Health Benefits of High Voltage Electrostatic Field Processing of Fruits and Vegetables.

High voltage electrostatic field processing (HVEF) is a food preservation procedure frequently used to produce healthy minimally processed fruits and vegetables (F&V) as it reduces the growth of microorganisms and activates or inhibits various enzymes, thus retarding their natural ripening while preserving and even enhancing native nutritional quality and sensory characteristics. HVEF is one of the various nonthermal processing technology (NTPT) regarded as abiotic stress that can activate the antioxidant system of F&V and can also inhibith spoilage enzymes as, polyphenol oxidase (PPO), lipoxygenase (LOX), pectin methylesterase (PME), polygalacturonase (PG), cellulase (Cel), ß-xylosidase, xyloglucan and endotransglycosylase/hydrolase, bringing positive effect on hardness, firmness, colour attributes, electric conductivity, antioxidant compounds, microstructure and decreasing electrolyte leakage (EL), malondialdehyde (MDA) contents and browning degree. This technique can also increase the contents of fructose, glucose, and sucrose and decrease the production of CO2 and H2O2. Additionally, it has been reported that HVEF could be used with other treatments, such as modified atmosphere packaging (MAP) and acidic electrolyzed water (AEW) treatment, to enhance its effects. Future works should deepen on elucidating the activation of the antioxidant systems by applying HVEF of critical enzymes related to the synthesis pathways of phenolic compounds (PC) and carotenoids (Car). Holistic approaches to the effects of HVEF on metabolism based on systems biology also need to be studied by considering the overall biochemical, physical, and process engineering related aspects of this technique.

Effect of Agavins and Agave Syrup Use in the Formulation of a Synbiotic Gelatin Gummy with Microcapsules of Saccharomyces Boulardii.

Agavins are reserve carbohydrates found in agave plants; they present texture-modifying properties and prebiotic capacity by increasing the viability of the intestinal microbiota. Through its hydrolysis, agave syrup (AS) can be obtained and can be used as a sweetener in food matrices. The objective of this work was to evaluate the effect of the variation in the content of agavins and AS on the physical, structural, and viability properties of Saccharomyces boulardii encapsulates incorporated into gelatin gummies. An RSM was used to obtain an optimized formulation of gelatin gummies. The properties of the gel in the gummy were characterized by a texture profile analysis and Aw. The humidity and sugar content were determined. A sucrose gummy was used as a control for the variable ranges. Alginate microcapsules containing S. boulardii were added to the optimized gummy formulation to obtain a synbiotic gummy. The viability of S. boulardii and changes in the structure of the alginate gel of the microcapsules in the synbiotic gummy were evaluated for 24 days by image digital analysis (IDA). The agavins and agave syrup significantly affected the texture properties (<1 N) and the Aw (>0.85). The IDA showed a change in the gel network and an increase in viability by confocal microscopy from day 18. The number of pores in the gel increased, but their size decreased with an increase in the number of S. boulardii cells. Agavins and cells alter the structure of capsules in gummies without affecting their viability.

A Biophysics Toolbox for Reliable Data Acquisition and Processing in Integrated Force-Confocal Fluorescence Microscopy.

Integrated single-molecule force-fluorescence spectroscopy setups allow for simultaneous fluorescence imaging and mechanical force manipulation and measurements on individual molecules, providing comprehensive dynamic and spatiotemporal information. Dual-beam optical tweezers (OT) combined with a confocal scanning microscope form a force-fluorescence spectroscopy apparatus broadly used to investigate various biological processes, in particular, protein:DNA interactions. Such experiments typically involve imaging of fluorescently labeled proteins bound to DNA and force spectroscopy measurements of trapped individual DNA molecules. Here, we present a versatile state-of-the-art toolbox including the preparation of protein:DNA complex samples, design of a microfluidic flow cell incorporated with OT, automation of OT-confocal scanning measurements, and the development and implementation of a streamlined data analysis package for force and fluorescence spectroscopy data processing. Its components can be adapted to any commercialized or home-built dual-beam OT setup equipped with a confocal scanning microscope, which will facilitate single-molecule force-fluorescence spectroscopy studies on a large variety of biological systems.

