Revisión por pares / joruiz@clinic.ub.es / Furazolidone-resistant mutants were obtained from four clinical isolates of diarrhoeagenic Escherichia coli. The stability of the resistance and the frequency of mutation were established. The minimal inhibitory concentration of furazolidone, nitrofurantoin, nalidixic acid, ampicillin, chloramphenicol and tetracycline was established both in the presence and absence of the efflux pump inhibitor Phe-Arg-β-Naphtylamyde. The presence of mutations in the nitroreductase genes nfsA and nfsB was analysed by PCR; sequencing and their enzymatic activity was assessed by a spectrophotometric assay. Alterations in outer membrane proteins were studied by SDS-PAGE. The frequency of mutation ranged from <9.6 × 10-10 to 9.59 × 10-7 . Neither an effect on efflux pumps inhibited by Phe-Arg-β-Naphtylamyde nor cross-resistance with the antibiotics studied was observed. Nineteen mutants (52.94%) presented mutations in the nitroreductase-encoding genes: 17 in the nfsA gene (15 mutants with an internal stop codon, 2 with amino acid changes), 2 in the nfsB (all amino acid changes). Alterations in the outer membrane proteins OmpA and OmpW were also observed. Although more studies are necessary to find other resistance mechanisms, present data showed the low potential of selecting furazolidone-resistant mutants, together with the lack of cross-resistance with unrelated antimicrobial agents.

Corn (Zea mays) is frequently infected by a soil fungal pathogen Aspergillus flavus. The fungus produces aflatoxins, which cause liver cancer. Maize inbred lines that are resistant to infection by A. flavus have been developed, and these inbred lines provide excellent models for studying molecular mechanisms of maize resistance to the fungus. MicroRNA-like RNAs (milRNAs) recently identified in A. flavus had been found to be correlated with aflatoxin production conditions, suggesting that the milRNAs might play a role in the regulation of aflatoxin production. In this research, small RNAs were isolated from kernels of maize (resistant Mp719 and susceptible Va35) inoculated with A. flavus NRRL 3357 (aflatoxigenic) and NRRL 21882 (nonaflatoxigenic) and then subjected to RNA sequencing. Sequencing had identified 69 A. flavus milRNAs and 691 Z. mays miRNAs. The differential expression of some maize miRNAs revealed their potential role in response to inoculation, A. flavus growth, and aflatoxin production.

The trimethoprim and sulfamethoxazole combination, co-trimoxazole, plays a vital role in the treatment of Burkholderia pseudomallei infections. Previous studies demonstrated that the B. pseudomallei BpeEF-OprC efflux pump confers widespread trimethoprim resistance in clinical and environmental isolates, but this is not accompanied by significant resistance to co-trimoxazole. Using the excluded select-agent strain B. pseudomallei Bp82, we now show that in vitro acquired trimethoprim versus cotrimoxazole resistance is mainly mediated by constitutive BpeEF-OprC expression due to bpeT mutations or by BpeEF-OprC overexpression due to bpeS mutations. Mutations in bpeT affect the carboxy-terminal effector-binding domain of the BpeT LysR-type activator protein. Trimethoprim resistance can also be mediated by dihydrofolate reductase (FolA) target mutations, but this occurs rarely unless BpeEF-OprC is absent. BpeS is a transcriptional regulator that is 62% identical to BpeT. Mutations affecting the BpeS DNA-binding or carboxy-terminal effector-binding domains result in constitutive BpeEF-OprC overexpression, leading to trimethoprim and sulfamethoxazole efflux and thus to cotrimoxazole resistance. The majority of laboratory-selected co-trimoxazole-resistant mutants often also contain mutations in folM, encoding a pterin reductase. Genetic analyses of these mutants established that both bpeS mutations and folM mutations contribute to co-trimoxazole resistance, although the exact role of folM remains to be determined. Mutations affecting bpeT, bpeS, and folM are common in co-trimoxazole-resistant clinical isolates, indicating that mutations affecting these genes are clinically significant. Cotrimoxazole resistance in B. pseudomallei is a complex phenomenon, which may explain why resistance to this drug is rare in this bacterium. IMPORTANCE Burkholderia pseudomallei causes melioidosis, a tropical disease that is difficult to treat. The bacterium's resistance to antibiotics limits therapeutic options. The paucity of orally available drugs further complicates therapy. The oral drug of choice is co-trimoxazole, a combination of trimethoprim and sulfamethoxazole. These antibiotics target two distinct enzymes, FolA (dihydrofolate reductase) and FolP (dihydropteroate synthase), in the bacterial tetrahydrofolate biosynthetic pathway. Although co-trimoxazole resistance is minimized due to two-target inhibition, bacterial resistance due to folA and folP mutations does occur. Co-trimoxazole resistance in B. pseudomallei is rare and has not yet been studied. Co-trimoxazole resistance in this bacterium employs a novel strategy involving differential regulation of BpeEF-OprC efflux pump expression that determines the drug resistance profile. Contributing are mutations affecting folA, but not folP, and folM, a folate pathway-associated gene whose function is not yet well understood and which has not been previously implicated in folate inhibitor resistance in clinical isolates.

