• Refine Query
  • Source
  • Publication year
  • to
  • Language
  • 113
  • 112
  • 33
  • 10
  • 10
  • 8
  • 5
  • 3
  • 2
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • Tagged with
  • 336
  • 75
  • 62
  • 52
  • 36
  • 33
  • 30
  • 26
  • 25
  • 23
  • 23
  • 23
  • 22
  • 21
  • 20
  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
201

Type-II Ribosome Inactivating Proteins From Abrus Precatorius : Cytotoxicity And Mechanism Of Cell Death

Surendranath, Kalpana 04 1900 (has links)
Type-II Ribosome Inactivating Proteins from Abrus precatorius: Cytotoxicity and Mechanism of Cell Death A/B toxins produced by bacteria and plants are among the deadliest molecules known. The plant type-II ribosome inactivating proteins (RIPs) are prototype of A/B toxins. They are two subunit proteins with a toxic A subunit that harbors an RNA N-glycosidase activity and a lectin like B subunit which allows toxin entry into cells. The toxicity of A chain is due to its RNA-N-glycosidase activity which cleaves the bond between the ribose sugar and the adenine at position 4324 as demonstrated in rat liver ribosomes. The B- chain, a lectin, binds to the cell surface receptors terminating in galactose sugars and allows toxin entry into cells. The seeds of the subtropical climber Abrus precatorius contain two RIPs: the potent toxic lectin abrin and the relatively less toxic Abrus agglutinin. The toxic property of RIPs has widespread applications in the field of agriculture and medicine. The cells of our body commit suicide in response to genetic or environmental cues by the process, apoptosis or programmed cell death which results in the safe clearance of the dead cells without affecting the extra-cellular milieu. Apoptosis is essential for development, tissue homeostasis, and defense against pathogens. It involves the interplay of multiple pathways that are initiated and executed by a family of proteases termed caspases. Several plant type-I and type-II RIPs as well as bacterial toxins have been shown to induce apoptosis in cultured cell lines. Though many agents that inhibit macromolecular synthesis in cells induce DNA fragmentation and morphological changes associated with apoptosis, the link between protein synthesis inhibition by these toxins and apoptosis remains elusive. Though extensive studies have been carried out on several RIPs for e.g. ricin and shiga toxin, only few reports are available in literature on the mechanisms of toxicity exhibited by abrin, a type-II RIP, of South-East Asian origin. Earlier studies from the laboratory have focused on the sensitivity and mechanism of abrin induced cell death in Jurkat, a cell line of haematopoietic lineage and its variants. In the same direction, the objectives of my study were: (1) To delineate the structure-function relationship of Abrus agglutinin-I in comparison with abrin, (2) To establish monoclonal antibodies to the A subunit of abrin, analyzing their neutralizing effect on abrin toxicity in vitro and in vivo and (3) To delineate the pathway and determine the kinetics of apoptosis induced by abrin on cell lines of epithelial lineage. The thesis will be presented in three four chapters. The first chapter, ‘Introduction’, begins with a brief history of RIPs, followed by the description of their distribution and classification. The transport of toxins which is a unique property of this class of proteins is discussed in detail and supported with appropriate figures. Also, information pertaining to the structure of abrin and apoptosis induced by RIPs is written in brief. In the second chapter of the thesis the structural and functional studies of Abrus agglutinin-I (APA-I) as compared to abrin are discussed. Abrin and APA-I share a high degree of homology, however, previous reports by Liu et al., indicate that APA-I is many fold less toxic in cell free systems as compared to abrin. In our studies, APA-I was found to be less toxic on cultured cell lines. The IC50 value of protein synthesis inhibition by abrin was