Editorial for the Special Issue "Food and Microbial Bioprocesses".

The aim of this Special Issue was to provide readers with a holistic, systematic, and integrative approach to microbial processes involved in the production of selected foods, nutraceuticals, and bioactive materials by using modern biotechnological tools [...].

Exploration adhesion properties of Liquorilactobacillus and Lentilactobacillus isolated from two different sources of tepache kefir grains.

Due to the distinctive characteristics of probiotics, it is essential to pinpoint strains originating from diverse sources that prove efficacious in addressing a range of pathologies linked to dysfunction of the intestinal barrier. Nine strains of lactic acid bacteria were isolated from two different sources of tepache kefir grains (KAS2, KAS3, KAS4, KAS7, KAL4, KBS2, KBS3, KBL1 and KBL3), and were categorized to the genus Lacticaseibacillus, Liquorilactobacillus, and Lentilactobacillus by 16S rRNA gene. Kinetic behaviors of these strains were evaluated in MRS medium, and their probiotic potential was performed: resistance to low pH, tolerance to pepsin, pancreatin, bile salts, antibiotic resistance, hemolytic activity, and adhesion ability. KAS7 strain presented a higher growth rate (0.50 h-1) compared with KAS2 strain, who presented a lower growth rate (0.29 h-1). KBS2 strain was the only strain that survived the in vitro stomach simulation conditions (29.3%). Strain KBL1 demonstrated significantly higher viability (90.6%) in the in vitro intestine simulation conditions. Strain KAS2 demonstrated strong hydrophilic character with chloroform (85.6%) and xylol (57.6%) and a higher percentage of mucin adhesion (87.1%). However, strains KBS2 (84.8%) and KBL3 (89.5%) showed the highest autoaggregation values. In terms of adhesion to the intestinal epithelium in rats, strains KAS2, KAS3 and KAS4 showed values above 80%. The growth of the strains KAS2, KAS3, KAS4, KBS2, and KBL3 was inhibited by cefuroxime, cefotaxime, tetracycline, ampicillin, erythromycin, and cephalothin. Strains KBS2 (41.9% and 33.5%) and KBL3 (42.5% and 32.8%) had the highest co-aggregation values with S. aureus and E. coli. The results obtained in this study indicate that lactic acid bacteria isolated from tepache can be considered as candidates for potentially probiotic bacteria, laying the foundations to evaluate their probiotic functionality in vivo and thus to be used in the formulation of functional foods.

Mobilizing faith-based COVID-19 health ambassadors to address COVID-19 health disparities among African American older adults in under-resourced communities: A hybrid, community-based participatory intervention.

INTRODUCTION: The COVID-19 pandemic disproportionately affected older adults, particularly those with pre-existing chronic health conditions. To address the health disparity and challenges faced by under-resourced African American older adults in South Los Angeles during this period, we implemented a hybrid (virtual/in-person), pre-post, community-based participatory intervention research project utilizing a faith-based lay health advisor model (COVID-19 Health Ambassador Program (CHAP)). We recruited COVID-19 Health Ambassadors (CHAs) and African American older adults (participants) from faith-based organizations who partook in CHA-led meetings and follow-ups that educated and supported the participants. This paper seeks to evaluate this intervention's implementation using the Consolidated Framework for Implementation Research (CFIR) as a reporting tool with an emphasis on fidelity, challenges, and adaptations based on data collected via stakeholder interviews and surveys. RESULTS: CHAP was delivered to 152 participants by 19 CHAs from 17 faith-based organizations. CHAs assisted with chronic disease management, resolved medication-related challenges, encouraged COVID-19 vaccination, reduced psychological stress and addressed healthcare avoidance behaviors such as COVID-19 vaccine hesitancy among the participants. Challenges encountered include ensuring participant engagement and retention in the virtual format and addressing technological barriers for CHAs and participants. Adaptations made to better suit the needs of participants included providing communication tools and additional training to CHAs to improve their proficiency in using virtual platforms in addition to adapting scientific/educational materials to suit our participants' diverse cultural and linguistic needs. CONCLUSION: The community-centered hybrid approach in addition to our partnership with faith-based organizations and their respective COVID-19 health ambassadors proved to be essential in assisting underserved African American older adults manage chronic health conditions and address community-wide health disparities during the COVID-19 pandemic. Adaptability, cultural sensitivity, and teamwork are key to implementing health interventions especially in underserved populations.

