Microbial aromatic amino acid metabolism is modifiable in fermented food matrices to promote bioactivity.

Ingestion of fermented foods impacts human immune function, yet the bioactive food components underlying these effects are not understood. Here, we interrogated whether fermented food bioactivity relates to microbial metabolites derived from aromatic amino acids, termed aryl-lactates. Using targeted metabolomics, we established the presence of aryl-lactates in commercially available fermented foods. After pinpointing fermented food-associated lactic acid bacteria that produce high levels of aryl-lactates, we identified fermentation conditions to increase aryl-lactate production in food matrices up to 5 × 103 fold vs. standard fermentation conditions. Using ex vivo reporter assays, we found that food matrix conditions optimized for aryl-lactate production exhibited enhanced agonist activity for the human aryl-hydrocarbon receptor (AhR) as compared to standard fermentation conditions and commercial products. Reduced microbial-induced AhR activity has emerged as a hallmark of many chronic inflammatory diseases, thus we envision strategies to enhance AhR bioactivity of fermented foods to be leveraged to improve human health.

Honey Added to Yogurt with Bifidobacterium animalis subsp. lactis DN-173 010/CNCM I-2494 Supports Probiotic Enrichment but Does Not Reduce Intestinal Transit Time in Healthy Adults: A Randomized, Controlled, Crossover Trial.

BACKGROUND: Honey improves probiotic survival in vitro. However, if this effect translates to humans has not been investigated. OBJECTIVES: We aimed to determine effects of honey + yogurt containing the probiotic Bifidobacterium animalis subsp. lactis DN-173 010/CNCM I-2494 (B. animalis) on intestinal transit time, probiotic enrichment, digestive health, mood, and cognition in adults. METHODS: Sixty-six healthy adults (34 female; 33.6±9.8 y; 24.6±3.0 kg/m2) in a crossover trial were randomized to 2-week yogurt conditions in a counterbalanced order with > 4-week washout: 1) Honey (HON): yogurt + honey and 2) Negative Control (NC): heat-treated yogurt + sugar. N=62 completed the trial and n=37 (17 female; 32.0±8.3 y; 25.0±2.9 kg/m2) elected to enroll in a third condition (a non-randomized study extension) after ≥ 4-week washout with a reference Positive Control (PC): yogurt + sugar. At baseline and end of each of the 3 conditions, intestinal transit time was measured with dye capsules; probiotic abundance with fecal sample 16S sequencing; digestive health with symptom/function records, Bristol stool consistency, Gastrointestinal Tolerability, and Gastrointestinal Quality of Life Index (GIQLI); mood with Positive and Negative Affect Schedule-Short Form (PANAS-SF), Depression Anxiety Stress Scales-42 (DASS-42), Patient-Reported Outcomes Measurement Information System (PROMIS) questionnaires, and emotional image task; and cognition with spatial reconstruction task. Data were analyzed using linear mixed-effects models (LMM) with significance at P ≤ 0.05. Baseline and end data were included in the LMM, with fixed effects being treatment, time, treatment by time interaction, and baseline covariate, and random effect being the participant. RESULTS: B. animalis was enriched in HON (d=3.54, P=0.0002) compared to controls with Linear Discriminant Analysis Effect Size (LEfSe). Intestinal transit time, gastrointestinal health, mood, and cognition did not differ between conditions (LMM: Ps>0.05). CONCLUSIONS: Yogurt + honey enriched B. animalis but did not reduce intestinal transit time or have other functional gastrointestinal, mood, or cognitive effects in adults. CLINICAL TRIAL REGISTRY: NCT04187950 and NCT04901390, www. CLINICALTRIALS: gov.

Fully Human Bifunctional Intrabodies Achieve Graded Reduction of Intracellular Tau and Rescue Survival of MAPT Mutation iPSC-derived Neurons.

