Risk for Waterborne Transmission and Environmental Persistence of Avian Influenza Virus in a Wildlife/Domestic Interface in Mexico.

Aquatic habitats provide a bridge for influenza transmission among wild and domestic species. However, water sources pose highly variable physicochemical and ecological characteristics that affect avian influenza virus (AIV) stability. Therefore, the risk of survival or transmissibility of AIV in the environment is quite variable and has been understudied. In this study, we determine the risk of waterborne transmission and environmental persistence of AIV in a wild/domestic bird interface in the Central Mexico plateau (North America) during the winter season using a multi-criteria decision analysis (MCDA). A total of 13 eco-epidemiological factors were selected from public-access databases to develop the risk assessment. The MCDA showed that the Atarasquillo wetland presents a higher persistence risk in January. Likewise, most of the backyard poultry farms at this wild-domestic interface present a high persistence risk (50%). Our results suggest that drinking water may represent a more enabling environment for AIV persistence in contrast with wastewater. Moreover, almost all backyard poultry farms evidence a moderate or high risk of waterborne transmission especially farms close to water bodies. The wildlife/domestic bird interface on the Atarasquillo wetland holds eco-epidemiological factors such as the presence of farms in flood-prone areas, the poultry access to outdoor water, and the use of drinking-water troughs among multiple animal species that may enhance waterborne transmission of AIV. These findings highlight the relevance of understanding the influence of multiple factors on AIV ecology for early intervention and long-term control strategies.

Chemical transesterification of coconut and corn oils using different metal hydroxides as catalysts to determine the chemical and physiochemical changes to the oils.

BACKGROUND: The transesterification of butteroil has been shown to alter its lipid chemistry and thus alter the crystallization of the fat. The reaction kinetics and resulting crystallization of the butteroil differ depending on the nature of the catalyst used. Modeling the reaction with vegetable oils is a simpler method for the analysis of resulting products to understand the chemical and physiochemical changes that occur based on catalyst selection. The objective of this work is to perform a chemical transesterification of coconut and corn oil using monovalent and divalent catalysts to investigate the chemical and crystal changes that occur. RESULTS: Coconut and corn oil were subjected to chemical transesterification using both Ca(OH)2 and KOH as catalysts. In both the coconut and corn oil samples, transesterification caused monoglycerides (MAGs) and diacylglycerides (DAGs) to form from the most abundant fatty acid found in each sample. Coconut oil's melting temperature, solid fat content (SFC), and storage modulus decreased as a result of the transesterification, and crystals began to form in the corn oil causing melting thermograms to be evident, higher SFC, and a more viscous oil as a result. Using Ca(OH)2 as a catalyst resulted in more MAG formation, and a higher SFC and melting temperature than when KOH was used as a catalyst. CONCLUSION: The results demonstrate that the chemical changes that result from transesterification of plant-based oils change the crystallization behavior of the oils and can therefore be used for different applications in the food industry. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Trypanosoma cruzi in Wild and Synanthropic Mammals in Two Regions of Mexico: A Fieldwork and Genetic Discrete Typing Unit Review.

Background: Marsupials and rodents are the most important wild and synanthropic hosts of Trypanosoma cruzi due to the high frequency of infection, maintenance of diverse genetic populations of the parasite, and their close proximity to interact with both transmission cycles, sylvatic and peridomestic. Our aim was to identify the discrete typing units (DTU) of T. cruzi from different wild and synanthropic hosts in two regions of Mexico and to carry out a review of historical data focusing on current knowledge on the diversity and T. cruzi DTUs of host species. Materials and Methods: One hundred fifteen samples were obtained from two areas in Tabasco and Nayarit state. The presence of T. cruzi was evaluated by PCR. Results: The 12.6% (12/95) of samples from Tabasco and 65% (13/20) from Nayarit were found to be positive for parasite DNA. All the sequences analyzed were grouped in T. cruzi DTU I; low nucleotide diversity was observed in Tabasco (π = 0.00566, and Ï´ = 0.00632), while high genetic diversity was observed in Nayarit sequences, up to 8.63 (π) to 11.10 (Ï´) times greater than Tabasco sequences. Genetic flow and migration between Tabasco, and Nayarit were scarce (FST = 0.37329 and Nm = 0.42), and genetic exchange was observed only between nearby areas. The bibliographic review of hosts in Mexico, together with our data, shows a heterogeneous T. cruzi prevalence in Chiroptera and domestic animals. For Atelidae and Canids, prevalence is generally below 25%. However, a high prevalence, greater than 25% and up to 100%, was recorded in Didelphimorphia, and Rodentia. Few studies in regions of Mexico have been described as infected with the parasite; in these, the genetic group with the highest prevalence is the DTU I. Conclusion: Marsupials and rodents are important reservoirs of T. cruzi; DTU I was frequently reported; however, recent genetic and reservoir studies have demonstrated the presence of greater diversity of genetic groups.