Differential DNA Methylation from Autistic Children Enriches Evidence for Genes Associated with ASD and New Candidate Genes.

The etiology of Autism Spectrum Disorders (ASD) is a result of the interaction between genes and the environment. The study of epigenetic factors that affect gene expression, such as DNA methylation, has become an important area of research in ASD. In recent years, there has been an increasing body of evidence pointing to epigenetic mechanisms that influence brain development, as in the case of ASD, when gene methylation dysregulation is present. Our analysis revealed 853 differentially methylated CpG in ASD patients, affecting 509 genes across the genome. Enrichment analysis showed five related diseases, including autistic disorder and mental disorders, which are particularly significant. In this work, we identified 64 genes that were previously reported in the SFARI gene database, classified according to their impact index. Additionally, we identified new genes that have not been previously reported as candidates with differences in the methylation patterns of Mexican children with ASD.

A chromatinized origin reduces the mobility of ORC and MCM through interactions and spatial constraint.

Chromatin replication involves the assembly and activity of the replisome within the nucleosomal landscape. At the core of the replisome is the Mcm2-7 complex (MCM), which is loaded onto DNA after binding to the Origin Recognition Complex (ORC). In yeast, ORC is a dynamic protein that diffuses rapidly along DNA, unless halted by origin recognition sequences. However, less is known about the dynamics of ORC proteins in the presence of nucleosomes and attendant consequences for MCM loading. To address this, we harnessed an in vitro single-molecule approach to interrogate a chromatinized origin of replication. We find that ORC binds the origin of replication with similar efficiency independently of whether the origin is chromatinized, despite ORC mobility being reduced by the presence of nucleosomes. Recruitment of MCM also proceeds efficiently on a chromatinized origin, but subsequent movement of MCM away from the origin is severely constrained. These findings suggest that chromatinized origins in yeast are essential for the local retention of MCM, which may facilitate subsequent assembly of the replisome.

Protective Effect of Alginate Microcapsules with Different Rheological Behavior on Lactiplantibacillus plantarum 299v.

Alginate encapsulation is a well-known technique used to protect microorganisms from adverse conditions. However, it is also known that the viscosity of the alginate is dependent on its composition and degree of polymerization and that thermal treatments, such as pasteurization and sterilization, can affect the structure of the polymer and decrease its protection efficiency. The goal of this study was to evaluate the protective effect of encapsulation, using alginates of different viscosities treated at different temperatures, on Lactiplantibacillus plantarum 299v under in vitro gastrointestinal conditions and cold storage at 4 °C and -15 °C, respectively. Steady- and dynamic-shear rheological tests were used to characterize the polymers. Thermal treatments profoundly affected the rheological characteristics of alginates with high and low viscosity. However, the solutions and gels of the low-viscosity alginate were more affected at a temperature of 117 °C. The capsules elaborated with high-viscosity alginate solution and pasteurized at 63 °C for 30 min provided better protection to the cells of L. plantarum 299v under simulated gastrointestinal and cold storage conditions.

Production of Reactive Oxygen (ROS) and Nitrogen (RNS) Species in Macrophages J774A.1 Activated by the Interaction between Two Escherichia coli Pathotypes and Two Probiotic Commercial Strains.