Tau protein aggregation is a hallmark of several neurodegenerative diseases, including Alzheimer's disease, frontotemporal dementia (FTD) and progressive supranuclear palsy (PSP), spurring development of tau-lowering therapeutic strategies. Here, we report fully human bifunctional anti-tau-PEST intrabodies that bind the mid-domain of tau to block aggregation and degrade tau via the proteasome using the ornithine decarboxylase (ODC) PEST degron. They effectively reduced tau protein in human iPSC-derived cortical neurons in 2D cultures and 3D organoids, including those with the disease-associated tau mutations R5L, N279K, R406W, and V337M. Anti-tau-hPEST intrabodies facilitated efficient ubiquitin-independent proteolysis, in contrast to tau-lowering approaches that rely on the cell's ubiquitination system. Importantly, they counteracted the proteasome impairment observed in V337M patient-derived cortical neurons and significantly improved neuronal survival. By serial mutagenesis, we created variants of the PEST degron that achieved graded levels of tau reduction. Moderate reduction was as effective as high reduction against tau V337M-induced neural cell death.

Effects of 2'-fucosyllactose on the viability of starter cultures and Bifidobacterium strains of human origin in yogurt during refrigerated storage.

2'-Fucosyllactose (2'-FL) is postulated to provide health benefits and promote the growth of probiotics. This work was undertaken to study the effects of 2'-FL on the viability of starter cultures and Bifidobacterium strains of human origin in yogurt during refrigerated storage. Yogurts were produced containing 2'-FL (0 or 2 g/L) and Bifidobacterium strains of human origin (Bifidobacterium longum subsp. longum BB536 or Bifidobacterium longum subsp. infantis ATCC 15697) at a concentration of at least 109 CFU/mL. All yogurts were stored at 4°C for 5 weeks. Results showed that 2'-FL was stable in yogurts for at least 5 weeks of cold storage, and the addition of 2'-FL did not significantly alter yogurt fermentation parameters, associated metabolites, and the viability of mixed yogurt starter cultures and Bifidobacterium strains (p > 0.05). The addition of bifidobacteria had a negative impact (p < 0.05) on the survival rate of starter cultures, Streptococcus thermophilus and Lactobacillus delbureckii subsp. bulgaricus. Meanwhile, it is difficult to maintain a high survival rate of bifidobacteria in final yogurt products, and the addition of 2'-FL could not enhance the viability of bifidobacteria. B. longum BB536 survived at a level higher than 106 CFU/g for 28 days, while B. infantis ATCC15697 maintained this level for only 7 days. In summary, this study has shown the impact of 2'-FL and bifidobacterial species on yogurt properties, and results suggest that it is promising to use 2'-FL in yogurt products as a prebiotic. PRACTICAL APPLICATION: Yogurt is known for its beneficial effects on human health and nutrition. This study reported the production of symbiotic yogurt containing bifidobacteria and 2'-fucosyllactose (2'-FL) as a functional food for specified health uses. The viability of yogurt starter cultures and probiotic bifidobacterial strains was analyzed in this study. Moreover, this research demonstrated that 2'-FL could be added to yogurt without affecting the characteristics of yogurt significantly.

Female-Specific Health Care of Military Female-Designated Service Members and Veterans: A Systematic Overview of Reviews.

