Sites et monuments : bulletin de la Société pour la protection des paysages et de l'esthétique générale de la France

juillet 19841984/07 (N106)-1984/09

Additional file 2: of Available, Bed-sided, Comprehensive (ABC) score to a diagnosis of Methicillin-resistant Staphylococcus aureus infection: a derivation and validation study

Comparison of the two patient cohorts. (PDF 149 kb

Novel Anti-Microbial Peptide SR-0379 Accelerates Wound Healing via the PI3 Kinase/Akt/mTOR Pathway

<div><p>We developed a novel cationic antimicrobial peptide, AG30/5C, which demonstrates angiogenic properties similar to those of LL-37 or PR39. However, improvement of its stability and cost efficacy are required for clinical application. Therefore, we examined the metabolites of AG30/5C, which provided the further optimized compound, SR-0379. SR-0379 enhanced the proliferation of human dermal fibroblast cells (NHDFs) via the PI3 kinase-Akt-mTOR pathway through integrin-mediated interactions. Furthermore SR-0379 promoted the tube formation of human umbilical vein endothelial cells (HUVECs) in co-culture with NHDFs. This compound also displays antimicrobial activities against a number of bacteria, including drug-resistant microbes and fungi. We evaluated the effect of SR-0379 in two different would-healing models in rats, the full-thickness defects under a diabetic condition and an acutely infected wound with full-thickness defects and inoculation with <i>Staphylococcus aureus</i>. Treatment with SR-0379 significantly accelerated wound healing when compared to fibroblast growth factor 2 (FGF2). The beneficial effects of SR-0379 on wound healing can be explained by enhanced angiogenesis, granulation tissue formation, proliferation of endothelial cells and fibroblasts and antimicrobial activity. These results indicate that SR-0379 may have the potential for drug development in wound repair, even under especially critical colonization conditions.</p></div

MICs of several compounds against <i>E. coli</i>, <i>P. aeruginosa</i> and <i>S. aureus</i>.

<p>The scores indicate the MICs (mg/ml) for <i>E. coli</i>, <i>P. aeruginosa and S. aureus</i>. MICs represent the individual data from two independent experiments.</p

Genotypes of carbapenem-resistant <i>A</i>. <i>baumannii</i> (CRAb) isolates from hospital patients in Bangkok.

<p>Total^a: Total number of isolates with each resistance gene</p><p>%^b: The proportion of isolates with each resistance gene</p><p>Genotypes of carbapenem-resistant <i>A</i>. <i>baumannii</i> (CRAb) isolates from hospital patients in Bangkok.</p

Clinical Specimen-Direct LAMP: A Useful Tool for the Surveillance of <i>bla</i>_OXA-23-Positive Carbapenem-Resistant <i>Acinetobacter baumannii</i>

<div><p>Healthcare-associated infections are a leading cause of morbidity and mortality worldwide. Treatment is increasingly complicated by the escalating incidence of antimicrobial resistance. Among drug-resistant pathogens, carbapenem-resistant <i>Acinetobacter baumannii</i> (CRAb) is of increasing concern because of the limited applicable therapies and its expanding global distribution in developed countries and newly industrialized countries. Therefore, a rapid detection method that can be used even in resource-poor countries is urgently required to control this global public health threat. Conventional techniques, such as bacterial culture and polymerase chain reaction (PCR), are insufficient to combat this threat because they are time-consuming and laborious. In this study, we developed a loop-mediated isothermal amplification (LAMP) method for detecting <i>bla</i>_OXA-23-positive CRAb, the most prevalent form of CRAb in Asia, especially in Thailand, and confirmed its efficacy as a surveillance tool in a clinical setting. Clinical samples of sputum and rectal swabs were collected from patients in a hospital in Bangkok and used for LAMP assays. After boiling and centrifugation, the supernatants were used directly in the assay. In parallel, a culture method was used for comparison purposes to evaluate the specificity and sensitivity of LAMP. As a first step, a total of 120 sputum samples were collected. The sensitivity of LAMP was 88.6% (39/44), and its specificity was 92.1% (70/76) using the culture method as the “gold standard”. When surveillance samples including sputum and rectal swabs were analyzed with the LAMP assay, its sensitivity was 100.0%. This method enables the direct analysis of clinical specimens and provides results within 40 minutes of sample collection, making it a useful tool for surveillance even in resource-poor countries.</p></div

<i>In vitro</i> activities of SR-0379 against Gram-positive and Gram-negative bacteria and fungi.

