Short-chain fatty acids (SCFAs) comprise the largest group of gut microbial fermentation products. While absorption of most nutrients occurs in the small intestine, indigestible dietary components, such as fiber, reach the colon and are processed by the gut microbiome to produce a wide array of metabolites that influence host physiology. Numerous studies have implicated SCFAs as key modulators of host health, such as in regulating irritable bowel syndrome (IBS). However, robust methods are still required for their detection and quantitation to meet the demands of biological studies probing the complex interplay of the gut-host-health paradigm. In this study, a sensitive, rapid-throughput, and readily expandible UHPLC-QqQ-MS platform using 2-PA derivatization was developed for the quantitation of gut-microbially derived SCFAs, related metabolites, and isotopically labeled homologues. The utility of this platform was then demonstrated by investigating the production of SCFAs in cecal contents from mice feeding studies, human fecal bioreactors, and fecal/bacterial fermentations of isotopically labeled dietary carbohydrates. Overall, the workflow proposed in this study serves as an invaluable tool for the rapidly expanding gut-microbiome and precision nutrition research field.

BackgroundVagal afferent neurons represent the key neurosensory branch of the gut-brain axis, which describes the bidirectional communication between the gastrointestinal system and the brain. These neurons are important for detecting and relaying sensory information from the periphery to the central nervous system to modulate feeding behavior, metabolism, and inflammation. Confounding variables complicate the process of isolating the role of the vagal afferents in mediating these physiological processes. Therefore, we developed a microfluidic model of the sensory branch of the gut-brain axis. We show that this microfluidic model successfully compartmentalizes the cell body and neurite terminals of the neurons, thereby simulates the anatomical layout of these neurons to more accurately study physiologically-relevant processes.MethodsWe implemented a primary rat vagal afferent neuron culture into a microfluidic platform consisting of two concentric chambers interconnected with radial microchannels. The microfluidic platform separated cell bodies from neurite terminals of vagal afferent neurons. We then introduced physiologically-relevant gastrointestinal effector molecules at the nerve terminals and assessed their retrograde transport along the neurite or capacity to elicit an electrophysiological response using live cell calcium imaging.ResultsThe angle of microchannel outlets dictated the probability of neurites growing into a chamber versus tracking along chamber walls. When the neurite terminals were exposed to fluorescently-labeled cholera toxin subunit B, the proteins were taken up and retrogradely transported along the neurites over the course of 24 h. Additionally, mechanical perturbation (e.g., rinsing) of the neurite terminals significantly increased intracellular calcium concentration in the distal soma. Finally, membrane-displayed receptor for capsaicin was expressed and trafficked along newly projected neurites, as revealed by confocal microscopy.ConclusionsIn this work, we developed a microfluidic device that can recapitulate the anatomical layout of vagal afferent neurons in vitro. We demonstrated two physiologically-relevant applications of the platforms: retrograde transport and electrophysiological response. We expect this tool to enable controlled studies on the role of vagal afferent neurons in the gut-brain axis.

Bifidobacteria represent an important gut commensal in humans, particularly during initial microbiome assembly in the first year of life. Enrichment of Bifidobacterium is mediated though the utilization of human milk oligosaccharides (HMOs), as several human-adapted species have dedicated genomic loci for transport and metabolism of these glycans. This results in the release of fermentation products into the gut lumen which may offer physiological benefits to the host. Synbiotic pairing of probiotic species with a cognate prebiotic delivers a competitive advantage, as the prebiotic provides a nutrient niche. To determine the fitness advantage and metabolic characteristics of an HMO-catabolizing Bifidobacterium strain in the presence or absence of 2′-fucosyllactose (2′-FL), conventionally colonized mice were gavaged with either Bifidobacterium pseudocatenulatum MP80 (B.p. MP80) (as the probiotic) or saline during the first 3 days of the experiment and received water or water containing 2′-FL (as the prebiotic) throughout the study. 16S rRNA gene sequencing revealed that mice provided only B.p. MP80 were observed to have a similar microbiota composition as control mice throughout the experiment with a consistently low proportion of Bifidobacteriaceae present. Using 1H NMR spectroscopy, similar metabolic profiles of gut luminal contents and serum were observed between the control and B.p. MP80 group. Conversely, synbiotic supplemented mice exhibited dramatic shifts in their community structure across time with an overall increased, yet variable, proportion of Bifidobacteriaceae following oral inoculation. Parsing the synbiotic group into high and moderate bifidobacterial persistence based on the median proportion of Bifidobacteriaceae, significant differences in gut microbial diversity and metabolite profiles were observed. Notably, metabolites associated with the fermentation of 2′-FL by bifidobacteria were significantly greater in mice with a high proportion of Bifidobacteriaceae in the gut suggesting metabolite production scales with population density. Moreover, 1,2-propanediol, a fucose fermentation product, was only observed in the liver and brain of mice harboring high proportions of Bifidobacteriaceae. This study reinforces that the colonization of the gut with a commensal microorganism does not guarantee a specific functional output.

