One-pot synthesis of alginate-antimicrobial peptide nanogel.

Nanosized alginate-based particles (NAPs) were obtained in a one-pot solvent-free synthesis procedure, achieving the design of a biocompatible nanocarrier for the encapsulation of IbM6 antimicrobial peptide (IbM6). IbM6 is integrated in the nascent nanosized hydrogel self-assembly guided by electrostatic interactions and by weak interactions, typical of soft matter. The formation of the nanogel is a dynamic and complex process, which presents an interesting temporal evolution. In this work, we optimized the synthesis conditions of IbM6-NAPs based on small-angle X-ray scattering (SAXS) measurements and evaluated its time evolution over several weeks by sensing the IbM6 environment in IbM6-NAPs from photochemical experiments. Fluorescence deactivation experiments revealed that the accessibility of different quenchers to the IbM6 peptide embedded in NAPs is dependent on the aging time of the alginate network. Lifetimes measurements indicate that the deactivation paths of the excited state of the IbM6 in the nanoaggregates are reduced when compared with those exhibited by the peptide in aqueous solution, and are also dependent on the aging time of the nanosized alginate network. Finally, the entrapment of IbM6 in NAPs hinders the degradation of the peptide by trypsin, increasing its antimicrobial activity against Escherichia coli K-12 in simulated operation conditions.

Caracterización de embarazos en adolescentes atendidas en el Hospital Regional de Talca: estudio retrospectivo (2017-2022) / Characterization of adolescent pregnancy treated at the Regional Hospital of Talca: retrospective study (2017-2022)

Teenage pregnancy is a health problem that, despite being in decline, continues to be an important component in maternal and fetal morbidity and mortality. The objective of this investigation is to examine the data on adolescent pregnancies obtained from the Regional Hospital of Talca, describe the results collected and compare them with information found in the literature, analyzing the most relevant aspects, in order to provide a greater perspective on the subject, and therefore, contribute to the generation of new prevention measures. In this retrospective and descriptive observational study, the data of all pregnant women under 20 years who gave birth during the period from February 2017 to November 2022 and were treated at the Regional Hospital of Talca were collected and analyzed. For the analysis of the data, a script was made in Python. Obtaining as a result 1,566 pregnant adolescents, with an average age of 17.67 years, 89.6% primigest, in terms of newborns, 13.6% were premature and 5.8% of them extremely premature. 1.2% of newborns (both live births and stillborns/stillborns) developed some malformation.

Immobilization Systems of Antimicrobial Peptide Ib-M1 in Polymeric Nanoparticles Based on Alginate and Chitosan.

The development of new strategies to reduce the use of traditional antibiotics has been a topic of global interest due to the resistance generated by multiresistant microorganisms, including Escherichia coli, as etiological agents of various diseases. Antimicrobial peptides are presented as an alternative for the treatment of infectious diseases caused by this type of microorganism. The Ib-M1 peptide meets the requirements to be used as an antimicrobial compound. However, it is necessary to use strategies that generate protection and resist the conditions encountered in a biological system. Therefore, in this study, we synthesized alginate and chitosan nanoparticles (Alg-Chi NPs) using the ionic gelation technique, which allows for the crosslinking of polymeric chains arranged in nanostructures by intermolecular interactions that can be either covalent or non-covalent. Such interactions can be achieved through the use of crosslinking agents that facilitate this binding. This technique allows for immobilization of the Ib-M1 peptide to form an Ib-M1/Alg-Chi bioconjugate. SEM, DLS, and FT-IR were used to determine the structural features of the nanoparticles. We evaluated the biological activity against E. coli ATCC 25922 and Vero mammalian cells, as well as the stability at various temperatures, pH, and proteases, of Ib-M1 and Ib-M1/Alg-Chi. The results showed agglomerates of nanoparticles with average sizes of 150 nm; an MIC of 12.5 µM, which was maintained in the bioconjugate; and cytotoxicity values close to 40%. Stability was maintained against pH and temperature; in proteases, it was only evidenced against pepsin in Ib-M1/Alg-Chi. The results are promising with respect to the use of Ib-M1 and Ib-M1/Alg-Chi as possible antimicrobial agents.

