Desarrollo de nanocompositos de cápside viral P22 para la terapia con profármacos enzimáticos en cáncer de mama

Development of P22 viral capsid nanocomposites for enzymatic prodrug therapy in breast cancer

Astrid Rebeca Luna Rios (2024, [Tesis de maestría])

Las estrategias actuales contra el cáncer de mama presentan limitaciones debido a la falta de selectividad, disminuyendo su efectividad. La mayoría de los fármacos, como el tamoxifeno, requieren la activación metabólica por parte de las enzimas de la familia citocromo P450 (CYP) para ejercer mayores efectos anticancerígenos. Sin embargo, la concentración del CYP es variable y baja en las células tumorales, especialmente en el cáncer de mama, lo que provoca efectos secundarios. En la terapia de activación enzimática de profármacos (EPT), las enzimas se dirigen al tumor para transformar el profármaco, lo que normalmente implica la administración de la enzima seguida del profármaco. Sin embargo, las diferencias entre la farmacocinética y farmacodinamia son un obstáculo para mejorar el tratamiento. Por lo tanto, la administración conjunta de profármaco y enzima es esencial para garantizar su interacción favorable en el tumor. Este trabajo reporta una nueva clase de nanocompositos terapéuticos basados en partículas tipo virus P22 confinando CYP, funcionalizadas superficialmente con glucosa oxidasa (GOx) que transforma la glucosa en D-glucono-δ-lactona produciendo H2O2, el aceptor final de electrones en la transformación de tamoxifeno mediado por CYP, y conjugadas con un derivado de tamoxifeno como profármaco y ligando dirigido utilizando polietilenglicol como conector. En un microambiente tumoral rico en glucosa, estos nanocompuestos pueden producir fármaco activo in situ. Se caracterizaron las propiedades fisicoquímicas y la catálisis secuencial mediada por glucosa de los nanocompositos. Los estudios in vitro demostraron una disminución en la viabilidad celular en líneas celulares de cáncer de mama ER+ y ER-. Sin embargo, la internalización celular en ausencia de glucosa mostró ser preferencial con las VLPs dirigidas en ambas líneas celulares. La administración conjunta de enzimas y profármacos con una localización mejorada de las VLPs desarrolladas después de la funcionalización del tamoxifeno, lo que sugiere el potencial de los nanocompositos desarrollados para superar los desafíos existentes de la EPT y mejorar los resultados terapéuticos con efectos secundarios reducidos.

Current treatment strategies against breast cancer have limitations due to lack in selectivity. Most drugs, such as tamoxifen, require metabolic activation by cytochrome P450 (CYP) enzymes to perform greater anticancer effects. However, the concentration of CYP varies and is low in tumor cells, especially in breast cancer, resulting in side-effects. In enzyme prodrug therapy (EPT), enzymes are targeted to the tumor cells for prodrug transformation, typically involving the sequential delivery of the enzyme followed by the prodrug. However, differences in pharmacokinetics and pharmacodynamics are a major hindrance for improving treatment. Therefore, co-delivery of prodrug and enzyme is essential to ensure their favorable interaction in tumor. This work reports a new class of therapeutic nanocomposites based on P22 virus like particles (VLPs) confining the CYP activity, surface functionalized with glucose oxidase (GOx) that transforms glucose into D-glucono-δ-lactone producing hydrogen peroxide, the final electron acceptor in the CYP-mediated transformation of tamoxifen, and together conjugated with a tamoxifen derivative as prodrug and targeting ligand using polyethylene glycol as a linker. In glucose-rich tumor microenvironment, these nanocomposites can produce active drug in situ. The physicochemical properties and sequential glucose-mediated catalysis of the nanocomposites were characterized. In vitro studies demonstrated a decrease in cell viability in both ER+ and ER- breast cancer cell lines. However, cellular internalization in the absence of glucose showed improved uptake of targeted VLPs in both cell lines. The co-delivery of enzymes and prodrug with improved localization of developed VLPs after tamoxifen functionalization, suggests the potential of developed nanocomposites to overcome the existing challenges of EPT and improve the therapeutic outcomes with reduced side effects.