The gut-brain axis embodies the bi-directional communication between the gastrointestinal tract and the central nervous system (CNS), where vagal afferent neurons (VANs) serve as sensors for a variety of gut-derived signals. The gut is colonized by a large and diverse population of microorganisms that communicate via small (effector) molecules, which also act on the VAN terminals situated in the gut viscera and consequently influence many CNS processes. However, the convoluted in vivo environment makes it difficult to study the causative impact of the effector molecules on VAN activation or desensitization. Here, we report on a VAN culture and its proof-of-principle demonstration as a cell-based sensor to monitor the influence of gastrointestinal effector molecules on neuronal behavior. We initially compared the effect of surface coatings (poly-L-lysine vs. Matrigel) and culture media composition (serum vs. growth factor supplement) on neurite growth as a surrogate of VAN regeneration following tissue harvesting, where the Matrigel coating, but not the media composition, played a significant role in the increased neurite growth. We then used both live-cell calcium imaging and extracellular electrophysiological recordings to show that the VANs responded to classical effector molecules of endogenous and exogenous origin (cholecystokinin serotonin and capsaicin) in a complex fashion. We expect this study to enable platforms for screening various effector molecules and their influence on VAN activity, assessed by their information-rich electrophysiological fingerprints.

The gastroenterology community lost a remarkable scientist with the death of Helen Cooke, PhD, on December 27, 2021. Helen was 78 and passed peacefully in Ann Arbor, Michigan, due to complications of Parkinson’s disease. Helen pioneered the field of neurobiology of the intestinal mucosa and in particular the enteric neural control of epithelial transport, including neuroimmune effects on epithelial function and the neural integration of secretion and motility. Helen was born on May 22, 1943, outside of Sunderland, Massachusetts, to Roger and Stella (Wolejko) Radowicz. Growing up in a farming town and doing the hard work of picking cucumbers each summer, Helen longed to see more of the world and strove to do so through academics, graduating valedictorian of her high school class. Helen’s curiosity about life and her passion to engage beyond the farm were evident early on; she once mentioned to one of us (H.V.C.) that her first career aspiration as a teenager was to be a spy. Helen received a BS degree in zoology from the University of Massachusetts, Amherst, in 1965 and an MS in physiology from the University of California, Los Angeles, in 1967, where she overlapped with Ernie Wright, Jared Diamond, and Morton Grossman. Shortly thereafter, she moved to Australia and obtained a PhD in physiology 1971 from the University of Sydney, studying electrolyte and amino acid transport in the kidney. Helen returned to the USA and accepted a part-time appointment as instructor and then Assistant Professor in the Physiology Department at the University of Iowa. After a brief time at the University of Kansas Medical School, Helen was recruited to the Department of Physiology at the University of Nevada, Reno. She relocated to the Physiology Department at Ohio State University together with several other faculty, including Jack Wood. It was at OSU that Helen spent many years, engaging in long and productive collaborations with several faculty and finishing her career in the Departments of Pharmacology and Neuroscience. Helen’s research program was marked by groundbreaking discoveries that revealed the pivotal role played by the enteric nervous system in regulating how nutrients and electrolytes are transported across the gut epithelium, and how this transport is integrated into gastrointestinal processes. Her exploration of this field began in the early 1980s with the demonstration that intrinsic nerves in the gut can regulate sodium and chloride (and thereby fluid) transport, which was first recognized by Ken Hubel a few years earlier. Through initial collaborations with the Wood laboratory, Helen showed that electrical field stimulation of isolated sections of guinea pig ileum mounted in Ussing chambers caused release of acetylcholine, activating muscarinic receptors on epithelial cells to induce active chloride secretion. Her team expanded on that early work to show that electrical field stimulation activates neurons located in the submucosal plexus and that enteric neural reflexes control neurogenic chloride secretion. With her trainees and collaborators, Helen went on to identify pathways by which neurotransmitters regulate epithelial ion transport, including acetylcholine, VIP, 5-HT, substance P, serotonin, histamine, and ATP. Her team was one