody (mAb) process development, where virus filtration is pivotal. In section 3, prefiltration studies were performed with an industrially relevant IgG1 type mAb using adsorptive and size-exclusion-based prefilters with different mechanisms of action. This mAb has an isoelectric point range of 7.1 to 8.0 and a molecular weight (MW) of 148 kDa. Decoupled prefiltration and virus filtration studies were conducted. We attempted to elute bound species from the membrane to identify them. Permeate fractions from the prefilters were introduced as feed fractions to a Planova BioEX (Asahi Kasei Medical, Tokyo, Japan) commercial virus filter for flux decay studies. Prefiltration and virus filtration studies were performed at different pH and ionic strength buffer conditions. By adjusting buffer conditions, and choosing prefilters with an appropriate mechanism of action, increased selectivity for foulant capture resulting in improved flux behavior during virus filtration could be achieved. Extensive characterization was also performed for the various filtration fractions to determine molecular species that increase fouling propensity in the virus filter and the efficacy of the different prefilters at removing these species. In section 4, prefiltration and flux decay studies on a Viresolve Pro (MilliporeSigma, Billerica, MA) as well as the Planova BioEX virus filter was performed with another industrially relevant mAb with an isoelectric point range of 5.95 - 6.55. The impact of excipients on mAb fouling behavior was determined. The impact of buffer pH was also evaluated with one pH condition below the isoelectric point (pI) of the mAb and another pH condition above the mAb pI. Decoupled prefiltration was performed to evaluate the impact of different types of prefilters on the filterability of this mAb. The pharmaceutical analysis system PA800 plus (SCIEX, Redwood City, CA) was also used to characterize the various mAb fractions from prefiltration and virus filtration. Dynamic light scattering (particle size analysis), size exclusion chromatography, SDS PAGE, capillary electrophoresis, and MALDI mass spectrometry were used for characterization. In section 5, a new technique of fractionating close molecular weight biomolecules was evaluated for virus clearance. The technique is known as internally staged ultrafiltration (ISUF), where layers of ultrafiltration membranes operate in stages to fractionate biomolecules based on differences in isoelectric points. The membranes of interest were the Pall Omega PES 300 kDa molecular weight cutoff (MWCO) flat sheet membrane, Pall Omega PES 100 kDa MWCO membrane, Millipore Ultracel 100kDa MWCO, and the Millipore Ultracel 30kDa MWCO. Virus clearance studies were performed using internally staged ultrafiltration membranes in skin and backing configurations. Section 6 is an overall conclusion for this work showing major findings and identifying areas for future study

Characterizing the Role of Chemo-Mechanical Couplings in the Activation Process of Proteins that Undergo Conformational Changes

Proteins are not static entities; rather, they are dynamic macromolecules that undergo conformational changes to perform their biological functions. These structural transitions are often coupled to chemical events such as lipid interactions, or changes in the cellular environment. Understanding the intricate interplay between chemical perturbations and the resulting mechanical response is crucial for elucidating the activation mechanisms of proteins. This dissertation aims to characterize the role of chemo-mechanical couplings in the activation process of two G protein-coupled receptors (GPCRs): Cannabinoid Receptor 1 (CB1) and Metabotropic Glutamate Receptor 1 (mGluR1). We employ molecular dynamics simulations to investigate the conformational landscapes of CB1 and mGluR1. By studying the active and inactive states of apo CB1, we uncover distinct conformational dynamics between these states. Our results reveal that the inactive state of CB1 explores a wider conformational space compared to the more restricted active state. We identify key structural features and residues, such as the pivotal role of transmembrane helix 7 (TM7), that mediate the state-dependent conformational changes in CB1. Furthermore, we investigate the role of cholesterol in modulating the conformational dynamics of mGluR1. Our simulations show that cholesterol influences the conformational changes of the receptor, particularly in the TM1 and TM2 regions that form the dimeric interface. Intriguingly, we find that low cholesterol concentrations induce more significant conformational changes in mGluR1 compared to higher cholesterol levels or the absence of cholesterol. Our findings highlight the importance of considering the dynamic nature of proteins, rather than just their static structures, in understanding activation mechanisms. The activation process of CB1 and mGluR1 involves a complex network of dynamic conformational changes that are coupled to chemical perturbations. These results not only advance our understanding of GPCR activation but also underscore the necessity of incorporating protein dynamics into the development and refinement of therapeutic agents targeting these receptors. The knowledge gained from this dissertation could provide a framework for structure-based drug discovery that targets the state-specific conformational dynamics of CB1, mGluR1 and GPCRs at large. Our findings have potential applications in drug discovery, biotechnology, and precision medicine, where a detailed understanding of protein dynamics is essential for developing effective and selective treatments

Characterizing the Role of Chemo-Mechanical Couplings in the Activation Process of Proteins that Undergo Conformational Changes