VLP-P22, Citocromo P45O, Glucosa oxidasa, Tamoxifeno, Terapia de activación enzimática de profármacos VLP-P22, Cytochrome P45O, Glucose oxidase, Tamoxifen, Enzyme prodrug therapy BIOLOGÍA Y QUÍMICA QUÍMICA QUÍMICA FARMACÉUTICA DISEÑO.SÍNTESIS Y ESTUDIO NUEVOS FÁRMACOS DISEÑO.SÍNTESIS Y ESTUDIO NUEVOS FÁRMACOS

Evaluación del uso de vibriófagos encapsulados en el alimento para el control de Vibrio parahaemolitycus causante de AHPND en el camarón Penaeus vannamei

Esther Imelda Ponce García (2023, [Tesis de maestría])

"Las enfermedades causadas por bacterias del género Vibrio en el cultivo de camarón son de gran importancia debido a su impacto negativo en la industria acuícola. Estas bacterias pueden causar diversas enfermedades en camarones, como la necrosis hepatopancreática aguda (AHPND). La fagoterapia es una alternativa para el biocontrol de estas bacterias patógenas que presenta importantes ventajas, como la alta especificidad a sus huéspedes, la replicación en el sitio de infección, y la capacidad de infectar a bacterias resistentes a antibióticos. Sin embargo, la estabilidad de los fagos en el sistema de cultivo es una preocupación latente, ya que los fagos presentan una estabilidad limitada en solución y experimentan una caída significativa en el título de fagos durante el procesamiento y almacenamiento. El método de encapsulación de bacteriófagos, un área que aún no se estudia del todo, se presenta como una potencial alternativa para atender esta problemática. En el presente estudio se evaluó la eficacia terapéutica de un cóctel de bacteriófagos (vB_Vp_PvVp05, vB_Vp_PvVp07 y vB_Vp_PvVp11), encapsulados en alginato de sodio comercial y de mediana viscosidad, pectina, carboxilmetilcelulosa, liposomas y liofilizado en pectina como agente de biocontrol de Vibrio parahaemolyticus para su uso en terapias fagicas pasiva y activa. Se observó que el encapsulado en alginato de sodio de mediana viscosidad presentó la mayor liberación de fagos activos y al momento de ser incorporado a alimento para camarón y horneados a 80 – 100°C, se mantuvo una concentración de 1.3 x105 UFP/g después de 100 días de almacenamiento. Para la evaluación de las terapias pasiva y activa se realizó una infección experimental en juveniles de Penaeus vannamei para conocer la efectividad de los encapsulados, utilizando para la terapia pasiva la cepa Vp M0904 y para la activa la cepa Vp M0605. El experimento de terapia pasiva mostró una mortalidad del 80.6% en los camarones a las 48 hpi (p > 0.05) y una concentración de 5 x102 UFC/mL de Vibrio spp. no fermentativos en TCBS. En el caso de la terapia activa no se presentó mortalidades, pero hubo una disminución de la concentración de Vibrio spp. no fermentativos en TCBS con 5.56 x101 UFC/mL a las 48 hpi (p < 0.05) y aumentó la densidad de vibriófagos en 1.4 x102 UFP/mL en el agua de los acuarios (p < 0.05)..."