Probiotics play an important role against infectious pathogens, such as Escherichia coli (E. coli), mainly through the production of antimicrobial compounds and their immunomodulatory effect. This protection can be detected both on the live probiotic microorganisms and in their inactive forms (paraprobiotics). Probiotics may affect different cells involved in immunity, such as macrophages. Macrophages are activated through contact with microorganisms or their products (lipopolysaccharides, endotoxins or cell walls). The aim of this work was the evaluation of the effect of two probiotic bacteria (Escherichia coli Nissle 1917 and Bifidobacterium animalis subsp. lactis HN019 on macrophage cell line J774A.1 when challenged with two pathogenic strains of E. coli. Macrophage activation was revealed through the detection of reactive oxygen (ROS) and nitrogen (RNS) species by flow cytometry. The effect varied depending on the kind of probiotic preparation (immunobiotic, paraprobiotic or postbiotic) and on the strain of E. coli (enterohemorrhagic or enteropathogenic). A clear immunomodulatory effect was observed in all cases. A higher production of ROS compared with RNS was also observed.

Assessing Food Access, Exercise, and Dietary History among Older African American Parishioners During the COVID-19 Pandemic (C-FED Study): Design, Opportunities, Challenges, and Lessons Learned.

OBJECTIVES: Unhealthy diets and inadequate exercise are associated with chronic health conditions and excess mortality. Older African Americans do not meet dietary and exercise guidelines, and this may have worsened during the COVID-19 pandemic due to individual and environmental factors, including food insecurity. Studies evaluating these dynamics are essential for developing interventions. This narrative details a study protocol and data collection experiences during the pandemic. METHODS: Participants > 55 years African American old completed detailed food frequency, exercise, and food access questionnaires between October 2020 and July 2021. Observations of the study administrators (authors of this manuscript) for the duration of the study are presented. Details on the study design and reflections on the opportunities, challenges, and lessons learned are summarized. Future manuscripts will report data analysis of study findings. RESULTS: A total of 123 older African American adults participated in the study, and 118 (70% female) completed all three questionnaires. More than 50% of the participants had at least two primary chronic conditions. About 85% were fully vaccinated against COVID-19. Applying community-based participatory approaches, leveraging partnerships, and exercising flexibility approaches were pivotal to successfully implementing the study protocol. CONCLUSIONS: Despite challenges related to the COVID-19 pandemic, detailed data on older African American adults' diet and exercise habits were obtained. Our study design and experiences will benefit future researchers. More importantly, results from our study will inform interventions and policies aimed at minimizing consequences associated with poor diet and exercise habits during the pandemic among this vulnerable population.

Pro-Apoptotic Activity and Cell Cycle Arrest of Caulerpa sertularioides against SKLU-1 Cancer Cell in 2D and 3D Cultures.

Cancer is a disease with the highest mortality and morbidity rate worldwide. First-line drugs induce several side effects that drastically reduce the quality of life of people with this disease. Finding molecules to prevent it or generate less aggressiveness or no side effects is significant to counteract this problem. Therefore, this work searched for bioactive compounds of marine macroalgae as an alternative treatment. An 80% ethanol extract of dried Caulerpa sertularioides (CSE) was analyzed by HPLS-MS to identify the chemical components. CSE was utilized through a comparative 2D versus 3D culture model. Cisplatin (Cis) was used as a standard drug. The effects on cell viability, apoptosis, cell cycle, and tumor invasion were evaluated. The IC50 of CSE for the 2D model was 80.28 µg/mL versus 530 µg/mL for the 3D model after 24 h of treatment exposure. These results confirmed that the 3D model is more resistant to treatments and complex than the 2D model. CSE generated a loss of mitochondrial membrane potential, induced apoptosis by extrinsic and intrinsic pathways, upregulated caspases-3 and -7, and significantly decreased tumor invasion of a 3D SKLU-1 lung adenocarcinoma cell line. CSE generates biochemical and morphological changes in the plasma membrane and causes cell cycle arrest at the S and G2/M phases. These findings conclude that C. sertularioides is a potential candidate for alternative treatment against lung cancer. This work reinforced the use of complex models for drug screening and suggested using CSE's primary component, caulerpin, to determine its effect and mechanism of action on SKLU-1 in the future. A multi-approach with molecular and histological analysis and combination with first-line drugs must be included.

