Validation of analytical methodology for quantification of cefazolin sodium pharmaceutical dosage form by high performance liquid chromatography to be applied for quality control in pharmaceutical industry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mechanisms of SnO2 Nanoparticles Formation and Growth in Acid Ethanol Solution Derived from SAXS and Combined Raman-XAS Time-Resolved Studies

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Nonlinear dynamics and control strategies: On a energy harvester vibrating system with a linear form to non-ideal motor torque

In this paper, we deal with the research of a vibrating model of an energy harvester device, including the nonlinearities in the model of the piezoelectric coupling and the non-ideal excitation. We show, using numerical simulations, in the analysis of the dynamic responses, that the harvested power is influenced by non-linear vibrations of the structure. Chaotic behavior was also observed, causing of the loss of energy throughout the simulation time. Using a perturbation technique, we find an approximate analytical solution for the non-ideal system. Then, we apply both two control techniques, to keep the considered system, into a stable condition. Both the State Dependent Ricatti Equation (SDRE) control as the feedback control by changing the energy of the oscillator, were efficient in controlling of the considered non-ideal system.

Effect of caffeine on larval mortality of Aedes aegypti: efficiency related to solution concentration and age

In two experiments, the duration of the effect of caffeine (CAF) solutions on larval mortality (LM) of Aedes aegypti was analyzed. In the first, LM was studied using solutions at 0.2, 0.5, 1.0 and 2.0 mg/mL aged from zero to five days in artificial breeding sites exposed to the laboratory environment (LE). In the second, the solutions aged at 1.0, 2.0 and 2.5 mg/mL closed flasks were stored in LE or in the refrigerator (R), and the effect on LM was tested in the experimental breeding sites at 30 days interval. In the first, the duration of the effect increased with the solution age in each CAF concentration. CAF at 1.0 and 2.0 mg/mL, without addition of food, produced 100% LM until 25 days after preparation; with food, at 11 and 18 days, respectively. In the second the effectiveness of CAF solutions lasted up to the seventh month, irrespective of whether they were stored in R or in LE. No adult emerged at any of the CAF concentrations used in second experiment.

Influence of beta-cyclodextrin on the properties of norfloxacin form A

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of temperature on salt-salt aqueous biphasic systems: manifestations of upper critical solution temperature

Water-miscible ionic liquids (ILs) may be salted out using kosmotropic salts such as potassium phosphate (K3PO4) to form salt-salt aqueous biphasic systems (ABS). The effect of temperature on these systems has been studied using phase diagrams and it is observed that the degree of binodal shift decreases (requiring lower IL and kosmotropic salt concentrations) with the increase of temperature following the trend [C(4)mim]Cl > [C(4)py]Cl > [C(4)mmim] Cl > [N-4444]Cl. This trend can be correlated with the decreasing hydrogen bonding abilities of each salt. The phase behavior was also interpreted on the basis of critical solution temperature behavior of pure aqueous ionic liquid solutions. Additionally, the distribution of alcohols in these systems was studied as a function of temperature and it was found that the distribution ratios did not change with changes in temperature. The Gibbs energy of transfer of a methylene group in these systems and correlation to tie-line length was also determined. It was concluded that while the miscibility of alcohols increases in the ILs with increasing temperature, phase divergence in the aqueous biphasic system decreases, and thus these competing forces tend to cancel each other out for small polar molecules. A comparison is provided for the response to temperature in the currently studied salt-salt systems and analogous ABS formed by the addition of hydrophilic polymers to kosmotropic salts (polymer-salt) or other polymers (polymer-polymer).

Effect of polymerization techniques and cleaning solution on flexural resistance of acrylic resin chemically activated

The purpose of this study was to evaluate the impact of different disinfection solutions on flexural resistance of chemically-activated acrylic resin. Test pieces were made of clear acrylic resin using a rectangular mold and employing two techniques: wet polymerization under pressure (n = 20) and dry polymerization under pressure (n = 20). Test pieces were subdivided into four equal groups: distilled water (control), sodium bicarbonate, 1% sodium hypochlorite and effervescent ats. The 30-day cycling technique consisted of immersing the test pieces in 100 ml of solution for 10 min three times a day and placing them in closed containers containing artificial saliva at 37°C. Subsequently, the flexural resistance of samples was tested. Data were analyzed using two-way analysis of variance (ANOVA) with forces serving as the dependent variables and the polymerization technique and cleaning agents as independent variables. Post hoc multiple comparisons were performed using Tukey’s test. There was no statistically significant difference in the flexural strength between the two polymerization techniques. The greatest flexural strength was observed for the effervescent tablets group followed by the control and 1% sodium hypochlorite groups which were statistically similar. Thus, the sodium bicarbonate solution caused the lowest flexural resistance of the test pieces.