of the first to demonstrate that physical stroking of the mucosa evokes chloride secretion by activating a neural reflex involving serotoninergic and muscarinic receptors in the submucosal plexus. She also demonstrated that enterochromaffin cells can act as “glucose sensors” during food ingestion, leading to release of 5-HT to induce multiple effects on epithelial, neural, and smooth muscle function in the GI tract and beyond. Another landmark contribution was the establishment of neuroimmune communication between submucosal plexus and immune cells under conditions of sensitization by antigens and infection. Helen’s collaboration with Fievos Christofi demonstrated the role of endogenous nucleotides interacting with purinergic receptors to transduce mechanically evoked chloride secretion. Notably, while she worked primarily with animal models, Helen also applied her preclinical findings to human tissues. The influence of Helen’s career on the scientific community went well beyond her own laboratory, touching the professional lives of many. She was a life-long proponent of mentoring in science, crediting Dr David Dawson at the University of Iowa as the key mentor who helped launch her career in intestinal transport physiology. In turn, many early-stage investigators benefited from Helen’s wisdom, encouragement, and career support provided through formal academic mentorship (eg, H.V.C.) and through informal interactions with junior colleagues in the GI community (eg, H.E.R.). Helen’s contributions to their lives were praised by many who attended a virtual celebration of her life and career in early 2021. In his tribute to Helen, Gary Mawe recalled that “When I was a postdoc in Mike [Gershon]’s lab, and after I came here to the University of Vermont, every time I saw you at a meeting you took the time to sit down with me and ask me how things were going—and you really listened to my answers. You asked me how my science was going, how the publications were coming, what I was thinking in terms of developing grants, how grants I submitted were doing, and also how my life balance was. I appreciated that so much … and I know you were treating many other investigators in the same way.” Helen’s mentorship of early career scientists was just one component of her dedication to service in scientific societies. Within the AGA she promoted the involvement of women in committee activities and leadership positions as a member of the Steering Committee on Women, which eventually was formalized as the AGA Women’s Committee. Helen was the first woman to chair the Section on Motility and Nerve-Gut Interactions and was the inaugural chair of the Task Force on the Status of PhD, DVM, and MD/PhD Members in the AGA, which eventually became a standing committee with an expanded mission. Helen engaged in similar activities within the American Physiological Society (APS). In the early 1980s she was part of the ad hoc Women’s Caucus that met informally to discuss the status of women in the Society; later, after the Women in Physiology Committee was established, Helen served as its second chair. Helen was also the first woman to chair the Gastrointestinal and Liver Physiology Section of the APS and in 1991 was elected to the APS Council, becoming the third woman to serve in that capacity in the Society’s 104-year history. Helen’s support of early career investigators went beyond the laboratory and her professional service and reflected the close friendships that she developed and maintained over her long and fruitful career. She was a founding member of the SWANS, a group of mostly early-stage basic and physician scientist women in GI that met at DDW to share experiences, aspirations, achievements, and much fun over great meals (Figure 1). Helen encouraged us to seek leadership opportunities and not be deterred when difficulties arose as our careers unfolded. Many of the SWANS went on to become leaders in the fields of gastroenterology, physiology, and neurogastroenterology in academia and clinical medicine. Helen was an elegant, stylish, quiet but strong presence who had a deep sense of the essential values of fairness and equity in science. Her laboratory and her collaborators were part of her extended family, and it is through her career that she found some of her closest friends and most treasured experiences. In Columbus, she indulged her love of OSU football, colorful artwork, and music, particularly jazz. But her greatest joy in life was her family, including her sons Ian (a computer scientist and software engineer at Sandia National Laboratories) and Colin (an Associate Professor of Pulmonary and Critical Care Medicine at the University of Michigan), and her 4 grandchildren. Helen is greatly missed by all of us who knew and worked with her and who called her friend, and her impact on our lives will not be forgotten.