Proteins are not static entities; rather, they are dynamic macromolecules that undergo conformational changes to perform their biological functions. These structural transitions are often coupled to chemical events such as lipid interactions, or changes in the cellular environment. Understanding the intricate interplay between chemical perturbations and the resulting mechanical response is crucial for elucidating the activation mechanisms of proteins. This dissertation aims to characterize the role of chemo-mechanical couplings in the activation process of two G protein-coupled receptors (GPCRs): Cannabinoid Receptor 1 (CB1) and Metabotropic Glutamate Receptor 1 (mGluR1). We employ molecular dynamics simulations to investigate the conformational landscapes of CB1 and mGluR1. By studying the active and inactive states of apo CB1, we uncover distinct conformational dynamics between these states. Our results reveal that the inactive state of CB1 explores a wider conformational space compared to the more restricted active state. We identify key structural features and residues, such as the pivotal role of transmembrane helix 7 (TM7), that mediate the state-dependent conformational changes in CB1. Furthermore, we investigate the role of cholesterol in modulating the conformational dynamics of mGluR1. Our simulations show that cholesterol influences the conformational changes of the receptor, particularly in the TM1 and TM2 regions that form the dimeric interface. Intriguingly, we find that low cholesterol concentrations induce more significant conformational changes in mGluR1 compared to higher cholesterol levels or the absence of cholesterol. Our findings highlight the importance of considering the dynamic nature of proteins, rather than just their static structures, in understanding activation mechanisms. The activation process of CB1 and mGluR1 involves a complex network of dynamic conformational changes that are coupled to chemical perturbations. These results not only advance our understanding of GPCR activation but also underscore the necessity of incorporating protein dynamics into the development and refinement of therapeutic agents targeting these receptors. The knowledge gained from this dissertation could provide a framework for structure-based drug discovery that targets the state-specific conformational dynamics of CB1, mGluR1 and GPCRs at large. Our findings have potential applications in drug discovery, biotechnology, and precision medicine, where a detailed understanding of protein dynamics is essential for developing effective and selective treatments

A versatile platform for three-dimensional dynamic suspension culture applications

In the last decades, the rapid upgrading in cell biological knowledge has bumped the interest in using cell-based therapeutic approaches as well as cell-based model systems for the treatment of diseases. Given the rapid translation towards cell-based clinical treatments and the consequent increasing demand of cell sources, three-dimensional (3D) suspension cultures have demonstrated to be an advantageous alternative to monolayer techniques for large scale expansion of cells and for the generation of three-dimensional model systems in a scale-up perspective. In this scenario, a versatile bioreactor platform suitable for 3D dynamic suspension cell culture under tuneable shear stress conditions is developed and preliminarily tested in two different biotechnological applications. By adopting simple technological solutions and avoiding rotating components, the bioreactor exploits a laminar hydrodynamics, enabling dynamic cell suspension in an environment favourable to mass transport. Technically, the bioreactor is conceived to produce dynamic suspension cell culture under tuneable shear stress conditions without the use of moving components (from ultralow to moderate shear stress). A multiphysics computational modelling strategy is applied for the development and optimization of the suspension bioreactor platform. The in silico modelling is used to support the design and optimization phase of the bioreactor platform, providing a comprehensive analysis of its operating principles, also supporting the development/optimization of culture protocols directly in silico, and thus minimizing preliminary laboratory tests. After the technical assessment of the functionality of the device and a massive number of in silico simulations for its characterization, the bioreactor platform has been employed for two preliminary experimental applications, in order to determine the suitability of the device for culturing human cells under dynamic suspension. In detail, the bioreactor platform has been used to culture lung cancer cells for spheroid formation (Calu-3 cell line) under ultralow shear stress conditions, and for human induced pluripotent stem cell (hiPSC) dynamic suspension culture. The use of the bioreactor platform for the formation of cancer cell spheroids under low shear stress conditions confirms the suitability of the device for its use as dynamic suspension bioreactor. In fact, compared to static cell suspension, after 5 days of dynamic suspension culture the bioreactor platform preserves morphological features, promotes intercellular connection, increases the number of cycling cells, and reduces double strand DNA damage. Calu-3 cells form functional 3D spheroids characterized by more functional adherence junctions between cells. Moreover, the computational model has been used as a tool for assisting the setup of the experimental framework with the extraction of the fluid dynamic features establishing inside the bioreactor culture chamber. As second proof of concept application, the bioreactor platform has been tested for the dynamic suspension of hiPSCs. Starting from the ‘a priori’ knowledge gained by the development of the in silico culture protocol, the agglomeration of human induced pluripotent stem cells has been modulated by means of the combination of moderate intermittent shear stress and free-fall transport within the bioreactor culture chamber. The inoculation of single cells suspensions inside the bioreactor chamber promotes cell-cell interaction and consequently the formation of human induced pluripotent stem cell aggregates. In conclusion, the impeller-free functioning principle characterizing the proposed bioreactor platform demonstrates to be promising for human cell dynamic suspension culture. In the future, this bioreactor platform will be further optimized for the realization of impeller-free dynamic suspension bioreactors dedicated and optimized to specific applications in stem cell and cancer cell culture