The power of computational proteomics platforms to decipher protein-protein interactions.

Adopting computational tools for analyzing extensive biological datasets has profoundly transformed our understanding and interpretation of biological phenomena. Innovative platforms have emerged, providing automated analysis to unravel essential insights about proteins and the complexities of their interactions. These computational advancements align with traditional studies, which employ experimental techniques to discern and quantify physical and functional protein-protein interactions (PPIs). Among these techniques, tandem mass spectrometry is notably recognized for its precision and sensitivity in identifying PPIs. These approaches might serve as important information enabling the identification of PPIs with potential pharmacological significance. This review aims to convey our experience using computational tools for detecting PPI networks and offer an analysis of platforms that facilitate predictions derived from experimental data.

Anti-bacterial, anti-biofilm and synergistic effects of phenazine-based ruthenium(II) complexes.

Antimicrobial resistance has become a global threat to human health, which is coupled with the lack of novel drugs. Metallocompounds have emerged as promising diverse scaffolds for the development of new antibiotics. Herein, we prepared some metal compounds mainly focusing on cis-[Ru(bpy)(dppz)(SO3)(NO)](PF6) (PR02, bpy = 2,2'-bipyridine, dppz = dipyrido[3,2-a:2',3'-c]phenazine), in which phenazinic and nitric oxide ligands along with sulfite conferred some key properties. This compound exhibited a redox potential for bound NO+/0 of -0.252 V (vs. Ag|AgCl) and a high pH for nitrosyl-nitro conversion of 9.16, making the nitrosyl ligand the major species. These compounds were still able to bind to DNA structures. Interestingly, reduced glutathione (GSH) was unable to promote significant NO/HNO release, an uncommon feature of many similar systems. However, this reducing agent was essential to generate superoxide radicals. Antimicrobial studies were carried out using six bacterial strains, where none or very low activity was observed for Gram-negative bacteria. However, PR02 and PR (cis-[Ru(bpy)(dppz)Cl2]) showed high antibacterial activity in some Gram-positive strains (MBC for S. aureus up to 4.9 µmol L-1), where the activity of PR02 was similar to or at least 4-fold better than that of PR. Besides, PR02 showed capacity to inhibit bacterial biofilm formation, a major health issue leading to bacterial tolerance to antibiotics. Interestingly, we also showed that PR02 can function in synergism with the known antibiotic ampicillin, improving their action up to 4-fold even against resistant strains. Altogether, these results showed that PR02 is a promising antimicrobial nitrosyl ruthenium compound combining features beyond its killing action, which deserves further biological studies.

Promising Dual Anticancer and Antimetastatic Action by a Cu(II) Complex Derived from Acylhydrazone on Human Osteosarcoma Models.

Osteosarcoma cancers are becoming more common in children and young adults, and existing treatments have low efficacy and a very high mortality rate, making it pressing to search for new chemotherapies with high efficacy and high selectivity index. Copper complexes have shown promise in the treatment of osteosarcoma. Here, we report the synthesis, characterization, and anticancer activity of [Cu(N-N-Fur)(NO3)(H2O)] complex where N-N-Fur is (E)-N'-(2-hydroxy-3-methoxybenzylidene)furan-2-carbohydrazide. The [Cu(N-N-Fur)(NO3)(H2O)] complex was characterized via X-ray diffraction and electron spin resonance (ESR), displaying a copper center in a nearly squared pyramid environment with the nitrate ligand acting as a fifth ligand in the coordination sphere. We observed that [Cu(N-N-Fur)(NO3)(H2O)] binds to DNA in an intercalative manner. Anticancer activity on the MG-63 cell line was evaluated in osteosarcoma monolayer (IC50 2D: 1.1 ± 0.1 µM) and spheroids (IC50 3D: 16.3 ± 3.1 µM). Selectivity assays using nontumoral fibroblast (L929 cell line) showed that [Cu(N-N-Fur)(NO3)(H2O)] has selectivity index value of 2.3 compared to cis-diamminedichloroplatinum(II) (CDDP) (SI = 0.3). Additionally, flow cytometry studies demonstrated that [Cu(N-N-Fur)(NO3)(H2O)] inhibits cell proliferation and conveys cells to apoptosis. Cell viability studies of MG-63 spheroids (IC50 = 16.3 ± 3.1 µM) showed that its IC50 value is 4 times lower than for CDDP (IC50 = 65 ± 6 µM). Besides, we found that cell death events mainly occurred in the center region of the spheroids, indicating efficient transport to the microtumor. Lastly, the complex showed dose-dependent reductions in spheroid cell migration from 7.5 to 20 µM, indicating both anticancer and antimetastatic effects.