INTRODUCTION: Since the War in Afghanistan began in 2001, service members have faced significant health effects related to service during war, with female-designated service members facing unique challenges. Numerous high-quality review articles have been published on the health and care of female-designated service members and veterans. Given the increasing volume of literature, we completed an overview of reviews on the health and health care of female-designated military populations. Our objective was to conduct an overview of reviews on the obstetrics and gynecologic health and health care of female-designated military populations since 2000 to understand female-specific health consequences of military service during war and make clinical recommendations. MATERIALS AND METHODS: On May 10, 2022, a medical librarian performed a comprehensive search across five databases (Ovid Medline, Embase, CINAHL, PsycINFO, Ovid All EBM Reviews, and Web of Science) for all relevant reviews published from 2000 to May 10, 2022. Results were limited to English language. After the removal of duplicates, 2,438 records were reviewed, and 69 studies were included in the final review. The search strategy and methods were registered with PROSPERO and are reported according to the Preferred Reporting Items for Overviews of Reviews (PRIOR) guidelines. Two independent reviewers conducted title and abstract screening and subsequent full text review using Covidence Systematic Review Software. Reviews addressing female-specific and obstetrics and gynecologic health of female-designated service members or veterans, utilizing a clear and systematic methodology, were eligible for inclusion. Quality assessment was conducted by teams of two reviewers. RESULTS: A total of 69 studies were included in the final review. Themes included mental health and impact of sexual assault on service members or veterans, veteran health care, issues of menstruation, pregnancy, and urogenital concerns. Areas with few reviews included occupational risks of military service and impact on obstetric outcomes, eating disorders, and menopause. There were insufficient or no reviews on the impact of military service on fertility, access to abortion care, reproductive health outcomes of lesbian, bisexual and transgender service members, surgical treatment of gynecologic conditions, and screening and treatment for breast, gynecologic, and non-pelvic organ cancers. CONCLUSIONS: Female-designated military populations serving during periods of war face unique health challenges that should be considered in screening practices and the delivery of trauma informed care. Further research and reviews are needed for female-specific oncology, fertility, abortion access, and sexual and non-binary and expansive gender identities to better capture female-designated service member and veteran health during wartime and beyond.

Precision fermentation for improving the quality, flavor, safety, and sustainability of foods.

Precision fermentation involves the rewiring of metabolic pathways in generally recognized as safe microorganisms, fermentation scale-up, and downstream processing to produce food ingredients from abundant and inexpensive substrates. Using precise genome editing of food-fermenting microorganisms, precision fermentation can also produce fermented foods with more desirable properties. These genetic tools allow for the manipulation of flavors and nutritional content in fermented foods, the economic production of functional food ingredients, and the sustainable production of otherwise-costly macronutrients. By introducing the metabolic designs, genetic modifications, and resulting products of engineered microorganisms developed through academic and industrial research, this review aims to provide insights into the potentials and challenges of precision fermentation for the economic, safe, and sustainable production of foods.

Honey Varietals Differentially Impact Bifidobacterium animalis ssp. lactis Survivability in Yogurt through Simulated In Vitro Digestion.

BACKGROUND: Bifidobacterium animalis ssp. lactis DN-173 010/CNCM I-2494 (B. animalis) is a probiotic strain commonly added to yogurt. Yogurt and honey are a popular culinary pairing. Honey improves bifidobacteria survival in vitro. However, probiotic survival in yogurt with honey during in vitro digestion has not been investigated. OBJECTIVES: The study aimed to evaluate the effects of different honey varietals and concentrations on B. animalis survivability in yogurt through in vitro digestion. METHODS: Yogurt with honey or control-treated samples underwent in vitro simulated oral, gastric, and intestinal digestion. B. animalis cells were enumerated on de Man Rogosa and Sharpe (MRS) medium followed by an overlay with a modified selective MRS medium; all underwent anaerobic incubation. B. animalis were enumerated predigestion and after oral, gastric, and intestinal digestion. There were 2 study phases: Phase 1 tested 4 honey varietals at 20% wt/wt per 170 g yogurt, and Phase 2 tested 7 dosages of clover honey (20, 14, 10, 9, 8, 6, and 4% wt/wt) per 170 g yogurt. RESULTS: Similar B. animalis counts were observed between all treatments after oral and gastric digestion (<1 Log colony forming units (CFU)/g probiotic reduction). Higher B. animalis survivability was observed in yogurt with clover honey after exposure to simulated intestinal fluids (∼3.5 Log CFU/g reduction; P < 0.05) compared to all control treatments (∼5.5 Log CFU/g reduction; P < 0.05). Yogurt with 10-20% wt/wt clover honey increased B. animalis survivability after simulated in vitro digestion (≤ ∼4.7 Log CFU/g survival; P < 0.05). CONCLUSIONS: Yogurt with added honey improves probiotic survivability during in vitro digestion. The effective dose of clover honey in yogurt was 10-20% wt/wt per serving (1-2 tablespoons per 170 g yogurt) for increased probiotic survivability during in vitro digestion.

