Phage types, virulence genes and PFGE profiles of Shiga toxin-producing Escherichia coli O157:H7 isolated from raw beef, soft cheese and vegetables in Lima (Peru)

The present study was conducted in Lima Metropolitana to evaluate the prevalence of Shiga toxin-producing Escherichia coli (STEC) O157:H7 in raw beef, raw ground beef, soft cheese and fresh vegetables, sampled at different markets in the city. Between October 2000 and February 2001, 407 food samples were collected from different markets in the 42 districts of Lima Metropolitana. Samples were assayed for E. coli O157 by selective enrichment in modified Tryptic Soy Broth containing novobiocin, followed by immunomagnetic separation (IMS) and plating onto sorbitol MacConkey agar supplemented with cefixime and potassium tellurite. Fifty (12.3%) of 407 food samples resulted positive for E. coli O157 isolation (23 of 102 ground beef; 15 of 102 beef meat; eight of 102 soft cheese and four of 101 fresh vegetables). Thirty-five E. coli O157 isolates were further analysed for the presence of virulence genes. All 35 were positive by PCR for O157 rfbE, fliCh7, eae-gamma1 and ehxA genes. In addition, genes encoding Shiga toxins were detected in 33 of 35 isolates, five isolates (14%) encoded stx(1), stx(2), and 28 (80%) stx2 only. The isolates were of seven different phage types (PT4, PT8, PT14, PT21, PT34, PT54, and PT87) with three phage types accounting for 80% of isolates: PT4 (15 isolates), PT14 (8 isolates), and PT21 (5 isolates). Interestingly, the majority (31 of 35; 89%) of E. coli O157:H7 isolates characterized in this study belonged mainly to the phage types previously found in STEC O157:H7 strains associated with severe human disease in Europe and Canada. Pulsed-field gel electrophoresis (PFGE) of 32 isolates revealed 14 XbaI-PFGE groups (I to XIV) of similarity >85%, with 23 (72%) isolates grouped in five clusters. Some isolates from different districts presented a high clonal relatedness. Thus, PFGE group VIII clustered eleven strains from nine different districts. The broad range of PFGE subtypes found in this study demonstrates the natural occurrence of many genetic variants among STEC O157:H7 spread in Lima.     

Evaluation of in vitro antioxidant properties of some traditional Sardinian medicinal plants: Investigation of the high antioxidant capacity of Rubus ulmifolius

The antioxidant capacities of 11 botanical species used in the tradition of Sardinia as teas beverages or as decoction for medicinal purposes were evaluated using different in vitro methods (BR, TEAC, DPPH and FC). Among the various species, Rubus ulmifolius, resulted the more active with all the used methods. Phytochemical investigation on the extract yields in the isolation of several phenolic compounds namely caffeic acid, ferulic acid, quercetin-3-O-glucuronide, kaempferol-3-O-glucuronide, kaempferol-3-O-(6″-p-coumaroyl)-β-d-glucopyranoside, kaempferol-3-O-(6″-caffeoyl)-β-d-glucopyranoside, chlorogenic acid, 4-caffeoylquinic acid and 5-caffeoylquinic acid. The antioxidant activity of isolated compounds was also evaluated.     

Optimization and Validation of a UV–HPLC Method for Vitamin C Determination in Strawberries (<Emphasis Type="Italic">Fragaria ananassa</Emphasis> Duch.), Using Experimental Designs

Vitamin C or total ascorbic acid (TAA) in fruits can be assumed as ascorbic acid (AA) plus dehydroascorbic acid (DHA) content. The aim of this work was to optimize and validate, using experimental designs, a new high-performance liquid chromatographic method for vitamin C determination in strawberries. The mobile phase (MP) consisted of a 0.03 M sodium acetate/acetic acid buffer, 5% methanol. For optimization, a Box–Behnken design was used (three factors at three levels: (a) pH of MP, 3.8–5.8; (b) wavelength, 240–270 nm; and (c) flow rate, 0.5–1.2 ml min⁻¹). Responses were: AA and TAA areas, peak widths, and retention times. A global optimization was performed using the Derringer desirability function, and a value of 0.84 was reached for the combination of design factors: A = 5.8, B = 251 nm, and C = 1.15 ml min⁻¹. Method validation, using AA standard solutions, included: linearity study, limits of detection and quantification, and calibration and analytical sensitivity quantifications. Precision and accuracy were studied in strawberry extracts. The coefficients of variation (percent) were: AA, 1.5%; TAA, 1.8%, and DHA, 4.9%. Accuracy was evaluated with AA standard spiked in 30–150% range of the expected amount of analyte in real samples. The joint confidence elliptical region test and t test were employed for the study of the difference between recoveries (percent) and the ideal 100%. The robustness was analyzed using a fractional factorial design (3⁴⁻²), and an AA recovery study after slight changes in operative variables was performed. The results indicate that the optimized method was linear, sensible, precise, accurate, and robust.     