N-glycosylation is a ubiquitous posttranslational modification that affects protein structure and function, including those of the central nervous system. N-glycans attached to cell membrane proteins play crucial roles in all aspects of biology, including embryogenesis, development, cell–cell recognition and adhesion, and cell signaling and communication. Although brain function and behavior are known to be regulated by the N-glycosylation state of numerous cell surface glycoproteins, our current understanding of brain glycosylation is limited, and glycan variations associated with functional brain regions remain largely unknown. In this work, we used a well-established cell surface glycomic nanoLC-Chip-Q-TOF platform developed in our laboratory to characterize the N-glycome of membrane fractions enriched in cell surface glycoproteins obtained from specific functional brain areas. We report the cell membrane N-glycome of two major developmental divisions of mice brain with specific and distinctive functions, namely the forebrain and hindbrain. Region-specific glycan maps were obtained with ∼120 N-glycan compositions in each region, revealing significant differences in “brain-type” glycans involving high mannose, bisecting, and core and antenna fucosylated species. Additionally, the cell membrane N-glycome of three functional regions of the forebrain and hindbrain, the cerebral cortex, hippocampus, and cerebellum, was characterized. In total, 125 N-glycan compositions were identified, and their region-specific expression profiles were characterized. Over 70 N-glycans contributed to the differentiation of the cerebral cortex, hippocampus, and cerebellum N-glycome, including bisecting and branched glycans with varying degrees of core and antenna fucosylation and sialylation. This study presents a comprehensive spatial distribution of the cell-membrane enriched N-glycomes associated with five discrete anatomical and functional brain areas, providing evidence for the presence of a previously unknown brain glyco-architecture. The region-specific molecular glyco fingerprints identified here will enable a better understanding of the critical biological roles that N-glycans play in the specialized functional brain areas in health and disease.

The gut microbiota is able to influence overall energy balance via effects on both energy intake and expenditure, and is a peripheral target for potential obesity therapies. However, the precise mechanism by which the gut microbiota influences energy intake and body weight regulation is not clear. Microbes use small molecules to communicate with each other; some of these molecules are ligands at mammalian receptors and this may be a mechanism by which microbes communicate with the host. Here we briefly review the literature showing beneficial effects of microbial metabolites on food intake regulation and examine the potential role for vagal afferent neurons, the gut-brain axis.

2 '-Fucosyllactose (2 '-FL) is one of the predominant oligosaccharides found in human milk and has several well-established beneficial effects in the host. It has previously been shown that 2 '-FL can improve the metabolic phenotype in high-fat (HF)-fed mice. Here we investigated whether dietary supplementation with 2 '-FL was associated with improved intestinal barrier integrity, signaling in the vagal afferent pathway and cognitive function. Mice were fed either a low-fat (LF, 10% fat per kcal) or HF (45% fat per kcal) diet with or without supplementation of 2 '-FL (10% w/w) in the diet for 8 weeks. Body weight, energy intake, fat and lean mass, intestinal permeability (ex vivo in Ussing chambers), lipid profiles, gut microbiome and microbial metabolites, and cognitive functions were measured. Vagal afferent activity was measured via immunohistochemical detection of c-Fos protein in the brainstem in response to peripheral administration of cholecystokinin (CCK). 2 '-FL significantly attenuated the HF-induced increase in fat mass and energy intake. 2 '-FL significantly reduced intestinal permeability and significantly increased expression of interleukin (IL)-22, a cytokine known for its protective role in the intestine. Additionally, 2 '-FL led to changes in the gut microbiota composition and in the associated microbial metabolites. Signaling in the vagal afferent pathway was improved but there was no effect on cognitive function. In conclusion, 2 '-FL supplementation improved the metabolic profiles, gut barrier integrity, lipid metabolism and signaling in the vagal afferent pathway. These findings support the utility of 2 '-FL in the control of gut barrier function and metabolic homeostasis under a metabolic challenge.

