Evidence for peripheral and central actions of codeine to dysregulate swallowing in the anesthetized cat.

Systemic administration of opioids has been associated with aspiration and swallow dysfunction in humans. We speculated that systemic administration of codeine would induce dysfunctional swallowing and that this effect would have a peripheral component. Experiments were conducted in spontaneously breathing, anesthetized cats. The animals were tracheotomized and electromyogram (EMG) electrodes were placed in upper airway and chest wall respiratory muscles for recording swallow related motor activity. The animals were allocated into three groups: vagal intact (VI), cervical vagotomy (CVx), and supra-nodose ganglion vagotomy (SNGx). A dose response to intravenous codeine was performed in each animal. Swallowing was elicited by injection of 3 mL of water into the oropharynx. The number of swallows after vehicle was significantly higher in the VI group than in SNGx. Codeine had no significant effect on the number of swallows induced by water in any of the groups. However, the magnitudes of water swallow-related EMGs of the thyropharyngeus muscle were significantly increased in the VI and CVx groups by 2-4 fold in a dose-related manner. In the CVx group, the geniohyoid muscle EMG during water swallows was significantly increased. There was a significant dose-related increase in spontaneous swallowing in each group from codeine. The spontaneous swallow number at the 10 mg/kg dose of codeine was significantly larger in the CVx group than that in the SNGx group. During water-evoked swallows, intravenous codeine increased upper airway motor drive in a dose-related manner, consistent with dysregulation. The data support the existence of both central and peripheral actions of codeine on spontaneous swallowing. At the highest dose of codeine, the reduced spontaneous swallow number in the SNGx group relative to CVx is consistent with a peripheral excitatory action of codeine either on pharyngeal/laryngeal receptors or in the nodose ganglion itself. The higher number of swallows in the CVx group than the VI group supports disinhibition of this behavior by elimination of inhibitory vagal sensory afferents.

What Is Chalky? Investigating Consumer Language and Perception of Fine Particles in Beverages Containing Pea and Potato Starch.

Despite its importance as an undesirable food texture, the phenomenon of chalkiness remains understudied. Chalky sensations presumably arise from fine particulates found in foods, but semantic overlap with other common descriptors of small particles, like gritty or sandy, is unclear. Here, we compare the usage of Chalky with related descriptors, and determine the effect of particle size, concentration, and xanthan content on Chalky ratings in a model beverage. A 23 factorial design with starch particle size (D90 = 33.8 and 64.6 µm), starch concentrations (10 and 20% w/v), and xanthan content (0.075 and 0.15% w/v) was used. Participants' salivary flow rate was also assessed. A multi-sip taste test was performed where naïve consumers (n = 82; 39% men, 60% women; age range = 18-79 years) rated the intensity of Chalky, Powdery, Gritty, Sandy, Mouthdrying, and Residual mouthcoating at 0, 30, and 60 s after each of three consecutive sips. All attribute ratings were highly correlated, with Chalky, Powdery, and Residual Mouthcoating being more closely correlated with each other than Gritty or Sandy. Although Chalky was still reported 60 s after consumption, no evidence of build-up was found with repeated sips. A larger size and higher concentration increased Chalky ratings, with the low-salivary-flow group reporting greater ratings for Chalky relative to the high-flow group. Our results suggest consumer percepts of small particles are overlapping but not entirely redundant. This suggests researchers and product developers should carefully distinguish between these descriptors when trying to understand consumer perception of food products containing fine particles.

Calculation method for novel upright CT scanner isodose curves.

PURPOSE: A computational method based on Monte-Carlo calculations is presented and used to calculate isodose curves for a new upright and tilting CT scanner useful for radiation protection purposes. METHODS: The TOPAS code platform with imported CAD files for key components was used to construct a calculation space for the scanner. A sphere of water acts as the patient would by creating scatter out of the bore. Maximum intensity dose maps are calculated for various possible tilt angles to make sure radiation protection for site planning uses the maximum possible dose everywhere. RESULTS: The resulting maximum intensity isodose lines are more rounded than ones for just a single tilt angle and so closer to isotropic. These maximum intensity curves are closer to the isotropic assumption used in CTDI or DLP based methods of site planning and radiation protection. The isodose lines are similar to those of a standard CT scanner, just tilted upwards. There is more metal above the beam that lessens the dose above versus below isocenter. CONCLUSION: Aside from the orientation, this upright scanner is very similar to a typical CT scanner, and nothing different for shielding needs to be done for this new upright tilting CT scanner, because an isotropic scatter source is often assumed for any CT scanner.

Differential Activation of TAS2R4 May Recover Ability to Taste Propylthiouracil for Some TAS2R38 AVI Homozygotes.