"Diseases caused by bacteria of the Vibrio genus in shrimp farming are of great importance due to their negative impact on the aquaculture industry. These bacteria can cause various diseases in shrimp, such as acute hepatopancreatic necrosis disease (AHPND). Phage therapy is an alternative for the biocontrol of these pathogenic bacteria, offering significant advantages, including high specificity to their hosts, replication at the infection site, and the ability to infect antibiotic-resistant bacteria. However, the stability of phages in the farming system is a persistent concern, as phages have limited stability in solution and undergo a significant drop in phage titer during processing and storage. The method of bacteriophage encapsulation, an area that is not yet fully explored, emerges as a potential alternative to address this issue. In this study, the therapeutic efficacy of a cocktail of bacteriophages (vB_Vp_PvVp05, vB_Vp_PvVp07, and vB_Vp_PvVp11), encapsulated in commercial medium-viscosity sodium alginate, pectin, carboxymethylcellulose, liposomes, and lyophilized pectin, was evaluated as a biocontrol agent against Vibrio parahaemolyticus for use in passive and active phage therapies. It was observed that encapsulation in medium-viscosity sodium alginate showed the highest release of active phages, maintaining a concentration of 1.3 x 105 PFU/g after 100 days of storage when incorporated into shrimp feed and baked at 80–100°C.For the evaluation of passive and active therapies, an experimental infection was conducted on Penaeus vannamei juveniles to assess the effectiveness of the encapsulated phages. The passive therapy experiment showed a mortality rate of 80.6% in shrimp at 48 hpi (p > 0.05) and a concentration of 5 x 102 CFU/mL of non-fermentative Vibrio spp. on TCBS agar. In the case of active therapy, no mortalities were observed, but there was a decrease in the concentration of non-fermentative Vibrio sp. on TCBS agar to 5.56 x 101 CFU/mL at 48 hpi (p < 0.05), and the density of vibriophages increased to 1.4 x 102 PFU/mL in the aquarium water (p < 0.05). It was determined that passive therapy offered no advantage for the control of Vibrio parahaemolyticus, unlike active therapy, which proved to be effective in controlling this bacterium."

vibriófagos, encapsulado, Penaeus vannamei, terapia activa, terapia pasiva vibriophages, encapsulated, active therapy, passive therapy BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA MICROBIOLOGÍA BACTERIÓFAGOS BACTERIÓFAGOS

Dual function of EDTA with silver nanoparticles for root canal treatment–A novel modification

JUAN MANUEL MARTINEZ ANDRADE (2018, [Artículo])

The chelating and antimicrobial capacity of a novel modification of 17% EDTA with silver nanoparticles (AgNPs) (EDTA-AgNPs) was evaluated in-vitro for root canal treatment (RCT). The EDTA-AgNPs solution was characterized by UV-Vis spectroscopy, ?-potential and high-resolution transmission electron microscopy (HRTEM). Antimicrobial capacity was evaluated against Candida albicans and Staphylococcus aureus in planktonic and biofilm cells by broth macrodilution (24 h) and XTT assays, (1, 10 and 30 min) respectively. The chelating capacity of EDTA-AgNPs was assessed indirectly (smear layer removal) and directly (demineralizing effect) in bovine dentin at two silver concentrations, 16 and 512 ?g/ ml at 1 and 10 minutes of exposure time. Smear layer removal was evaluated by atomic force microscopy (AFM) and scanning electron microscopy (SEM). The demineralizing effect was determined by atomic absorption spectroscopy (AAS), microhardness test (MH) and X-ray diffractometer (XRD). Synthesized AgNPs were quasi-spherical in shape with an average size of 13.09 ± 8.05 nm. 17% EDTA-AgNPs was effective to inhibit C. albicans and S. aureus in planktonic and biofilm cultures. The smear layer removal and demineralizing effect were similar between 17% EDTA-AgNPs and 17% EDTA treatments. The 17% EDTA-AgNPs solution proved to be an effective antimicrobial agent, and has a similar chelating capacity to 17% EDTA alone. These in-vitro studies strongly suggest that EDTA-AgNPs could be used for effective smear layer removal, having an antimicrobial effect at the same time during RCT. © 2018 Martinez-Andrade et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

edetic acid, silver nanoparticle, edetic acid, metal nanoparticle, silver, antibacterial activity, antibiotic sensitivity, antifungal activity, antimicrobial activity, Article, atomic absorption spectrometry, atomic force microscopy, biofilm, bovine, BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA MICROBIOLOGÍA MICROBIOLOGÍA

Evaluación de indicadores de calidad y funcionalidad en islas de recursos, de mezquite (Prosopis articulata) y cardón (Pachycereus pringlei) en una zona restaurada de Baja California Sur

Geovanna Zárate Camargo (2024, [Tesis de maestría])