Reactivity of a nitrosyl ruthenium complex and its potential impact on the fate of DNA - An in vitro investigation.

The role of metal complexes on facing DNA has been a topic of major interest. However, metallonitrosyl compounds have been poorly investigated regarding their reactivities and interaction with DNA. A nitrosyl compound, cis-[Ru(bpy)2(SO3)(NO)](PF6)(A), showed a variety of promising biological activities catching our attention. Here, we carried out a series of studies involving the interaction and damage of DNA mediated by the metal complex A and its final product after NO release, cis-[Ru(bpy)2(SO3)(H2O](B). The fate of DNA with these metal complexes was investigated upon light or chemical stimuli using electrophoresis, electronic absorption spectroscopy, circular dichroism, size-exclusion resin, mass spectrometry, electron spin resonance (ESR) and viscometry. Since many biological disorders involve the production of oxidizing species, it is important to evaluate the reactivity of these compounds under such conditions as well. Indeed, the metal complex B exhibited important reactivity with H2O2 enabling DNA degradation, with detection of an unusual oxygenated intermediate. ESR spectroscopy detected mainly the DMPO-OOH adduct, which only emerges if H2O2 and O2 are present together. This result indicated HOO• as a key radical likely involved in DNA damage as supported by agarose gel electrophoresis. Notably, the nitrosyl ruthenium complex did not show evidence of direct DNA damage. However, its aqua product should be carefully considered as potentially harmful to DNA deserving further in vivo studies to better address any genotoxicity.

Bacteria from gut microbiota associated with diarrheal infections in children promote virulence of Shiga toxin-producing and enteroaggregative Escherichia coli pathotypes.

Background: Diarrheagenic E. coli (DEC) pathogenicity relies on the interaction of bacteria with the host's gut environment, which is regulated by the resident microbiota. Previously, we identified indicative bacterial species of gut microbiota in DEC-positive stool samples from children. Here, we evaluated the role of two indicative species, Citrobacter werkmanii (CW) and Escherichia albertii (EA), in the virulence of two DEC pathotypes, Shiga toxin-producing (STEC) and enteroaggregative (EAEC) Escherichia coli. Methods: We determined the effect of supernatants obtained from CW and EA cultures on the gene expression of STEC strain 86-24 and EAEC strain 042 by RNA-seq analysis. We evaluated IL-8 secretion from T84 cells infected with these DEC strains in the presence or absence of the supernatant from EA. The effect of the supernatant from EA on the growth and adherence of STEC and EAEC to cells was also evaluated. Finally, we studied the effect of the EA supernatant on the STEC-induced inflammation mediated by the long polar fimbriae (Lpf) in T84 cells and the expression of plasmid-encoded toxin (Pet) in EAEC. Results: RNA-seq analysis revealed that several virulence factors in STEC and EAEC were upregulated in the presence of supernatants from CW and EA. Interestingly, an increase in the secretion of IL-8 was observed in cells infected with STEC or EAEC in the presence of a supernatant from EA. Similar results were observed with the supernatants obtained from clinical strains of E. albertii. The supernatant from EA had no effect on the growth of STEC and EAEC, or on the ability of these DEC strains to adhere to cells. We found that Pet toxin in EAEC was upregulated in the presence of a supernatant from EA. In STEC, using mutant strains for Lpf fimbriae, our data suggested that these fimbriae might be participating in the increase in IL-8 induced by STEC in cells in the presence of a supernatant from EA. Conclusion: Supernatant obtained from an indicative species of DEC-positive diarrhea could modulate gene expression in STEC and EAEC, and IL-8 secretion induced by these bacteria. These data provide new insights into the effect of gut microbiota species in the pathogenicity of STEC and EAEC.

