Клінічні особливості генетично детермінованих гіпобета-ліпопротеїнемії та гіпохолестеролемії

Familial hypobetalipoproteinemia (FHBL) is an autosomal dominant disorder of lipid metabolism characterized by extremely low plasma levels of apolipoprotein B, total cholesterol and low-density lipoprotein cholesterol. Heterozygotes for FHBL are often asymptomatic. Clinical features of homozygous FHBL can include acanthocytosis, deficiencies in fat-soluble vitamins secondary to malabsorption, atypical retinitis pigmentosa and neuromuscular abnormalities

Influence of antibiotic pressure on bacterial bioluminescence, with emphasis on Staphylococcus aureus

Bioluminescence imaging is used for longitudinal evaluation of bacteria in live animals. Clear relations exist between bacterial numbers and their bioluminescence. However, bioluminescence images of Staphylococcus aureus Xen29, S. aureus Xen36 and Escherichia coli Xen14 grown on tryptone soy agar in Etests demonstrated increased bioluminescence at sub-MICs of different antibiotics. This study aimed to further evaluate the influence of antibiotic pressure on bioluminescence in S. aureus Xen29. Bioluminescence of S. aureus Xen29, grown planktonically in tryptone soy broth, was quantified in the absence and presence of different concentrations of vancomycin, ciprofloxacin, erythromycin or chloramphenicol and was related to expression of the luxA gene under antibiotic pressure measured using real-time PCR. In the absence of antibiotics, staphylococcal bioluminescence increased over time until a maximum after ca. 6 h of growth, and subsequently decreased to the detection threshold after 24 h of growth owing to reduced bacterial metabolic activity. Up to MICs of the antibiotics, bioluminescence increased according to a similar pattern up to 6 h of growth, but after 24 h bioluminescence was higher than in the absence of antibiotics. Contrary to expectations, bioluminescence per organism (CFU) after different growth periods in the absence and at MICs of different antibiotics decreased with increasing expression of luxA. Summarising, antibiotic pressure impacts the relation between CFU and bioluminescence. Under antibiotic pressure, bioluminescence is not controlled by luxA expression but by co-factors impacting the bacterial metabolic activity. This conclusion is of utmost importance when evaluating antibiotic efficacy in live animals using bioluminescent bacterial strains. (C) 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved

Unveiling the Antifouling Performance of Different Marine Surfaces and Their Effect on the Development and Structure of Cyanobacterial Biofilms

Since biofilm formation by microfoulers significantly contributes to the fouling process, it is important to evaluate the performance of marine surfaces to prevent biofilm formation, as well as understand their interactions with microfoulers and how these affect biofilm development and structure. In this study, the long-term performance of five surface materials-glass, perspex, polystyrene, epoxy-coated glass, and a silicone hydrogel coating-in inhibiting biofilm formation by cyanobacteria was evaluated. For this purpose, cyanobacterial biofilms were developed under controlled hydrodynamic conditions typically found in marine environments, and the biofilm cell number, wet weight, chlorophyll a content, and biofilm thickness and structure were assessed after 49 days. In order to obtain more insight into the effect of surface properties on biofilm formation, they were characterized concerning their hydrophobicity and roughness. Results demonstrated that silicone hydrogel surfaces were effective in inhibiting cyanobacterial biofilm formation. In fact, biofilms formed on these surfaces showed a lower number of biofilm cells, chlorophyll a content, biofilm thickness, and percentage and size of biofilm empty spaces compared to remaining surfaces. Additionally, our results demonstrated that the surface properties, together with the features of the fouling microorganisms, have a considerable impact on marine biofouling potential

Biofilm formation behaviour of marine filamentous cyanobacterial strains in controlled hydrodynamic conditions