New PEPTIR-2.0 Peptide Designed for Use as Recognition Element in Electrochemical Biosensors with Improved Specificity towards E. coli O157:H7.

The detection of pathogens through alternative methodologies based on electrochemical biosensors is being studied. These devices exhibit remarkable properties, such as simplicity, specificity, and high sensitivity in monitoring pathogens. However, it is necessary to continue conducting studies that adequately improve these characteristics, especially the recognition molecule. This work aims to design and evaluate a new peptide, named PEPTIR-2.0, as a recognition molecule in electrochemical biosensors to detect E. coli O157:H7 in water. PEPTIR-2.0 was obtained from modifications of the PEPTIR-1.0 peptide sequence, which was previously reported and exhibited excellent properties for detecting and quantifying this pathogenic microorganism. PEPTIR-1.0 is a peptide analogous to the TIR (Translocated Intimin Receptor) protein capable of interacting with the Intimin outer membrane. The basis of this study was to obtain, by using bioinformatics tools, a molecule analogous to PEPTIR-1.0 that maintains its three-dimensional structure but increases the hydrophobic interactions between it and Intimin, since these intermolecular forces are the predominant ones. The designed PEPTIR-2.0 peptide was immobilized on screen-printed electrodes modified with gold nanoparticles. The detection capacity of E. coli O157:H7 in water was evaluated using electrochemical impedance spectroscopy in the presence of other microorganisms, such as P. aeruginosa, S. aureus, and non-pathogenic E. coli. The results showed that PEPTIR-2.0 confers remarkable specificity to the biosensor towards detecting E. coli, even higher than PEPTIR-1.0.

A Bioinspired Peptide in TIR Protein as Recognition Molecule on Electrochemical Biosensors for the Detection of E. coli O157:H7 in an Aqueous Matrix.

Currently, the detection of pathogens such as Escherichia coli through instrumental alternatives with fast response and excellent sensitivity and selectivity are being studied. Biosensors are systems consisting of nanomaterials and biomolecules that exhibit remarkable properties such as simplicity, portable, affordable, user­friendly, and deliverable to end­users. For this, in this work we report for the first time, to our knowledge, the bioinformatic design of a new peptide based on TIR protein, a receptor of Intimin membrane protein which is characteristic of E. coli. This peptide (named PEPTIR­1.0) was used as recognition element in a biosensor based on AuNPs­modified screen­printed electrodes for the detection of E. coli. The morphological and electrochemical characteristics of the biosensor obtained were studied. Results show that the biosensor can detect the bacteria with limits of detection and quantification of 2 and 6 CFU/mL, respectively. Moreover, the selectivity of the system is statistically significant towards the detection of the pathogen in the presence of other microorganisms such as P. aeruginosa and S. aureus. This makes this new PEPTIR­1.0 based biosensor can be used in the rapid, sensitive, and selective detection of E. coli in aqueous matrices.

Cambios en el perfil protéico de E. coli O157:H7 frente al tratamiento con Ib-M1 e IONP@Ib-M1 / Changes in the protein profile of E. coli O157: H7 against treatment with Ib-M1 and IONP@Ib-M1 / Identificação de alterações no perfil protetor de E. coli O157: H7 contra o tratamento com Ib-M1 e IONP@Ib-M1

Resumen Escherichia coli 0157:H7 es una bacteria patógena reconocida por su capacidad de resistencia a diversos antibióticos; razón por la cual, se generan complicaciones en el tratamiento de infecciones producidas por esta bacteria. El péptido Ib-M1 y el bioconjugado I0NP@Ib-M1 han surgido como una nueva alternativa antimicrobiana contra E. coli 0157:H7. El mecanismo de acción de Ib-Mi e I0NP@Ib-M1 contra esta bacteria aún es desconocido; por lo tanto, el objetivo de esta investigación fue identificar el cambio en el perfil de proteínas de E. coli 0157:H7 luego del tratamiento con Ib-M1 e I0NP@ Ib-M1 como primer paso para determinar su mecanismo de acción. Para esto, se llevó a cabo la obtención de proteínas, posteriormente se realizó una electroforesis bidimensional para finalmente realizar la determinación de la variabilidad de los perfiles proteicos. Una vez obtenidos estos perfiles, se llevó a cabo un análisis de varianza (AN0VA). Se identificaron 72 proteínas expresadas diferencialmente, las cuales pueden relacionarse con el efecto sobre el crecimiento de la bacteria en presencia de Ib-M1 e I0NP@Ib-M. Estas proteínas se encuentran involucradas en procesos de transferencia de grupos acilo (proteína Yhbs), translocación de lipoproteínas (proteína LolA) y transporte de aminoácidos (proteína GpmA), entre otros.