Synergy of DNA intercalation and catalytic activity of a copper complex towards improved polymerase inhibition and cancer cell cytotoxicity.

Improving the binding of metal complexes to DNA to boost cancer cell cytotoxicity requires fine tuning of their structural and chemical properties. Copper has been used as a metal center in compounds containing intercalating ligands due to its ability to catalytically generate reactive oxygen species (ROS), such as hydroxyl radicals (OH˙). We envision the synergy of DNA binding and ROS generation in proximity to target DNA as a powerful chemotherapy treatment. Here, we explore the use of [Cu(2CP-Bz-SMe)]2+ (2CP-Bz-SMe = 1,3-bis(1,10-phenanthrolin-2-yloxy)-N-(4-(methylthio)benzylidene)propan-2-amine) for this purpose by characterizing its cytotoxicity, DNA binding, and ability to affect DNA replication through the polymerase chain reaction - PCR and nuclease assays. We determined the binding (Kb) and Stern-Volmer constants (KSV) for complex-DNA association of 5.8 ± 0.14 × 104 and 1.64 (±0.08), respectively, through absorption titration and competitive fluorescence experiments. These values were superior to those of other Cu-complex intercalators. We hypothesize that the distorted trigonal bipyramidal geometry of [Cu(2CP-Bz-SMe)]2+ allows the phenanthroline fragments to be better accommodated into the DNA double helix. Moreover, the aromaticity of these fragments increases the local hydrophobicity thus increasing the affinity for the hydrophobic domains of DNA. Nuclease assays in the presence of common reducing agents ascorbic acid, nicotinamide adenine dinucleotide, and glutathione showed the effective degradation of DNA due to the in situ generation of OH˙. The [Cu(2CP-Bz-SMe)]2+ complex showed cytotoxicity against the following human cancer cells lines A549, MCF-7, MDA-MB-231 and MG-63 with half maximal inhibitory concentration (IC50) values of 4.62 ± 0.48, 5.20 ± 0.76, 5.70 ± 0.42 and 2.88 ± 0.66 µM, respectively. These low values of IC50, which are promising if compared to that of cisplatin, are ascribed to the synergistic effect of ROS generation with the intercalation ability into the DNA minor grooves and blocking DNA replication. This study introduces new principles for synergizing the chemical and structural properties of intercalation compounds for improved drug-DNA interactions targeting cancer.

Comparación del confort de dos protectores bucales en jugadores de fútbol americano / Comparing the comfort of two mouth protectors in football players