Spectrophotometric determination of ciprofloxacin hydrochloride in ophthalmic solution

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Low-Resolution Molecular Models Reveal the Oligomeric State of the PPAR and the Conformational Organization of Its Domains in Solution

The peroxisome proliferator-activated receptors (PPARs) regulate genes involved in lipid and carbohydrate metabolism, and are targets of drugs approved for human use. Whereas the crystallographic structure of the complex of full length PPAR gamma and RXR alpha is known, structural alterations induced by heterodimer formation and DNA contacts are not well understood. Herein, we report a small-angle X-ray scattering analysis of the oligomeric state of hPPAR gamma alone and in the presence of retinoid X receptor (RXR). The results reveal that, in contrast with other studied nuclear receptors, which predominantly form dimers in solution, hPPAR gamma remains in the monomeric form by itself but forms heterodimers with hRXR alpha. The low-resolution models of hPPAR gamma/RXR alpha complexes predict significant changes in opening angle between heterodimerization partners (LBD) and extended and asymmetric shape of the dimer (LBD-DBD) as compared with X-ray structure of the full-length receptor bound to DNA. These differences between our SAXS models and the high-resolution crystallographic structure might suggest that there are different conformations of functional heterodimer complex in solution. Accordingly, hydrogen/deuterium exchange experiments reveal that the heterodimer binding to DNA promotes more compact and less solvent-accessible conformation of the receptor complex.

Synthesis of self-assembled tubes in direct conjugated porphyrin-gold nanohybrid solution

We report on the formation of self-assembled meso-tetrakis (p-sulfonatofenyl) porphyrin (H2 TPP'S POT. 4-''IND. 4') tubes stabilized by gold nanoparticles (NPs) in basic solution and on their spectroscopic chareterization. The role of the gold NPs in the aggregation dynamics of free-base sulfonated porphyrin (H2TPP'S POT. 4-''IND. 4') is also investigated. The direct conjugation of the gold NPs to the H2TPPS4 molecule quenches the fluorescence intensity, while absorption peaks are blue-shifted, indicating predominant H-type aggregation. It is observed that porphyrin molecules adsorbed on the surface of the gold NP interact and form tubes of maximum diameter ∼1.5 μm and length >100 μm. Steady-state and time-resolved spectroscopic techniques confirm nonradiative energy transfer from porphyrin to gold NP.

Structure determination of proteins and peptides in solution: simulation, chirality and NMR studies

The study of protein fold is a central problem in life science, leading in the last years to several attempts for improving our knowledge of the protein structures. In this thesis this challenging problem is tackled by means of molecular dynamics, chirality and NMR studies. In the last decades, many algorithms were designed for the protein secondary structure assignment, which reveals the local protein shape adopted by segments of amino acids. In this regard, the use of local chirality for the protein secondary structure assignment was demonstreted, trying to correlate as well the propensity of a given amino acid for a particular secondary structure. The protein fold can be studied also by Nuclear Magnetic Resonance (NMR) investigations, finding the average structure adopted from a protein. In this context, the effect of Residual Dipolar Couplings (RDCs) in the structure refinement was shown, revealing a strong improvement of structure resolution. A wide extent of this thesis is devoted to the study of avian prion protein. Prion protein is the main responsible of a vast class of neurodegenerative diseases, known as Bovine Spongiform Encephalopathy (BSE), present in mammals, but not in avian species and it is caused from the conversion of cellular prion protein to the pathogenic misfolded isoform, accumulating in the brain in form of amiloyd plaques. In particular, the N-terminal region, namely the initial part of the protein, is quite different between mammal and avian species but both of them contain multimeric sequences called Repeats, octameric in mammals and hexameric in avians. However, such repeat regions show differences in the contained amino acids, in particular only avian hexarepeats contain tyrosine residues. The chirality analysis of avian prion protein configurations obtained from molecular dynamics reveals a high stiffness of the avian protein, which tends to preserve its regular secondary structure. This is due to the presence of prolines, histidines and especially tyrosines, which form a hydrogen bond network in the hexarepeat region, only possible in the avian protein, and thus probably hampering the aggregation.