Development of probiotic yogurt products incorporated with Lactobacillus kefiranofaciens OSU-BSGOA1 in mono- and co-culture with Kluyveromyces marxianus.

The bacterium Lactobacillus kefiranofaciens OSU-BDGOA1 and yeast Kluyveromyces marxianus bdgo-ym6 were previously isolated from kefir grains and have shown probiotic traits in mono- and coculture. This research evaluates the effect of introducing probiotic kefir microorganisms in monoculture and in coculture alongside yogurt starter cultures on the physicochemical and rheological properties, volatile flavor compounds, survival of the microorganisms during simulated digestion, and sensory attributes of the final fermented products. The incorporation of Lactobacillus kefiranofaciens OSU-BDGOA1 in monoculture showed promising outcomes, resulting in a final product showing more solid-like characteristics and potentially improving the texture of the product. There was also a significant increase in the concentration of desirable volatile flavor compounds in the yogurt with the monoculture, particularly 2,3-butanedione, displaying a positive correlation with buttery flavor in the sensory analysis. The inclusion of L. kefiranofaciens in monoculture also promoted better sensory attributes and was significantly better than the yogurt with the coculture with the yeast showing promising results for the incorporation of this probiotic bacterium into functional fermented dairy products.

Exploring Tau Fibril-Disaggregating and Antioxidating Molecules Binding to Membrane-Bound Amyloid Oligomers Using Machine Learning-Enhanced Docking and Molecular Dynamics.

Intracellular tau fibrils are sources of neurotoxicity and oxidative stress in Alzheimer's. Current drug discovery efforts have focused on molecules with tau fibril disaggregation and antioxidation functions. However, recent studies suggest that membrane-bound tau-containing oligomers (mTCOs), smaller and less ordered than tau fibrils, are neurotoxic in the early stage of Alzheimer's. Whether tau fibril-targeting molecules are effective against mTCOs is unknown. The binding of epigallocatechin-3-gallate (EGCG), CNS-11, and BHT-CNS-11 to in silico mTCOs and experimental tau fibrils was investigated using machine learning-enhanced docking and molecular dynamics simulations. EGCG and CNS-11 have tau fibril disaggregation functions, while the proposed BHT-CNS-11 has potential tau fibril disaggregation and antioxidation functions like EGCG. Our results suggest that the three molecules studied may also bind to mTCOs. The predicted binding probability of EGCG to mTCOs increases with the protein aggregate size. In contrast, the predicted probability of CNS-11 and BHT-CNS-11 binding to the dimeric mTCOs is higher than binding to the tetrameric mTCOs for the homo tau but not for the hetero tau-amylin oligomers. Our results also support the idea that anionic lipids may promote the binding of molecules to mTCOs. We conclude that tau fibril-disaggregating and antioxidating molecules may bind to mTCOs, and that mTCOs may also be useful targets for Alzheimer's drug design.

Preventative Effects of Milk Fat Globule Membrane Ingredients on DSS-Induced Mucosal Injury in Intestinal Epithelial Cells.