"Las temperaturas extremas, radiación solar intensa, vientos fuertes, humedad limitada y la baja fertilidad de los suelos desérticos dificultan la recuperación de zonas degradadas en zonas áridas. Uno de los fenómenos naturales más importantes de revegetación en zonas áridas son las islas de recursos, conformadas por algunos tipos de plantas que actúan como nodrizas, las cuales, bajo su dosel, desarrollan un microhábitat favorable para el establecimiento de otras especies que trabajan en conjunto con una larga comunidad de microorganismos que se encuentran en el suelo. Entre las plantas nodriza que se observan con más frecuencia en el Desierto de Sonora, se encuentran las especies del género Prosopis. Las islas de recursos conformadas por mezquites han demostrado tener un efecto positivo como planta nodriza en zonas áridas. Las contribuciones del mezquite son el aumento del contenido de N en el suelo, temperaturas moderadas en el suelo y la superficie, altos niveles de humedad, mayor actividad microbiana, entre otros aspectos que hacen posible los efectos de las islas de recursos. En este estudio se evaluaron diferentes indicadores de calidad de suelo en islas de recursos establecidas hace 19 años, como parte de un proyecto de revegetación de suelos degradados, distintivo del Desierto Sonorense. El objetivo de este estudio fue determinar si existe un gradiente con respecto a la distancia y profundidad, en la actividad microbiana y enzimática presente en suelos de islas de recursos, resultado de la interacción de mezquite amargo (Prosopis articulata), como planta nodriza y cardón (Pachycereus pringlei), como planta objetivo. Los resultados demostraron que existe un aumento la actividad enzimática y carbono asociado a biomasa microbiana del suelo perteneciente a estas islas, con mayores resultados en zonas próximas a la planta nodriza que en zonas sin su influencia, también se encontró un aumento en la capa superficial del suelo mostrando mayores valores en los indicadores de calidad y funcionalidad bajo el dosel de las islas de recursos. Lo anterior sugiere que el uso de estas islas de recursos es una potencial alternativa en la restauración de suelos degradados, favoreciendo el ciclaje de nutrientes en suelos degradados."

"At desert, the high temperatures, intense solar radiation, strong winds, limited water, and low fertility, determine how challenging will be the recovery of degraded soils in arid lands. One of the most important natural phenomena of revegetation in arid lands are the “resource islands”, that consist of some trees or bushes acting as nurse plants, which under their canopy, develop a beneficial microhabitat for the establishment of other plant species the work with an extensive community of microorganisms found in the soil. Among the most common nurse plants observed in the Sonoran Desert, is the genus Prosopis. The resource islands formed with mesquite have shown positive effects as nurse plant in arid lands. The principal contributions of mesquite are the increase of N in the soil, moderate temperatures on soil and topsoil, high moisture levels, increased microbial activity, among other aspects that make possible the effect of resource islands. In this study, different quality indicators were evaluated in resource islands established 19 years ago, as part of a project for revegetation in degraded soils, distinctive from the Sonoran Desert. The aim of this research was to explore whether there is a correlation between distance, depth, microbial activity, and enzymatic activity in the soil surrounding resource islands formed by the interaction between mesquite amargo (Prosopis articulata) as the nurse plant and cardon (Pachycereus pringlei) as the objective plant. The results demonstrated that there is an increase in the enzymatic activity and microbial biomass carbon from the soil under the canopy of the resource islands, with higher results in zones near the nurse plant than zones without its influence, furthermore, there was found an increase in the topsoil showing higher values in the soil quality and functionality indicators under the canopy of the resource islands. The above indicates that the use of these resource islands is a potential alternative in the degraded soil restoration, supporting the nutrient cycling in degraded soils."

suelo, ecología, revegetación, actividad enzimática, ciclaje de nutrientes soil, ecology, revegetation, enzymatic activity, nutrient cycling CIENCIAS AGROPECUARIAS Y BIOTECNOLOGÍA CIENCIAS AGRARIAS AGRONOMÍA FERTILIDAD DEL SUELO FERTILIDAD DEL SUELO

XYZ Micropositioning System Based on Compliance Mechanisms Fabricated by Additive Manufacturing

CARLOS ANDRES FERRARA BELLO Pedro Vargas Chable José Gerardo Vera Dimas Margarita Tecpoyotl Torres (2021, [Artículo])