Understanding how exogenous microbes stably colonize the animal gut is essential to reveal mechanisms of action and tailor effective probiotic treatments. Bifidobacterium species are naturally enriched in the gastrointestinal tract of breast-fed infants. Human milk oligosaccharides (HMOs) are associated with this enrichment. However, direct mechanistic proof of the importance of HMOs in this colonization is lacking given milk contains additional factors that impact the gut microbiota. This study examined mice supplemented with the HMO 2ʹfucosyllactose (2ʹFL) together with a 2ʹFL-consuming strain, Bifidobacterium pseudocatenulatum MP80. 2ʹFL supplementation creates a niche for high levels of B.p. MP80 persistence, similar to Bifidobacterium levels seen in breast-fed infants. This synergism impacted gut microbiota composition, activated anti-inflammatory pathways and protected against chemically-induced colitis. These results demonstrate that bacterial-milk glycan interactions alone drive enrichment of beneficial Bifidobacterium and provide a model for tunable colonization thus facilitating insight into mechanisms of health promotion by bifidobacteriain neonates.

The efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) is limited by acute and chronic graft-versus-host disease (GVHD). The impact of obesity on allo-HSCT outcomes is poorly understood. Here, we report that obesity had a negative and selective impact on acute gut GVHD after allo-HSCT in mice with diet-induced obesity (DIO). These animals exhibited increased gut permeability, endotoxin translocation across the gut, and radiation-induced gastrointestinal damage after allo-HSCT. After allo-HSCT, both male and female DIO mouse recipients showed increased proinflammatory cytokine production and expression of the GVHD marker ST2 (IL-33R) and MHC class II molecules; they also exhibited decreased survival associated with acute severe gut GVHD. This rapid-onset, obesity-associated gut GVHD depended on donor CD4+ T cells and occurred even with a minor MHC mismatch between donor and recipient animals. Retrospective analysis of clinical cohorts receiving allo-HSCT transplants from unrelated donors revealed that recipients with a high body mass index (BMI, >30) had reduced survival and higher serum ST2 concentrations compared with nonobese transplant recipients. Assessment of both DIO mice and allo-HSCT recipients with a high BMI revealed reduced gut microbiota diversity and decreased Clostridiaceae abundance. Prophylactic antibiotic treatment protected DIO mouse recipients from endotoxin translocation across the gut and increased inflammatory cytokine production, as well as gut pathology and mortality, but did not protect against later development of chronic skin GVHD. These results suggest that obesity-induced alterations of the gut microbiota may affect GVHD after allo-HSCT in DIO mice, which could be ameliorated by prophylactic antibiotic treatment.

2'-fucosyllactose (2'-FL), the most predominant oligosaccharide found in human milk, acts as a prebiotic with beneficial effects on the host. The aim of this study was to determine the beneficial effect of 2'-FL on intestinal barrier integrity and metabolic functions in low-fat (LF)- and high-fat (HF)-fed mice. Male C57/BL6 mice (n = 32, 8/group; 6 weeks old, JAX, CA) were counter-balanced into four weight-matched groups and fed either a low-fat (LF; 10% kcal fat with 7% kcal sucrose) or HF (45% kcal fat with 17% kcal sucrose) with or without supplementation of 2'-FL in the diet [10% (w/w), 8 weeks; LF/2'-FL or HF/2'-FL; BASF, Germany]. General phenotypes (body weight, energy intake, fat and lean mass), intestinal permeability (ex vivo in Ussing chambers), lipid profiles, and microbial metabolites were assessed. 2'-FL significantly attenuated the HF-induced increase in body fat mass with a trend to decrease body weight gain. 2'-FL significantly decreased intestinal permeability in LF-fed mice with a trend for a decrease in HF-fed mice. This was associated with a significant increase in interleukin-22, a cytokine known to have a protective role in intestinal barrier function. Visceral adipocyte size was significantly decreased by 2'-FL in both LF- and HF-fed mice. 2'-FL suppressed HF-induced upregulation of adipogenic transcription factors peroxisome proliferator-activated receptor gamma and sterol regulatory element binding protein-1c in the liver. Lastly, 2'-FL supplementation led to a significant elevation of lactic acid concentration in the cecum of HF-fed mice, which is known to be a product from beneficial microbes. 2'-FL supplementation improved gut barrier integrity and lipid metabolism in mice with and without the metabolic challenge of HF feeding. These findings support the use of 2'-FL in the control of gut barrier function and metabolic homeostasis under normal and abnormal physiological conditions. BASF (Germany).