found to be 0.4 ng/ml for both Jurkat and MCF-7 cell lines. A 20-1000 fold difference was observed in the sensitivity of these cell lines to APA-I. The extent of apoptosis induced by APA-I in A3I9.2 a caspases-8 mutant Jurkat variant cell line was comparable to abrin indicating that the apoptosis induction by APA-I might not be through the extrinsic pathway. instead, our studies showed that APA-I induced apoptosis followed the mitochondrial pathway of cell death, in a caspase dependent manner similar to that of abrin. Unlike other agglutinins like wheat germ agglutinin, the agglutinating ability of the agglutinin-I had no role in the apoptosis induced. Protein synthesis inhibition appeared to be mandatory for the apoptosis induced by APA-I. The reason for the decreased toxicity of agglutinin-I became apparent on the analysis of the crystal structure of agglutinin-I obtained by us in comparison to that of the reported structure of abrin. The substitution of Asn200 in abrin with Pro199 in agglutinin-I seems to be a major cause for the decreased toxicity. This perhaps is not a consequence of any kink formation by Pro residue in the helical segment, as reported by others earlier but due to fewer interactions that proline can possibly have with the bound substrate. Passive immuno-neutralization by administration of neutralizing antibodies is widely used as therapy against poisoning by various toxins. In case of type-II RIPs like ricin, antibodies to the toxic subunit were proven to have better protective efficacy than those to the lectin subunit. Neutralizing antibodies to abrin are not reported in literature. Therefore, a panel of monoclonal antibodies (mAbs) to the recombinant A chain of abrin was developed in our laboratory and characterized, which is presented in the third chapter of the thesis. Of these, D6F10 a high affinity antibody, exhibited neutralizing effect on abrin induced cytotoxicity on different cell lines tested. Antibodies may neutralize biological toxins in multiple ways; our studies suggested that mAb D6F10 interferes in the earliest event i.e. attachment of the toxin to the cell surface. Significantly, with the administration of mice with mAb D6F10 the prophylactic effect of the mAb could be demonstrated. In chapter 4, the sensitivity, kinetics of proteins synthesis inhibition and the mechanism of abrin induced cell death in cell lines of epithelial lineage is presented. Both sensitivity and kinetics of MCF-7/pv, Ovcar3, and T47D cells appeared comparable while, a variant culture of MCF-7 over-expressing caspases-3 was 50 times more sensitive to abrin. There was no significant difference in the binding of abrin between MCF-7/pv and MCF-7/C3+ cells. Previous studies in our laboratory indicated that abrin induced apoptosis is a caspases-3 dependent process. Also, in several systems it has been shown that caspases-3 is an indispensable molecule for apoptotic cell death. To test the absolute requirement of caspase-3, we examined abrin-induced apoptosis in a human breast cancer cell line MCF-7/pv reportedly deficient in caspases-3. Unlike other molecules like cisplatin, apoptosis induced by abrin in the MCF- 7/pv cells was found to be caspase -3 independent. However faster kinetics of apoptosis is observed, indicating that there is amplification of the apoptotic signals in the presence of caspases-3 resulting in an early onset of DNA fragmentation. The kinetics of protein synthesis inhibition and apoptosis follows similar kinetics in Jurkat cells while there is a time lapse between the two events in epithelial cells. Even with very high concentrations of abrin no detectable apoptosis was observed within 24 h in epithelial cells. The onset of fragmentation occurs after 24 h in the cell lines tested as opposed to Jurkat where it is observed as early as 6 h. Inhibition of caspases rescued the toxins from DNA fragmentation suggesting that the toxin does not cause direct nuclear damage in the cell line which does not involve the activation of caspases.
202