This article presents the design and implementation of a micropositioning system actuated

by three piezoelectric stacks to control its displacements on XYZ axes. The use of conventional

piezoelectric buzzers allows us to reduce fabrication costs. The working or mobile platform is the base

for objects that will be manipulated, for example, in automated assembling. The micropositioner can

be integrated into a microgripper to generate a complete manipulation system. For micropositioner

fabrication, at first, Polylactic Acid (PLA) was chosen as the structural material, but after simulation

and some experimental tests performed with a micropositioner made of Acrylonitrile Butadiene

Styrene (ABS), it showed larger displacement (approx. 20%) due to its lower stiffness. A third test

was performed with a positioner made with Polyethylene Terephthalate Glycol (PETG), obtaining an

intermediate performance. The originality of this work resides in the geometrical arrangement based

on thermoplastic polymer compliance mechanisms, as well as in the use of additive manufacturing

to fabricate it. An experimental setup was developed to carry out experimental tests. ANSYS™ was

used for simulation.

INGENIERÍA Y TECNOLOGÍA CIENCIAS TECNOLÓGICAS PETG, MEMS, micromanipulation, piezoelectric actuation, hysteresis effect linearization

Procesamiento emocional ante estímulos música-color y su respuesta electrofisiológica en personas con depresión

PABLO VALDÉS ALEMÁN (2024, [Tesis de maestría])

Dado el fenómeno del vínculo emocional que existe entre música y color, este

proyecto exploró los posibles efectos terapéuticos en personas con depresión, al

presentarles estímulos combinados de música con color que, en conjunto, evocan

emociones positivas y placenteras. El proyecto se dividió en tres partes, con el fin

de subsanar los vacíos en la literatura. Primero se evaluaron los 37 colores del

Berkeley Color Project como estímulos capaces de evocar emociones y placer,

distinguibles mediante los modelos de asimetría frontal de alfa (FAA) y de theta con

electroencefalografía (EEG). El segundo estudio exploró los posibles efectos

sinérgicos al combinar música y color de emoción congruente –i.e., misma

emoción–, mediante respuestas subjetivas y EEG. Por último, se probó el impacto

de estímulos música-color positivos y placenteros sobre el estado de ánimo de

personas con depresión, a partir de sus respuestas subjetivas y EEG. Se encontró

que los colores brillantes, alegres y placenteros se asocian con una mayor

activación –supresión de alfa– frontal izquierda, de acuerdo con el modelo de FAA.

En lugar de un efecto sinérgico, los hallazgos mostraron un predominio emocional

hacia la música, reafirmando su poder emocional por encima del color. Por último,

los resultados del tercer estudio demostraron una mejora en el estado de ánimo en

las personas con sintomatología depresiva tras la estimulación. Estos resultados

podrían informar a profesionales de la salud mental sobre nuevos métodos

terapéuticos para ayudar a pacientes con depresión, al conjuntar la musicoterapia

con la cromoterapia.

Given the phenomenon of the emotional link between music and color, this project

explored the possible therapeutic effects on people with depression by presenting

them with combined music-color stimuli that collectively evoke positive and pleasant

emotions. The project was divided into three parts to address gaps in the literature.

Firstly, the 37 colors from the Berkeley Color Project were evaluated as stimuli

capable of evoking emotions and pleasure, distinguishable through the frontal alpha

(FAA) and theta asymmetry models using electroencephalography (EEG). The

second study explored the potential synergic effects by combining music and color

of congruent emotion –i.e., the same emotion– using subjective responses and EEG.

Lastly, the impact of positive and pleasant music-color stimuli on the mood of

individuals with depression was tested based on their subjective responses and

EEG. It was found that bright, joyful, and pleasant colors are associated with

increased activation –alpha suppression– in the left frontal region of the scalp,

according to the FAA model. Instead of a synergic effect, the results showed an

emotional predominance towards music, reaffirming its emotional power over color.

Finally, the results of the third study demonstrated an improvement in the mood of

individuals with depressive symptoms following the stimulation. These findings could

inform mental health professionals about new therapeutic methods to assist patients

with depression by combining music therapy with chromotherapy.

MEDICINA Y CIENCIAS DE LA SALUD CIENCIAS MÉDICAS música, color y emoción; patrones de actividad EEG; Berkeley Color Project; musico-cromoterapia; depresión. music, color, and emotion; EEG activity patterns; Berkeley Color Project; music-chromotherapy; depression.