Streptococcus pneumoniae is a leading cause of pneumonia, bacteremia, meningitis, otitis media and sinusitis, and is responsible for significant morbidity and mortality worldwide. The burden of pneumococcal disease has been greatly impacted by the high prevalence of HIV, especially in developing countries. Macrolides are commonly used for the treatment of pneumococcal infections with the resulting effect of increasing resistance. Pneumococci develop resistance to macrolides predominantly by two mechanisms; target modification and drug efflux. Target modification occurs through the acquisition of an erm(B) gene (MLSB phenotype) or through ribosomal mutation, and drug efflux occurs through the acquisition of a mef(A) gene (M phenotype). Alternative protein synthesis-inhibiting antibiotics such as linezolid and telithromycin have been developed in response to the increasing level of antibiotic resistance. In this study, novel mechanisms of resistance to protein synthesis-inhibiting antibiotics, and the current prevalence and epidemiology of macrolide resistance in South Africa were investigated. Two clinical isolates of S. pneumoniae resistant to macrolides, linezolid and chloramphenicol were identified in the PROTEKT surveillance study and the ABCs program of the CDC. The isolates were found to each contain a 6 bp deletion, resulting in the deletion of two amino acids from a highly conserved region of ribosomal protein L4 (64PWRQ67 to 64P_Q67 and 67QKGT70 to 67Q_T70). The genes encoding the mutant ribosomal proteins transformed susceptible strain R6 to macrolide, linezolid and chloramphenicol resistance, proving that the ii deletions conferred the resistance on the isolates, and indicating that these antibiotics share a common binding site. Growth studies of the R6 transformants showed increased mass doubling times, suggesting that the L4 mutations were associated with a fitness cost, but the original strains showed evidence of fitness compensation. The L4 mutations in these isolates represent a novel mechanism of cross-resistance to macrolides, linezolid and chloramphenicol. A macrolide-resistant clinical isolate of S. pneumoniae with mutations in 23S rRNA showed a heterogeneous phenotype and genotype. A mutant gene encoding 23S rRNA from this isolate transformed susceptible strain R6 to resistance. Transformants displayed similar heterogeneity to the isolate. Culture of resistant strain R6 in the presence of antibiotic maintained resistance, however culture of the strain in the absence of antibiotic pressure resulted in a reversion to susceptibility. By DNA sequencing, gene conversion was shown to occur between the wild-type and mutant 23S rRNA alleles. Growth studies indicated that the resistant phenotype was associated with a fitness cost. Therefore, under antibiotic selective pressure alleles converted to the mutant form, and in the absence of selective pressure alleles reverted to wild-type, in order to regain fitness. Through gene conversion the pneumococcus has the ability to rapidly adapt to the environment, with implications for susceptibility testing and patient treatment. A rare clinical isolate of S. pneumoniae, highly resistant to telithromycin, was received from the Canadian Bacterial Surveillance Network and was investigated for the mechanism of resistance. The isolate was found to contain an erm(B) gene iii with a truncated control peptide, as well as a mutant ribosomal protein L4, containing a number of mutations. Transformation of susceptible strain PC13, containing a wild-type erm(B) gene, with the mutant erm(B) gene decreased the