Bitterness from phenylthiocarbamide and 6-n-propylthiouracil (PROP) varies with polymorphisms in the TAS2R38 gene. Three SNPs form two common (AVI, PAV) and four rare haplotypes (AAI, AAV, PVI, and PAI). AVI homozygotes exhibit higher detection thresholds and lower suprathreshold bitterness for PROP compared to PAV homozygotes and heterozygotes, and these differences may influence alcohol and vegetable intake. Within a diplotype, substantial variation in suprathreshold bitterness persists, and some AVI homozygotes report moderate bitterness at high concentrations. A second receptor encoded by a gene containing a functional polymorphism may explain this. Early work has suggested that PROP might activate TAS2R4 in vitro, but later work did not replicate this. Here, we identify three TAS2R4 SNPs that result in three diplotypes-SLN/SLN, FVS/SLN, and FVS/FVS-which make up 25.1%, 44.9%, and 23.9% of our sample. These TAS2R4 haplotypes show minimal linkage disequilibrium with TAS2R38, so we examined the suprathreshold bitterness as a function of both. The participants (n = 243) rated five PROP concentrations in duplicate, interleaved with other stimuli. As expected, the TAS2R38 haplotypes explained ~29% (p < 0.0001) of the variation in the bitterness ratings, with substantial variation within the haplotypes (AVI/AVI, PAV/AVI, and PAV/PAV). Notably, the TAS2R4 diplotypes (independent of the TAS2R38 haplotypes) explained ~7-8% of the variation in the bitterness ratings (p = 0.0001). Given this, we revisited if PROP could activate heterologously expressed TAS2R4 in HEK293T cells, and calcium imaging indicated 3 mM PROP is a weak TAS2R4 agonist. In sum, our data are consistent with the second receptor hypothesis and may explain the recovery of the PROP tasting phenotype in some AVI homozygotes; further, this finding may potentially help explain the conflicting results on the TAS2R38 diplotype and food intake.

Orthorexia Nervosa and Healthy Orthorexia in a Physically Active North American Population.

The Teruel Orthorexia Scale (TOS) defines two related but distinct constructs: Orthorexia Nervosa (OrNe), a pathological fixation on a healthy diet, and Healthy Orthorexia (HeOr), an interest in a healthy diet independent of psychopathology. Here, we (a) assessed both types of Orthorexia in a large North American sample using the TOS and (b) explored if engaging in regular physical activity was associated with a greater risk of Orthorexia. A cohort of physically active adults (n = 927; 41% men) completed the TOS, as well as the Rapid Assessment of Physical Activity (RAPA), to broadly assess aerobic physical activity level and participation in strength and/or flexibility training. As expected, scores for HeOr and OrNe differed between participants, with lower scores for Orthorexia Nervosa in our physically active non-clinical sample. Higher HeOr scores were associated with lower BMI, and this was true for both men and women. We also found that measures of Orthorexia were associated with self-reported physical activity: active adults reporting more aerobic physical activity had higher HeOr scores, with the most active men having the highest scores. Notably, adults who reported regular strength training had higher scores for both HeOr and OrNe, with men who strength trained showing higher OrNe scores than women. Here, those who participate in regular strength training are more likely to exhibit orthorexic behaviors, and this effect was more pronounced for men than women. Prior work has validated the TOS in young, primarily female samples of non-English speakers outside the United States: present data from an age-diverse, physically active, gender balanced sample support the use of TOS for measurement of Orthorexia Nervosa and Healthy Orthorexia in English speakers and suggest that more work is needed to assess potential gender differences in these constructs.

Distinct Sensory Hedonic Functions for Sourness in Adults.

Over the last half-century, variable responses to sweetness have repeatedly been shown to fall into a small number of hedonic responses, implying that looking only at group means may can obfuscate meaningfully different response patterns. Comparative data for sourness is quite sparse, especially in adults. While increased liking with higher acid concentration has been reported for some children, in adults, sourness is classically assumed to be aversive, with a monotonic drop in liking with increasing sourness. Here, we test this assumption using a simple model system or experimental beverage in convenience samples of adults from the United States (increasing citric acid in water) and Italy (increasing citric acid in pear juice). Participants rated intensity and liking of sampled stimuli. For both cohorts, we find clear evidence of three distinct patterns of responses: a strong negative group where liking dropped with increased sourness, an intermediate group who showed a more muted drop in liking with more sourness, and a strong positive group where liking increased with more sourness. Strikingly, both cohorts showed similar proportions of response patterns, with ~63-70% in the strong negative group, and 11-12% in the strong positive group, suggesting these proportions may be stable across cultures. Notably, the three groups did not differ by age or gender. These data support the existence of different hedonic response profiles to sour stimuli in adults, once again highlighting the importance of looking at individual differences and potential consumer segments, rather than merely averaging hedonic responses across all individuals within a group.

Immunotherapy and Radiotherapy Combinations for Sarcoma.

