Glyconanoparticles for colorimetric bioassays

Carbohydrate molecules are involved in many of the cellular processes that are important for life. By combining the specific analyte targeting of carbohydrates with the multivalent structure and change of solution colour as a consequence of plasmonic interactions with the aggregation of metal nanoparticles, glyconanoparticles have been used extensively for the development of bioanalytical assays. The noble metals used to create the nanocore, the methodologies used to assemble the carbohydrates on the nanoparticle surface, the carbohydrate chosen for each specific target, the length of the tether that separates the carbohydrate from the nanocore and the density of carbohydrates on the surface all impact on the structural formation of metal based glyconanoparticles. This tutorial review highlights these key components, which directly impact on the selectivity and sensitivity of the developed bioassay, for the colorimetric detection of lectins, toxins and viruses

Quantifying the cellular uptake of semiconductor quantum dot nanoparticles by analytical electron microscopy

Semiconductor quantum dot nanoparticles are in demand as optical biomarkers yet the cellular uptake process is not fully understood; quantification of numbers and the fate of internalized particles are still to be achieved. We have focussed on the characterization of cellular uptake of quantum dots using a combination of analytical electron microscopies because of the spatial resolution available to examine uptake at the nanoparticle level, using both imaging to locate particles and spectroscopy to confirm identity. In this study, commercially available quantum dots, CdSe/ZnS core/shell particles coated in peptides to target cellular uptake by endocytosis, have been investigated in terms of the agglomeration state in typical cell culture media, the traverse of particle agglomerates across U-2 OS cell membranes during endocytosis, the merging of endosomal vesicles during incubation of cells and in the correlation of imaging flow cytometry and transmission electron microscopy to measure the final nanoparticle dose internalized by the U-2 OS cells. We show that a combination of analytical transmission electron microscopy and serial block face scanning electron microscopy can provide a comprehensive description of the internalization of an initial exposure dose of nanoparticles by an endocytically active cell population and how the internalized, membrane bound nanoparticle load is processed by the cells. We present a stochastic model of an endosome merging process and show that this provides a data-driven modelling framework for the prediction of cellular uptake of engineered nanoparticles in general

Contact-facilitated drug delivery with Sn2 lipase labile prodrugs optimize targeted lipid nanoparticle drug delivery

Sn2 lipase labile phospholipid prodrugs in conjunction with contact-facilitated drug delivery offer an important advancement in Nanomedicine. Many drugs incorporated into nanosystems, targeted or not, are substantially lost during circulation to the target. However, favorably altering the pharmacokinetics and volume of distribution of systemic drug delivery can offer greater efficacy with lower toxicity, leading to new prolonged-release nanoexcipients. However, the concept of achieving Paul Erhlich's inspired vision of a ‘magic bullet’ to treat disease has been largely unrealized due to unstable nanomedicines, nanosystems achieving low drug delivery to target cells, poor intracellular bioavailability of endocytosed nanoparticle payloads, and the substantial biological barriers of extravascular particle penetration into pathological sites. As shown here, Sn2 phospholipid prodrugs in conjunction with contact-facilitated drug delivery prevent premature drug diffusional loss during circulation and increase target cell bioavailability. The Sn2 phospholipid prodrug approach applies equally well for vascular constrained lipid-encapsulated particles and micelles the size of proteins that penetrate through naturally fenestrated endothelium in the bone marrow or thin-walled venules of an inflamed microcirculation. At one time Nanomedicine was considered a ‘Grail Quest’ by its loyal opposition and even many in the field adsorbing the pains of a long-learning curve about human biology and particles. However, Nanomedicine with innovations like Sn2 phospholipid prodrugs has finally made ‘made the turn’ toward meaningful translational success

Guiding the Design of Synthetic DNA-Binding Molecules with Massively Parallel Sequencing

Genomic applications of DNA-binding molecules require an unbiased knowledge of their high affinity sites. We report the high-throughput analysis of pyrrole-imidazole polyamide DNA-binding specificity in a 10^(12)-member DNA sequence library using affinity purification coupled with massively parallel sequencing. We find that even within this broad context, the canonical pairing rules are remarkably predictive of polyamide DNA-binding specificity. However, this approach also allows identification of unanticipated high affinity DNA-binding sites in the reverse orientation for polyamides containing β/Im pairs. These insights allow the redesign of hairpin polyamides with different turn units capable of distinguishing 5′-WCGCGW-3′ from 5′-WGCGCW-3′. Overall, this study displays the power of high-throughput methods to aid the optimal targeting of sequence-specific minor groove binding molecules, an essential underpinning for biological and nanotechnological applications

Sequence-selective detection of double-stranded DNA sequences using pyrrole-imidazole polyamide microarrays

We describe a microarray format that can detect double-stranded DNA sequences with a high degree of sequence selectivity. Cyclooctyne-derivatized pyrrole-imidazole polyamides were immobilized on azide-modified glass substrates using microcontact printing and a strain-promoted azide-alkyne cycloaddition (SPAAC) reaction. These polyamide-immobilized substrates selectively detected a seven-base-pair binding site incorporated within a double-stranded oligodeoxyribonucleotide sequence even in the presence of an excess of a sequence with a single-base-pair mismatc

CREDIT POLICY OF IMEX BANKA D.D.