susceptibility of PC13 to telithromycin, but did not confer high-level resistance. Transformation of PC13 with the mutant L4 gene or a fragment of the L4 gene containing only the 69GTG71 to TPS mutation, conferred high-level resistance on PC13. In contrast, transformation of R6, which did not contain an erm(B) gene, with the L4 gene or L4 fragment only conferred reduced telithromycin susceptibility. High-level telithromycin resistance was therefore conferred by a combination of an erm(B) gene with a 69GTG71 to TPS mutation in a highly conserved region of ribosomal protein L4. The combination of mechanisms inhibited the binding of telithromycin to the ribosome, whereas neither mechanism individually was sufficient. A telithromycin-resistant clinical isolate of S. pneumoniae was received from the PROTEKT surveillance study and was investigated for the resistance mechanism. The isolate was found to contain a 136 bp deletion in the regulatory region of erm(B). This mutant gene was shown, by transformation studies, to confer resistance on susceptible strain PC13. Expression of erm(B) on the transcriptional level was quantified by real-time reverse transcription PCR. In the presence of erythromycin and telithromycin, erm(B) expression was significantly higher in the mutant PC13 strain than the wild-type strain. Growth studies showed that the mutant PC13 strain had a shorter lag phase than the wild-type strain in the presence of erythromycin. Telithromycin resistance was conferred by the mutant iv erm(B) gene that was expressed at a higher level than the wild-type gene, most likely resulting in higher ribosomal methylation levels sufficient to hinder telithromycin binding. Macrolide resistance in invasive pneumococcal disease in South Africa for the period 2000 to 2005 was investigated through a national laboratory-based surveillance system. Viable isolates (n=15982) collected during the six-year period were phenotypically characterised, by determination of MICs and serotyping. Two hundred and sixty random isolates from 2005 were genotypically screened for the presence of erm(B) and mef(A). Macrolide resistance increased significantly from 9% in 2000 to 14% in 2005. Resistant isolates were received from all provinces of South Africa, with Gauteng and the Western Cape having the highest incidence. Serotype 14 was the most common macrolide-resistant serotype and 96% of macrolide-resistant isolates in 2005 were serotypes included in the 7-valent pneumococcal conjugate vaccine and serotype 6A. Macrolide resistance was significantly higher in children <5 than in individuals 5 years and older. The majority of strains (75%) over the six-year period displayed the MLSB phenotype. Of the 260 strains genotypically screened, 57% were positive for erm(B), 27% were positive for mef(A), 15% contained both erm(B) and mef(A), and 1% were negative for both genes and were found to contain ribosomal mutations. Eighty percent of isolates containing both erm(B) and mef(A) were serotype 19F and were found to be clonal by PFGE and MLST. These multidrug-resistant isolates were related to the Taiwan19F-14 global clone. v Many protein synthesis-inhibiting antibiotics share overlapping binding sites on the large ribosomal subunit. Alterations in 23S rRNA and ribosomal proteins L4 and L22, within the binding pocket, confer resistance and often cross-resistance to many of these antibiotics. The ability of the pneumococcus to develop resistance and the global spread of resistant strains highlights the importance of monitoring resistance levels and understanding resistance mechanisms.