Los deportes de contacto aumentan el riesgo de traumatismo causando alteraciones funcionales, estéticas y psicológicas. Los protectores bucales son parte del equipo de seguridad que protegen al deportista de estos accidentes, es importante que brinde confort para lograr el mejor rendimiento físico. Objetivo: Comparar el confort de los jugadores de fútbol americano con el uso de protectores prefabricados y protectores personalizados. Metodología: Se realizó un estudio cuasiexperimental, analítico y prospectivo donde se incluyeron 15 jugadores de futbol americano de la Facultad de Estudios Superiores Acatlán, UNAM, que firmaron consentimiento informado. La autora principal de este proyecto fabricó los protectores bucales superiores con hoja de copolímero de etilvinilacetato (EVA) #35. El protector prefabricado fue de silicón. Los jugadores realizaron dos prácticas de entrenamientos donde hicieron 30 minutos de carrera, 15 de estiramiento, 60 de fuerza y destreza en donde requerían de estar en constante comunicación verbal y 30 de cardiovascular en gimnasio (usaron un protector en cada entrenamiento). Se aplicó un cuestionario ­previamente validado por consenso de expertos­ para evaluar el confort en la respiración, lenguaje, náuseas, adaptación, comodidad e hidratación bucal con cada tipo de protector. La información se analizó en el programa SPSS v19.0; se usaron proporciones para las variables cualitativas y medidas de tendencia central y dispersión para las cuantitativas. Para comparar el confort con el uso de ambos protectores se empleó la prueba χ2, con un nivel de significancia p < 0.05. Resultados: La media de edad fue de 20 ± 2 años. Con el protector personalizado 11 jugadores (0.73) refirieron que respiraron y hablaron mejor, sintieron menos náusea 12 (0.80), mejor adaptación 11 (0.73) y mayor comodidad 12 (0.80) que, con el uso de protector bucal prefabricado, encontrando diferencias estadísticamente significativas (p < 0.05) y ocho (0.53) refirió menos hidratación bucal (p > 0.05). Doce (0.80) de los jugadores mencionaron tener mayor confort con el protector personalizado. Conclusiones: Con la metodología empleada y en la muestra estudiada, el uso de un protector bucal personalizado fabricado con hoja de copolímero de etilvinilacetato (EVA) #35 ofrece una mejor adaptación y comodidad, después de dos prácticas de entrenamiento (AU)

The contact sports, increase the risk of trauma causing functional, aesthetic and psychological alterations, mouth guards are part of the safety equipment that protects the athlete from this kind of accidents, it is also important that the mouth guard had to provides comfort, for achieve a best physical performance. Objective: Make the comparison with the comfort of American football players with the use of prefabricated protectors and customized protectors. Method: A quasi-experimental, analytical and prospective study was carried out, including 15 American football players from the Facultad de Estudios Superiores Acatlán, UNAM, who signed an informed consent. The lead author of this project manufactured the upper mouth guards with ethylvinylacetate (EVA) #35 copolymer foil. The prefabricated protector was made of silicone. The players performed two practice sessions where they did 30 minutes of running, 15 of stretching, 60 of strength and skill where they required being in constant verbal communication and 30 of cardiovascular in a gymnasium (they used a protector in each training). A questionnaire ­previously validated by expert consensus­ was applied to evaluate comfort in breathing, language, nausea, adaptation, comfort and oral hydration with each type of protector. The information was analyzed in the program SPSS v19.0; proportions were used for the qualitative variables and measures of central tendency and dispersion for the quantitative ones. To compare the comfort with the use of both protectors the test χ2 was used, with a level of signifi cance p < 0.05. Results: The mean age was 20 ± 2 years. With the personalized protector, 11 players (0.73) reported that they breathed and spoke better, feeling less nausea 12 (0.80), better adaptation 11 (0.73) and greater comfort 12 (0.80) than with the use of prefabricated mouth guards, fi nding statistically signifi cant diff erences (p < 0.05) and 8 (0.53) reported less oral hydration (p > 0.05). 12 (0.80) of players mentioned having more comfort with the custom protector. Conclusions: The use of a personalized mouth guard off ers better fi t and comfort made with sheet copolymer of ethylvinylacetate, after two training practices (AU)

La monitorización neuromuscular y su importancia en el uso de los bloqueantes neuromusculares / Neuromuscular monitoring and its importance in neuromuscular blockade