Coronin is a component of the endocytic collar of hyphae of neurospora crassa and is necessary for normal growth and morphogenesis

RAMON OSVALDO ECHAURI ESPINOSA (2012, [Artículo])

Coronin plays a major role in the organization and dynamics of actin in yeast. To investigate the role of coronin in a filamentous fungus (Neurospora crassa), we examined its subcellular localization using fluorescent proteins and the phenotypic consequences of coronin gene (crn-1) deletion in hyphal morphogenesis, Spitzenkörper behavior and endocytosis. Coronin-GFP was localized in patches, forming a subapical collar near the hyphal apex; significantly, it was absent from the apex. The subapical patches of coronin colocalized with fimbrin, Arp2/3 complex, and actin, altogether comprising the endocytic collar. Deletion of crn-1 resulted in reduced hyphal growth rates, distorted hyphal morphology, uneven wall thickness, and delayed establishment of polarity during germination; it also affected growth directionality and increased branching. The Spitzenkörper of Δcrn-1 mutant was unstable; it appeared and disappeared intermittently giving rise to periods of hyphoid-like and isotropic growth respectively. Uptake of FM4-64 in Δcrn-1 mutant indicated a partial disruption in endocytosis. These observations underscore coronin as an important component of F-actin remodeling in N. crassa. Although coronin is not essential in this fungus, its deletion influenced negatively the operation of the actin cytoskeleton involved in the orderly deployment of the apical growth apparatus, thus preventing normal hyphal growth and morphogenesis. © 2012 Echauri-Espinosa et al.

actin related protein 2-3 complex, F actin, fimbrin protein, fluorescent dye, fungal protein, fungal protein coronin, green fluorescent protein, unclassified drug, actin binding protein, coronin proteins, fungal protein, article, cell polarity, contr BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA MICROBIOLOGÍA MICROBIOLOGÍA

Octopus maya white body show sex-specific transcriptomic profiles during the reproductive phase, with high differentiation in signaling pathways

Oscar Juárez (2019, [Artículo])

White bodies (WB), multilobulated soft tissue that wraps the optic tracts and optic lobes, have been considered the hematopoietic organ of the cephalopods. Its glandular appearance and its lobular morphology suggest that different parts of the WB may perform different functions, but a detailed functional analysis of the octopus WB is lacking. The aim of this study is to describe the transcriptomic profile of WB to better understand its functions, with emphasis on the difference between sexes during reproductive events. Then, validation via qPCR was performed using different tissues to find out tissue-specific transcripts. High differentiation in signaling pathways was observed in the comparison of female and male transcriptomic profiles. For instance, the expression of genes involved in the androgen receptor-signaling pathway were detected only in males, whereas estrogen receptor showed higher expression in females. Highly expressed genes in males enriched oxidation-reduction and apoptotic processes, which are related to the immune response. On the other hand, expression of genes involved in replicative senescence and the response to cortisol were only detected in females. Moreover, the transcripts with higher expression in females enriched a wide variety of signaling pathways mediated by molecules like neuropeptides, integrins, MAPKs and receptors like TNF and Toll-like. In addition, these putative neuropeptide transcripts, showed higher expression in females’ WB and were not detected in other analyzed tissues. These results suggest that the differentiation in signaling pathways in white bodies of O. maya influences the physiological dimorphism between females and males during the reproductive phase. © 2019 Juárez et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

androgen receptor, integrin, mitogen activated protein kinase, neuropeptide, transcriptome, tumor necrosis factor, argonaute protein, corticotropin releasing factor receptor, corticotropin releasing factor receptor 2, DEAD box protein, estradiol 17be BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA BIOLOGÍA ANIMAL (ZOOLOGÍA) BIOLOGÍA ANIMAL (ZOOLOGÍA)

Enhancement of antibiotics antimicrobial activity due to the silver nanoparticles impact on the cell membrane

Roberto Vazquez-Munoz (2019, [Artículo])