The goblet cells of the gastrointestinal tract (GIT) produce glycoproteins called mucins that form a protective barrier from digestive contents and external stimuli. Recent evidence suggests that the milk fat globule membrane (MFGM) and its milk phospholipid component (MPL) can benefit the GIT through improving barrier function. Our objective was to compare the effects of two digested MFGM ingredients with or without dextran sodium sulfate (DSS)-induced barrier stress on mucin proteins. Co-cultured Caco-2/HT29-MTX intestinal cells were treated with in vitro digests of 2%, 5%, and 10% (w/v) MFGM or MPL alone for 6 h or followed by challenge with 2.5% DSS (6 h). Transepithelial electrical resistance and fluorescein isothiocyanate (FITC)-dextran (FD4) permeability measurements were used to measure changes in barrier integrity. Mucin characterization was performed using a combination of slot blotting techniques for secreted (MUC5AC, MUC2) and transmembrane (MUC3A, MUC1) mucins, scanning electron microscopy (SEM), and periodic acid Schiff (PAS)/Alcian blue staining. Digested MFGM and MPL prevented a DSS-induced reduction in secreted mucins, which corresponded to the prevention of DSS-induced increases in FD4 permeability. SEM and PAS/Alcian blue staining showed similar visual trends for secreted mucin production. A predictive bioinformatic approach was also used to identify potential KEGG pathways involved in MFGM-mediated mucosal maintenance under colitis conditions. This preliminary in silico evidence, combined with our in vitro findings, suggests the role of MFGM in inducing repair and maintenance of the mucosal barrier.

Reduction in the antigenicity of beta-lactoglobulin in whole milk powder via supercritical CO2 treatment.

Cow milk allergy is a common phenomenon experienced in early childhood (<5 yr of age) with an average occurrence rate of roughly 2.5%. The most prevalent allergen in cow milk is believed to be ß-LG. The objective of this study was to evaluate the use of hydrophobic supercritical CO2 (ScCO2) to modify the chemical structure ß-LG, thus impairing its recognition by antibodies. Whole milk powder (WMP) was selected because of its closest compositional resemblance to bovine fluid milk and its applications in reconstitution and in the beverage (infant, toddler, and adult), confectionary, bakery, and meat industries. For this study, WMP was treated with food-grade CO2 at temperatures of 50, 63, and 75°C under operating pressures of 100, 150, 200, 250, and 300 bar. Proteins in WMP were examined using SDS-PAGE, western blot, and ELISA. Orbitrap Fusion liquid chromatography-tandem MS (LC-MS/MS) and periodic staining was performed to confirm post-translational modifications in ß-LG. Functional properties of WMP before and after treatment were assessed by its solubility index, oil holding capacity, emulsion capacity and stability, zeta potential, particle size, and color analysis. SDS-PAGE of treated samples yielded fuzzy bands (variable mobility of molecules due to different molecular weights results in ill-defined bands) indicative of an increase in molecular weight, presumably due to chemical change in the protein, and demonstrated a maximum of 71.13 ± 0.29% decrease in the band intensity of ß-LG under treatment conditions of 75°C/300 bar for 30 min. These changes were small with samples treated with heat only. Lighter, diffused bands were observed using western blot analysis. The ELISA tests proved that ScCO2 treatment specifically and significantly affected the antigenicity of ß-LG with a reduction of 42.9 ± 2.83% and 54.75 ± 2.43% at 63°C/200 bar and 75°C/300 bar, respectively. Orbitrap fusion detected the presence of fatty acids and sugar moieties bound to ß-LG and the latter was confirmed by periodic staining. Functional properties of ScCO2-treated milk powder yielded a decrease in solubility index and an increase in emulsion capacity of WMP was observed under ScCO2 treatment at 75°C/300 bar, with small and insignificant changes at other treatments producing a decrease in antigenicity. Color changes were small for most samples, except at 63°C/200 bar, where a significant increase in yellowness was observed. Zeta potential and particle size measurements indicated that most changes were temperature driven. This study demonstrates 2 approaches to mitigate ß-LG antigenicity via fatty acid binding and lactosylation using hydrophobic ScCO2.

Canine parvovirus type 2 (CPV-2) serological and molecular patterns in dogs with viral gastroenteritis from southern Brazil.