Achieving the Sustainable Development Goals: improving water services in cities affected by extreme weather events

Abstract

This article discusses how key risks from extreme weather events might affect progress towards meeting Sustainable Development Goals 6 and 11 in cities in developing countries. It outlines the magnitude of the existing shortfall in safe water and sanitation services, and how climate change will exacerbate existing problems. It argues that the performance of many governments thus far has lacked urgency and purpose. Unless governments in particular become more committed, with redoubled effort, the goals are unlikely to be achieved.     

Design and Characterization of Myristoylated and Non-Myristoylated Peptides Effective against Candida spp. Clinical Isolates

The increasing resistance of fungi to antibiotics is a severe challenge in public health, and newly effective drugs are required. Promising potential medications are lipopeptides, linear antimicrobial peptides (AMPs) conjugated to a lipid tail, usually at the N-terminus. In this paper, we investigated the in vitro and in vivo antifungal activity of three short myristoylated and non-myristoylated peptides derived from a mutant of the AMP Chionodracine. We determined their interaction with anionic and zwitterionic membrane-mimicking vesicles and their structure during this interaction. We then investigated their cytotoxic and hemolytic activity against mammalian cells. Lipidated peptides showed a broad spectrum of activity against a relevant panel of pathogen fungi belonging to Candida spp., including the multidrug-resistant C. auris. The antifungal activity was also observed vs. biofilms of C. albicans, C. tropicalis, and C. auris. Finally, a pilot efficacy study was conducted on the in vivo model consisting of Galleria mellonella larvae. Treatment with the most-promising myristoylated peptide was effective in counteracting the infection from C. auris and C. albicans and the death of the larvae. Therefore, this myristoylated peptide is a potential candidate to develop antifungal agents against human fungal pathogens.     

Structural Analysis of the Black-Legged Tick Saliva Protein Salp15

Salp15 is one of the proteins in the saliva of the tick Ixodes scapularis. Together with other biomolecules injected into the mammalian host at the biting site, it helps the tick to sustain its blood meal for days. Salp15 interferes with the cellular immune response of the mammalian host by inhibiting the activation of CD4⁺ T-lymphocytes. This function is co-opted by pathogens that use the tick as a vector and invade the host when the tick bites, such as Borrelia burgdorferi, the causative agent of Lyme borreliosis. Because of the immunity-suppressing role of Salp15, it has been proposed as a candidate for therapeutic applications in disorders of the immune system. The protein is produced as a 135-residue long polypeptide and secreted without its N-terminal signal 1–21 sequence. Detailed structural studies on Salp15 are lacking because of the difficulty in producing large amounts of the folded protein. We report the production of Salp15 and its structural analysis by NMR. The protein is monomeric and contains a flexible N-terminal region followed by a folded domain with mixed α + β secondary structures. Our results are consistent with a three-dimensional structural model derived from AlphaFold, which predicts the formation of three disulfide bridges and a free C-terminal cysteine.     

Crystallization of Honey at −20°C

Honeys from different regions of the province of Buenos Aires were stored at ?20°C, and factors that affect crystallization were analyzed. Crystals were observed by light microscopy. Firmness, adhesivity and viscosity of the samples were measured. Honey was characterized by determining the water activity, turbidity, moisture, fructose, and glucose contents. Results show that the viscous characteristics of the samples depend on the number, size, and disposition of crystals. Various honey samples exhibited Newtonian, pseudoplastic, and thixotropic behaviors. Crystallization was favored at higher moisture contents, suggesting that the parameters that affect honey crystallization at room temperature have a different effect at freezing temperatures. Honey that presented higher values of firmness had a moisture content lower than 17%, and a linear inverse relationship was observed between the adhesivity and firmness of honey samples.     

Linking Contradictions and Laws of Engineering System Evolution within the TRIZ Framework

Tools and methods developed during the era of quality and optimization have shown their limitations and become inappropriate in the context of the requirements of innovation. Nowadays the need to rebuild design practices in enterprises is strongly felt both in terms of human skills and methodological expertise. In part, a way to face the innovation era's difficulties has been provided through the theory of inventive problem solving. This theory represents a significant breakthrough in driving problem statement and solving in a direction that is expressed through the idea that technical systems are driven by objective laws. A second postulate concerns the notion of contradiction, but so far only few contributions have addressed the relations between laws and contradictions. This paper, through a qualitative approach, presents a solution to this limitation and proposes a possible use of laws within the choice of the appropriate conflicting pair, prior to the use of any TRIZ solving techniques. Tests to observe the impact of the proposed approach were conducted in a French engineering ‘grande école’ during three semesters with 180 engineers. The contribution of this paper is twofold. On the one hand, there is a theoretical contribution to the theory of inventive problem solving. In addition, the proposed method offers especially TRIZ practitioners new ways for problem understanding and problem formulation.