Fates of attached E. coli o157:h7 on intact leaf surfaces revealed leafy green susceptibility.

Leafy greens, especially lettuce, are repeatedly linked to foodborne outbreaks. This paper studied the susceptibility of different leafy greens to human pathogens. Five commonly consumed leafy greens, including romaine lettuce, green-leaf lettuce, baby spinach, kale, and collard, were selected by their outbreak frequencies. The behavior of E. coli O157:H7 87-23 on intact leaf surfaces and in their lysates was investigated. Bacterial attachment was positively correlated with leaf surface roughness and affected by the epicuticular wax composition. At room temperature, E. coli O157:H7 had the best growth potentials on romaine and green-leaf lettuce surfaces. The bacterial growth was positively correlated with stomata size and affected by epicuticular wax compositions. At 37 °C, E. coli O157:H7 87-23 was largely inhibited by spinach and collard lysates, and it became undetectable in kale lysate after 24 h of incubation. Kale and collard lysates also delayed or partially inhibited the bacterial growth in TSB and lettuce lysate at 37 °C, and they sharply reduced the E. coli O157:H7 population on green leaf lettuce at 4 °C. In summary, the susceptibility of leafy greens to E. coli O157:H7 is determined by a produce-specific combination of physiochemical properties and temperature.

Medication Safety Counseling Practices of Pediatric Primary Care Clinicians.

Medication exposures and poisonings are a major cause of pediatric morbidity and mortality. Unsafe patient practices are well documented despite the American Academy of Pediatrics recommending that pediatric primary care clinicians discuss medication safety with patients. Current clinician counseling practices for pediatric patients are unknown. Studies of adult patients suggest that physician counseling practices often focus on administration but not storage or disposal. To address this gap, we administered a web-based survey to clinically active pediatric primary care clinicians in two mid-Atlantic health care systems. Survey content focused on characteristics of medication safety counseling practices by age group, including safe medication storage, administration, and disposal. Of 151 clinicians emailed, 40 (26.5%) responded. The majority were physicians (93.5%), female (87.1%), and completed residency/clinical training in pediatrics >15 years ago (58.1%). Most (82.5%) reported having >1 pediatric patient (aged < 19 years) in their practice who experienced an unintentional or intentional medication exposure or poisoning event. Reported practices for medication safety counseling often varied by patient age but safe disposal was rarely addressed for any age group. Respondents generally felt less knowledgeable and less comfortable with providing counseling on safe disposal in comparison to safe storage and safe administration. Nearly all respondents (97%) would like to provide more counseling about medication safety, and the majority (81.3%) wanted additional educational resources. In this survey, we identified several modifiable deficits in pediatric medical counseling practices and a need for additional clinician training and resources, most notably in the content area of safe disposal.

Human cytomegalovirus modulates mTORC1 to redirect mRNA translation within quiescently infected monocytes.