Canine Parvovirus type 2 (CPV-2) is a highly contagious virus that can cause severe systemic disease with gastroenteric symptoms in dogs, particularly in young puppies. Originating from the feline parvovirus in the late 1970s, it swiftly propagated globally, instigating a pandemic in dogs. Despite vaccination advancements, CPV-2 remains a substantial challenge for veterinary professionals and pet owners. This study aimed to contribute knowledge about the current situation of CPV-2 among dogs in southern Brazil. In this study, the sera of 125 dogs (mostly with gastroenteritis symptoms) were screened for antibodies against CPV-2 and their faeces for the virus itself. The results showed that 40% (50/125) of dogs were infected with CPV-2. Most animals (65.5%) had previously been exposed to CPV-2 (with serotitres equal or above 1:40), and only 37.6% had protective antibody titres equal or above 1:80. The findings have also demonstrated that vaccination against CPV-2 significantly reduced the risk of infection, with positive cases decreasing from 56.9% (unvaccinated) to 2.0% (fully vaccinated). Furthermore, the prevalence of CPV-2 decreased as dogs aged, with younger dogs and those with an incomplete or non-existent vaccination history at the highest risk of infection. In conclusion, this study provides valuable insight into the prevalence and risk factors associated with CPV-2 infection in dogs in southern Brazil, thereby providing valuable knowledge for the improvement of veterinary care and pet health.

Evaluation of kefir grain microbiota, grain viability, and bioactivity from fermenting dairy processing by-products.

Four dairy foods processing by-products (acid whey permeate [AWP], buttermilk [BM], sweet whey permeate [SWP], and sweet whey permeate with added milk fat globule ingredient [SWP+MFGM]) were fermented for 4 wk and compared with traditional kefir milks for production of novel kefir-like dairy products. Sweet whey permeates and SWP supplemented with 1.5% milk fat globule membrane (MFGM) proved to be the most viable by-products for kefir grain fermentation, exhibiting diverse abundance of traditional kefir microorganisms and positive indicators of bioactive properties. Grain viability was assessed with shotgun metagenomics, texture profile analysis, live cell counts, and scanning electron microscopy. Assessed bioactivities of the kefir-like products included antibacterial, antioxidant, potential anticancerogenic properties, and membrane barrier effects on human colorectal adenocarcinoma Caco-2 cells. All kefir grains were most abundant in Lactobacillus kefiranofaciens when analyzed with shotgun metagenomics. When analyzed with live cell counts on selective media, AWP kefir-like product had no countable Lactococcus spp., indicating suboptimal conditions for kefir grain microbiota survival and application for fermented dairy starter culture bacterium. Live cell counts were affirmed with kefir grain surface scanning electron microscopy images. The SWP treatment had the most adhesive kefir grain surface, and SWP+MFGM had the largest exopolysaccharide yield from grain extraction. All kefir and kefir-like products were able to achieve a 6-log reduction against Listeria innocua and Escherichia coli. Traditional milk kefirs had the highest antioxidant capacity for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid; ABTS) assay. The AWP formulation had a significantly higher DPPH antioxidant activity compared with the other kefir and kefir-like products, and SWP had the lowest Trolox equivalence concentration in the ABTS assay. Sweet whey and supplemented milk fat sweet whey had upregulation of Cldn-1 and Ocln-1 gene expression, which correspond with a significant increase in transepithelial electrical resistance.

Tonic noradrenergic input to neurons in the dorsal raphe nucleus mediates food intake in male mice.

The dorsal raphe nucleus (DRN) is essential for the control of food intake. Efferent projections from the DRN extend to several forebrain regions that are involved in the control of food intake. However, the neurotransmitters released in the DRN related to the control of food intake are not known. We have previously demonstrated that a tonic α1 action on DRN neurons contributes to satiety in the fed rats. In this study we investigated the participation of norepinephrine (NE) signaling in the DRN in the satiety response. Intra-DRN administration of NE causes an increase in the 2-hour food intake of sated mice, an effect that was blocked by previous administration of yohimbine, an α2 antagonist. Similarly, Intra-DRN administration of clonidine, an α2 agonist, increases food intake in sated mice. This result indicates that in the satiated mice exogenous NE acts on α2 receptors to increase food intake. Furthermore, administration of phenylephrine, an α1 agonist, decreases food intake in fasted mice and prazosin, an α1 antagonist, increases food intake in the sated mice. Taken together these results indicate that, in a satiated condition, a tonic α1 adrenergic action on the DRN neurons inhibits food intake and that exogenous NE administered to the DRN acts on α2 adrenergic receptors to increase food intake. These data reinforce the intricate neuronal functioning of the DRN and its effects on feeding.