Inactivation of <i>Ascaris suum</i> by Ammonia in Feces Simulating the Physical-Chemical Parameters of the Solar Toilet Under Laboratory Conditions

Cruz Espinoza, Ligia Maria 10 November 2010 (has links)
Access to sustainable sanitation systems is a determining factor in human health and economic development. However, more than a third of the world’s population lives without access to improved sanitation facilities. To meet the sanitation United Nations Millennium Development target, "halve, by 2015, the proportion of people without sustainable access to safe drinking water and basic sanitation", a wide range of non conventional sanitation technologies have been implemented in developing countries, including waterless systems. These systems function by diverting urine away from feces and collecting, storing, and dehydrating the fecal material in watertight dehydration vaults. From a public health perspective, adequate inactivation of fecal pathogens in a sanitation system is essential before any use or disposal of fecal material. In rural areas of El Salvador, the solar toilet is capable of inactivating fecal pathogens and reducing the prevalence of parasitic infections in its users when compared to other waterless systems. Nevertheless, not all solar toilets are able to inactivate completely Ascaris spp. ova after the recommended storage period. Un-ionized ammonia (NH3) has the potential to inactivate pathogens in solutions and sludge, including Ascaris spp. ova. This study hypothesized that adding ammonia to the solar toilet will improve the technology since pathogen inactivation with ammonia could be potentiated by the alkaline medium and high temperatures achieved inside the toilet vaults. To evaluate this approach, a series of experiments in solution and biosolid were performed in a laboratory environment using physical and chemical parameters similar to those achieved by the solar toilet. Eggs of the swine Ascaris species, Ascaris suum, were used as model in all experiments. In ammonia solution, the parasite ova were stored for a period of three days and; in biosolid, the parasite ova were stored for two months. Urea was used as the source of ammonia in biosolid. In addition to the experiments with ammonia, normal viability and morphological changes within the parasite ova during incubation in vitro at 28 C° were investigated and described to complement current literature published. Results from the experiments in ammonia solution indicated that addition of ammonia (1% and 2%) could improve the system since the critical parameters that significantly reduced A. suum ova viability to zero in three days could be achieved by the solar toilet: temperature of 35°C or higher and pH value of 9.3. Results from the experiments in biosolid further showed that inactivation of A. suum ova was faster in samples exposed to urea and to temperatures higher than 28°C. All samples exposed to urea achieved 100% inactivation after 14 days (28°C), 3 days (35°C) and 24 hours (40°C and 45°C). Survival analysis of the data showed that there was a significant difference (p value <.0001) between the inactivation achieved in the samples exposed to urea (1% and 2%) and the samples not exposed to urea. A logistic regression analysis estimated the effect of Urea (Treatment, OR: 25.9), Temperature (OR: 1.8), and Storage (OR: 1.17) on inactivation. Results from the experiment with A. suum ova in normal incubation solution showed that the ova went through clearly identified morphological changes at different speed of development. Two new additional stages of development were identified (Pre-larva 1 and Pre-larva 2) and no significant statistical difference was observed among the viability reported early in incubation and the one reported after three weeks of in vitro incubation, indicating that early stages of development may be use as an alternative to reduce the time to report viability. The results of this study suggest that inactivation of Ascaris spp ova by ammonia is possible in fecal material stored in the solar toilet or any other dry toilet, if the following physical and chemical conditions are met: a closed vault with a minimum temperature of 28°C; an initial pH of 8.3, minimum moisture of 27.5%, and addition of 1% urea to the biosolid. At 28°C longer storage time would be required for 100% inactivation while at higher temperatures less time of storage would be necessary. A community intervention is recommended to include field conditions and human behavior as other predictors for Ascaris spp. inactivation by ammonia.
203

Genome-wide Analysis of Ctcf-RNA Interactions

Kung, Johnny Tsun-Yi January 2014 (has links)
Ctcf is a "master regulator" of the genome that plays a role in a variety of gene regulatory functions as well as in genome architecture. Evidence from studying the epigenetic process of X-chromosome inactivation suggests that, in certain cases, Ctcf might carry out its functions through interacting with RNA. Using mouse embryonic stem (ES) cells and a modified protocol for UV-crosslinking and immunoprecipitation followed by high-throughput sequencing (CLIP-seq), Ctcf is found to interact with a multitude of transcripts genome-wide, both protein-coding mRNA (or noncoding transcripts therein) as well as many long-noncoding RNA (lncRNA). Examples of the latter include both well-characterized species from imprinted loci and previously unannotated transcripts from intergenic space. RNA binding targets of Ctcf are validated by a variety of biochemical methods, and Ctcf is found to interact with RNA through its C-terminal domain, distinct from its DNA-binding zinc-finger domain. Ctcf chromatin immunoprecipitation (ChIP)-seq done in parallel reveals distinct but correlated binding of Ctcf to DNA and RNA. In addition, allelic analysis of Ctcf ChIP pattern reveals significant differences between Ctcf binding to the presumptive inactive and active X chromosomes. Together, the current work reveals a further layer of complexity to Ctcf biology by implicating a role for Ctcf-RNA interactions in its recruitment to genomic binding sites.
204

