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The EPI VITRAKVI study is a retrospective study designed to place the results of the single-arm Phase I/II larotrectinib SCOUT trial into context by comparison with external historical controls. Its primary objective is to compare the time to medical treatment failure between larotrectinib and the historical standard of care (chemotherapy) in patients with infantile fibrosarcoma. External historical cohorts have been selected by using objective criteria. The Inverse Probability of Treatment Weighting method will be used to adjust for potential confounding. The current publication illustrates how an external control arm study can complement data from a single-arm trial and addresses uncertainties encountered in the assessment of therapies targeting rare abnormalities where randomized controlled trials are considered not feasible. Clinical Trial Registration: NCT05236257 (ClinicalTrials.gov) Infantile fibrosarcoma (IFS) is a rare type of childhood cancer that commonly affects the legs and arms. In IFS cancers, the units which carry the information that determines your traits (genes), typically have specific changes which leads to the creation of an altered fusion protein, a protein which is created by joining parts of two different genes. This altered fusion protein can cause cancer cells to survive and to grow. Larotrectinib works by blocking the altered fusion protein and is already available in Europe and in many other countries. It is approved for prescription to patients with the altered fusion protein, whose cancer has spread to nearby tissues and/or lymph nodes or to other parts of the body. Since IFSs a rare disease, previous studies did not compare larotrectinib with the standard of care, which is chemotherapy. The main purpose of our study is to collect more results on how well larotrectinib works compared with chemotherapy taken from real world evidence data. The present publication explains how such a comparison can be made and how such a study can help in the assessment of treatments that target rare diseases. [ABSTRACT FROM AUTHOR]

In this work, we present the in-silico design of a new aptamer (named Apt917) capable of interacting with E. coli and its evaluation as a recognition element in electrochemical biosensors. A search and bioinformatic analysis of various aptamers reported in the literature was carried out. Parameters such as low dissociation constant and short sequences were considered to design and model new hybrid sequences. The designed Apt917 has a minimum free energy and high percent to frequency, making it promising compared to the initial ones. Apt917 was synthesized and immobilized on gold nanoparticles-modified screen-printed electrodes. The evaluation of the obtained biosensor shows a promising detection of E. coli O157:H7 with limits of detection and quantification of 8 and 27 CFU/mL, respectively. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Copyright of Revista Latinoamericana de Hipertension is the property of Revista Latinoamericana de Hipertension and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Latinoamericana de Hipertension is the property of Revista Latinoamericana de Hipertension and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Latinoamericana de Hipertension is the property of Revista Latinoamericana de Hipertension and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Latinoamericana de Hipertension is the property of Revista Latinoamericana de Hipertension and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Latinoamericana de Hipertension is the property of Revista Latinoamericana de Hipertension and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Salmonella Typhimurium is a causative agent of nontyphoidal salmonellosis, for which there is a lack of a clinically approved vaccine in humans. As an intracellular pathogen, Salmonella impacts many cellular pathways. However, the intercellular communication mechanism facilitated by host-derived small extracellular vesicles (EVs), such as exosomes, is an overlooked aspect of the host responses to this infection. We used a comprehensive proteome-based network analysis of exosomes derived from Salmonella-infected macrophages to identify host molecules that are trafficked via these EVs. This analysis predicted that the host-derived small EVs generated during macrophage infection stimulate macrophages and promote activation of T helper 1 (Th1) cells. We identified that exosomes generated during infection contain Salmonella proteins, including unique antigens previously shown to stimulate protective immune responses against Salmonella in murine studies. Furthermore, we showed that host EVs formed upon infection stimulate a mucosal immune response against Salmonella infection when delivered intranasally to BALB/c mice, a route of antigen administration known to initiate mucosal immunity. Specifically, the administration of these vesicles to animals stimulated the production of anti-Salmonella IgG antibodies, such as anti-OmpA antibodies. Exosomes also stimulated antigen-specific cell-mediated immunity. In particular, splenic mononuclear cells isolated from mice administered with exosomes derived from Salmonella-infected antigen-presenting cells increased CD4+ T cells secreting Th1-type cytokines in response to Salmonella antigens. These results demonstrate that small EVs, formed during infection, contribute to Th1 cell bias in the anti-Salmonella responses. Collectively, this study helps to unravel the role of host-derived small EVs as vehicles transmitting antigens to induce Th1-type immunity against Gram-negative bacteria. Understanding the EV-mediated defense mechanisms will allow the development of future approaches to combat bacterial infections. Author summary: Salmonella Typhimurium is a causative agent of nontyphoidal salmonellosis, and we still lack a clinically approved vaccine against this infection. The design of efficacious vaccines against Salmonella can benefit from studying overlooked aspects of host immune responses to this pathogen. The cell-to-cell communication facilitated by nano-sized extracellular vesicles (EVs) can affect the immune response. Our earlier work showed that Salmonella infection leads to the generation of immunogenic small EVs by macrophages. Here, we performed a proteomic and metabolomic analysis of the EV cargo, which indicated that EVs generated by Salmonella-infected macrophages stimulate macrophages and promote T-helper 1 (Th1) cell activation. We confirmed in vivo that mucosal administration of these EVs promotes a mucosal immune response against Salmonella infection in BALB/c mice. EVs from infected macrophages increased the CD4+ T cells secreting IL-2, IFN-γ, and TNF-α in response to Salmonella antigens, contributing to Th1 cell bias. These EVs not only stimulated antigen-specific cell-mediated immunity, but also contributed to anti-Salmonella antibody production, which included IgGs recognizing bacterial proteins present in EVs. Our study unraveled a novel role of EVs as vehicles transmitting antigens to induce Th1-type immunity against Gram-negative bacteria, thus supporting the future development of preventative approaches to combat Salmonella infection. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Purpose: Hereditary Breast and Ovarian Cancer (HBOC) syndrome is responsible for ~5–10% of all diagnosed breast and ovarian cancers. Breast cancer is the most common malignancy and the leading cause of cancer-related mortality among women in Latin America (LA). The main objective of this study was to develop a comprehensive understanding of the genomic epidemiology of HBOC throughout the establishment of The Latin American consortium for HBOC-LACAM, consisting of specialists from 5 countries in LA and the description of the genomic results from the first phase of the study. Methods: We have recruited 403 individuals that fulfilled the criteria for HBOC from 11 health institutions of Argentina, Colombia, Guatemala, Mexico and Peru. A pilot cohort of 222 individuals was analyzed by NGS gene panels. One hundred forty-three genes were selected on the basis of their putative role in susceptibility to different hereditary cancers. Libraries were sequenced in MiSeq (Illumina, Inc.) and PGM (Ion Torrent-Thermo Fisher Scientific) platforms. Results: The overall prevalence of pathogenic variants was 17% (38/222); the distribution spanned 14 genes and varied by country. The highest relative prevalence of pathogenic variants was found in patients from Argentina (25%, 14/57), followed by Mexico (18%, 12/68), Guatemala (16%, 3/19), and Colombia (13%, 10/78). Pathogenic variants were found in BRCA1 (20%) and BRCA2 (29%) genes. Pathogenic variants were found in other 12 genes, including high and moderate risk genes such as MSH2, MSH6, MUTYH , and PALB2. Additional pathogenic variants were found in HBOC unrelated genes such as DCLRE1C, WRN, PDE11A , and PDGFB. Conclusion: In this first phase of the project, we recruited 403 individuals and evaluated the germline genetic alterations in an initial cohort of 222 patients among 4 countries. Our data show for the first time in LA the distribution of pathogenic variants in a broad set of cancer susceptibility genes in HBOC. Even though we used extended gene panels, there was still a high proportion of patients without any detectable pathogenic variant, which emphasizes the larger, unexplored genetic nature of the disease in these populations. [ABSTRACT FROM AUTHOR]

