Helicobacter pylori is a Gram-negative bacterium that is the major cause of many upper gastrointestinal diseases such as gastritis and peptic ulcer disease. It has the unique ability of colonising the human gastric mucosa. Failure in complete eradication of H. pylori in infected patients, mainly due to antibiotic resistance, has necessitated the development of better therapeutics, especially from natural sources. In this study, extract of Maris piper potatoes were obtained and evaluated for antibacterial activity against H. pylori in vitro. Antibacterial activity was carried out against antibiotic-sensitive and clinical antibiotic-resistant H. pylori strains, as well as a range of Gram-negative bacteria including Helicobacter and Campylobacter species, using the viable count method. Result of the antibacterial assays indicated that potato extract is bactericidal against H. pylori lab strain as well as clinical antibiotic-resistant strains, with minimum inhibitory concentration at 15.6 mg/ml. Potato extract also showed minimal antibacterial activity against other Gram- negative bacteria tested, with minimum inhibitory concentration at 250 mg/ml. The effect of the extract on the morphology of H. pylori was also observed by transmission electron microscopy (TEM). TEM analysis of potato extract-treated H. pylori cells showed disruption of the morphology of H. pylori, characterized by separation of the outer membrane from the inner membrane and loss of cell shape. Potato extract also caused hyperpolarisation of H. pylori plasma membrane; however it is unclear whether the membrane active pumping activity is affected. Mutants of H. pylori that are resistant to potato extract were generated as a means to identify the target of potato extract within the H. pylori genome. Genome sequence analysis led to the discovery of a hypothetical protein, encoded by HP0603 gene, which may be involved in inducing resistance to potato extract. The results obtained in this study provide great insights into the anti-H. pylori activity of potato extract. Overall, this study suggests the potential use of potato extract as a source of anti-H. pylori agents; and stimulates further studies into identifying its mechanism of action.

No description available.

Antibacterial agent delivery is of great importance in medicine and dentistry since the bacterial infections are still one of the major reasons for hospitalization and mortality. Despite of the development of technique and pharmacy, more antimicrobial agents are optimized and utilized to treat infections, and their action of principal is better understood which lay a foundation for developing strategies for infection treatment. Over the last decades, many delivery systems have been established to deliver bacterial agents and maintain a sustained activity against them. However, the bacteria are always developing and finding a way to defend themselves. A more responsive antibacterial agent delivery system, which can release the active substances on demand to match the stages of diseases, is highly desirable. Therefore, it motivates us to carry out the work to develop a multifunctional delivery system for antibacterial particle formulation and encapsulation based on the layer-by-layer self-assembly technique and electrospinning, to manipulate the release with external triggers, such as near-infrared (NIR) light and alternating magnetic field (AMF). Strategically, two different kinds of antibacterial agents, chlorhexidine and doxycycline, were studied. Chlorhexidine was fabricated into spherical particles and functionalized with both gold and magnetite nanoparticles, and doxycycline was encapsulated within microcapsules which were also functionalized with magnetite nanoparticles. Their release kinetics and possibilities to trigger the release with either a NIR light or AMF was explored. The first two chapters of the thesis give a general introduction and literature review on the current use of antibacterial agents and the problems concerned, strategies already developed for antibacterial agent delivery, and the potential triggers to induce a smart release. In chapter 3, a brief description of materials and methods, and instruments is presented. Chapter 4 is about chlorhexidine particle formulation. Firstly, particulation of chlorhexidine and mechanism of 4 spherical interconnected structure formation was explored, and then the chlorhexidine particles are encapsulated either by LbL assembly or spray-drying. The chlorhexidine spheres were also functionalized with gold nanorods and Fe3O4 nanoparticles to achieve NIR light and magnetic field manipulated release, and the effect of nanoparticles on the formation of chlorhexidine spheres was also studied. When the chlorhexidine particles were incorporated into electrospun fibers, a sustained antibacterial activity was demonstrated. Chapter 5 is about the delivery of doxycycline to cells with microcapsules and the sustained intracellular doxycycline activity was demonstrated via EGFP expression when the cells were engineered with a tetracycline regulated gene expression system. Intracellular triggered release and upregulation of EGFP expression was achieved by an AMF. The results successfully demonstrated the possibility of chlorhexidine and doxycycline delivery and NIR light/AMF triggered release, which is promising for a future application in medicine and dentistry.