Customized Raptor Code Designs for Finite Lengths and Practical Settings

Mahdaviani, Kaveh Unknown Date
No description available.
205

Endogenous and exogenous factors affecting lipoprotein lipase activity

Larsson, Mikael January 2014 (has links)
Individuals with high levels of plasma triglycerides are at high risk to develop cardiovascular disease (CVD), currently one of the major causes of death worldwide. Recent epidemiological studies show that loss-of-function mutations in the APOC3 gene lower plasma triglyceride levels and reduce the incidence of coronary artery disease. The APOC3 gene encodes for apolipoprotein (APO) C3, known as an inhibitor of lipoprotein lipase (LPL) activity. Similarly, a common gain-of-function mutation in the LPL gene is associated with reduced risk for CVD. LPL is central for the metabolism of lipids in blood. The enzyme acts at the endothelial surface of the capillary bed where it hydrolyzes triglycerides in circulating triglyceride-rich lipoproteins (TRLs) and thereby allows uptake of fatty acids in adjacent tissues. LPL activity has to be rapidly modulated to adapt to the metabolic demands of different tissues. The current view is that LPL is constitutively expressed and that the rapid modulation of the enzymatic activity occurs by some different controller proteins. Angiopoietin-like protein 4 (ANGPTL4) is one of the main candidates for control of LPL activity. ANGPTL4 causes irreversible inactivation through dissociation of the active LPL dimer to inactive monomers. Other proteins that have effects on LPL activity are the APOCs which are surface components of the substrate TRLs. APOC2 is a well-known LPL co-factor, whereas APOC1 and APOC3 independently inhibit LPL activity. Given the important role of LPL for triglyceride homeostasis in blood, the aim of this thesis was to find small molecules that could increase LPL activity and serve as lead compounds in future drug discovery efforts. Another aim was to investigate the molecular mechanisms for how APOC1 and APOC3 inhibit LPL activity. Using a small molecule screening library we have identified small molecules that can protect LPL from inactivation by ANGPTL4 during incubations in vitro. Following a structure-activity relationship study we have synthesized lead compounds that more efficiently protect LPL from inactivation by ANGPTL4 in vitro and also have dramatic triglyceride-lowering properties in vivo. In a separate study we show that low concentrations of fatty acids possess the ability to prevent inactivation of LPL by ANGPTL4 under in vitro conditions. With regard to APOC1 and APOC3 we demonstrate that when bound to TRLs, these apolipoproteins prevent binding of LPL to the lipid/water interface. This results in decreased lipolysis and in an increased susceptibility of LPL to inactivation by ANGPTL4. We demonstrate that hydrophobic amino acid residues that are centrally located in the APOC3 molecule are critical for attachment of this protein to lipid emulsion particles and consequently for inhibition of LPL activity. In summary, this work has identified a lead compound that protects LPL from inactivation by ANGPTL4 in vitro and lowers triglycerides in vivo. In addition, we propose a molecular mechanism for inhibition of LPL activity by APOC1 and APOC3.
206

Cell autonomous and cell non-autonomous effects of mosaic Mecp2 expression on layer V pyramidal cell morphology in a mouse model of Rett Syndrome

Rietveld, Leslie A. 19 December 2012 (has links)
Rett Syndrome (RTT) is a neurodevelopmental disorder primarily caused by mutations in the X-linked gene methyl-CpG-binding protein 2 (MECP2). The mosaic brain environment in heterozygous (MECP2+/-) females consists of both MeCP2-wildtype (MeCP2+) and Mecp2-mutant (MeCP2-) neurons. To separate possible cell autonomous and cell non-autonomous effects three-dimensional morphological analysis was performed on individually genotyped layer V pyramidal neurons in the primary motor cortex of heterozygous (Mecp2+/-) and wild-type (Mecp2+/+) mature female mice (>8 months old) from the Mecp2tm1.1Jae line. Mecp2+/+ neurons and Mecp2+ were found to be indistinguishable while Mecp2- neurons have significantly reduced basal dendritic length (p<0.05), predominantly in the region 70-130 μm from the cell body, culminating in a total reduction of 15%. Mecp2- neurons have three (17%) fewer total branch points, lost specifically at the second and third branch orders. Thus the reduced total dendritic length in Mecp2- neurons is a result of fewer higher-order branches. Soma and nuclear areas of 30 Mecp2+/- female mice (5-21 months) with X chromosome inactivation (XCI) ratios ranging from 12% to 56% were analyzed. On average Mecp2- somata and nuclei were 15% and 13% smaller than Mecp2+ neurons respectively. The variation observed in the soma and nuclear sizes of Mecp2- neurons was not due to age, but was found to be correlated with the XCI ratio. Animals with a balanced XCI ratio (approximately 50% Mecp2-) were found to have Mecp2- neurons with a less severe cellular phenotype (11-17% smaller than Mecp2+). Animals with a highly skewed XCI ratio favouring expression of the wild-type allele (less than 30% Mecp2-) were found to have a more severe Mecp2- cellular phenotype (17-22% smaller than Mecp2+). These data support indicate that mutations in Mecp2 exert both cell autonomous and cell non- autonomous effects on neuronal morphology. / Graduate
207