Nucleotide binding halts diffusion of the eukaryotic replicative helicase during activation.

The eukaryotic replicative helicase CMG centrally orchestrates the replisome and leads the way at the front of replication forks. Understanding the motion of CMG on the DNA is therefore key to our understanding of DNA replication. In vivo, CMG is assembled and activated through a cell-cycle-regulated mechanism involving 36 polypeptides that has been reconstituted from purified proteins in ensemble biochemical studies. Conversely, single-molecule studies of CMG motion have thus far relied on pre-formed CMG assembled through an unknown mechanism upon overexpression of individual constituents. Here, we report the activation of CMG fully reconstituted from purified yeast proteins and the quantification of its motion at the single-molecule level. We observe that CMG can move on DNA in two ways: by unidirectional translocation and by diffusion. We demonstrate that CMG preferentially exhibits unidirectional translocation in the presence of ATP, whereas it preferentially exhibits diffusive motion in the absence of ATP. We also demonstrate that nucleotide binding halts diffusive CMG independently of DNA melting. Taken together, our findings support a mechanism by which nucleotide binding allows newly assembled CMG to engage with the DNA within its central channel, halting its diffusion and facilitating the initial DNA melting required to initiate DNA replication.

Study of Candelilla Wax Concentrations on the Physical Properties of Edible Nanocoatings as a Function of Support Polysaccharides.

Solid lipid nanoparticles (SLN) based on candelilla wax were prepared using the hot homogenization technique. The resulting suspension had monomodal behavior with a particle size of 809-885 nm; polydispersity index < 0.31, and zeta potential of -3.5 mV 5 weeks after monitoring. The films were prepared with SLN concentrations of 20 and 60 g/L, each with a plasticizer concentration of 10 and 30 g/L; the polysaccharide stabilizers used were either xanthan gum (XG) or carboxymethyl cellulose (CMC) at 3 g/L. The effects of temperature, film composition, and relative humidity on the microstructural, thermal, mechanical, and optical properties, as well as the water vapor barrier, were evaluated. Higher amounts of SLN and plasticizer gave the films greater strength and flexibility due to the influence of temperature and relative humidity. The water vapor permeability (WVP) was lower when 60 g/L of SLN was added to the films. The arrangement of the SLN in the polymeric networks showed changes in the distribution as a function of the concentrations of the SLN and plasticizer. The total color difference (ΔE) was greater when the content of the SLN was increased, with values of 3.34-7.93. Thermal analysis showed an increase in the melting temperature when a higher SLN content was used, whereas a higher plasticizer content reduced it. Edible films with the most appropriate physical properties for the packaging, shelf-life extension, and improved quality conservation of fresh foods were those made with 20 g/L of SLN, 30 g/L of glycerol, and 3 g/L of XG.

Pro-Apoptotic Activity of Bioactive Compounds from Seaweeds: Promising Sources for Developing Novel Anticancer Drugs.

The process by which cancer cells evade or inhibit apoptosis is considered one of the characteristics of cancer. The ability of cancer cells to escape apoptosis contributes to tumor proliferation and promotes metastasis. The discovery of new antitumor agents is essential for cancer treatment due to the lack of selectivity of drugs and cellular resistance to anticancer agents. Several studies showed that macroalgae produce various metabolites with different biological activities among marine organisms. This review discusses multiple metabolites extracted from macroalgae and their pro-apoptotic effects through regulating apoptosis signaling pathway target molecules and the structure-activity relationship. Twenty-four promising bioactive compounds have been reported, where eight of these compounds exhibited values of maximum inhibitory concentration (IC50) of less than 7 µg/mL. Fucoxanthin was the only carotenoid reported that induced apoptosis in HeLa cells with an IC50 below 1 µg/mL. Se-PPC (a complex of proteins and selenylated polysaccharides) is the magistral compound because it is the only one with an IC50 of 2.5 µg/mL which regulates the primary proteins and critical genes of both apoptosis pathways. Therefore, this review will help provide the basis for further studies and the development of new anticancer drugs, both as single agents and adjuvants, decreasing the aggressiveness of first-line drugs and offering patients better survival and quality of life.