Human cytomegalovirus (HCMV) utilizes peripheral blood monocytes as a means to systemically disseminate throughout the host. Following viral entry, HCMV stimulates non-canonical Akt signaling leading to the activation of mTORC1 and the subsequent translation of select antiapoptotic proteins within infected monocytes. However, the full extent to which the HCMV-initiated Akt/mTORC1 signaling axis reshapes the monocyte translatome is unclear. We found HCMV entry alone was able to stimulate widescale changes to mRNA translation levels and that inhibition of mTOR, a component of mTORC1, dramatically attenuated HCMV-induced protein synthesis. Although monocytes treated with normal myeloid growth factors also exhibited increased levels of translation, mTOR inhibition had no effect, suggesting HCMV activation of mTOR stimulates the acquisition of a unique translatome within infected monocytes. Indeed, polyribosomal profiling of HCMV-infected monocytes identified distinct prosurvival transcripts that were preferentially loaded with ribosomes when compared to growth factor-treated cells. Sirtuin 1 (SIRT1), a deacetylase that exerts prosurvival effects through regulation of the PI3K/Akt pathway, was found to be highly enriched following HCMV infection in an mTOR-dependent manner. Importantly, SIRT1 inhibition led to the death of HCMV-infected monocytes while having minimal effect on uninfected cells. SIRT1 also supported a positive feedback loop to sustain Akt/mTORC1 signaling following viral entry. Taken together, HCMV profoundly reshapes mRNA translation in an mTOR-dependent manner to enhance the synthesis of select factors necessary for the survival of infected monocytes.IMPORTANCEHuman cytomegalovirus (HCMV) infection is a significant cause of morbidity and mortality among the immunonaïve and immunocompromised. Peripheral blood monocytes are a major cell type responsible for disseminating the virus from the initial site of infection. In order for monocytes to mediate viral spread within the host, HCMV must subvert the naturally short lifespan of these cells. In this study, we performed polysomal profiling analysis, which demonstrated HCMV to globally redirect mRNA translation toward the synthesis of cellular prosurvival factors within infected monocytes. Specifically, HCMV entry into monocytes induced the translation of cellular SIRT1 to generate an antiapoptotic state. Defining the precise mechanisms through which HCMV stimulates survival will provide insight into novel anti-HCMV drugs able to target infected monocytes.

Attributes of Provider Referrals for Digital Mental Health Applications in an Integrated Health System, 2019-2021.

OBJECTIVE: This article describes trends and attributes associated with digital mental health application (DMHA) referrals from December 2019 through December 2021. METHODS: In total, 43,842 DMHA referrals for 25,213 unique patients were extracted from the electronic health record of a large, diverse, integrated health system. DMHAs were aggregated by type (cognitive-behavioral therapy [CBT] or mindfulness and meditation [MM]). Monthly referral patterns were described and categorized into mutually exclusive clusters (MM, CBT, or MM and CBT). Multinomial logistic regression and post hoc predicted probabilities were used to profile patient, clinical, and encounter attributes among referral clusters. RESULTS: DMHA referrals increased, reached equilibrium, and then began to decline over the 25-month observation period. Compared with the referral cluster average, MM-alone referrals were more likely to occur for patients who were ages ≥65, who were Hispanic or Asian, whose reason for visit concerned mental health, and who had a primary diagnosis of other anxiety disorders. CBT-alone referrals were more likely to occur for patients with a primary diagnosis of depression and less likely to occur for Hispanic patients. Combined MM and CBT referrals were more likely to occur for patients who were ages 18-30, whose reason for visit was "other," and who had a primary diagnosis of depression and were less likely to occur for Hispanic patients and those ages ≥65. CONCLUSIONS: Although this study demonstrates readiness to integrate DMHA referral into clinical workflows, observed variations in attributes of referral clusters support the need to further investigate provider decision making and whether referral patterns are optimal and sustainable.

Milk fat globule membrane protects Bifidobacterium longum ssp. infantis ATCC 15697 against bile stress by modifying global transcriptional responses.