Genomics and Transcriptomics of Hybrid Male Sterility Assessed in Multiple Interspecies Feline Breeds

Davis, Brian W 03 October 2013 (has links)
Hybrid male sterility (HMS) is typically the first mechanism fortifying reproductive isolation resulting from genomic incompatibilities. Three interspecies feline breeds derived from domestic cat crosses to wild cat species (Asian leopard cat and African serval) manifest HMS through several generations of backcrossing before eventually regaining fertility. This work utilized 199 hybrid individuals with varying fertilities in a genome wide association study (GWAS) comprising 63,000 genome wide SNPs. Leveraging these results with whole-testis transcriptome sequencing and quantitative real-time PCR data facilitated the comparison of transcripts in sterile and fertile hybrids. This dissertation describes four loci with highly significant and fifty with moderately significant association to sterility within each individual hybrid domestic breed and combinations of breeds. These associations help identify epistatic targets for hybrid incompatibility contributing to sterility. Comparative QTL mapping between pairs of species provides a framework to describe the accumulation of clade-specific reproductive isolating loci. Detailed exploration of gene misregulation between domestic and hybrid individuals, as well as between littermate hybrids of varying fertilities outlines a pattern of expression consistent with a meiotic sex-chromosome inactivation failure in early generations and apoptotic failure in later hybrid generations. Combining comparative genomic association and transcriptomic characterization among hybrid felids of varying divergence, new insight is gained into the mechanisms of mammalian reproductive isolation.
208

Segregation within afferent pathways in primate vision

Roy, Sujata January 2009 (has links)
The current knowledge of the visual pathways in primates includes the patterns of projection from the retina through the dorsal lateral geniculate nucleus (dLGN) to the striate cortex (V1) and the extra-striate projections towards the dorsal and ventral streams. Cells with short wavelength sensitive cone (S-cone) inputs in the dLGN have been studied extensively in New World marmosets but not in Old World macaques. This thesis presents results from studies in the macaque monkey which are more relevant to humans since humans are closer in evolution to Old World than New World monkeys. / The spatial, temporal, chromatic and orientation preferences of neurons in the dLGN of the macaque were investigated by electrophysiological methods. The physiological findings of cells with S-cone inputs were compared to cells with opponent inputs from the long and medium wavelength sensitive cones (L-cones & M-cones, respectively). The cells receiving S-cone inputs (blue-yellow or B-Y cells) preferred lower spatial frequencies than the cells with opponent L-cone and M-cone inputs (red-green or R-G cells). Orthodromic latencies from optic chiasm stimulation were measured where possible to distinguish differences in conduction velocity between the cell groups. Although the B-Y cells usually had longer latencies than R-G cells, there wasconsiderable overlap between the cell groups. / The recorded cells were localised through histological reconstruction of dLGN sections stained for Nissl substance. The distribution of B-Y cells within the dLGN was compared to the distribution of R-G cells. The majority of B-Y cells were located within the intercalated koniocellular layers as well as the koniocellular bridges (extensions of the koniocellular layers into the adjacent parvocellular layers). The B-Y cells were also largely segregated within the middle dLGN layers (K3, P3, K4 & P4). The R-G cells were mainly concentrated within the parvocellular layers (P3, P4, P5 & P6) and were evenly distributed throughout the middle and outer layers of the dLGN. / The study also included recordings from the extra-striate middle temporal area (MT) to determine whether a fast S-cone input exists from the dLGN to area MT which bypasses V1. The pattern of cone inputs to area MT neurons was investigated before and during inactivation of V1. The inactivation was done through reversible cooling with a Peltier thermocouple device or focal inactivation with y-amino butyric acid (GABA) iontophoresis. Precise inactivation of V1 to the topographically matching visual fields of the recording sites in area MT revealed a preservation of all three coneinputs in many cells. The subcortical sources of these preserved inputs are discussed with their relevance to blindsight, which is the limited retention of visual perception after V1 damage. Analysis of the latencies of area MT cells revealed a rough segregation into latencies faster or slower than 70 ms. Cells both with and without a significant change in response during V1 inactivation were present in each group. The findings reported in this thesis indicate that some of the preserved inputs in area MT during V1 inactivation may be carried by a direct input from the dLGN which bypasses V1.
209