The vagus nerve conveys gastrointestinal cues to the brain to control eating behavior. In obesity, vagally mediated gut-brain signaling is disrupted. Here, we show that the cocaine- and amphetamine-regulated transcript (CART) is a neuropeptide synthesized proportional to the food consumed in vagal afferent neurons (VANs) of chow-fed rats. CART injection into the nucleus tractus solitarii (NTS), the site of vagal afferent central termination, reduces food intake. Conversely, blocking endogenous CART action in the NTS increases food intake in chow-fed rats, and this requires intact VANs. Viral-mediated Cartpt knockdown in VANs increases weight gain and daily food intake via larger meals and faster ingestion rate. In obese rats fed a high-fat, high-sugar diet, meal-induced CART synthesis in VANs is blunted and CART antibody fails to increase food intake. However, CART injection into the NTS retains its anorexigenic effect in obese rats. Restoring disrupted VAN CART signaling in obesity could be a promising therapeutic approach.

Background Bifidobacterium longum subsp. infantis ( B. infantis ) is a commensal bacterium that colonizes the gastrointestinal tract of breast-fed infants. B. infantis can efficiently utilize the abundant supply of oligosaccharides found in human milk (HMO) to help establish residence. We hypothesized that metabolites from B. infantis grown on HMO produce a beneficial effect on the host. Results In a previous study, we demonstrated that B. infantis routinely dominated the fecal microbiota of a breast fed Bangladeshi infant cohort (1). Characterization of the fecal metabolome of binned samples representing high and low B. infantis populations from this cohort revealed higher amounts of the tryptophan metabolite indole-3-lactic acid (ILA) in feces with high levels of B. infantis . Further in vitro analysis confirmed that B. infantis produced significantly greater quantities of the ILA when grown on HMO versus lactose, suggesting a growth substrate relationship to ILA production. The direct effects of ILA were assessed in a macrophage cell line and intestinal epithelial cell lines. ILA (1-10 mM) significantly attenuated lipopolysaccharide (LPS)-induced activation of NF-kB in macrophages. ILA significantly attenuated TNF-α- and LPS-induced increase in the pro-inflammatory cytokine IL-8 in intestinal epithelial cells. ILA increased mRNA expression of the aryl hydrogen receptor (AhR)-target gene CYP1A1 and nuclear factor erythroid 2–related factor 2 (Nrf2)-targeted genes glutathione reductase 2 (GPX2), superoxide dismutase 2 (SOD2), and NAD(P) H dehydrogenase (NQO1). Pretreatment with either the AhR antagonist or Nrf-2 antagonist inhibited the response of ILA on downstream effectors. Conclusions These findings suggest that ILA, a predominant metabolite from B. infantis grown on HMO and elevated in infant stool high in B. infantis , and protects gut epithelial cells in culture via activation of the AhR and Nrf2 pathway.

BackgroundObesity is a major public health issue afflicting over 93 million people in the US and is projected to continue increasing. Impairments of cognitive abilities such as memory loss and mood disorders such as depression and anxiety are debilitating conditions associated with obesity. N‐glycosylation, an essential posttranslational modification of proteins is required for hippocampal long‐term potentiation and memory consolidation. However, whether altered N‐glycosylation is a potential mechanism involved in impaired hippocampal functioning in obesity remains unknown.AimOur goal was to develop a multi‐omics platform including glycomics, proteomics, and transcriptomics to determine the impact of high‐fat‐diet (HFD) consumption on hippocampal N‐glycosylation and expression of cell surface receptors in the context of diet‐induced obesity.HypothesisWe hypothesize that in HFD‐induced obese mice abnormal hippocampal N‐glycosylation occurs, affecting cell surface expression and signaling of key receptors.MethodsC57BL/6J male mice were fed a HFD (45% fat) for 8 weeks to induce obesity or a low‐fat diet (LFD, 10% fat) as the lean control group. Mice were euthanized, and brain tissues collected. Hippocampi were surgically dissected and processed for glycomics, proteomics, glycoproteomics, and RT‐qPCR analysis. Hippocampal cell membrane fractions (MF) were obtained, and N‐glycans released and analyzed by nano‐Liquid Chromatography‐Mass Spectrometry (nanoLC‐MS). Peptides and glycopeptides derived from MF were analyzed by nano‐LC‐FusionOrbitrap MS. Expression level of glycosyltransferase genes were obtained by qPCR.ResultsThe N‐glycome of LFD‐fed mice was dominated by highly branched fucosylated and sialylated N‐glycans (61%) while in HFD‐fed obese mice these N‐glycans were significantly decreased (51%). Correspondingly, transcript analysis of the glycosyltransferases showed that MGAT2 gene, which encodes the enzyme responsible for initiating the synthesis of complex branched N‐glycans, was significantly decreased by 50% in hippocampi of HFD‐fed obese mice compared to lean controls. Our findings show a decrease in complex branched glycans on the hippocampal cell surface of HFD‐obese mice that may be due to the downregulation of the MGAT2 gene. Proteomics and glycoproteomic analysis revealed site‐specific occupancy of over hundreds of hippocampal cell surface receptors.ConclusionsOur results suggest that chronic consumption of HFD and induced obesity results in abnormal hippocampal cell surface N‐glycosylation, and a possible enzyme associated with the altered glycosylation phenotype was identified. Aberrant N‐glycosylation may contribute to altered neuronal or glial cell function. This approach may be useful to identify novel targets for the development of new treatments for obesity‐associated mood and cognitive deficits.Support or Funding InformationFunding from National Institutes of Health, NIDDK (award 5R21DK118379) to M.B.