<p>The scores indicate the MICs (mg/ml) for gram-positive and gram-negative bacteria and fungi. MICs represent the individual data from two independent experiments.</p><p>NT: Not tested.</p

Cellular function of SR-0379.

<p>A) Effect of SR-0379 on NHDFs proliferation. NHDFs were treated with SR-0379 (1, 3 and 10 μg/ml) or FGF2 (0.1 μg/ml). N = 4 per group. *P<0.05 vs. control. B) The upper panel shows representative pictures of tube formation in a co-culture of HUVECs and NHDFs (Control, FGF2: 0.2 μg/ml) and SR-0379 (10 μg/ml). The lower panel shows the effects of SR-0379 on tube formation in a co-culture of HUVECs and NHDFs. N = 5 per group. *P<0.05, **P<0.01 vs. control. C) The upper panel shows representative pictures of the migration induced by FGF2 (0.1 μg/ml) and SR-0379 (10 μg/ml). The lower panel shows the effects of FGF2 (0.1 μg/ml) and SR-0379 (1 and 10 μg/ml) on migration. N = 4 per group. *P<0.05, **P<0.01 vs. control, #P<0.01 vs. SR-0379 (1 μg/ml). D) The upper panel shows representative pictures of the fibroblast-collagen-matrix contraction assay with FGF2 (0.3 μg/ml) and SR-0379 (1, 3, 10 and 30 μg/ml). The lower panel shows the effects of FGF2 (0.3 μg/ml) and SR-0379 (1, 3, 10 and 30 μg/ml) on fibroblast-collagen matrix contraction. N = 3 per group. *P<0.05, **P<0.01 vs. control.</p

Lead optimization from the angiogenic peptide AG30/5C.

<p>A) Major metabolites of AG30/5C determined by MALDI-TOF MS. The parent compound (AG30/5C) was incubated with rat serum <i>in vitro</i> 60 minutes. The metabolites were identified by comparison with the pre-incubation peptide. B) Sequences and net charges of AG30/5C and AG30/5C-derived peptides (SR-0007 and SR-0379). The lysine (K) of SR-007 was replaced with D-lysine in SR-0379. C) Effect of AG30/5C (10 μg/ml) and SR-0007 (10 μg/ml) on HUVECs proliferation. N = 3 per group. *P<0.05 vs. control. D) Effect of AG30/5C (10 μg/ml) and SR-0007 (10 μg/ml) on tube formation. The formation of capillary-like structures was observed in co-cultures of HUVECs and NHDFs. N = 5-12 per group. *P<0.05 vs. control. E) Stability of SR-0007 and SR-0379 in rat and human sera. SR-0007 and SR-0379 were quantified before or after incubation <i>in vitro</i> with rat and human sera for either 3 or 10 minutes. N = 2.</p

Real-time turbidity assays under various conditions using a turbidimeter.

<p>(A) To determine the optimal reaction conditions, a LAMP assay was performed on extracted bacterial DNA at temperatures ranging from 62°C to 67°C. At 65°C, the reaction finished within the shortest period of time, and the negative control remained transparent after 60 minutes of incubation. (B) To determine the detection limit, the extracted DNA templates were serially diluted 10 times (from 2 pg to 2×10⁻⁶ pg) and used in the LAMP assay. The turbidity was evaluated with a turbidimeter every 5 minutes.</p