The role of gamma-aminobutyric acid (GABA) in adrenal medulla chromaffin cell (CC) function is just beginning to unfold. GABA is stored in catecholamine (CA)-containing dense core granules and is presumably released together with CA, ATP, and opioids in response to physiological stimuli, playing an autocrine-paracrine role on CCs. The reported paradoxical 'dual action' of GABA-R activation (enhancement of CA secretion and inhibition of synaptically evoked CA release) is only one aspect of GABA's multifaceted actions. In this review, we discuss recent physiological experiments on rat CCs in situ which suggest that GABA regulation of CC function may depend on the physiological context: During non-stressful conditions, GABA-R activation by endogenous GABA tonically inhibits acetylcholine release from splanchnic nerve terminals and decreases spontaneous Ca fluctuations in CCs, preventing unwanted CA secretion. During intense stress, splanchnic nerve terminals release acetylcholine, which depolarizes CCs and allows the Ca influx that triggers the release of CA and GABA. With time, CA secretion declines, due to voltage-independent inhibition of Cachannels and desensitization of cholinergic nicotinic receptors. Nonetheless, acute activation of GABA-R is depolarizing in about 50% of CCs, and thus GABA, acting as an autocrine/paracrine mediator, could help to maintain CA exocytosis under stress. GABA-R activation is not excitatory in about half of CCs' population because it hyperpolarizes them or elicits no response. This percentage possibly varies, depending on functional demands, since GABA-R-mediated actions are determined by the intracellular chloride concentration ([Cl]) and therefore on the activity of cation-chloride co transporters, which is functionally regulated. These findings underscore a potential importance of a novel and complex GABA-mediated regulation of CC function and of CA secretion. [ABSTRACT FROM AUTHOR]

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, user‑friendly, and deliverable to end‑users. 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 PEPTIR‑1.0) was used as recognition element in a biosensor based on AuNPs‑modified screen‑printed 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 PEPTIR‑1.0 based biosensor can be used in the rapid, sensitive, and selective detection of E. coli in aqueous matrices. [ABSTRACT FROM AUTHOR]

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.

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., Competing Interests: The authors declare no competing financial interest.