Factors Associated with Telehealth Utilization among Older African Americans in South Los Angeles during the COVID-19 Pandemic.

BACKGROUND: The COVID-19 pandemic transformed healthcare delivery with the expansive use of telemedicine. However, health disparities may result from lower adoption of telehealth among African Americans. This study examined how under-resourced, older African Americans with chronic illnesses use telehealth, including related sociodemographic and COVID-19 factors. METHODS: Using a cross-sectional design, 150 middle-aged and older African Americans were recruited from faith-based centers from March 2021 to August 2022. Data collected included sociodemographics, comorbidities, technological device ownership, internet usage, and attitudes toward COVID-19 disease and vaccination. Descriptive statistics and multivariable regression models were conducted to identify factors associated with telehealth use. RESULTS: Of the 150 participants, 32% had not used telehealth since the COVID-19 pandemic, with 75% reporting no home internet access and 38% having no cellular/internet network on their mobile device. Age, access to a cellular network on a mobile device, and wireless internet at home were significantly associated with the utilization of telehealth care. Higher anxiety and stress with an increased perceived threat of COVID-19 and positive attitudes toward COVID-19 vaccination were associated with telehealth utilization. DISCUSSION: Access and integration of telehealth services were highlighted as challenges for this population of African Americans. To reduce disparities, expansion of subsidized wireless internet access in marginalized communities is necessitated. Education outreach and training by healthcare systems and community health workers to improve uptake of telehealth currently and post-COVID-19 should be considered.

[Association between telomere length and cognitive impairment in older adults]. / Asociación entre la longitud de los telómeros y deterioro cognitivo en adultos mayores.

INTRODUCTION: Cognitive impairment is a transition stage between normal aging and dementia, the prevalence of last one increases with age; the damage of the functions and physical integrity, places the older adult in a greater susceptibility to get sick. Telomere length is a hallmark of aging to characterize this phenotype, as well as a biomarker that reflects the underlying state of the cell. In this work, the relative length of telomeres in older adults with cognitive impairment was correlated. MATERIAL AND METHODS: Observational-analytical study, in samples of adult patients older than 65 years with and without cognitive impairment, in whom the relative length of telomeres was measured. RESULTS: Ninety samples of older adults were included in the study and in the association analysis according to multivariate logistic models, cognitive impairment showed almost five times more risk for telomere shortening in relation to the presence of the diagnosis of cognitive impairment (Odds ratio 4.88, p=0.027). CONCLUSIONS: When correlating the relative length of telomeres in older adults diagnosed with cognitive impairment, this association was confirmed for shorter.

Effects of comorbidity on executive functions among children with ADHD, finding trends.

There is still no basic overview about the effect of various types of comorbidity in executive functions due to two main reasons: (1) the type and number of comorbidities in ADHD is significantly varied, (2) EFs are very diverse and have different neuropsychological properties. Our objective was to determine the effect of comorbid disorders (number and type) on the performance in a wide range (seven) of executive functions in a sample of children with ADHD. Fifty-five male children aged seven to nine years with ADHD were divided into six groups: G1 = ADHD only (ADHD-O), G1 = Oppositional defiant disorder (ODD), G3 = (anxiety/depressive disorder (ADD), G4 = ODD + ADD, G5 = ODD + learning disorder (LD), G6 = ODD + LD + conduct disorder (CD). The six groups exhibited different number of deficits in EFs; G1 showed only 1 deficit in contrast, G6 presented 11. Statistical analysis (ANOVA and logistic regression) identified three most affected EFs: Working memory, generation/classification of semantic categories, and inhibitory control. Alterations in EFs increased mainly in relation to the increase of the specific number and type of comorbidity. To date, no studies have addressed comorbidity from this perspective. A wide range approach of EF confirms the need to further study comorbidity in ADHD from a wide range/variety perspective and determine all possible combinations (number/type) to clarify its contribution to the complex neuropsychology functioning in ADHD.