U ovom radu analizirane su i obrazložene procedure koje banke koriste prilikom procjene kreditne sposobnosti klijenata. Iznesene su teorijske postavke o kreditnoj politici, kreditnom riziku, kao i načini procjene kreditne sposobnosti. Nadalje, prikazan je cjelokupan proces kreditne aktivnosti banke, od zaprimanja obrasca kreditnog zahtjeva i ostale dokumentacije, do postupaka analize financijskih izvještaja putem pokazatelja, te donošenja odluke o odobravanju ili odbijanju kreditnog zahtjeva. Doneseni su zaključci kako se banke koriste formaliziranim pravilima i procedurama koje smanjuju rizik odobravanja po banku loših kredita. Utvrđeno je da su financijski pokazatelji temelj za donošenje pozitivne ili negativne odluke te da dugoročno dobri odnosi banke i klijenta predstavljaju vrlo bitan faktor kod donošenja odluke o kreditiranju.In this thesis, procedures used by banks upon the assessment of the creditworthiness of clients have been analysed and explained. Theoretical assumptions regarding credit policy and credit risk have been presented, as well as the multiple methods of the assessment of creditworthiness. Furthermore, the entire process of the bank lending has been shown, from the receiving of the loan application form and other documentation to the methods of analysis of financial statements by means of indicators, and the making of the decision on approving or rejecting the loan application. Conclusions were made about the bank using formalized rules and procedures which reduce the risk of granting loans which do not benefit the bank. It was found that the financial indicators form the basis for making a positive or negative decision and that good relations between a bank and a client represent a very important factor when making a decision on lending in the long term

Evaluation Boards for 8-bit and 32-bit Microcontrollers

U okviru ovog rada razvijena su dva edukacijska razvojna sustava za rad s 8-bitnim i 32-bitnim mikrokontrolerima. Kao temelj sustava za rad s 8-bitnim mikrokontrolerima odabran je ATmega4809 tvrtke Microchip, a za rad s 32-bitnim mikrokontrolerima STM32F407VGT6 tvrtke STMicroelectronics. Obje pločice opremljene su identičnim skupom periferija koji uključuje svjetleće diode, tipke, 7-segmentne pokaznike, zujalicu, grijač, I2C termometar, potenciometar, RS232 i USB sučelje. Obje pločice sadrže i konektor za buduća proširenja. Napajanje je izvedeno putem USB priključka i ugrađenog linearnog regulatora. Programiranje i uhodavanje mikrokontrolera izvodi se vanjskim uređajima putem ugrađenog konektora. U radu je opisana sklopovska implementacija, skup programskih funkcija za rad s ugrađenim periferijama i jednostavna korisnička aplikacija koja demonstrira rad svih ugrađenih komponenti. Zbog odabranog skupa periferija i svoje jednostavnosti, opisani razvojni sustavi pogodni su za uvod u rad s ugradbenim računalima.This thesis demonstrates the development of two educational development systems for working with 8-bit and 32-bit microcontrollers. The 8-bit system is based on Microchip ATmega4809 platform, and the 32-bit system is based on STMicroelectronics STM32F407VGT6 platform. Both PC boards are equipped with the identical set of peripheral devices that includes Light emitting diodes, 7-segment display, buzzer, heater, I2C thermometer, potentiometer, RS232 and USB interface. Both development boards also contain the connector for the future extensions. Power supply is implemented via USB port and built-in linear regulator. Microcontroller programming and debugging is accomplished with external devices via the built-in connector. This thesis describes the hardware implementation, set of software functions, and simple user application that demonstrates the functionalities of the built-in components. Due to the well-selected peripheral devices and simplicity, these systems are especially appropriate as introduction into the field of embedded systems