Some yeast strains secrete extracellular polypeptide toxins known to have potential growth inhibitory activity on sensitive yeast cells. These yeast strains are known as killer yeasts and their toxins are named as killer toxins or killer proteins. Yeast killer proteins are found inhibitory to Gram-positive bacteria in several studies which were based on microbial interactions of the producer strains tested with sensitive strains. K5 type yeast killer protein produced by Pichia anomala NCYC 434 was previously purified and characterized in our laboratory. The protein is glycosilated and has a pI value of 3,7 and molecular mass of 49 kDa, with exo &amp / #946 / -1,3-glucanase activity. Antibacterial activity of the pure K5 type yeast killer protein was tested against 19 clinical isolates of gram-positive bacteria causing skin infections and 2 quality control strains and found to have inhibitory activity on the isolates of Methicillin-sensitive Staphylococcus aureus (MSSA) and Enterococcus faecium. Toxin MIC and MBC ranges were 32 - 256 &micro / g/ml and 64 - &gt / 512 &micro / g/ml respectively. Cell killing analysis revealed that toxin has a bacteriostatic activity and the inhibitory effect starts between 8. and 12. hours. Regrowth of the bacteria is retarded with the increased dose of the toxin. K5 type yeast killer protein might be used as a topical antibacterial agent with its bacteriostatic activity for skin and wound infections caused by MSSA and Enterococcus faecium with appropriate formulation studies upon the antibacterial spectrum determination of the toxin in this study.

Nowdays, an emphasis is plased on an antibacterial properties of various products, including floor and coating materials. This work is about an testing of effectivity of the chosen antibacterial agents in the water born and solvent free floor systems which are epoxide or polyurethane based. As first step, the most suitable representative of each floor system was chosen, the silver based agents (respectively silver anion Ag+ based) which should have an antibacterial effect when incorporated into the floor system was chosen. The selected ones are Ag/SiO2, Ag/TiO2, Ag/Al2O3, Ag/ZnO, Ag/BaO, Ag/zeolite, AgI. The necessary was a creating of the test method which has to offer exact and reproducable outputs. At the begining, the method came out from the norms ČSN EN ISO 20645 Squared textiles – Determination of antibacterial activity – Test of diffusion throught the agar plate, ČSN 79 3880 Testing of antimicrobial properties of material and product of boot industry, Antifugal and antibacterial properties. Testing by this way did not bring repriducable outputs. Only the bacterial strains Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus was used according to the norms. The method of recultivation was used for testing. The process is: an application of the coating on the bottom of the Petri´s dish, curing, appplication of the bacterial solution, cultivation for 24 hours at 37 °C, inoculation onto the agar in the Petri´s dish, cultivation for 24 hours at 37 °C, interpretation of the grow/inhibition of the bacteries. The results offer the overview about the advantages and disadvantages of each antibacterial agent in the chosen systems.

Syftet med denna studie var att undersöka den antibakteriella effekten hos ett polyestertyg med tunna invävda koppartrådar. Frågeställningen som skulle besvaras var ifall koppartyget hade en baktericid eller bakteristatisk effekt. Koppartyget är ännu i prototypstadie och om det visar sig ha antibakteriella egenskaper är det tänkt att användas inom klinisk verksamhet för att förhindra bakterietillväxt i sår och andra känsliga lokaler. Koppar är ett essentiellt spårämne, men har också antimikrobiella egenskaper som utövas genom ett brett spektra av mekanismer där skador på cellmembranet är en av de viktigare. Metoderna som användes för att inokulera bakterier på tyget var absorptionsmetoden, där en näringsbuljong innehållande Staphylococcus aureus ATCC 6538 pipetterades på tygproverna, och transfermetoden där tygproverna trycktes mot en agar som racklats med peptonsaltlösning innehållande S. aureus. Totalantalet viabla bakterier per tygprov beräknades efter kort kontakt (<1min) och inkubering (18-24 h vid 37±2°C) genom att räkna viable count. Resultaten efter inkubering visade signifikant skillnad i totalantal bakterier mellan koppartyget och negativ kontroll i tre av fyra försök. Kort kontakt visade tendens till viss antibakteriell effekt. Slutsatsen är att koppartyget skadade och dödade bakterier då de fick inkubera på tyget, medan fler försök behövs för att säkerställa effekten av kort kontakt med koppartyget. / The purpose of this study was to test the antibacterial effect of thin copper treads woven into a polyester fabric. The investigation was done by inoculation of Staphylococcus aureus strain ATCC 6538 to the fabric and evaluation of the number of viable cells post exposure by viable count. The issue to be answered was whether the copper fabric had a bactericide or bacteriostatic effect? The fabric is still in prototype stage, and if proven to have antibacterial properties the aim is to use it to prevent bacterial growth in wounds and other