Failure to repair endogenous DNA damage in ß-cells causes adult-onset diabetes in mice.

Age is the greatest risk factor for the development of type 2 diabetes mellitus (T2DM). Age-related decline in organ function is attributed to the accumulation of stochastic damage, including damage to the nuclear genome. Islets of T2DM patients display increased levels of DNA damage. However, whether this is a cause or consequence of the disease has not been elucidated. Here, we asked if spontaneous, endogenous DNA damage in ß-cells can drive ß-cell dysfunction and diabetes, via deletion of Ercc1, a key DNA repair gene, in ß-cells. Mice harboring Ercc1-deficient ß-cells developed adult-onset diabetes as demonstrated by increased random and fasted blood glucose levels, impaired glucose tolerance, and reduced insulin secretion. The inability to repair endogenous DNA damage led to an increase in oxidative DNA damage and apoptosis in ß-cells and a significant loss of ß-cell mass. Using electron microscopy, we identified ß-cells in clear distress that showed an increased cell size, enlarged nuclear size, reduced number of mature insulin granules, and decreased number of mitochondria. Some ß-cells were more affected than others consistent with the stochastic nature of spontaneous DNA damage. Ercc1-deficiency in ß-cells also resulted in loss of ß-cell function as glucose-stimulated insulin secretion and mitochondrial function were impaired in islets isolated from mice harboring Ercc1-deficient ß-cells. These data reveal that unrepaired endogenous DNA damage is sufficient to drive ß-cell dysfunction and provide a mechanism by which age increases the risk of T2DM.

High-Performance 3D Nanostructured Silver Electrode for Micro-Supercapacitor Application.

In the current energy crisis scenario, the development of renewable energy forms such as energy storage systems among the supercapacitors is an urgent need as a tool for environmental protection against increasing pollution. In this work, we have designed a novel 3D nanostructured silver electrode through an antireplica/replica template-assisted procedure. The chemical surface and electrochemical properties of this novel 3D electrode have been studied in a 5 M KOH electrolyte. Microstructural characterization and compositional analysis were studied by SEM, energy-dispersive X-ray spectroscopy, XRD technique, and Kripton adsorption at -198 °C, together with cyclic voltammetry and galvanostatic charge-discharge cycling measurements, Coulombic efficiency, cycle stability, and their leakage current drops, in addition to the self-discharge and electrochromoactive behavior, were performed to fully characterize the 3D nanostructured electrode. Large areal capacitance value of 0.5 F/cm2 and Coulombic efficiency of 97.5% are obtained at a current density of 6.4 mA/cm2 for a voltage window of 1.2 V (between -0.5 and 0.8 V). The 3D nanostructured silver electrode exhibits excellent capacitance retention (95%) during more than 2600 cycles, indicating a good cyclic stability. Additionally, the electrode delivers a high energy density of around 385.87 µWh/cm2 and a power density value of 3.82 µW/cm2 and also displays an electrochromoactive behavior. These experimental results strongly support that this versatile combined fabrication procedure is a suitable strategy for improving the electrochemical performances of 3D nanostructured silver electrodes for applications as micro-supercapacitors or in electrochemical devices.

[Depression in health personnel associated with exposure to COVID-19]. / Depresión en personal de salud asociado a la exposición de la COVID-19.