Educational Evaluation Board for 8-bit and 32-bit Microcontrollers

U ovom radu razvijen je edukacijski sustav za rad s mikrokontrolerima. Kao temelj sustava odabrane su razvojne pločice ATmega4809 Curiosity Nano i STM32F4 Discovery. U okviru sustava, ove pločice su proširene svjetlećim diodama, 7-segmentnim pokaznicima, zujalicom, grijačem, tipkama, rotacijskim enkoderom, potenciometrom, analognim pretvornikom temperature, RS232 sučeljem te I2C termometrom i I2C sklopom za proširenje. Razvojni sustav je realiziran na tiskanoj pločici dimenzija 100 mm x 100 mm na koju se priključuju spomenute razvojne pločice. U radu je opisana sklopovska implementacija. Nadalje, razvijen je skup funkcija za rad s periferijama na pločici te korisnička aplikacija koja demonstrira rad cijelog sustava. Razvijeni sustav je jednostavan, cjenovno povoljan i bogat periferijama što ga čini pogodnim za prvi susret s ugradbenim računalnim sustavima.In this thesis, an educational system for work with microcontrollers has been developed. Development boards ATmega4809 Curiosity Nano and STM32F4 Discovery were selected for the basis of the system. In the system framework, these boards were then further extended with LEDs, 7-segment LED displays, a buzzer, a heater, keys, a rotary encoder, a potentiometer, an analog temperature sensor, an RS232 interface, an I2C temperature sensor and an I2C port expander. The educational system was built on a printed circuit board (dimensions 100 mm x 100 mm) that has enabled the attaching of the aforementioned development boards. This thesis describes the hardware implementation. Furthermore, a set of functions for utilizing the peripherals and a demo user application have been developed. The presented system is simple, cost-effective and rich in peripherals, which makes it ideal for the introduction to the embedded computer systems

Educational Evaluation Board for 8-bit and 32-bit Microcontrollers

U ovom radu razvijen je edukacijski sustav za rad s mikrokontrolerima. Kao temelj sustava odabrane su razvojne pločice ATmega4809 Curiosity Nano i STM32F4 Discovery. U okviru sustava, ove pločice su proširene svjetlećim diodama, 7-segmentnim pokaznicima, zujalicom, grijačem, tipkama, rotacijskim enkoderom, potenciometrom, analognim pretvornikom temperature, RS232 sučeljem te I2C termometrom i I2C sklopom za proširenje. Razvojni sustav je realiziran na tiskanoj pločici dimenzija 100 mm x 100 mm na koju se priključuju spomenute razvojne pločice. U radu je opisana sklopovska implementacija. Nadalje, razvijen je skup funkcija za rad s periferijama na pločici te korisnička aplikacija koja demonstrira rad cijelog sustava. Razvijeni sustav je jednostavan, cjenovno povoljan i bogat periferijama što ga čini pogodnim za prvi susret s ugradbenim računalnim sustavima.In this thesis, an educational system for work with microcontrollers has been developed. Development boards ATmega4809 Curiosity Nano and STM32F4 Discovery were selected for the basis of the system. In the system framework, these boards were then further extended with LEDs, 7-segment LED displays, a buzzer, a heater, keys, a rotary encoder, a potentiometer, an analog temperature sensor, an RS232 interface, an I2C temperature sensor and an I2C port expander. The educational system was built on a printed circuit board (dimensions 100 mm x 100 mm) that has enabled the attaching of the aforementioned development boards. This thesis describes the hardware implementation. Furthermore, a set of functions for utilizing the peripherals and a demo user application have been developed. The presented system is simple, cost-effective and rich in peripherals, which makes it ideal for the introduction to the embedded computer systems

Evaluation Boards for 8-bit and 32-bit Microcontrollers

U okviru ovog rada razvijena su dva edukacijska razvojna sustava za rad s 8-bitnim i 32-bitnim mikrokontrolerima. Kao temelj sustava za rad s 8-bitnim mikrokontrolerima odabran je ATmega4809 tvrtke Microchip, a za rad s 32-bitnim mikrokontrolerima STM32F407VGT6 tvrtke STMicroelectronics. Obje pločice opremljene su identičnim skupom periferija koji uključuje svjetleće diode, tipke, 7-segmentne pokaznike, zujalicu, grijač, I2C termometar, potenciometar, RS232 i USB sučelje. Obje pločice sadrže i konektor za buduća proširenja. Napajanje je izvedeno putem USB priključka i ugrađenog linearnog regulatora. Programiranje i uhodavanje mikrokontrolera izvodi se vanjskim uređajima putem ugrađenog konektora. U radu je opisana sklopovska implementacija, skup programskih funkcija za rad s ugrađenim periferijama i jednostavna korisnička aplikacija koja demonstrira rad svih ugrađenih komponenti. Zbog odabranog skupa periferija i svoje jednostavnosti, opisani razvojni sustavi pogodni su za uvod u rad s ugradbenim računalima.This thesis demonstrates the development of two educational development systems for working with 8-bit and 32-bit microcontrollers. The 8-bit system is based on Microchip ATmega4809 platform, and the 32-bit system is based on STMicroelectronics STM32F407VGT6 platform. Both PC boards are equipped with the identical set of peripheral devices that includes Light emitting diodes, 7-segment display, buzzer, heater, I2C thermometer, potentiometer, RS232 and USB interface. Both development boards also contain the connector for the future extensions. Power supply is implemented via USB port and built-in linear regulator. Microcontroller programming and debugging is accomplished with external devices via the built-in connector. This thesis describes the hardware implementation, set of software functions, and simple user application that demonstrates the functionalities of the built-in components. Due to the well-selected peripheral devices and simplicity, these systems are especially appropriate as introduction into the field of embedded systems