Com o objetivo de caracterizar a resistência ao glyphosate em Lolium multiflorum (azevém) foi desenvolvida a pesquisa para: (a) testar a efetividade dos testes de dose-resposta em placas de Petri e em vasos, de acumulação in vivo de ácido shiquímico e de comportamento da fotossíntese líquida, como metodologia de detecção da resistência ao glyphosate em Lolium spp.; (b) caracterizar os mecanismos que conferem a resistência ao glyphosate em azevém e; (c) modelar a relação entre os biótipos susceptível (S) e resistente (R) ao glyphosate por meio de duas variáveis independentes (doses do herbicida e estádios fenológicos) e propor um modelo de predição da resistência ao glyphosate. A avaliação dos testes de detecção permitiu concluir que todos os testes foram adequados para identificar os biótipos resistentes. Na caracterização dos mecanismos de resistência, experimentos foram instalados para analisar o padrão de acúmulo de ácido shiquímico, o comportamento das variáveis fisiológicas e a composição das ceras epicuticulares, assim como, caracterizar a superfície foliar, a absorção e a translocação do 14Cglyphosate. As conclusões foram: as variáveis caracterização da absorção e quantificação e composição química das ceras epicuticulares, não apresentaram distinção entre R e S; todavia, o padrão de distribuição dos tricomas em pares em R difere da distribuição individualizada em S, assim como a translocação do glyphosate foi maior em direção as regiões que não receberam o tratamento no biótipo R. O movimento do glyphosate pelo floema segue a mesma rota dos produtos fotoassimilados sendo diretamente proporcional ao transporte dos carbonos assimilados, como se constatou em R que manteve a taxa de fotossíntese líquida constante durante o experimento. Os baixos níveis de acúmulo de ácido shiquímico constatados em R indicam que o glyphosate não é totalmente inibido de atuar na EPSPs ou que a EPSPs pode ser parcialmente inibida pelo herbicida, permanecendo na forma ativa. Desta forma, a resistência do azevém ao glyphosate, provavelmente, não é atribuída a um único mecanismo de resistência, assim como a maior translocação do herbicida no biótipo R representa um dos prováveis mecanismos que confere resistência. Nos estudos de modelagem, o modelo não-linear que proporcionou uma descrição precisa da relação entre a variável dependente e as variáveis independentes foi a seguinte equação: z = a/{[1+[(x-b)/c]2].[1+[(y-d)/c]2]}, onde z representa a fitomassa fresca relativa (%) na dose x do herbicida; y é o estádio fenológico (número de perfilhos, Np) e a, b, c e d são parâmetros empíricos. Para cada biótipo, ocorre um decréscimo generalizado em relação a fitomassa fresca relativa, comparativamente a respectiva testemunha, com o aumento da dose de glyphosate e com o decréscimo do estádio fenológico (GR50R/S = - 0,0148Np3 + 0,2986Np2 - 1,7758Np + 3,439; R2 = 1,0). O modelo proposto: Z = 100 - {100/[1+[(ax)b]c]}, onde Z representa a porcentagem de controle na dose x do herbicida e a, b, c são parâmetros empíricos que proporcionam um ajuste biológico apropriado aos dados; apresentou o menor quadrado médio do resíduo, comparativamente aos outros modelos utilizados neste tipo de estudo. As informações geradas neste trabalho auxiliam na compreensão da característica da resistência e na interpretação dos efeitos desta nas estratégias de manejo. / With the objective of characterizing the glyphosate resistance in Lolium multiflorum (Italian ryegrass) it was developed this research in order to: (a) test the effectiveness of the doseresponse tests in Petri dishes and in whole-plant, of in vivo test of the shikimic acid accumulation and of net photosynthesis behavior test, as a methodology to detection glyphosate resistance in Lolium spp.; (b) characterize the resistance mechanisms in Italian ryegrass and; (c) modeling the relationship among the susceptible (S) and the resistant (R) biotypes to glyphosate, with two independent variables (doses of herbicide and plant growth stages) and performing a predictive model of the glyphosate resistance in Italian ryegrass. The evaluation of the detection tests allowed concluded that all tests were appropriate to identify the resistant biotypes. In the characterization of the resistance mechanisms, experiments were installed to analyze the pattern of shikimic acid accumulation, the behavior of physiological processes affected by glyphosate and the epicuticular waxes composition, as well as, characterize the foliar surface, the 14C-glyphosate uptake and translocation. The conclusion were: the variables characterization of the uptake and epicuticular waxes quantification and chemical composition, didn\'t present distinction between R and S; though, the pattern of trichomes distribution on pairs in R biotype differs of the alone distribution in the case of S one, likewise, the glyphosate translocation was greater in direction to the areas that didn\'t receive the treatment with glyphosate in R biotype. The movement of glyphosate in the phloem follows the same pathway of the photosynthetic products being directly proportional to the transport of carbon fixed, as it was verified in R biotype that maintained the net photosynthesis constant during the experiment. The low levels of shikimic acid accumulation verified in R indicated that either glyphosate is not totally excluded from acting in EPSPs or that EPSPs can be partially inhibited by glyphosate, staying in the active form. Likewise, the glyphosate resistance in Italian ryegrass, probably, it is not attributed to a single resistance mechanism, as well as, the greater translocation of the herbicide in the R biotype probable represents the resistance mechanism. In the modeling studies, the non-linear model that provided a precise description of the relationship among the dependent variable and the independent variables was the following equation: z = a/{[1+[(x-b)/c]2]. [1+[(y-d)/c]2]}, where z represents the shoot biomass (%) at herbicide dose x; y is the growth stage (number of tillers, Nt) and a, b, c and d are empiric parameters. For each biotype, there was a general decreasing in relative shoot biomass in comparison to its respective untreated controls as glyphosate dose increased and growth stage decreased (GR50R/S = - 0.0148Nt3 + 0.2986Nt2 - 1.7758Nt + 3.439; R2 = 1.0). The proposed model: Z = 100 - {100/[1+[(ax)b]c]}, where Z represents the percentage of control at the herbicide dose x and a, b, c are empiric parameters that provide an appropriate biological fit to the data. It presented the smallest mean square error, comparatively to the other models used in this type of study. The information generated in this research help in the understanding of the resistance characteristic and in the interpretation of the effects of the management strategies.