The milk fat globule membrane (MFGM) can protect probiotic bacteria from bile stress. However, its potential mechanism has not been reported. In this study, the viability, morphology and gene transcriptional response of Bifidobacterium longum ssp. infantis ATCC 15697 (BI_15697) stressed by bile salts with or without MFGM were investigated. It was shown that MFGM alleviated the reduction in BI_15697 population induced by 0.2% porcine bile stress and restored the population to the control levels. MFGM ameliorated the shrunken, fragmented appearance and irregular morphology of BI_15697 and maintained cell integrity disrupted by bile stress. RNA-sequencing results showed that MFGM increased transport of glucose and raffinose and decreased that of branched-chain amino acids (BCAA) in the presence of bile salts. MFGM stimulated the expression of genes involved in the synthesis of raffinose in galactose metabolism and the metabolism of BCAA, suggesting that MFGM stimulated the accumulation of raffinose and BCAA in the presence of bile. In addition, MFGM stimulated the expression of 2 bile efflux transporters under bile stress. Together, the multifactorial response helps BI_15697 excrete bile salts and maintain cellular integrity in response to bile stress. This study proposes a mechanism for the protection of BI_15697 against bile salt stress by MFGM, thereby providing a molecular basis for its application in incorporation of probiotics.

Image Fusion Technology in Interventional Radiology.

Image fusion technology aims to improve patient outcomes for image-guided interventions by leveraging the strengths of multimodality imaging datasets. This most commonly involves the overlay or co-display of advanced cross-sectional imaging permitting freedom of device placement via conventional image guidance such as ultrasound, fluoroscopy, and computed tomography. This can allow the interventionalist to target and treat lesions that would otherwise be difficult or impossible to visualize and access using conventional imaging guidance. Furthermore, the use of image fusion can allow for procedures traditionally performed with cross-sectional imaging to be performed under ultrasound or fluoroscopy, by importing the data from preacquired cross-sectional imaging into the interventional procedure. This manuscript provides an overview of image fusion technologies used for interventional radiology (IR) guidance, with an emphasis on technical considerations.

Use of Doehlert Matrix as a Tool for High-Throughput Screening of Organic Acids and Essential Oils on Miniaturized Pork Loins, Followed by Lab-Scale Validation That Confirmed Tested Compounds Do Not Show Synergistic Effects against Salmonella Typhimurium.

The many possible treatments and continuously changing consumer trends present a challenge when selecting antimicrobial interventions during pork processing. Thirty-five potential antimicrobials were screened at commercial working concentrations by individually adding them to miniaturized (69 cm3) disks of pork loin ends, followed by inoculation with Salmonella Typhimurium ATCC 19585. Two organic acids and nine essential oils significantly inhibited Salmonella counts on pork (p < 0.05). However, six compounds that represent different levels of significance (p < 0.05-p < 0.0001) were selected as independent variables to build a Response Surface Methodology model based on a Doehlert matrix (Doehlert Matrix-RSM): lactic acid 1.25%, formic acid 0.25%, cumin 0.25%, clove 0.25%, peppermint 0.5%, and spearmint 0.5%. The goal of the Doehlert Matrix-RSM was to study single and paired effects of these antimicrobials on the change in Salmonella over 24 h. The Doehlert Matrix-RSM model predicted that lactic acid, formic acid, cumin, peppermint, and spearmint significantly reduced Salmonella when added alone, while no significant interactions between these antimicrobials were found. A laboratory-scale validation was carried out on pork loin end slices, which confirmed the results predicted by the model. While this screening did not identify novel synergistic combinations, our approach to screening a variety of chemical compounds by implementing a miniaturized pork loin disk model allowed us to identify the most promising antimicrobial candidates to then formally design experiments to study potential interactions with other antimicrobials.

Migration Behavior of Anguilla celebesensis Silver Eels within their Tomini Bay Spawning Area.