Expressão dos genes ALS3, HWP1, BCR1, TEC1, CPH1 e EFG1 de Candida albicans em biofilmes após inativação fotodinâmica. / Expression of the Candida albicans genes ALS3, HWP1, BCR1, TEC1, CPH1 and EFG1 in biofilms after photodynamic inactivation.

Freire, Fernanda [UNESP] 30 November 2017 (has links)
Submitted by Fernanda Freire null (fernanda.freire@fosjc.unesp.br) on 2017-12-13T19:59:30Z No. of bitstreams: 1 Tese - Fernanda Freire.pdf: 1272262 bytes, checksum: 11df7d222fe968088750e08ceb6fa488 (MD5) / Submitted by Fernanda Freire null (fernanda.freire@fosjc.unesp.br) on 2017-12-14T11:25:01Z No. of bitstreams: 1 Tese - Fernanda Freire.pdf: 1272262 bytes, checksum: 11df7d222fe968088750e08ceb6fa488 (MD5) / Submitted by Fernanda Freire null (fernanda.freire@fosjc.unesp.br) on 2017-12-14T13:50:30Z No. of bitstreams: 1 Tese - Fernanda Freire.pdf: 1272262 bytes, checksum: 11df7d222fe968088750e08ceb6fa488 (MD5) / Approved for entry into archive by Silvana Alvarez null (silvana@ict.unesp.br) on 2017-12-14T18:37:35Z (GMT) No. of bitstreams: 1 freire_f_dr_sjc.pdf: 1272262 bytes, checksum: 11df7d222fe968088750e08ceb6fa488 (MD5) / Made available in DSpace on 2017-12-14T18:37:35Z (GMT). No. of bitstreams: 1 freire_f_dr_sjc.pdf: 1272262 bytes, checksum: 11df7d222fe968088750e08ceb6fa488 (MD5) Previous issue date: 2017-11-30 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Os micro-organismos estão se tornando cada vez mais resistentes aos antimicrobianos e cepas de Candida albicans resistentes aos antifúngicos tem sido isoladas, assim, torna-se importante e necessário a realização de pesquisas que avaliem os efeitos de novos métodos terapêuticos, como a inativação fotodinâmica antimicrobiana (aPDI). Assim, o objetivo deste estudo foi verificar os efeitos da inativação fotodinâmica sobre biofilmes de Candida albicans, avaliando seus efeitos sobre a expressão dos genes TEC1 (fator de transcrição), HWP1 (proteína de parede celular das hifas), EFG1 (regulador transcricional relacionado com a morfogênese), BCR1 (regulador da formação de biofilme e da parede celular), CPH1 (regulador transcricional envolvido na morfogênese) e ALS3 (adesina) de C. albicans. Foram avaliadas 30 amostras isoladas de pacientes portadores de HIV e 30 amostras de pacientes com estomatite protética, quanto a produção de biofilme, peso seco e filamentação. Destas, foram selecionadas as amostras mais virulentas de cada grupo que apresentaram melhor capacidade de formação de biofilme e filamentação. Assim, foi utilizada uma amostra clínica de C. albicans isolada de paciente portador de HIV, uma amostra clínica de C. albicans isolada de paciente com estomatite protética e uma cepa padrão ATCC 18804. A quantificação da expressão dos genes foi relacionada à produção desses genes nas amostras clínicas e na cepa de referência utilizando-se ensaio de PCR em tempo real. Para a aPDI, foram utilizados os fotossensibilizadores azul de metileno a 300 μM e eritrosina a 400 μM sensibilizados com laser de Índio-Gálio-Alumínio-Fósforo de baixa potência (vermelho visível, 660 nm) e LED verde (532 ± 10 nm), respectivamente. Foram avaliados quatro grupos experimentais para a aPDI: a) F+L+: sensibilização com o corante e irradiação com luz; b) F+L-: somente tratamento com o fotossensibilizador; c) F-L+: somente irradiação