Marine biofouling has severe economic impacts and cyanobacteria play a significant role as early surface colonizers. Despite this fact, cyanobacterial biofilm formation studies in controlled hydrodynamic conditions are scarce. In this work, computational fluid dynamics was used to determine the shear rate field on coupons that were placed inside the wells of agitated 12-well microtiter plates. Biofilm formation by three different cyanobacterial strains was assessed at two different shear rates (4 and 40 s-1 ) which can be found in natural ecosystems and using different surfaces (glass and perspex). Biofilm formation was higher under low shear conditions, and differences obtained between surfaces were not always statistically significant. The hydrodynamic effect was more noticeable during the biofilm maturation phase rather than during initial cell adhesion and optical coherence tomography showed that different shear rates can affect biofilm architecture. This study is particularly relevant given the cosmopolitan distribution of these cyanobacterial strains and the biofouling potential of these organisms.This work was financially supported by project UID/EQU/ 00511/2019 – Laboratory for Process Engineering, Environment, Biotechnology and Energy – LEPABE funded by national funds through FCT/MCTES (PIDDAC) and by the CVMAR+i – Industrial Innovation and Marine Biotechnology Valorisation, funded by INTERREG V A Espanha Portugal (POCTEP) [0302_CVMAR_I_1_P]. The authors also acknowledge support from the EU COST Actions iPROMEDAI (TD1305) and ENBA (CA15216), and M.J.R. acknowledges a PhD grant from FCT (SFRH/BD/140080/2018)

Salt shock-inducible Photosystem I cyclic electron transfer in Synechocystis PCC6803 relies on binding of ferredoxin:NADP+ reductase to the thylakoid membranes via its CpcD phycobilisome-linker homologous N-terminal domain

AbstractRelative to ferredoxin:NADP+ reductase (FNR) from chloroplasts, the comparable enzyme in cyanobacteria contains an additional 9 kDa domain at its amino-terminus. The domain is homologous to the phycocyanin associated linker polypeptide CpcD of the light harvesting phycobilisome antennae. The phenotypic consequences of the genetic removal of this domain from the petH gene, which encodes FNR, have been studied in Synechocystis PCC 6803. The in frame deletion of 75 residues at the amino-terminus, rendered chloroplast length FNR enzyme with normal functionality in linear photosynthetic electron transfer. Salt shock correlated with increased abundance of petH mRNA in the wild-type and mutant alike. The truncation stopped salt stress-inducible increase of Photosystem I-dependent cyclic electron flow. Both photoacoustic determination of the storage of energy from Photosystem I specific far-red light, and the re-reduction kinetics of P700+, suggest lack of function of the truncated FNR in the plastoquinone–cytochrome b6f complex reductase step of the PS I-dependent cyclic electron transfer chain. Independent gold-immunodecoration studies and analysis of FNR distribution through activity staining after native polyacrylamide gelelectrophoresis showed that association of FNR with the thylakoid membranes of Synechocystis PCC 6803 requires the presence of the extended amino-terminal domain of the enzyme. The truncated ΔpetH gene was also transformed into a NAD(P)H dehydrogenase (NDH1) deficient mutant of Synechocystis PCC 6803 (strain M55) (T. Ogawa, Proc. Natl. Acad. Sci. USA 88 (1991) 4275–4279). Phenotypic characterisation of the double mutant supported our conclusion that both the NAD(P)H dehydrogenase complex and FNR contribute independently to the quinone cytochrome b6f reductase step in PS I-dependent cyclic electron transfer. The distribution, binding properties and function of FNR in the model cyanobacterium Synechocystis PCC 6803 will be discussed

The use of biomimetic surfaces to reduce single- and dual-species biofilms of Escherichia coli and Pseudomonas putida

The ability of bacteria to adhere to and form biofilms on food contact surfaces poses serious challenges, as these may lead to the cross-contamination of food products. Biomimetic topographic surface modifications have been explored to enhance the antifouling performance of materials. In this study, the topography of two plant leaves, Brassica oleracea var. botrytis (cauliflower, CF) and Brassica oleracea capitate (white cabbage, WC), was replicated through wax moulding, and their antibiofilm potential was tested against single- and dual-species biofilms of Escherichia coli and Pseudomonas putida. Biomimetic surfaces exhibited higher roughness values (SaWC = 4.0 ± 1.0 μm and SaCF = 3.3 ± 1.0 μm) than the flat control (SaF = 0.6 ± 0.2 μm), whilst the CF surface demonstrated a lower interfacial free energy (ΔGiwi) than the WC surface (−100.08 mJ m−2 and −71.98 mJ m−2, respectively). The CF and WC surfaces had similar antibiofilm effects against single-species biofilms, achieving cell reductions of approximately 50% and 60% for E. coli and P. putida, respectively, compared to the control. Additionally, the biomimetic surfaces led to reductions of up to 60% in biovolume, 45% in thickness, and 60% in the surface coverage of single-species biofilms. For dual-species biofilms, only the E. coli strain growing on the WC surface exhibited a significant decrease in the cell count. However, confocal microscopy analysis revealed a 60% reduction in the total biovolume and surface coverage of mixed biofilms developed on both biomimetic surfaces. Furthermore, dual-species biofilms were mainly composed of P. putida, which reduced E. coli growth. Altogether, these results demonstrate that the surface properties of CF and WC biomimetic surfaces have the potential for reducing biofilm formation