Abstract Escherichia coli 0157: H7 is a pathogenic bacterium which is recognized for the ability to resist multiple antibiotics; accordingly, complications occur in the treatment of infections caused by this bacterium. The Ib-M1 peptide and the I0NP @ Ib-M1 bioconjugate have emerged as a new antimicrobial alternatives against E. coli 0157: H7. The mechanism of action of Ib-M1 and I0NP @ Ib-M1 against this bacterium is still unknown; therefore, the goal of this research was to identify the change in the proteins profile of E. coli 0157: H7 after treatment with Ib-M1 and I0NP @ Ib-M1 as a first step to determine its mechanism of action. For this, the proteins were obtained first, and then a two-dimensional electrophoresis was performed to finally determine the variability of the protein profiles. 0nce the protein profiles were obtained, an analysis of variance (AN0VA) was carried out. 72 differentially expressed proteins were identified, which can be connected to the effect on the bacterium's growth in the presence of Ib-M1 and I0NP @ Ib-M. These proteins are involved in acyl groups transfer processes (Yhbs protein), lipoprotein translocation (LolA protein) and amino acid transport (GpmA protein), among others.

Resumo Escherichia coli O157: H7 é uma bactéria patogênica reconhecida por sua capacidade de resistir a vários antibióticos; razão pela qual, complicações são geradas no tratamento de infecções produzidas por essa bactéria. O peptídeo Ib-M1 livre e imobilizado em nanopartículas magnéticas de óxido de ferro (IONP @ Ib-M1) surgiu como uma nova alternativa antimicrobiana contra E. coli O157: H7 e isolados clínicos desta bactéria. O mecanismo de ação de Ib-M1 e IONP @ Ib-M1 contra E. coli O157: H7 ainda é desconhecido; Portanto, o objetivo desta pesquisa foi identificar a alteração no perfil proteico de E. coli O157: H7 após o tratamento com Ib-M1 e IONP @ Ib-M1 como um primeiro passo para determinar seu mecanismo de ação. Para isso, foi realizada a obtenção das proteínas, posteriormente foi realizada uma eletroforese bidimensional para finalmente determinar a variabilidade dos perfis protéicos. Uma vez obtidos os perfis de proteínas, foi realizada uma análise de variância (ANOVA). Os resultados mostram a identificação de proteínas expressas diferencialmente e que estão envolvidas em processos de transferência de grupos acila (proteína Yhbs), translocação de lipoproteínas (proteína LolA) e transporte de aminoácidos (proteína GpmA), entre outros.

Antimicrobial activity of Ib-M peptides against Escherichia coli O157: H7.

The development of new antimicrobial peptides has become an attractive alternative to conventional antibiotics due to the increasing rates of microbial drug resistance. Ib-M corresponds to a family of cationic synthetic peptides, 20 amino acids in length, that have shown inhibitory effect against the non-pathogenic strain Escherichia coli K-12. This work evaluated the antimicrobial potential of Ib-M peptides against the pathogenic E. coli O157: H7 using a reference strain and a clinical isolate. The Ib-M peptides showed antibacterial activity against both strains of E. coli O157: H7; the minimum inhibitory concentration of Ib-M peptides ranged from 1.6 to 12.5 µM and the minimum bactericidal concentration ranged from 3.7 to 22.9 µM, being Ib-M1 and Ib-M2 the peptides that presented the highest inhibitory effect. Time-kill kinetics assay showed a reduction of the bacterial population by more than 95% after 4 hours of exposure to 1xMIC of Ib-M1. Low cytotoxicity was observed in VERO cells with 50% cytotoxic concentration in the range from 197.5 to more than 400 µM. All peptides showed a random structure in hydrophilic environments, except Ib-M1, and all of them transitioned to an α-helical structure when the hydrophobicity of the medium was increased. In conclusion, these findings support the in vitro antimicrobial effect of Ib-M peptides against the pathogenic bacteria E. coli O157: H7 and prove to be promising molecules for the development of new therapeutic alternatives.