Introducción: La necesidad por incorporar nuevas guías o estrategias en la buena práctica de uso de los bloqueantes neuromusculares no es un hecho de obligado cumplimento en la actualidad dentro de la anestesiología. Solo existen recomendaciones de actuación con el propósito de convencer que la monitorización neuromuscular es una herramienta muy útil para el buen uso racional de los bloqueantes neuromusculares. Metodología: Las complicaciones surgen, y la parálisis residual es un evento destacado. Por esta razón, los autores propugnamos que la monitorización del bloqueo neuromuscular puede ser un factor determinante en la mejora del cuidado de nuestros pacientes, disminuyendo tanto la morbilidad como la mortalidad. Esta revisión y su metodología en base a la experiencia de los autores solo pretende exponer de forma sencilla conocimientos que consideramos básicos para su utilización sistemática en nuestra práctica rutinaria. Resultados y conclusiones: Esta actualización describe los principios fundamentales de los métodos que disponemos en la actualidad, priorizando las medidas cuantitativas de registro. Y también demuestra el diferente comportamiento de la musculatura al efecto de los bloqueantes neuromusculares, fundamentos relevantes que es preciso conocer. La monitorización neuromuscular es una práctica que debe utilizarse siempre que un bloqueo neuromuscular sea necesario. Somos conscientes que generar una recomendación explícita es difícil. Pero nuestro entusiasmo parte del beneficio de una experiencia personal con estos métodos que son conocidos desde antiguo. Debido a la potencial morbilidad asociada con bloqueos neuromusculares residuales, la monitorización perioperatoria de la función neuromuscular es esencial.

Introduction: The incorporation of new guidelines or strategies as part of good practices in the use of muscle relaxants is not a requirement at present in the practice of anesthesia. There are only action recommendations designed to persuade clinicians of the fact that neuromuscular monitoring is a very useful tool for the rational use of muscle relaxants. Methodology:Complications occur, and residual paralysis is a significant event. For this reason, the authors advocate that monitoring neuromuscular block may be a determining factor in improving patient care and reducing morbidity and mortality. This review and its methodology based on the experience of the authors is designed to present, in a simple format, the knowledge that considered fundamental for the systematic use of neuromuscular monitoring in every day practice. Results and conclusions: This update describes the fundamental principles of the methods available at present, emphasizing quantitative recording measurements. It then describes the different ways in which muscles respond to the effect of neuromuscular blockade, as these are critical fundamental principles that have to be known. Neuromuscular monitoring is a practice that should be implemented every time a neuromuscular block is required. We are aware of the difficulty of generating an explicit recommendation, but our enthusiasm is derived from the benefits we have personally experienced when applying these methods that have been known for a long time. Due to the potential morbidity associated with residual muscle relaxation, perioperative monitoring of neuromuscular function is essential.

Hemosiderosis pulmonar primaria con glomerulonefritis progresiva, síndrome de Goodpasture: presentación de 1 caso / Primary pulmonary hemosiderosis with progressive glomerulonephritis, Goodpasture`s syndrome: report of a case

Se presenta el caso de un lactante de 18 meses de edad, del sexo masculino, de la raza blanca y eutrófico, al que se le diagnostica al mes de ingresado una hemosiderosis pulmonar primaria. Como hallazgo anatomopatológico encontramos, además de la hemosiderosis pulmonar, una hipertrofia cardíaca, riñones difíciles de descapsular con superficie granular, un estudio hístico compatible con una glomerulonefritis crónica. En la radiografía torácica se observaron infiltrados difusos parabiliares. El diagnóstico se basó fundamentalmente en la presencia de macrófagos cargados de hemosiderina en secreciones bronquiales. El paciente falleció a los 4 meses de efectuado el diagnóstico

Hemosiderosis pulmonar primaria con glomerulonefritis progresiva, síndrome de Goodpasture: presentación de 1 caso / Primary pulmonary hemosiderosis with progressive glomerulonephritis, Goodpasture`s syndrome: report of a case

Se presenta el caso de un lactante de 18 meses de edad, del sexo masculino, de la raza blanca y eutrófico, al que se le diagnostica al mes de ingresado una hemosiderosis pulmonar primaria. Como hallazgo anatomopatológico encontramos, además de la hemosiderosis pulmonar, una hipertrofia cardíaca, riñones difíciles de descapsular con superficie granular, un estudio hístico compatible con una glomerulonefritis crónica. En la radiografía torácica se observaron infiltrados difusos parabiliares. El diagnóstico se basó fundamentalmente en la presencia de macrófagos cargados de hemosiderina en secreciones bronquiales. El paciente falleció a los 4 meses de efectuado el diagnóstico