vulnerable locations in clinical care. Copper is an essential trace element, but also has antimicrobial properties through a wide range of mechanisms where cell membrane damage is one of the more important. Methods used for inoculation was the absorption method, where a nutrient broth containing S. aureus was pipetted on to the fabric specimens, and the transfer method where the fabric specimens were pressed onto an agar plate that had previously been spread with peptone salt solution containing S. aureus. Total number of bacteria per fabric specimen after short contact (<1 min) and incubation (18-24 h at 37±2°C) was calculated. Incubation showed significant difference in total number of bacteria between the copper fabric and negative control in three of four tests. Short contact showed a tendency of antibacterial effect. The conclusion was that the copper fabric harmed and killed bacteria during incubation but that more records would be needed to be sure about the effects of short contact on bacteria.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O gÃnero Guettarda (Rubiaceae) compreende plantas extensamente distribuÃdas em Ãreas tropicais. Com relaÃÃo à espÃcie Guettarda sericea, a literatura demonstra que existe uma ampla carÃncia acerca de estudos botÃnicos e fitoquÃmicos. Assim, o presente trabalho buscou avaliar o efeito antibacteriano do extrato etanÃlico das folhas de G. sericea (EEFGS) e suas fraÃÃes sobre o crescimento de Streptococcus oralis ATCC 10557 e S. salivarius ATCC 7073, nas formas planctÃnicas e de biofilmes. Diferentes metodologias foram empregadas para a verificaÃÃo do potencial antimicrobiano. Dentre estas estÃo a determinaÃÃo da concentraÃÃo inibitÃria mÃnima (MIC), a determinaÃÃo da curva de morte e a avaliaÃÃo da concentraÃÃo bactericida mÃnima (MBC). AlÃm disso, a quantificaÃÃo da biomassa e do nÃmero de cÃlulas viÃveis do biofilme foi realizada, respectivamente, atravÃs da coloraÃÃo pelo cristal violeta e contagem de unidades formadoras de colÃnia (UFC). Os controles, negativo e positivo utilizados em todos os ensaios foram, respectivamente, DMSO 4% e Gluconato de Clorexidina com concentraÃÃo ajustada de acordo com os dados da CIM de cada micro-organismo. Para a determinaÃÃo da toxicidade do EEFGS utilizou-se o ensaio de toxicidade sobre nÃuplios de Artemia. Os dados mostraram que o extrato e as subfraÃÃes 13 a 17 da fraÃÃo clorofÃrmio apresentaram um marcante efeito antimicrobiano, sendo capazes de inibir o crescimento planctÃnico, bem como o desenvolvimento de biofilmes da cepa de S. oralis atà a concentraÃÃo de 62,5 Âg.mL-1. Com relaÃÃo a S. salivarius, apenas a subfraÃÃo 12 interferiu sobre o crescimento bacteriano. No tocante à toxicidade, foi observado que a morte dos nÃuplios de Artemia ocorreu em concentraÃÃes mais elevadas do que aquelas que apresentaram efeito antibacteriano. A partir de tais resultados pode-se concluir que EEFGS e as subfraÃÃes 13 a 17 podem ser utilizados como insumos para o controle da formaÃÃo de biofilmes de S. oralis. Em adiÃÃo, metodologias complementares que busquem a purificaÃÃo dos compostos ativos e seus efeitos citotÃxicos sobre cÃlulas eucariÃticas necessitam ser realizadas, visando sua utilizaÃÃo como um agente fitoterÃpico. / The genus Guettarda (Rubiaceae) comprises plants widely distributed in tropical areas. Regarding the Guettarda sericea species, the literature shows that there is a lack of botanical and phytochemicals studies. Thus, the present study aimed to evaluate the antibacterial effect of ethanol extract of leaves of G. sericea (EEFGS) and its fractions on the growth of Streptococcus oralis ATCC 10557 and S. salivarius ATCC 7073 in both the planktonic and biofilms states. Different methods were employed to verify the antimicrobial potential. Among these are the determination of minimum inhibitory concentration (MIC) determination of the death curve and evaluation of minimum bactericidal concentration (MBC). Furthermore, quantification of biomass and the number of viable cells of the biofilm were performed, respectively, by staining with crystal violet and counting colony forming units (CFU). The negative and positive controls used in all assays were respectively 4% DMSO and chlorhexidine gluconate with concentration adjusted according to the data of the MIC of each microorganism. To determine the toxicity of EEFGS, it was used the toxicity test on Artemia nauplii. The data showed that the extract and subfractions 13 to 17 of the chloroform fraction show a remarkable antimicrobial effect, able to inhibit the growth of planktonic and development of biofilms of S. oralis strain until the concentration of 62.5 μg.mL-1. With respect to S. salivarius, only subfraction 12 interfered on bacterial growth. Regarding the toxicity, it was observed that death of Artemia nauplii occurred at higher concentrations than those that exhibited antibacterial effect. From these results it can be concluded that EEFGS and subfractions 13-17 can be used as agents for the control of biofilm formation of S. oralis. In addition, complementary methodologies that seek purification of the active compounds and their cytotoxic effects on eukaryotic cells need to be held, aiming its use as an herbal agent.