Highly Structured Polyvinyl Alcohol Porous Carriers: Tuning Inherent Stability and Release Kinetics in Water.

Polyvinyl alcohol (PVA) porous carriers were prepared by means of ice templating of aqueous solutions containing of 90 kD and/or 16 kD PVA. The carriers were loaded with traces of a colored probe (methyl orange) to screen their release properties, once immersed in water. The carriers prepared from solutions containing 90 kD and 16 kD PVA resulted in intimate polymer mixtures, exhibiting physical properties that stand in between those of the bare 90 kD or 16 kD PVA end members. The freezing protocols employed were adapted to prepare carriers textured in the form of either monolithic scaffolds (directional constant freezing rate) or millimetric pellets (flash-freeze). Monolithic carriers remain stable in aqueous solution, and the probe release is governed by a swelling-diffusion mechanism. The kinetics of probe release can be tuned from minutes to hours by either increasing the total PVA content or the 90 kD-to-16 kD PVA ratio in the parent solution. In contrast, pellets (millimetric carriers) immersed in water release the probe on the scale of minutes, irrespective of the PVA content or composition. However, the PVA content and the 90 kD-to-16 kD PVA ratio dramatically affect the stability of the carriers. Depending on the formulation, these small carriers can develop swelling, erosion, or eventually massive dissolution.

Carcinoma basoescamoso na cidade de Loja, Equador / Basosquamous carcinoma in the city of Loja, Equador

Introdução: O Equador encontra-se na latitude 0 grau, 0’0’’, posição terrestre exposta a maior quantidade de radiação solar do planeta, especialmente aos raios UVA e UVB que predispõe a dano solar e aparecimento de câncer de pele. Objetivos: Nesse contexto, decidimos descrever a presença de carcinoma basoescamoso em uma cidade ao sul do Equador. Casuística e Métodos: Realizamos uma revisão de prontuários dos pacientes com diagnóstico de carcinoma basocelular da região cérvico-facial submetidos a cirurgia como tratamento, entre 2009 e 2013 no único hospital oncológico da cidade. Resultados: Encontramos 186 carcinomas basocelulares e 17 desses tumores eram basoescamosos. A maior porcentagem desse tipo de tumor afetaram principalmente mulheres; a idade média foi 54,9 anos. A raça branca foi mais afetada (58,8%), com elevada taxa relativa de não-brancos (41,2%). Nariz, lábio e couro cabeludo foram as áreas mais acometidas. Dez casos apareceram em zona de alto risco (58,8%). A invasão perineural e angiolinfática esteve presente em mais da metade dos casos. Nossos resultados mostram uma frequência superior de carcinomas basoescamosos entre os carcinomas basocelulares em relação a maior parte das publicações internacionais. As razões para tal achado são multifatoriais e podem incluir viés de seleção, distribuição étnica diferenciada da população equatoriana e maior incidência de radiação solar nesta região.

Introduction: Equador is at 0, 0’0’ degrees latitude, therefore is exposed to a largest amount of solar radiation the planet, especially to UVA and UVB rays what predisposes to cell damage and the onset of skin cancer. Objectives: In this context, we decide to describe the presence of the basosquamous carcinoma in one city at the south of Equador. Casuistics and Methods: We realize a review from medical charts of every patient with the diagnosis of cervicofacial basal cell carcinoma and that went to surgery for treatment, during the period from 2009 and 2013 at the only oncological hospital in the city. Results: We found 186 basal cell carcinomas and 17 of these tumors were basosquamous carcinomas. The highest percentage of this kind of tumor affected mostly women, the mean age was 54,9 years. The white people was the most affected (58,8%), with a relative high rate of nonwhites (41,2%). The nose, lips and scalp were the areas that arise this tumor. Ten of the cases appear in the high risk zone (58,8%). Characteristics such as perineural and angiolymphatic were present in more than half of the patients.