Asociación entre la longitud de los telómeros y deterioro cognitivo en adultos mayores / Association between telomere length and cognitive impairment in older adults

Introducción: El deterioro cognitivo es una etapa de transición entre el envejecimiento normal y la demencia; la prevalencia de este aumenta con la edad. El deterioro de las funciones e integridad física colocan al adulto mayor en una mayor susceptibilidad de enfermar. La longitud de los telómeros se considera un pilar del envejecimiento para caracterizar este fenotipo; así mismo, es un biomarcador que refleja el estado subyacente de la célula. En este trabajo se correlacionó la longitud relativa de los telómeros en adultos mayores con deterioro cognitivo. Material y métodos: Estudio observacional-analítico en muestras de pacientes de adultos mayores de 65 años sin deterioro cognitivo y con deterioro cognitivo, a los cuales se midió la longitud relativa de los telómeros. Resultados: Se incluyeron en el estudio 90 muestras de adultos mayores, y en el análisis de asociación de acuerdo a modelos logísticos multivariados el deterioro cognitivo mostró casi 5 veces más riesgo para acortamiento de los telómeros en relación con la presencia del diagnóstico de deterioro cognitivo (razón de momios 4,88, p=0,027). Conclusiones: Se realizó la correlación entre la longitud relativa de los telómeros en adultos mayores con diagnóstico de deterioro cognitivo, y se confirmó esta asociación para una menor longitud. (AU)

Introduction: Cognitive impairment is a transition stage between normal aging and dementia, the prevalence of last one increases with age; the damage of the functions and physical integrity, places the older adult in a greater susceptibility to get sick. Telomere length is a hallmark of aging to characterize this phenotype, as well as a biomarker that reflects the underlying state of the cell. In this work, the relative length of telomeres in older adults with cognitive impairment was correlated. Material and methods: Observational-analytical study, in samples of adult patients older than 65 years with and without cognitive impairment, in whom the relative length of telomeres was measured. Results: Ninety samples of older adults were included in the study and in the association analysis according to multivariate logistic models, cognitive impairment showed almost five times more risk for telomere shortening in relation to the presence of the diagnosis of cognitive impairment (Odds ratio 4.88, p=0.027). Conclusions: When correlating the relative length of telomeres in older adults diagnosed with cognitive impairment, this association was confirmed for shorter. (AU)

Single-Cell Infection of Influenza A Virus Using Drop-Based Microfluidics.

Drop-based microfluidics has revolutionized single-cell studies and can be applied toward analyzing tens of thousands to millions of single cells and their products contained within picoliter-sized drops. Drop-based microfluidics can shed insight into single-cell virology, enabling higher-resolution analysis of cellular and viral heterogeneity during viral infection. In this work, individual A549, MDCK, and siat7e cells were infected with influenza A virus (IAV) and encapsulated into 100-µm-size drops. Initial studies of uninfected cells encapsulated in drops demonstrated high cell viability and drop stability. Cell viability of uninfected cells in the drops remained above 75%, and the average drop radii changed by less than 3% following cell encapsulation and incubation over 24 h. Infection parameters were analyzed over 24 h from individually infected cells in drops. The number of IAV viral genomes and infectious viruses released from A549 and MDCK cells in drops was not significantly different from bulk infection as measured by reverse transcriptase quantitative PCR (RT-qPCR) and plaque assay. The application of drop-based microfluidics in this work expands the capacity to propagate IAV viruses and perform high-throughput analyses of individually infected cells. IMPORTANCE Drop-based microfluidics is a cutting-edge tool in single-cell research. Here, we used drop-based microfluidics to encapsulate thousands of individual cells infected with influenza A virus within picoliter-sized drops. Drop stability, cell loading, and cell viability were quantified from three different cell lines that support influenza A virus propagation. Similar levels of viral progeny as determined by RT-qPCR and plaque assay were observed from encapsulated cells in drops compared to bulk culture. This approach enables the ability to propagate influenza A virus from encapsulated cells, allowing for future high-throughput analysis of single host cell interactions in isolated microenvironments over the course of the viral life cycle.