NMR studies on supramolecular aggregates in solution and the solid state

The aim of this work presented here is the characterization of structure and dynamics of different types of supramolecular systems by advanced NMR spectroscopy. One of the characteristic features of NMR spectroscopy is based on its high selectivity. Thus, it is desirable to exploit this technique for studying structure and dynamics of large supramolecular systems without isotopic enrichment. The observed resonance frequencies are not only isotope specific but also influenced by local fields, in particular by the distribution of electron density around the investigated nucleus. Barbituric acid are well known for forming strongly hydrogen-bonded complexes with variety of adenine derivatives. The prototropic tautomerism of this material facilitates an adjustment to complementary bases containing a DDA(A = hydrogen bond acceptor site, D = hydrogen bond donor site) or ADA sequences, thereby yielding strongly hydrogen-bonded complexes. In this contribution solid-state structures of the enolizable chromophor "1-n-butyl-5-(4-nitrophenyl)-barbituric acid" that features adjustable hydrogen-bonding properties and the molecular assemblies with three different strength of bases (Proton sponge, adenine mimetic 2,6-diaminopyridine (DAP) and 2,6-diacetamidopyridine (DAC)) are studied. Diffusion NMR spectroscopy gives information over such interactions and has become the method of choice for measuring the diffusion coefficient, thereby reflecting the effective size and shape of a molecular species. In this work the investigation of supramolecular aggregates in solution state by means of DOSY NMR techniques are performed. The underlying principles of DOSY NMR experiment are discussed briefly and more importantly two applications demonstrating the potential of this method are focused on. Calix[n]arenes have gained a rather prominent position, both as host materials and as platforms to design specific receptors. In this respect, several different capsular contents of tetra urea calix[4]arenes (benzene, benzene-d6, 1-fluorobenzene, 1-fluorobenzene-d5, 1,4-difluorobenzene, and cobaltocenium) are studied by solid state NMR spectroscopy. In the solid state, the study of the interaction between tetra urea calix[4]arenes and guest is simplified by the fact that the guests molecule remains complexed and positioned within the cavity, thus allowing a more direct investigation of the host-guest interactions.

Stable isotope fractionation of selenium by biomethylation in soil

Selen ist in geringen Mengen ein essentielles Nährelement, das aber in höheren Gehalten toxisch wird. Der Se-Kreislauf in der Umwelt ist eng mit Redoxreaktionen wie der Reduktion von Se-Oxyanionen zu Methylselenid verknüpft. Flüchtige Methylselenide sind weit verbreitet und stellen einen wichtigen Se-Fluss in der Umwelt dar. Das übergeordnete Ziel meiner Dissertation war, die Stabilisotopenfraktionierung von Se durch Biomethylierung verschiedener oxidierter Se-Spezies (Se[IV] und Se[VI]) im Boden aufzuklären. Zunächst wurde eine Methode entwickelt, die es erlaubte flüchte Methylselenide quantitativ zurückzuhalten. Es zeigte sich, dass alkalische Peroxid-Lösung dafür geeignet war. Mit alkalischer Peroxid-Lösung wurde eine Wiederfindung von 95,6 ± Standardabweichung 5,4% in Verflüchtigungsexperimenten mit Methylselenid-Standards erreicht. Bei Einsatz von alkalischer Peroxid-Lösung in geschlossenen Mikrokosmos-Experimenten kam es zu keinen Se-Verlusten und ausgeglichenen Se-Isotopenbilanzen. Die massengewichteten δ82/76Se-Werte lagen für Se(IV) und Se(VI) am Ende der Mikrokosmos-Inkubationen bei -0,31 ± 0,05‰ (n = 3) und -0,76 ± 0,07‰ (n = 3) verglichen mit -0,20 ± 0,05‰ und -0,69 ± 0,07‰ im jeweils zugegebenen Se. Im zweiten Teil meiner Dissertation wurde die Pilzart Alternaria alternata mit Se(VI) und Se(IV) in geschlossenen Mikrokosmen für 11-15 und Se(IV) zusätzlich für 3-5 Tage bei 30°C inkubiert. In 11-15 Tagen wurden 2,9-11% des Se(VI) und 21-29% des Se(IV) und in 3-5 Tagen, 3-5% des Se(IV) methyliert. Die anfänglichen δ82/76Se-Werte von Se(VI) und Se(IV) lagen bei -0,69 ± 0,07‰, und -0,20 ± 0,05‰. Die δ82/76Se-Werte der Methylselenide unterschieden sich nach 11-15 Tagen Inkubation signifikant zwischen Se(VI) (-3,97 bis -3,25 ‰) und Se(IV) (-1,44 bis -0,16‰) als Quellen. Die δ82/76Se-Werte der Methylselenide zeigen also die Quellen der Biomethylierung von Se an. Die kürzere Inkubation von Se(IV) für 3-5 Tage führte zu einer ausgeprägten Se-Isotopenfraktonierung von mindestens -6‰, bevor ein Fließgleichgewicht erreicht wurde. Im dritten Teil bestimmte ich die Bindungsformen von Se mit drei operativ definierten sequentiellen Extraktionen und die δ82/76S-Werte des gesamten Selens in zehn urbanen Oberböden mit 0,09-0,52 mg/kg Se, die fünf verschiedene Landnutzungstypen repräsentierten (Überschwemmungsgrünland, Garten, Park, Straßenrand und Wald). Nur ein kleiner Teil des Seleniums lag in austauschbarer und damit direkt bioverfügbarer und in residualer, wenig reaktiver Form vor. Das meiste Se war an die organische Substanz und Fe-(Hydr-)Oxide gebunden (42-77% des gesamten Selens). Der mittlere δ82/76Se-Wert des gesamten Selens in den Oberböden lag mit -0,03 ± 0,38‰ nahe beim Mittelwert der gesamten Erde. Geringfügig niedrigere Se-Isotopensignale von -0,59 bis -0,35‰ v.a. in Waldböden und geringfügig höhere von 0,26 to 0,45‰ in Überschwemmungsgrünland wurden vermutlich durch Boden-Pflanze-Recycling und Se-Kontaminationen durch das Flusswasser verursacht. Der vierte Teil umfasste ein “Natural Attenuation”-Experiment und Mikrokosmos-Inkubationen von Bodenproben mit A. alternata. Die Equilibrierung von zum Boden gegebenem Se(IV) und Se(VI) für drei Tage führte zu abnehmenden wasserlöslichen Se-Gehalten um 32-44% bzw. 8-14, die mit kleinen Isotopenfraktionierung (ε = -0,045 bis -0,12 ‰ and -0,05 to -0,07‰ verbunden waren. In zwei der inkubierten Böden mit mäßig sauren pH-Werten wurden zwischen 9,1 und 30% des zugefügten Se(IV) und 1,7% des zugefügten Se(VI) methyliert während in einem stark sauren Boden keine Methylierung auftrat. Das aus Se(IV) entstandene Methylselenid war deutlich gegenüber dem zugegebenen Se-Standard (0,20‰) an 82Se verarmt (δ82/76Se = -3,3 bis -4,5‰). Meine Ergebnisse zeigen, dass die stabilen Isotopenverhältnisse von Se neue Einblicke in Se-Transformationsprozesse erlauben.rn