Submitted by Neusa Fagundes (neusa.fagundes@unioeste.br) on 2018-07-12T19:16:44Z No. of bitstreams: 2 Roberta_Rodrigues2016.pdf: 550322 bytes, checksum: 270469fe680e8ab3e1299ebd0c7bcd19 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-07-12T19:16:44Z (GMT). No. of bitstreams: 2 Roberta_Rodrigues2016.pdf: 550322 bytes, checksum: 270469fe680e8ab3e1299ebd0c7bcd19 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-12-13 / Adhesive technology has been developed quickly since its introduction about fifty years ago. Its goal is to produce a close contact within enamel and dentin. Failures in this process could result in microleakage, and allow the infiltration of bacteria, fluids and ions. Recent studies have been shown the use of chlorhexidine associated with adhesive systems can be able to inhibit the bacteria action. The aim of this study was to evaluate the addiction of different percentages of chlorhexidine to a self-etch adhesive. The solution of digluconate chlorhexidine was increased to the primer of the two pass self-etch adhesive to create a 0,5%, 1,0% and 2,0% chlorhexidine primer solution and after they were distributed in four groups (G1, G2, G3 and G4). Saliva samples (N=10) were used to test bacteria activity. They were spread in a blood medium with filter paper disks containing the different treatments. After the incubation, the inhibitions halos formation were evaluated. This study demonstrated that, in vitro, the addition of different percentages of chlorhexidine digluconate to the self-etch adhesive induced inhibited halos at bacteria of saliva samples, independent from their concentration. / A tecnologia adesiva vem se desenvolvendo rapidamente desde que foi introduzida há mais de 50 anos. Seu principal objetivo é alcançar um íntimo contato entre a estrutura dental e o material restaurador e fornecer adequada adesão entre o esmalte e a dentina. Falhas nesse processo adesivo podem resultar em microinfiltração marginal com passagem de bactérias, fluidos ou íons entre a parede cavitária e o material restaurador. Recentes estudos têm demonstrado que a utilização da clorexidina associada ao sistema adesivo pode ser capaz de inibir a ação das bactérias. O objetivo do presente estudo foi avaliar o efeito antibacteriano de um sistema adesivo auto-condicionante de dois passos associado a diferentes concentrações de solução de digluconato de clorexidina. A solução de digluconato de clorexidina a 20% foi adicionada ao primer do sistema adesivo nas concentrações de 0,5, 1,0 e 2,0% e distribuídas em quatro grupos (G1, G2, G3 e G4). Para o teste de atividade antibacteriana foram utilizadas amostras de saliva (N=10) semeadas em meio de ágar sangue e discos de papel filtro contendo os diferentes grupos. Após incubação, as amostras foram inspecionadas quanto à formação de halo de inibição dos microrganismos através da utilização do paquímetro digital de precisão(0,0001mm) (Mitutoyou Sul Americana Ltda/ Starret Tools®). Este trabalho demonstrou que, in vitro, a adição de diferentes concentrações de digluconato de clorexidina ao sistema adesivo produziu halo de inibição dos microrganismos presentes nas amostras de saliva, independente da concentração utilizada.