Com o objetivo de caracterizar a resistência ao glyphosate em Lolium multiflorum (azevém) foi desenvolvida a pesquisa para: (a) testar a efetividade dos testes de dose-resposta em placas de Petri e em vasos, de acumulação in vivo de ácido shiquímico e de comportamento da fotossíntese líquida, como metodologia de detecção da resistência ao glyphosate em Lolium spp.; (b) caracterizar os mecanismos que conferem a resistência ao glyphosate em azevém e; (c) modelar a relação entre os biótipos susceptível (S) e resistente (R) ao glyphosate por meio de duas variáveis independentes (doses do herbicida e estádios fenológicos) e propor um modelo de predição da resistência ao glyphosate. A avaliação dos testes de detecção permitiu concluir que todos os testes foram adequados para identificar os biótipos resistentes. Na caracterização dos mecanismos de resistência, experimentos foram instalados para analisar o padrão de acúmulo de ácido shiquímico, o comportamento das variáveis fisiológicas e a composição das ceras epicuticulares, assim como, caracterizar a superfície foliar, a absorção e a translocação do 14Cglyphosate. As conclusões foram: as variáveis caracterização da absorção e quantificação e composição química das ceras epicuticulares, não apresentaram distinção entre R e S; todavia, o padrão de distribuição dos tricomas em pares em R difere da distribuição individualizada em S, assim como a translocação do glyphosate foi maior em direção as regiões que não receberam o tratamento no biótipo R. O movimento do glyphosate pelo floema segue a mesma rota dos produtos fotoassimilados sendo diretamente proporcional ao transporte dos carbonos assimilados, como se constatou em R que manteve a taxa de fotossíntese líquida constante durante o experimento. Os baixos níveis de acúmulo de ácido shiquímico constatados em R indicam que o glyphosate não é totalmente inibido de atuar na EPSPs ou que a EPSPs pode ser parcialmente inibida pelo herbicida, permanecendo na forma ativa. Desta forma, a resistência do azevém ao glyphosate, provavelmente, não é atribuída a um único mecanismo de resistência, assim como a maior translocação do herbicida no biótipo R representa um dos prováveis mecanismos que confere resistência. Nos estudos de modelagem, o modelo não-linear que proporcionou uma descrição precisa da relação entre a variável dependente e as variáveis independentes foi a seguinte equação: z = a/{[1+[(x-b)/c]2].[1+[(y-d)/c]2]}, onde z representa a fitomassa fresca relativa (%) na dose x do herbicida; y é o estádio fenológico (número de perfilhos, Np) e a, b, c e d são parâmetros empíricos. Para cada biótipo, ocorre um decréscimo generalizado em relação a fitomassa fresca relativa, comparativamente a respectiva testemunha, com o aumento da dose de glyphosate e com o decréscimo do estádio fenológico (GR50R/S = - 0,0148Np3 + 0,2986Np2 - 1,7758Np + 3,439; R2 = 1,0). O modelo proposto: Z = 100 - {100/[1+[(ax)b]c]}, onde Z representa a porcentagem de controle na dose x do herbicida e a, b, c são parâmetros empíricos que proporcionam um ajuste biológico apropriado aos dados; apresentou o menor quadrado médio do resíduo, comparativamente aos outros modelos utilizados neste tipo de estudo. As informações geradas neste trabalho auxiliam na compreensão da característica da resistência e na interpretação dos efeitos desta nas estratégias de manejo. / With the objective of characterizing