Potential Analysis for Hypoelliptic Second Order PDEs with Nonnegative Characteristic Form

In this Thesis we consider a class of second order partial differential operators with non-negative characteristic form and with smooth coefficients. Main assumptions on the relevant operators are hypoellipticity and existence of a well-behaved global fundamental solution. We first make a deep analysis of the L-Green function for arbitrary open sets and of its applications to the Representation Theorems of Riesz-type for L-subharmonic and L-superharmonic functions. Then, we prove an Inverse Mean value Theorem characterizing the superlevel sets of the fundamental solution by means of L-harmonic functions. Furthermore, we establish a Lebesgue-type result showing the role of the mean-integal operator in solving the homogeneus Dirichlet problem related to L in the Perron-Wiener sense. Finally, we compare Perron-Wiener and weak variational solutions of the homogeneous Dirichlet problem, under specific hypothesis on the boundary datum.

Improving the sustainability of an appropriate technology solution; bone char defluoridation filters in Arusha, Tanzania

High concentrations of fluoride naturally occurring in the ground water in the Arusha region of Tanzania cause dental, skeletal and non-skeletal fluorosis in up to 90% of the region’s population [1]. Symptoms of this incurable but completely preventable disease include brittle, discolored teeth, malformed bones and stiff and swollen joints. The consumption of high fluoride water has also been proven to cause headaches and insomnia [2] and adversely affect the development of children’s intelligence [3, 4]. Despite the fact that this array of symptoms may significantly impact a society’s development and the citizens’ ability to perform work and enjoy a reasonable quality of life, little is offered in the Arusha region in the form of solutions for the poor, those hardest hit by the problem. Multiple defluoridation technologies do exist, yet none are successfully reaching the Tanzanian public. This report takes a closer look at the efforts of one local organization, the Defluoridation Technology Project (DTP), to address the region’s fluorosis problem through the production and dissemination of bone char defluoridation filters, an appropriate technology solution that is proven to work. The goal of this research is to improve the sustainability of DTP’s operations and help them reach a wider range of clients so that they may reduce the occurrence of fluorosis more effectively. This was done first through laboratory testing of current products. Results of this testing show a wide range in uptake capacity across batches of bone char emphasizing the need to modify kiln design in order to produce a more consistent and high quality product. The issue of filter dissemination was addressed through the development of a multi-level, customerfunded business model promoting the availability of filters to Tanzanians of all socioeconomic levels. Central to this model is the recommendation to focus on community managed, institutional sized filters in order to make fluoride free water available to lower income clients and to increase Tanzanian involvement at the management level.