Orientador: Pedro Paulo Corbi / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-25T01:00:34Z (GMT). No. of bitstreams: 1 Paiva_RaphaelEnoqueFerrazde_M.pdf: 15807547 bytes, checksum: e13f0a8a3bbde08ba558ac7cdb13c9a5 (MD5) Previous issue date: 2014 / Resumo: Complexos metálicos têm sido estudados quanto as suas propriedades medicinais há décadas. Neste trabalho, dois complexos inéditos de Ag(I) e Pt(II) foram sintetizados com o anti-inflamatório nimesulida (NMS), e avaliados como agentes antibacterianos e antitumorais. O complexo Ag-NMS (AgC13H11N2O5S) apresenta o ligante em uma coordenação bidentada à prata pelos átomos de N e O do grupo sulfonamida. A estrutura proposta foi confirmada por DFT. Devido à baixa solubilidade em água, foi preparado um complexo de inclusão de Ag-NMS em b-CD, pelo método de co-evaporação. Utilizando o método de Scatchard, foi determinado o valor de Ka = 370 2 L mol. Estudos de RMN por correlação H-H através do espaço mostram que a inclusão ocorre pelo grupo fenoxi da NMS. Já o complexo Pt-NMS (PtC26H22N4O10S2) apresenta dois ligantes, coordenados pelos átomos de N e O do grupo sulfonamida, para cada Pt(II). A DFT indica que o isômero N, O trans é o mais estável. O complexo Ag-NMS apresentou valores de MIC na faixa de 15,0-120 mmol L sobre cepas de Pseudomonas aeruginosa, Escherichia col e Staphylococcus aureus. O CE-[(Ag-NMS)·b-CD], embora mais solúvel em água do que o Ag-NMS, não apresentou atividade antibacteriana nas concentrações testadas. Os complexos Ag-NMS e Pt-NMS mostraram-se citotóxicos sobre células normais (Balb/c 3T3) e tumorais (SK-Mel 103 e Panc-1), porém o Pt-NMS foi significativamente mais seletivo contra as linhagens tumorais. Os ensaios de intercalação com EtBr e a avaliação da estrutura do DNA por dicroísmo circular indicam que o DNA não é um alvo biológico para o complexo Pt-NMS, indicando um mecanismo de ação diferente da cisplatina / Abstract: Metal complexes have been studied regarding its medicinal properties for decades. In this work, novel complexes of Ag(I) and Pt(II) with the anti-inflammatory nimesulide were synthesized and evaluated regarding their antibacterial and antitumoral properties. The Ag-NMS complex (AgC13H11N2O5S) shows the ligand in a bidentate coordination mode, bound to silver through the N and O atoms of the sulfonamide group. The proposed structure was confirmed by DFT. Due to its poor solubility in water, the Ag-NMS complex was included in b-CD, by co-evaporation. The Ka = 370 2 L mol was determined using the Scatchard method. Studies by H-H NMR correlation through space shows the inclusion of NMS by the fenoxi group. The Pt-NMS complex (PtC26H22N4O10S2) contain two ligands, coordinated through the N and O atoms of the sulfonamide group, for each Pt(II). DFT studies indicate that the N,O trans isomer is the most stable. The Ag-NMS complex presents MIC values in the range 15.0-120 mmol L over Pseudomonas aeruginosa, Escherichia coli e Staphylococcus aureus. The inclusion complex CE-[(Ag-NMS)·b-CD], although more soluble in water than Ag-NMS, shows no antibacterial activity in the tested concentrations. Both complexes were cytotoxic against normal (Balb/c 3T3) and tumor (SK-Mel 103 and Panc-1) cells, but the Pt-NMS complex was significantly more selective against tumor cells. The EtBr competitive intercalation assay and the evaluation of CT-DNA structure using circular dichroism show that DNA is not a biological target for the Pt-NMS complex, indicating a mechanism of action different of the cisplatin one / Mestrado / Quimica Inorganica / Mestre em Química

No / The use of matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry on intact cell microorganisms, Intact Cell MALDI (ICM), has been shown by numerous workers to yield effective species level identification. Early work highlighted the significant effect that variation in culture media, incubation conditions and length of incubation had on the spectra produced. Therefore, in order to achieve reliable and reproducible species level identification and sub-typing of microorganisms from ICM fingerprints, it has been essential to develop standardised methods. For methicillin-resistant Staphylococcus aureus (MRSA), a major nosocomial pathogen, we have developed such a standardised method. In this paper we present the experimental parameters, namely, the incubation period, the number of passages required from lyophilised or stored isolates, the method of deposition of the bacterial cells, the concentration of matrix solution, the drying time of bacterial cells prior to the addition of the matrix solution, the time between preparation of the bacterial/matrix sample and analysis and the MALDI pulsed extraction setting, which were considered during the development of defined methods.