The tropical Celebes eel, Anguilla celebesensis, has a short migration between its spawning and growth habitats. Its spawning areas were hypothesized to be in Tomini Bay and the Celebes Sea after collecting their small leptocephali. However, there is no information about the silver eel oceanic spawning migration behavior of A. celebesensis. To better understand their short-distance spawning migration behavior, four large female silver eels (Eel 1-4) were equipped with pop-up satellite archival tags (PSATs) and released near the mouth of the Poso River in Tomini Bay of Sulawesi Island on 22 February (Eel 1-3) and 11 March 2010 (Eel 4). All PSATs ascended in Tomini Bay and transmitted their data. Eel 3 and 4 provided clear records of consistent diel vertical migration (DVM: eight days-Eel 3, 13 days-Eel 4) with daytime dives to mean depths of 444.7 m (Eel 3) and 539.0 m (Eel 4), where mean temperatures were 9.1°C (Eel 3) and 7.7°C (Eel 4), and nighttime ascents to mean depths of 132.8 m (Eel 3) and 112.4 m (Eel 4), where mean temperatures were 20.6°C (Eel 3) and 23.4°C (Eel 4). Eel 3 and 4 started to dive to deeper water around nautical dawn and swam up to shallower water around sunset. During nighttime, both eels swam in deeper and colder water during nights with moonlight than during nights without moonlight, and there was a negative linear relationship between experienced water temperatures with the moon in the sky and the lunar age for the eels. The A. celebesensis daily rhythm of DVM behaviors was similar to spawning-migration DVM behaviors of other anguillid species. Essential life history characteristics of A. celebesensis appear to be a short migration between freshwater growth habitat and ocean spawning habitat, and high GSI values with advanced gonadal development in downstream-migrating silver eels.

Engineered Secretory Immunoglobulin A provides insights on antibody-based effector mechanisms targeting Clostridiodes difficile.

Secretory (S) Immunoglobin (Ig) A is the predominant mucosal antibody, which mediates host interactions with commensal and pathogenic microbes, including Clostridioides difficile. SIgA adopts a polymeric IgA structure that is bound by secretory component (SC). Despite significance, how SIgA supports diverse effector mechanisms is poorly characterized and SIgA-based therapies nonexistent. We engineered chimeric (c) SIgAs, in which we replaced SC domain D2 with a single domain antibody or a monomeric fluorescent protein, allowing us to investigate and enhance SIgA effector mechanisms. cSIgAs exhibited increased neutralization potency against C. difficile toxins, promoted bacterial clumping and cell rupture, and decreased cytotoxicity. cSIgA also allowed us to visualize and/or quantify C. difficile morphological changes and clumping events. Results reveal mechanisms by which SIgA combats C. difficile infection, demonstrate that cSIgA design can modulate these mechanisms, and demonstrate cSIgA's adaptability to modifications that might target a broad range of antigens and effector mechanisms.

Virion-associated US28 rapidly modulates Akt activity to suppress HCMV lytic replication in monocytes.

Establishing a non-productive quiescent/silent infection within monocytes is essential for spread of human cytomegalovirus (HCMV). Yet, how HCMV establishes a quiescent infection in monocytes remains unclear. US28 is a viral G protein-coupled receptor (GPCR) essential for silent infections within cells of the myeloid lineage. We found virion-associated US28 was rapidly delivered to monocytes, while de novo synthesized US28 was delayed for several days. A recombinant mutant virus lacking US28 (US28Δ) was unable to establish a quiescent infection, resulting in a fully productive lytic replication cycle. Mechanistically, viral entry of US28Δ phosphorylated Akt at both serine 473 (S473) and threonine 308 (T308), which contrasted with the site-specific phosphorylation of Akt at S473 following WT infection. Preventing Akt bi-phosphorylation prevented lytic replication of US28Δ, and ectopic expression of a constitutively phosphorylated Akt variant triggered lytic replication of WT infection. Our data demonstrate that virion-delivered US28 fine-tunes Akt activity to permit HCMV infection to enter a quiescent state following primary infection of monocytes.

Effects of commercial and traditional kefir supplementation on apparent total tract macronutrient digestibility and the fecal characteristics, metabolites, and microbiota of healthy adult dogs.