Production and Characterization of Graphene Oxide Surfaces against Uropathogens

Graphene and its functionalized derivatives have been increasingly applied in the biomedi-cal field, particularly in the production of antimicrobial and anti-adhesive surfaces. This study aimed to evaluate the performance of graphene oxide (GO)/polydimethylsiloxane (PDMS) composites against Staphylococcus aureus and Pseudomonas aeruginosa biofilms. GO/PDMS composites containing different GO loadings (1, 3, and 5 wt.%) were synthesized and characterized regarding their morphol-ogy, roughness, and hydrophobicity, and tested for their ability to inhibit biofilm formation under conditions that mimic urinary tract environments. Biofilm formation was assessed by determining the number of total and culturable cells. Additionally, the antibacterial mechanisms of action of GO were investigated for the tested uropathogens. Results indicated that the surfaces containing GO had greater roughness and increased hydrophobicity than PDMS. Biofilm analysis showed that the 1 wt.% GO/PDMS composite was the most effective in reducing S. aureus biofilm formation. In oppo-sition, P. aeruginosa biofilms were not inhibited by any of the synthesized composites. Furthermore, 1% (w/v) GO increased the membrane permeability, metabolic activity, and endogenous reactive oxygen species (ROS) synthesis in S. aureus. Altogether, these results suggest that GO/PDMS com-posites are promising materials for application in urinary catheters, although further investigation is required

CLSM as Quantitative Method to Determine the Size of Drug Crystals in a Solid Dispersion

PURPOSE: To test whether confocal laser scanning microscopy (CLSM) can be used as an analytical tool to determine the drug crystal size in a powder mixture or a crystalline solid dispersion. METHODS: Crystals of the autofluorescent drug dipyridamole were incorporated in a matrix of crystalline mannitol by physical mixing or freeze-drying. Laser diffraction analysis and dissolution testing were used to validate the particle size that was found by CLSM. RESULTS: The particle size of the pure drug as determined by laser diffraction and CLSM were similar (D(50) of approximately 22 μm). CLSM showed that the dipyridamole crystals in the crystalline dispersion obtained by freeze-drying of less concentrated solutions were of sub-micron size (0.7 μm), whereas the crystals obtained by freeze-drying of more concentrated solutions were larger (1.3 μm). This trend in drug crystal size was in agreement with the dissolution behavior of the tablets prepared from these products. CONCLUSION: CLSM is a useful technique to determine the particle size in a powder mixture. Furthermore, CLSM can be used to determine the drug crystal size over a broad size distribution. A limitation of the method is that the drug should be autofluorescent

Production and Characterization of Graphene Oxide Surfaces against Uropathogens

Graphene and its functionalized derivatives have been increasingly applied in the biomedical field, particularly in the production of antimicrobial and anti-adhesive surfaces. This study aimed to evaluate the performance of graphene oxide (GO)/polydimethylsiloxane (PDMS) composites against Staphylococcus aureus and Pseudomonas aeruginosa biofilms. GO/PDMS composites containing different GO loadings (1, 3, and 5 wt.%) were synthesized and characterized regarding their morphology, roughness, and hydrophobicity, and tested for their ability to inhibit biofilm formation under conditions that mimic urinary tract environments. Biofilm formation was assessed by determining the number of total and culturable cells. Additionally, the antibacterial mechanisms of action of GO were investigated for the tested uropathogens. Results indicated that the surfaces containing GO had greater roughness and increased hydrophobicity than PDMS. Biofilm analysis showed that the 1 wt.% GO/PDMS composite was the most effective in reducing S. aureus biofilm formation. In opposition, P. aeruginosa biofilms were not inhibited by any of the synthesized composites. Furthermore, 1% (w/v) GO increased the membrane permeability, metabolic activity, and endogenous reactive oxygen species (ROS) synthesis in S. aureus. Altogether, these results suggest that GO/PDMS composites are promising materials for application in urinary catheters, although further investigation is required.</p