the glyphosate resistance in Lolium multiflorum (Italian ryegrass) it was developed this research in order to: (a) test the effectiveness of the doseresponse tests in Petri dishes and in whole-plant, of in vivo test of the shikimic acid accumulation and of net photosynthesis behavior test, as a methodology to detection glyphosate resistance in Lolium spp.; (b) characterize the resistance mechanisms in Italian ryegrass and; (c) modeling the relationship among the susceptible (S) and the resistant (R) biotypes to glyphosate, with two independent variables (doses of herbicide and plant growth stages) and performing a predictive model of the glyphosate resistance in Italian ryegrass. The evaluation of the detection tests allowed concluded that all tests were appropriate to identify the resistant biotypes. In the characterization of the resistance mechanisms, experiments were installed to analyze the pattern of shikimic acid accumulation, the behavior of physiological processes affected by glyphosate and the epicuticular waxes composition, as well as, characterize the foliar surface, the 14C-glyphosate uptake and translocation. The conclusion were: the variables characterization of the uptake and epicuticular waxes quantification and chemical composition, didn\'t present distinction between R and S; though, the pattern of trichomes distribution on pairs in R biotype differs of the alone distribution in the case of S one, likewise, the glyphosate translocation was greater in direction to the areas that didn\'t receive the treatment with glyphosate in R biotype. The movement of glyphosate in the phloem follows the same pathway of the photosynthetic products being directly proportional to the transport of carbon fixed, as it was verified in R biotype that maintained the net photosynthesis constant during the experiment. The low levels of shikimic acid accumulation verified in R indicated that either glyphosate is not totally excluded from acting in EPSPs or that EPSPs can be partially inhibited by glyphosate, staying in the active form. Likewise, the glyphosate resistance in Italian ryegrass, probably, it is not attributed to a single resistance mechanism, as well as, the greater translocation of the herbicide in the R biotype probable represents the resistance mechanism. In the modeling studies, the non-linear model that provided a precise description of the relationship among the dependent variable and the independent variables was the following equation: z = a/{[1+[(x-b)/c]2]. [1+[(y-d)/c]2]}, where z represents the shoot biomass (%) at herbicide dose x; y is the growth stage (number of tillers, Nt) and a, b, c and d are empiric parameters. For each biotype, there was a general decreasing in relative shoot biomass in comparison to its respective untreated controls as glyphosate dose increased and growth stage decreased (GR50R/S = - 0.0148Nt3 + 0.2986Nt2 - 1.7758Nt + 3.439; R2 = 1.0). The proposed model: Z = 100 - {100/[1+[(ax)b]c]}, where Z represents the percentage of control at the herbicide dose x and a, b, c are empiric parameters that provide an appropriate biological fit to the data. It presented the smallest mean square error, comparatively to the other models used in this type of study. The information generated in this research help in the understanding of the resistance characteristic and in the interpretation of the effects of the management strategies.