There is growing evidence to support the beneficial effects of supplementing direct-fed microbials (DFM) on performance, health status, and immune responses of weaned pigs. Therefore, the objective of this study was to investigate dietary supplementation of Bacillus subtilis (DSM 25841) on growth performance, diarrhea, gut permeability, immunity and metabolomic profiles of weaned pigs experimentally infected with a pathogenic F18 Escherichia coli (E. coli). E. coli infection reduced (P < 0.05) growth performance and intestinal villi height, whereas increased (P < 0.05) diarrhea and permeability in the jejunum compared with non-challenged control. Supplementation of Bacillus subtilis linearly enhanced average daily gain of E. coli infected pigs (d 0 to 5 post-inoculation (PI), P < 0.05; d 0 to 11 PI, P = 0.058). Inclusion of high dose Bacillus subtilis reduced (P < 0.05) jejunal permeability on d 5 and d 11 PI compared with the E. coli challenged control. E. coli challenged control pigs up-regulated (P < 0.05) the mRNA expression of SLC5A10 and MUC2 on d 5 PI, but down-regulated (P < 0.05) expression of SLC5A10, MUC2, and CLDN1 on d 11 PI in jejunal mucosa. Supplementation of Bacillus subtilis linearly up-regulated (P < 0.05) the mRNA expression of CFTR and ZO1 on d 5 PI and SLC5A10 and MUC2 on d 11 PI in jejunal mucosa of E. coli infected pigs. E. coli infection increased (P < 0.05) the mRNA expression of several immune genes in the ileal mucosa, while inclusion of Bacillus subtilis linearly down-regulated gene expression of IL1A on d 5 PI (P = 0.07) and IL6 on d 11 PI (P < 0.05) in ileal mucosa of E. coli infected pigs. Supplementation of Bacillus subtilis modified (Fold change > 1.5; FDR < 0.20) metabolomic profiles in colon digesta, related to pathogenesis and amino acid metabolism. In conclusion, supplementation of Bacillus subtilis enhanced growth rate, improved gut health, and modified metabolomic profiles of weaned pigs experimentally infected with a pathogenic E. coli.

BackgroundConsumption of high-fat diet (HF) leads to hyperphagia and increased body weight in male rodents. Female rodents are relatively resistant to hyperphagia and weight gain in response to HF, in part via effects of estrogen that suppresses food intake and increases energy expenditure. However, sex differences in energy expenditure and activity levels with HF challenge have not been systemically described. We hypothesized that, in response to short-term HF feeding, female mice will have a higher energy expenditure and be more resistant to HF-induced hyperphagia than male mice.MethodsSix-week-old male and female C57BL/6 J mice were fed either low fat (LF, 10% fat) or moderate HF (45% fat) for 5 weeks, and energy expenditure, activity and meal pattern measured using comprehensive laboratory animal monitoring system (CLAMS).ResultsAfter 5 weeks, HF-fed male mice had a significant increase in body weight and fat mass, compared with LF-fed male mice. HF-fed female had a significant increase in body weight compared with LF-fed female mice, but there was no significant difference in fat mass. HF-fed male mice had lower energy expenditure compared to HF-fed female mice, likely due in part to reduced physical activity in the light phase. HF-fed male mice also had increased energy intake in the dark phase compared to LF-fed male mice and a reduced response to exogenous cholecystokinin-induced inhibition of food intake. In contrast, there was no difference in energy intake between LF-fed and HF-fed female mice.ConclusionsThe data show that female mice are generally protected from short-term HF-induced alterations in energy balance, possibly by maintaining higher energy expenditure and an absence of hyperphagia. However, HF-feeding in male mice induced weight and fat mass gain and hyperphagia. These findings suggest that there is a sex difference in the response to short-term HF-feeding in terms of both energy expenditure and control of food intake.