Background: There are many working factors to do with depression. Objective: To determine the association between the exposure to COVID-19 and depression in physicians and nurses from the four hospitals at "Centro Médico Nacional Siglo XXI" (CMN SXXI) took part in: Oncology, Specialties, Cardiology and Pediatrics. Material and Methods: A cross-sectional study of 856 participants took place in January 2022, excluding workers, such as physicians and nurses from the CMN SXXI, disabled workers, staff with a union agreement, support staff and/or staff with less than one year of work labour. Through a self-administered questionnaire, the degree of occupational exposure to patients care with COVID-19, a history of COVID-19 infection, and the patient's health quiz were asked (PHQ-9). The analysis included simple frequency measurements, odds ratio (OR), Chi squared and multiple logistics regression with p ≤ 0.05. Results: The prevalence of depression in the medical and nursing staff at CMN SXXI was 32.2% (moderate to severe degree); In the multiple regression, an association was identified with not identifying support by the Institute (OR: 1.60, CI95%: 1.08-2.39, p = 0.02), high occupational exposure (OR: 8.35, CI95%: 3.02-23.09, p ˂ 0.0001), and more than 5 working days a week serving the COVID-19 patients (OR: 2.51, CI95%: 1.35-4.49, p = 0.004) and as a protective factor the fact that they have never had the COVID-19 (OR: 0.61, CI95%: 0.43-0.86, p = 0.01). Conclusions: The prevalence of depression was higher than expected being associated with the degree of occupational exposure in the COVID-19 patients´ assistance.

Introducción: existen factores laborales que influyen en la presencia de la depresión. Objetivo: determinar la asociación entre la exposición a la COVID-19 y la depresión en médicos y enfermeras de los cuatro hospitales del Centro Médico Nacional Siglo XXI (CMN SXXI): Oncología, Especialidades, Cardiología y Pediatría. Material y métodos: se realizó un estudio transversal, en enero del 2022, con 870 participantes pertenecientes a la población de médicos y enfermeras que laboraban en el CMN SXXI, se excluyó a los trabajadores que se encontraban con incapacidad, al personal temporal y/o con menos de un año de antigüedad laboral. A través de un cuestionario autoaplicable se interrogó sobre el grado de exposición laboral a la atención de pacientes con COVID-19, antecedentes de infección por la COVID-19 y el cuestionario de salud del paciente (PHQ-9). El análisis incluyo medidas de frecuencia simple y razón de Momios (RM), Chi cuadrada y regresión logística múltiple con p ≤ 0.05. Resultados: la prevalencia de depresión en personal médico y de enfermería del CMN SXXI fue del 32.2% (grado moderado a severo). En la regresión múltiple se encontró asociación con no identificar apoyo por parte del Instituto (RM: 1.60, IC95%: 1.08-2.39, p = 0.02), alta exposición laboral (RM: 8.35, IC95%: 3.02-23.09, p ˂ 0.0001) y más de 5 días laborados a la semana atendiendo pacientes con la COVID-19 (RM: 2.51, IC95%: 1.35-4.49, p = 0.004) y como factor protector el que nunca hayan enfermado de la COVID-19 (RM: 0.61, IC95%: 0.43-0.86, p = 0.01). Conclusiones: la prevalencia de depresión fue mayor a la esperada y se asoció con el grado de exposición laboral en la atención de pacientes con la COVID-19.

Milk phospholipids protect Bifidobacterium longum subsp. infantis during in vitro digestion and enhance polysaccharide production.

Bifidobacterium longum subsp. infantis is associated with the gut microbiota of breast-fed infants. Bifidobacterium infantis promotes intestinal barrier and immune function through several proposed mechanisms, including interactions between their surface polysaccharides, the host, and other gut microorganisms. Dairy foods and ingredients are some of the most conspicuous food-based niches for this species and may provide benefits for their delivery and efficacy in the gut. Milk phospholipid (MPL)-rich ingredients have been increasingly recognized for their versatile benefits to health, including interactions with the gut microbiota and intestinal cells. Therefore, our objective was to investigate the capacity for MPL to promote survival of B. infantis during simulated digestion and to modulate bacterial polysaccharide production. To achieve these aims, B. infantis was incubated with or without 0.5% MPL in de Man, Rogosa, and Sharpe (MRS) media at 37°C under anaerobiosis. Survival across the oral, gastric, and intestinal phases using in vitro digestion was measured using plate count, along with adhesion to goblet-like intestinal cells. MPL increased B. infantis survival at the end of the intestinal phase by at least 7% and decreased adhesion to intestinal cells. The bacterial surface characteristics, which may contribute to these effects, were assessed by ζ-potential, changes in surface proteins using comparative proteomics, and production of bound polysaccharides. MPL decreased the surface charge of the bifidobacteria from -17 to -24 mV and increased a 50 kDa protein (3-fold) that appears to be involved in protection from stress. The production of bound polysaccharides was measured using FTIR, HPLC, and TEM imaging. These techniques all suggest an increase in bound polysaccharide production at least 1.7-fold in the presence of MPL. Our results show that MPL treatment increases B. infantis survival during simulated digestion, induces a stress resistance surface protein, and yields greater bound polysaccharide production, suggesting its use as a functional ingredient to enhance probiotic and postbiotic effects.