Bacterial contamination of platelet concentrates (PCs) represents the highest post-transfusion infectious risk. The skin flora bacterium Staphylococcus epidermidis has been reported to be the predominant aerobic contaminant of PCs. The Ramirez' group has shown that S. epidermidis can form surface-attached bacterial aggregates known as biofilms, and can outcompete other coagulase-negative staphylococci, such as Staphylococcus capitis, in PCs. The ability of S. epidermidis to form biofilms has been linked to increased pathogenicity and missed detection during PC screening with an automated culture system (BacT/ALERT). This thesis aimed at investigating the proliferative advantage and resistance mechanisms displayed by S. epidermidis in the PC milieu. Furthermore, in an effort to enhance PC safety for transfusion patients, I studied the anti-biofilm properties of essential oils and antimicrobial peptides (AMPs). My studies aimed at improving PC safety by focussing on both the point of introduction of bacterial contaminants (blood collection), and the stage at which bacterial contaminants can form biofilms and proliferate (PC storage). S. epidermidis can be found in the skin of blood donors as biofilms, which are resistant to the blood donor skin disinfectant currently used by Canadian Blood Services, chlorhexidine-gluconate and isopropyl alcohol (CHG-IPA). Here, several plant-extracted essential oils were evaluated for their ability to enhance the anti-biofilm activity of CHG-IPA. Data revealed that the Lavandula multifida oil and its main component (linalool) greatly enhanced the activity of CHG-IPA against S. epidermidis biofilms. Furthermore, the ability of a combination of three synthetic AMPs to inhibit S. epidermidis biofilm formation during PC storage was assessed These results showed that the combination of AMPs could inhibit biofilm formation but was ineffective against pre-formed S. epidermidis biofilms. The accumulation associated protein (Aap) encoded by the aap gene, found in most S. epidermidis strains and absent in S. capitis, plays a role in biofilm formation. When S. epidermidis aap is transformed into S. capitis, this bacterium displayed increased biofilm formation and proliferated to higher concentrations compared to untransformed S. capitis and to a S. epidermidis aap deletion mutant. Based on these results, aap appears to play a role in providing S. epidermidis a proliferative advantage in PCs by enhancing biofilm formation. Lastly, the GraRS system and SepA were studied for their role in S. epidermidis resistance to platelet-derived AMPs using the synthetic AMP PD4 as a model molecule. Results indicate that the GraS mechanism is involved in resistance towards PD4. The work presented in my thesis provides further insights into why S. epidermidis has a proliferative advantage in the PC storage environment and allows for the proposal of alternative methods to enhance PC safety for transfusion patients.

No description available.

Glioblastoma (GBM) is the most malignant primary brain tumor without effective therapies. Since bevacizumab was FDA approved for targeting vascular endothelial growth factor receptor 2 (VEGFR2) in adult patients with recurrent GBM, targeted therapy against receptor tyrosine kinases (RTKs) has become a new avenue for GBM therapeutics. In addition to VEGFR, the epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), hepatocyte growth factor receptor (HGFR/MET), and fibroblast growth factor receptor (FGFR) are major RTK targets. However, results from clinical Phase II/III trials indicate that most RTK-targeting therapeutics including tyrosine kinase inhibitors (TKIs) and neutralizing antibodies lack clinical efficacy, either alone or in combination. The major challenge is to uncover the genetic RTK alterations driving GBM initiation and progression, as well as to elucidate the mechanisms toward therapeutic resistance. In this review, we will discuss the genetic alterations in these 5 commonly targeted RTKs, the clinical trial outcomes of the associated RTK-targeting therapeutics, and the potential mechanisms toward the resistance. We anticipate that future design of new clinical trials with combination strategies, based on the genetic alterations within an individual patient's tumor and mechanisms contributing to therapeutic resistance after treatment, will achieve durable remissions and improve outcomes in GBM patients.

Yes / In this study, anti-corrosion concrete for sewage system was developed with nano TiO2 (NT) and reactive powder concrete (RPC). The corrosion resistances of NT modified RPC (NTMRPC) in high concentration enhanced sewage were investigated from the perspectives of biological, physical and chemical corrosion resistances, respectively. In addition, mechanical properties of NTMRPC after sewage corrosion were also studied. Research results indicated that NT can endow RPC with antimicrobial property through their microorganism biodegradation properties. The inhibition and elimination rates of NTMRPC to its surface microorganisms were 37.35% and 80.93%, respectively. After sewage corrosion, the surface roughness, mass loss and deterioration depth of RPC were decreased by 62.57%, 15.48% and 18.44% due to the NT inclusion, respectively. In addition, the pH values of RPC in the deterioration depth ranges of 0-3 mm and 3-6 mm were increased by 11.45% and 23.62%, respectively. NT can restrain the strength deterioration of RPC in high concentration enhanced sewage. This may be due to the improved sewage biological anti-corrosion performances of RPC by inhibiting/eliminating the microorganisms on the surface of RPC as well as the enhanced sewage physical/chemical anti-corrosion performances of RPC by improving the compactness of RPC. / The authors thank the funding provided by the National Science Foundation of China 513 (51978127 and 51908103), and National Key Research and Development Program of China 514 (2018YFC070560 and 2017YFC0703410).