com luz e d) F-L-: sem sensibilização com o corante e ausência de luz. Os resultados foram analisados por t-test, com um nível de significância de 5%. Após a análise fenotípica, as amostras Ca30 e 39S foram selecionadas para a realização da aPDI. Como esperado, apenas para o grupo F+L+, quando comparado com o grupo F-L-, todos os genes analisados foram sub expressos após a aPDI. O fold-decrease para os genes ALS3, HWP1, BCR1, TEC1, CPH1 e EFG1 foram 0,73; 0,39; 0,77; 0,71; 0,67 e 0,60; para laser, respectivamente, e 0,66; 0,61; 0,50; 0,43; 0,54 e 0,66; para LED, respectivamente. Pode-se concluir que a aPDI mostrou uma redução na expressão dos genes de C. albicans, sugerindo a diminuição de sua virulência. / Micro-organisms are becoming increasingly resistant to antimicrobial agents and Candida albicans resistant strains to antifungal has been isolated, so it is important and necessary to carry out studies that evaluates the effects of new therapeutic methods, such as antimicrobial photodynamic inactivation (aPDI). The objective of this study was verify the effects of aPDI on C. albicans biofilms, evaluating its effects on genes expression: TEC1 (transcription factor), HWP1 (cell wall protein hyphae), EFG1 (transcriptional regulator related to morphogenesis), BCR1 (regulator of biofilm formation and cell wall), CPH1 (transcriptional regulator involved in morphogenesis) and ALS3 (adhesin) of C. albicans. Were evaluated 30 samples isolated from patients with HIV and 30 samples from patients with denture stomatitis, as the production of biofilm, dry weight and filamentation. Of these, the most virulent strains of each group that presented better biofilm formation capacity and filamentation were selected. Therefore, were used a clinical sample of C. albicans isolated from HIV positive patient, a clinical sample of C. albicans isolated from patient with denture stomatitis and a standard strain ATCC 18804. The quantification of gene expression was related to the production of these genes in clinical samples and in the reference strain using PCR assay in real time. For aPDI, were used the photosensitizer methylene blue at 300 uM and erythrosine at 400 uM, sensitized with low power laser Indium-Gallium-AluminumPhosphorus (visible red, 660 nm) and green LED (532 ± 10 nm), respectively. Were evaluated four groups for aPDI: a) P+L+: sensitization with the photosensitizer and irradiation with light; b) P+L-: only treatment with the photosensitizer; c) P-L+: only irradiation with light and d) P-L-: without sensitization with the dye and absence of light. The results were analyzed by t-test, with a significance level of 5%. After the phenotypic analysis, the samples Ca30 and 39 S were selected for aPDI . As expected, only in the group P+L+ when compared with the group P-L-, all analyzed genes were downregulated after aPDI. The fold-decrease for the genes ALS3, HWP1, BCR1, TEC1, CPH1 and EFG1, were 0.73, 0.39, 0.77, 0.71, 0.67 and 0.60, for laser, respectively, and 0.66, 0.61, .050, 0.43, 0.54 and 0.66, for LED, respectively. It could be concluded that aPDI showed a reduction in the expression of C. albicans genes, suggesting its virulence decrease. / 2013/22897-2
210

Compostos fenólicos e atividade antibacteriana em acessos de ipomoea batatas (l.) lam (batata-doce)