Phenylacetyl-/Trolox- Amides: Synthesis, Sigma-1, HDAC-6, and Antioxidant Activities.

In search of novel multi-mechanistic approaches for treating Alzheimer's disease (AD), we have embarked on synthesizing single small molecules for probing contributory roles of the following combined disease targets: sigma-1 (σ-1), class IIb histone deacetylase-6 (HDAC-6), and oxidative stress (OS). Herein, we report the synthesis and partial evaluation of 20 amides (i.e., phenylacetic and Trolox or 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid derivatives). Target compounds were conveniently synthesized via amidation by either directly reacting acyl chlorides with amines or condensing acids with amines in the presence of coupling agents 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo [4,5-b] pyridinium 3-oxide hexafluorophosphate (HATU) or 1,1'-carbonyldiimidazole (CDI). Overall, this project afforded compound 8 as a promising lead with σ-1 affinity (Ki = 2.1 µM), HDAC-6 (IC50 = 17 nM), and antioxidant (1.92 Trolox antioxidant equivalents or TEs) activities for optimization in ensuing structure-activity relationship (SAR) studies.

Invited review: Properties of ß-galactosidases derived from Lactobacillaceae species and their capacity for galacto-oligosaccharide production.

ß-galactosidase (enzymatic class 3.2.1.23) is one of the dairy industry's most important and widely used enzymes. The enzyme is part of a large family known to catalyze hydrolysis and transglycosylation reactions. Its hydrolytic activity is commonly used to decrease lactose content in dairy products, while its transglycosylase activity has recently been used to synthesize galacto-oligosaccharides (GOS). During the past couple of years, researchers have focused on studying ß-galactosidase isolated and purified from lactic acid bacteria. This review will focus on ß-galactosidase purified and characterized from what used to be the Lactobacillus genera. Furthermore, particular emphasis is given to its kinetics, biochemical characteristics, GOS production, market, and utilization by Lactobacilllaceae species.

Partition of milk phospholipids during ice cream manufacturing.

The distribution of phospholipids (PL) within the fat and serum phase of ice cream manufacturing was evaluated through partition coefficients (KPL) after mixing, pasteurization, freezing, and hardening. Ice creams containing about 40.41 ± 3.45 (± standard deviation; control formulation) and 112.29 ± 9.06 (enriched PL formulation) mg of PL per g of fat were formulated with nonfat dry milk and ß-serum, respectively. Overall, the KPL were lower than 1, indicating that the PL were predominantly found in the fat phase, and only a small amount was left in the serum and sediment. Confocal micrographs visually confirmed this generalization. The addition of PL significantly increased the viscosity of the mixes between 4- and 9-fold, depending on the shear rate. Additionally, mixes containing high PL exhibited higher yield stress than those formulated with low PL (0.15 ± 0.09 and 0.016 ± 0.08 Pa, respectively). Ice creams with high PL delayed the onset of meltdown and exhibited a slower rate of a meltdown than low-PL ice creams (18.53 ± 0.57 and 14.83 ± 0.85 min, and 1.01 ± 0.05 and 0.71 ± 0.04% min-1, respectively). This study provides useful guidelines for manufacturing ice cream enriched in milk PL. Additionally, the use of ß-serum, a byproduct stream, as a source of PL is illustrated. The development will require studying the sensorial description of the product as well as consumer acceptance.

Enhanced probiotic potential of Lactobacillus kefiranofaciens OSU-BDGOA1 through co-culture with Kluyveromyces marxianus bdgo-ym6.