The ability of microorganisms to generate resistance outcompetes with the generation of new and efficient antibiotics; therefore, it is critical to develop novel antibiotic agents and treatments to control bacterial infections. An alternative to this worldwide problem is the use of nanomaterials with antimicrobial properties. Silver nanoparticles (AgNPs) have been extensively studied due to their antimicrobial effect in different organisms. In this work, the synergistic antimicrobial effect of AgNPs and conventional antibiotics was assessed in Gram-positive and Gram-negative bacteria. AgNPs minimal inhibitory concentration was 10–12 μg mL-1 in all bacterial strains tested, regardless of their different susceptibility against antibiotics. Interestingly, a synergistic antimicrobial effect was observed when combining AgNPs and kanamycin according to the fractional inhibitory concentration index, FICI: &lt;0.5), an additive effect by combining AgNPs and chloramphenicol (FICI: 0.5 to 1), whereas no effect was found with AgNPs and β-lactam antibiotics combinations. Flow cytometry and TEM analysis showed that sublethal concentrations of AgNPs (6–7 μg mL-1) altered the bacterial membrane potential and caused ultrastructural damage, increasing the cell membrane permeability. No chemical interactions between AgNPs and antibiotics were detected. We propose an experimental supported mechanism of action by which combinatorial effect of antimicrobials drives synergy depending on their specific target, facilitated by membrane alterations generated by AgNPs. Our results provide a deeper understanding about the synergistic mechanism of AgNPs and antibiotics, aiming to combat antimicrobial infections efficiently, especially those by multi-drug resistant microorganisms, in order to mitigate the current crisis due to antibiotic resistance. © 2019 Vazquez-Muñoz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

ampicillin, antibiotic agent, aztreonam, beta lactam antibiotic, biapenem, chloramphenicol, kanamycin, silver nanoparticle, silver nitrate, antiinfective agent, metal nanoparticle, silver, antibiotic sensitivity, antimicrobial activity, Article, bact BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA BIOFÍSICA BIOFÍSICA

Comparative live-cell imaging analyses of SPA-2, BUD-6 and BNI-1 in Neurospora crassa reveal novel features of the filamentous fungal polarisome

Alexander Lichius (2012, [Artículo])

A key multiprotein complex involved in regulating the actin cytoskeleton and secretory machinery required for polarized growth in fungi, is the polarisome. Recognized core constituents in budding yeast are the proteins Spa2, Pea2, Aip3/Bud6, and the key effector Bni1. Multicellular fungi display a more complex polarized morphogenesis than yeasts, suggesting that the filamentous fungal polarisome might fulfill additional functions. In this study, we compared the subcellular organization and dynamics of the putative polarisome components BUD-6 and BNI-1 with those of the bona fide polarisome marker SPA-2 at various developmental stages of Neurospora crassa. All three proteins exhibited a yeast-like polarisome configuration during polarized germ tube growth, cell fusion, septal pore plugging and tip repolarization. However, the localization patterns of all three proteins showed spatiotemporally distinct characteristics during the establishment of new polar axes, septum formation and cytokinesis, and maintained hyphal tip growth. Most notably, in vegetative hyphal tips BUD-6 accumulated as a subapical cloud excluded from the Spitzenkörper (Spk), whereas BNI-1 and SPA-2 partially colocalized with the Spk and the tip apex. Novel roles during septal plugging and cytokinesis, connected to the reinitiation of tip growth upon physical injury and conidial maturation, were identified for BUD-6 and BNI-1, respectively. Phenotypic analyses of gene deletion mutants revealed additional functions for BUD-6 and BNI-1 in cell fusion regulation, and the maintenance of Spk integrity. Considered together, our findings reveal novel polarisome-independent functions of BUD-6 and BNI-1 in Neurospora, but also suggest that all three proteins cooperate at plugged septal pores, and their complex arrangement within the apical dome of mature hypha might represent a novel aspect of filamentous fungal polarisome architecture. © 2012 Lichius et al.

fungal protein, protein BNI 1, protein BUD 6, protein SPA 2, protein Spk, unclassified drug, actin binding protein, cytoskeleton protein, fungal protein, article, cell fusion, cellular distribution, comparative study, conidium, controlled study, cyto BIOLOGÍA Y QUÍMICA CIENCIAS DE LA VIDA MICROBIOLOGÍA MICROBIOLOGÍA