José, António Elísio January 2012 (has links)
No período compreendido entre os meses de Maio de 2010 e Dezembro de 2011 realizou-se, nos laboratórios de Higiene e Qualidade de Alimentos, de Microbiologia e Bromatologia e de Bioquímica e Microbiologia Aplicada do Instituto de Ciências e Tecnologia de Alimento (ICTA) da Universidade Federal do Rio Grande do Sul (UFRGS), experimentos com vista à quantificação de compostos fenólicos e à avaliação da atividade antibacteriana de dois acessos de batata-doce (Ipomoea batatas L. (Lam)) com objetivo de (i) avaliar a atividade antibacteriana nestes acessos perante padrões bacterianos de interesse em alimentos, (ii) determinar a quantidade de compostos fenólicos e (iii) estabelecer a relação entre a composição e quantidade de compostos fenólicos e a atividade antibacteriana nestes diferentes acessos. Com o material colhido em Porto Alegre, Santo Antônio de Patrulha, Palmares do Sul e Cerro grande do Sul, utilizou-se o método de diluição em sistema de tubos múltiplos para avaliar a intensidade de atividade antibacteriana dos diferentes extratos expressa como intensidade de atividade de inibição bacteriana (IINIB) e intensidade de atividade de inativação bacteriana (IINAB); o teor de polifenóis totais foi determinado pelo método de Folin & Ciocalteu, enquanto que a determinação do conteúdo de antocianinas foi efetuada usando o método de pH diferencial. O coeficiente de correlação de Pearson foi usado para estabelecer a relação entre os teores de polifenóis e a atividades antibacteriana. Os extratos alcoólicos inibiram e/ ou inativaram inóculos de Salmonella Enteritidis (ATCC 11076), Enterococcus faecalis (ATCC 19433), Escherichia coli (ATCC 11229) e Staphylococcus aureus (ATCC 25923), não obstante esta úlima ser significativamente mais resistente. A intensidade de atividade de inibição e/ ou inativação está positivamente relacionada à concentração de compostos fenólicos, ou seja, os compostos fenólicos e/ ou antocianinas seriam responsáveis pelo menos por parte da atividade antibacteriana dos extratos testados nas condições deste experimento. Extratos tidos por decocção e infusão, além de apresentarem concentrações muito baixas de antocianinas, não apresentaram atividade antibacteriana e mostraram menores teores de polifenóis do que os seus correspondentes extratos alcoólicos, sugerindo que o calor da infusão e/ ou decocção provavelmente teria degradado as substâncias essenciais que tomam parte no processo de atividade antimicrobiana. / In the period between the months of May 2010 and December 2011 was held in the laboratories of Higiene and Food Quality, Microbiology and Bromatology and Biochemistry and Applied Microbiology of the Institute for Food Science and Technology (ICTA) of the Federal University of Rio Grande do Sul (UFRGS), experiments for the evaluation of antibacterial activity and the quantification of phenolic compounds in two accessions of sweet potato (Ipomoea batatas L. (Lam) in order to (i) evaluate the antibacterial activity in these accessios against interest food bacteria, (ii) determine the quantity of phenolic (iii) establish the relationship between the composition and quantity of phenolic compounds and the antibacterial activity in these different accessions. With material collected in Porto Alegre, Santo Antônio de Patrulha, Palmares do Sul and Cerro Grande do Sul, the method of dilution in system of multiple-tube tests was used to evaluate the intensity of antibacterial activity of different extracts express as intensity of bacterial inhibition activity (IINIB) and intensity of bacterial inativation activity (IINAB); the total phenolic content was determined by the method of Folin & Ciocalteu, while the content of antocianinas was performed using the differential pH method. The Pearson correlation coefficient was used to establish the relationship between the levels of polyphenols and antibacterial activity. The alcoholic extracts inhibited and/or inactivated inocula of Salmonella Enteritidis (ATCC 11076), Enterococcus faecalis (ATCC 19433), Escherichia coli (ATCC 11229) and Staphylococcus aureus (ATCC 25923), although this this last was significantly tougher. The intensity of inhibition and/or inactivation activity is positively related to the concentration of phenolic compounds, ie, phenolic compounds and/or anthocyanins are responsible for at least, part of the antibacterial activity of the extracts tested in this experiment. Extracts got from decoction and infusion besides having very low concentrations of anthocyanins, showed no antibacterial activity and had lower levels of polyphenols than their corresponding alcoholic extracts, suggesting that the higher temperature used for infusion and/or decoction, would probably degraded essential substances taking part in the process of antimicrobial activity.

Page generated in 0.097 seconds