Kefir is a fermented dairy beverage that has been consumed by humans for centuries, but poorly studied in pets. The objective of this study was to determine the effects of commercial or traditional kefir supplementation on apparent total tract macronutrient digestibility (ATTD) and fecal characteristics, microbiota populations, and metabolite and immunoglobulin (Ig) A concentrations of healthy adult dogs. Twelve healthy adult dogs (5.67 ±â€…1.72 yr, 7.27 ±â€…1.15 kg) were used in a replicated 3 × 3 Latin square design (n = 12/group). All dogs were fed a commercial diet and allotted to 1 of 3 treatments (60 mL/d): 2% reduced-fat milk treated with lactase [CNTL; 4.57E + 03 lactic acid bacteria (LAB) colony-forming units (CFU)/mL], commercial kefir (C-Kefir; 6.95E + 04 LAB CFU/mL), or traditional kefir brewed daily from 2% reduced-fat milk and kefir grains (T-Kefir; 1.79E + 09 LAB CFU/mL). The experiment was composed of three 28-d periods, with each consisting of a 22-d transition phase, a 5-d fecal collection phase, and 1 d for blood collection. Fecal samples were collected for determination of ATTD and fecal pH, dry matter, microbiota, and metabolite, and IgA concentrations. Data were analyzed using the Mixed Models procedure of SAS 9.4. The main effects of treatment were tested, with significance set at P ≤ 0.05 and trends set at P ≤ 0.10. Kefir products differed in microbial density and profile, but fecal microbiota populations were weakly impacted. Bacterial alpha diversity tended to be greater (P = 0.10) in dogs fed T-Kefir than those fed CNTL. Bacterial beta diversity analysis identified a difference (P < 0.0004) between dogs-fed CNTL and those fed C-Kefir. Dogs-fed C-Kefir tended to have a greater (P = 0.06) relative abundance of Fusobacteriota than those fed CNTL or T-Kefir. Dogs-fed T-Kefir had a greater (P < 0.0001) relative abundance of Lactococcus than those fed CNTL or C-Kefir. Dogs-fed T-Kefir also tended to have a lower (P = 0.09) relative abundance of Escherichia Shigella and greater (P = 0.09) relative abundance of Candidatus stoquefichus than dogs-fed CNTL or C-Kefir. Dogs-fed C-Kefir tended to have lower (P = 0.08) fecal valerate concentrations than those fed CNTL or T-Kefir. All other measures were unaffected by kefir treatments. Our results suggest that kefir supplementation had minor effects on the fecal microbiota populations and fecal metabolite concentrations of healthy adult dogs without impacting ATTD, fecal characteristics, or fecal IgA concentrations.

Kefir is a fermented dairy beverage that has been consumed by humans for centuries, but poorly studied in pets. Our objective was to determine the effects of commercial or traditional kefir supplementation on apparent total tract macronutrient digestibility and fecal characteristics, microbiota populations, and metabolite and immunoglobulin A concentrations of healthy adult dogs. Using a replicated Latin square design, milk (control; CNTL), commercial kefir (C-Kefir), and traditional kefir (T-Kefir) treatments were tested. Kefir products differed in microbial density and profile, but fecal microbiota populations were weakly impacted. Bacterial alpha diversity tended to be greater in T-Kefir than CNTL. Bacterial beta diversity analysis identified differences between CNTL and C-Kefir. Dogs-fed C-Kefir tended to have greater relative abundance of Fusobacteriota than those fed CNTL or T-Kefir. Dogs-fed T-Kefir had a greater relative abundance of Lactococcus than those fed CNTL or C-Kefir. Dogs-fed T-Kefir tended to have a lower relative abundance of Escherichia-Shigella and greater relative abundance of Candidatus stoquefichus than dogs-fed CNTL or C-Kefir. Dogs-fed C-Kefir tended to have lower fecal valerate than those fed CNTL or T-Kefir. Our results suggest that kefir supplementation had minor effects on the fecal microbiota and metabolites of healthy adult dogs.