Introduction: Due to the increasing consumer demand for the development and improvement of functional foods containing probiotics, new probiotic candidates need to be explored as well as novel means to enhance their beneficial effects. Lactobacillus kefiranofaciens OSU-BDGOA1 is a strain isolated from kefir grains that has demonstrated probiotic traits. This species is the main inhabitant of kefir grains and is responsible for the production of an exopolysaccharide (EPS) whit vast technological applications and potential bioactivities. Research has shown that interkingdom interactions of yeast and lactic acid bacteria can enhance metabolic activities and promote resistance to environmental stressors. Methods: Comparative genomic analyses were performed to distinguish OSU-BDGOA1 from other strains of the same species, and the genome was mined to provide molecular evidence for relevant probiotic properties. We further assessed the cumulative effect on the probiotic properties of OSU-BDGOA1 and Kluyveromyces marxianus bdgo-ym6 yeast co-culture compared to monocultures. Results: Survival during simulated digestion assessed by the INFOGEST digestion model showed higher survival of OSU-BDGOA1 and bdgo-ym6 in co-culture. The adhesion to intestinal cells assessed with the Caco-2 intestinal cell model revealed enhanced adhesion of OSU-BDGOA1 in co-culture. The observed increase in survival during digestion could be associated with the increased production of EPS during the late exponential and early stationary phases of co-culture that, by enhancing co-aggregation between the yeast and the bacterium, protects the microorganisms from severe gastrointestinal conditions as observed by SEM images. Immune modulation and barrier function for recovery and prevention of flagellin-mediated inflammation by Salmonella Typhimurium heat-killed cells (HKSC) in Caco-2 cells were also measured. OSU-BDGOA1 in mono- and co-culture regulated inflammation through downregulation of pro-inflammatory cytokine expression and increased membrane barrier integrity assessed by TEER, FD4 permeability, and expression of tight junctions. Discussion: The results of the study warrant further research into the application of co-cultures of yeast and LAB in functional probiotic products and the potential to increase EPS production by co-culture strategies.

Draft genome sequence of Lactobacillus helveticus OSU-BDGOAK2 and Lactobacillus kefiranofaciens OSU-BDGOA1, kefir grain isolates with potential antibacterial activity.

We present the draft genome sequence and assembly of Lactobacillus helveticus OSU-BDGOAK2 and Lactobacillus kefiranofaciens OSU-BDGOA1 isolated from kefir grains that exhibited in vitro antibacterial activity against Escherichia coli ATCC 25922, Listeria innocua ATCC 51742, and Staphylococcus epidermidis ATCC 1222. Genome analysis of both strains revealed gene clusters encoding bacteriocins.

Milk-Fat-Globule-Membrane-Enriched Dairy Milk Compared with a Soy-Lecithin-Enriched Beverage Did Not Adversely Affect Endotoxemia or Biomarkers of Gut Barrier Function and Cardiometabolic Risk in Adults with Metabolic Syndrome: A Randomized Controlled Crossover Trial.

Full-fat dairy milk may protect against cardiometabolic disorders, due to the milk fat globule membrane (MFGM), through anti-inflammatory and gut-health-promoting activities. We hypothesized that a MFGM-enriched milk beverage (MEB) would alleviate metabolic endotoxemia in metabolic syndrome (MetS) persons by improving gut barrier function and glucose tolerance. In a randomized crossover trial, MetS persons consumed for two-week period a controlled diet with MEB (2.3 g/d milk phospholipids) or a comparator beverage (COMP) formulated with soy phospholipid and palm/coconut oil. They then provided fasting blood and completed a high-fat/high-carbohydrate test meal challenge for evaluating postprandial metabolism and intestinal permeability. Participants had no adverse effects and achieved high compliance, and there were no between-trial differences in dietary intakes. Compared with COMP, fasting endotoxin, glucose, incretins, and triglyceride were unaffected by MEB. The meal challenge increased postprandial endotoxin, triglyceride, and incretins, but were unaffected by MEB. Insulin sensitivity; fecal calprotectin, myeloperoxidase, and short-chain fatty acids; and small intestinal and colonic permeability were also unaffected by MEB. This short-term study demonstrates that controlled administration of MEB in MetS persons does not affect gut barrier function, glucose tolerance, and other cardiometabolic health biomarkers, which contradicts observational evidence that full-fat milk heightens cardiometabolic risk. Registered at ClinicalTrials.gov (NCT03860584).