Natural Variant of Collagen-Like Protein A in Serotype M3 Group A Streptococcus Increases Adherence and Decreases Invasive Potential

Group A Streptococcus (GAS) predominantly exists as a colonizer of the human oropharynx that occasionally breaches epithelial barriers to cause invasive diseases. Despite the frequency of GAS carriage, few investigations into the contributory molecular mechanisms exist. To this end, we identified a naturally occurring polymorphism in the gene encoding the streptococcal collagen-like protein A (SclA) in GAS carrier strains. All previously sequenced invasive serotype M3 GAS possess a premature stop codon in the sclA gene truncating the protein. The carrier polymorphism is predicted to restore SclA function and was infrequently identified by targeted DNA sequencing in invasive strains of the same serotype. We demonstrate that a strain with the carrier sclA allele expressed a full-length SclA protein, while the strain with the invasive sclA allele expressed a truncated variant. An isoallelic mutant invasive strain with the carrier sclA allele exhibited decreased virulence in a mouse model of invasive disease and decreased multiplication in human blood. Further, the isoallelic invasive strain with the carrier sclA allele persisted in the mouse nasopharynx and had increased adherence to cultured epithelial cells. Repair of the premature stop codon in the invasive sclA allele restored the ability to bind the extracellular matrix proteins laminin and cellular fibronectin. These data demonstrate that a mutation in GAS carrier strains increases adherence and decreases virulence and suggest selection against increased adherence in GAS invasive isolates.     

Spread of old and new clones of epidemic methicillin-resistant Staphylococcus aureus in Poland

Objective: To study the relatedness among methicillin-resistant Staphylococcus aureus (MRSA) isolates originating from two regions of Poland using different epidemiologic typing methods.
Methods: Forty-five MRSA isolates (19 from Warsaw and 26 from the Grajewo region) were collected between 1995 and 1996. For phenotypic epidemiologic analysis, antimicrobial susceptibility testing (AST) with a panel of 19 antibiotics was performed. For genotypic epidemiologic analysis, pulsed-field gel electrophoresis (PFGE) of Smal-digested chromosomal DNA, restriction endonuclease analysis of plasmid (REAP) DNA digested by Hin dIII, random amplification of polymorphic DNA (RAPD) and binary typing (BT) of genomic DNA by hybridization with five different RAPD-generated strain-specific DNA probes, were used.
Results: Six clusters of clonally related strains were found among the MRSA isolates analyzed. Three of these, identified in both regions, were related to previously described Polish epidemic clones, designated HeEMRSA-Pol1 (heterogeneously methicillin resistant—18 isolates) and HoEMRSA-Pol1 (homogeneously resistant—two clones, six isolates each). The remaining three clones, identified in the Grajewo region only, are previously undescribed. One of these, represented by 11 isolates, appears to be new epidemic heterogeneous MRSA clone (HeEMRSA-Pol2). Results of PFGE and BT in general showed good correlation, and, in some cases, RAPD using AP1 and AP7 primers could discriminate between isolates belonging to single PFGE or BT types. Broad AST and REAP can provide useful additional information concerning relatedness.
Conclusion: Evidence for the spread of previously recognized epidemic MRSA clones in Poland and the presence of a new epidemic heterogeneously resistant clone of MRSA in hospitals outside Warsaw is documented.     

Effects of old age on vascular complexity and dispersion of the hepatic sinusoidal network

OBJECTIVES: In old age, there are marked changes in both the structure of the liver sinusoidal endothelial cell and liver perfusion. The objective of this study was to determine whether there are also aging changes in the microvascular architecture and vascular dispersion of the liver that might influence liver function. METHODS: Vascular corrosion casts and light micrographs of young (4 months) and old (24 months) rat livers were compared. Fractal and Fourier analyses and micro-computed tomography were used. Vascular dispersion was determined from the dispersion number for sucrose and 100-nm microspheres in impulse response experiments. RESULTS: Age did not affect sinusoidal dimensions, sinusoidal density, or dispersion number. There were changes in the geometry and complexity of the sinusoidal network as determined by fractal dimension and degree of anisotropy. CONCLUSIONS: There are small, age-related changes in the architecture of the liver sinusoidal network, which may influence hepatic function and reflect broader aging changes in the microcirculation. However, sinusoidal dimensions and hepatic vascular dispersion are not markedly influenced by old age.     

Mechanical Properties of Different Nanopatterned TiO2 Substrates and Their Effect on Hydrothermally Synthesized Bioactive Hydroxyapatite Coatings

Nanotechnology is a very attractive tool for tailoring the surface of an orthopedic implant to optimize its interaction with the biological environment. Nanostructured interfaces are promising, especially for orthopedic applications. They can not only improve osseointegration between the implant and the living bone but also may be used as drug delivery platforms. The nanoporous structure can be used as a drug carrier to the surrounding tissue, with the intention to accelerate tissue–implant integration as well as to reduce and treat bacterial infections occurring after implantation. Titanium oxide nanotubes are promising for such applications; however, their brittle nature could be a significantly limiting factor. In this work, we modified the topography of commercially used titanium foil by the anodization process and hydrothermal treatment. As a result, we obtained a crystalline nanoporous u-shaped structure (US) of anodized titanium oxide with improved resistance to scratch compared to TiO₂ nanotubes. The US titanium substrate was successfully modified with hydroxyapatite coating and investigated for bioactivity. Results showed high bioactivity in simulated body fluid (SBF) after two weeks of incubation.     

Use of a tourniquet with and without adrenaline reduces blood loss during liposuction for lymphoedema of the arm

Sixty-two patients with lymphoedema of the arm after mastectomy and with hypertrophy of the adipose tissue were consecutively treated by liposuction in three different ways. The first group was operated on without the use of a tourniquet. In the second group, liposuction extended up to the distal edge of the tourniquet, and then into the proximal upper arm previously covered by the tourniquet using the ‘dry’ technique. Treatment of the third group was identical to that of the second one, but the area covered by the tourniquet was treated by the tumescent technique. Eighteen patients who did not have lymphoedema either treated or not treated with adrenaline served as a reference group to see how blood transfusions varied with various volumes of aspirate. Using a tourniquet significantly reduced blood loss and the number of transfusions, which was further reduced by tumescence. In the historical reference group, the number of blood transfusions increased as the volume of aspirate increased, and further if no adrenaline was added.