This review details the proceedings of a Pennington Biomedical scientific symposium titled, “What Should I Eat and Why? The Environmental, Genetic, and Behavioral Determinants of Food Choice.” The symposium was designed to review the literature about energy homeostasis, particularly related to food choice and feeding behaviors, from psychology to physiology. This review discusses the intrinsic determinants of food choice, including biological mechanisms (genetics), peripheral and central signals, brain correlates, and the potential role of the microbiome. This review also address the extrinsic determinants (environment) of food choice within our physical and social environments. Finally, this review reports the current treatment practices for the clinical management of eating‐induced overweight and obesity. An improved understanding of these determinants will inform best practices for the clinical treatment and prevention of obesity. Strategies paired with systemic shifts in our public health policies and changes in our “obesogenic” environment will be most effective at attenuating the obesity epidemic.

The gut microbiota and associated metabolites have emerged as potential modulators of pathophysiological changes in obesity and related metabolic disorders. Butyrate, a product of bacterial fermentation, has been shown to have beneficial effects in obesity and rodent models of diet-induced obesity. Here, we aimed to determine the beneficial effects of butyrate (as glycerol ester of butyrate monobutyrin, MB) supplementation on metabolic phenotype, intestinal permeability and inflammation, feeding behavior, and the gut microbiota in low-fat (LF)- and high-fat (HF)-fed mice. Two cohorts (separated by 2 weeks) of male C57BL/6J mice (n = 24 in each cohort, 6/group/cohort; 6 weeks old) were separated into four weight-matched groups and fed either a LF (10 % fat/kcal) or HF (45% fat/kcal) with or without supplementation of MB (LF/MB or HF/MB) at 0.25% (w/v) in drinking water for 6 weeks. Metabolic phenotypes (body weight and adiposity), intestinal inflammation, feeding behavior, and fecal microbiome and metabolites were measured. Despite identical genetic and experimental conditions, we found marked differences between cohorts in the response (body weight gain, adiposity, and intestinal permeability) to HF-diet and MB. Notably, the composition of the gut microbiota was significantly different between cohorts, characterized by lower species richness and differential abundance of a large number of taxa, including subtaxa from five phyla, including increased abundance of the genera Bacteroides, Proteobacteria, and Parasutterella in cohort 2 compared to cohort 1. These differences may have contributed to the differential response in intestinal permeability to the HF diet in cohort 2. MB supplementation had no significant effect on metabolic phenotype, but there was a trend to protect from HF-induced impairments in intestinal barrier function in cohort 1 and in sensitivity to cholecystokinin (CCK) in both cohorts. These data support the concept that microbiota composition may have a crucial effect on metabolic responses of a host to dietary interventions and highlight the importance of taking steps to ensure reproducibility in rodent studies.

Estrogens modulate different physiological functions, including reproduction, inflammation, bone formation, energy expenditure, and food intake. In this review, we highlight the effect of estrogens on food intake regulation and the latest literature on intracellular estrogen signaling. In addition, gut satiety hormones, such as cholecystokinin, glucagon-like peptide 1 and leptin are essential to regulate ingestive behaviors in the postprandial period. These peripheral signals are sensed by vagal afferent terminals in the gut wall and transmitted to the hindbrain axis. Here we 1. review the role of the vagus-hindbrain axis in response to gut satiety signals and 2. consider the potential synergistic effects of estrogens on gut satiety signals at the level of vagal afferent neurons and nuclei located in the hindbrain. Understanding the action of estrogens in gut-brain axis provides a potential strategy to develop estrogen-based therapies for metabolic diseases and emphasizes the importance of sex difference in the treatment of obesity.