Sarcomas are a heterogeneous group of bone and soft tissue tumors. Survival outcomes for advanced (unresectable or metastatic) disease remain poor, so therapeutic improvements are needed. Radiotherapy plays an integral role in the neoadjuvant and adjuvant treatment of localized disease as well as in the treatment of metastatic disease. Combining radiotherapy with immunotherapy to potentiate immunotherapy has been used in a variety of cancers other than sarcoma, and there is opportunity to further investigate combining immunotherapy with radiotherapy to try to improve outcomes in sarcoma. In this review, we describe the diversity of the tumor immune microenvironments for sarcomas and describe the immunomodulatory effects of radiotherapy. We discuss studies on the timing of radiotherapy relative to immunotherapy and studies on the radiotherapy dose and fractionation regimen to be used in combination with immunotherapy. We describe the impact of radiotherapy on the tumor immune microenvironment. We review completed and ongoing clinical trials combining radiotherapy with immunotherapy for sarcoma and propose future directions for studies combining immunotherapy with radiotherapy in the treatment of sarcoma.

Inhibition of glycogen synthase kinase-3 enhances NRF2 protein stability, nuclear localisation and target gene transcription in pancreatic beta cells.

Accumulation of reactive oxygen species (i.e., oxidative stress) is a leading cause of beta cell dysfunction and apoptosis in diabetes. NRF2 (NF-E2 p45-related factor-2) regulates the adaptation to oxidative stress, and its activity is negatively regulated by the redox-sensitive CUL3 (cullin-3) ubiquitin ligase substrate adaptor KEAP1 (Kelch-like ECH-associated protein-1). Additionally, NRF2 is repressed by the insulin-regulated Glycogen Synthase Kinase-3 (GSK3). We have demonstrated that phosphorylation of NRF2 by GSK3 enhances ß-TrCP (beta-transducin repeat-containing protein) binding and ubiquitylation by CUL1 (cullin-1), resulting in increased proteasomal degradation of NRF2. Thus, we hypothesise that inhibition of GSK3 activity or ß-TrCP binding upregulates NRF2 and so protects beta cells against oxidative stress. We have found that treating the pancreatic beta cell line INS-1 832/13 with the KEAP1 inhibitor TBE31 significantly enhanced NRF2 protein levels. The presence of the GSK3 inhibitor CT99021 or the ß-TrCP-NRF2 protein-protein interaction inhibitor PHAR, along with TBE31, resulted in prolonged NRF2 stability and enhanced nuclear localisation (P < 0.05). TBE31-mediated induction of NRF2-target genes encoding NAD(P)H quinone oxidoreductase 1 (Nqo1), glutamate-cysteine ligase modifier (Gclm) subunit and heme oxygenase (Hmox1) was significantly enhanced by the presence of CT99021 or PHAR (P < 0.05) in both INS-1 832/13 and in isolated mouse islets. Identical results were obtained using structurally distinct GSK3 inhibitors and inhibition of KEAP1 with sulforaphane. In summary, we demonstrate that GSK3 and ß-TrCP/CUL1 regulate the proteasomal degradation of NRF2, enhancing the impact of KEAP1 regulation, and so contributes to the redox status of pancreatic beta cells. Inhibition of GSK3, or ß-TrCP/CUL1 binding to NRF2 may represent a strategy to protect beta cells from oxidative stress.

Unit size influences ad libitum intake in a snacking context via eating rate.

Geometric and textural properties of food, like unit size, have previously been shown to influence energy intake. While mechanism(s) driving this effect are unclear, unit size may relate to intake by affecting eating microstructure (e.g., eating rate, bite size). In a randomized crossover study, we investigated relationships between unit size, eating microstructure, and intake. Adults (n = 75, 75% women) consumed an ad libitum snack three times in our laboratory. This snack was a 70-g portion (∼2.5 servings) of one of three sizes of pretzel (small, medium, large). Intake was measured in grams by difference in weight before and after the snack. Each session was video recorded to measure eating microstructure; snack duration (min) and number of bites were annotated and used to calculate mean eating rate (g/min) and mean bite size (g/bite). Results revealed unit size influenced intake (grams and kcal; both p's ≤ 0.001), such that participants consumed 31% and 22% more of the large pretzels (16.9 ± 2.3 g) compared to the small (12.9 ± 2.3 g) and medium sizes (13.8 ± 2.3 g), respectively. Unit size also influenced eating rate and bite size (both p's < 0.001); the largest pretzel size yielded the fastest eating rate and largest mean bite size. Further analysis revealed that after accounting for eating microstructure, the effects of unit size on intake were no longer significant, suggesting eating microstructure was driving these effects. Together, these findings indicate that unit size influences intake by affecting eating microstructure and that food properties like unit size can be leveraged to moderate snack intake.

Transcriptomic profiling of sciatic nerves and dorsal root ganglia reveals site-specific effects of prediabetic neuropathy.

Peripheral neuropathy (PN) is a severe and frequent complication of obesity, prediabetes, and type 2 diabetes characterized by progressive distal-to-proximal peripheral nerve degeneration. However, a comprehensive understanding of the mechanisms underlying PN, and whether these mechanisms change during PN progression, is currently lacking. Here, gene expression data were obtained from distal (sciatic nerve; SCN) and proximal (dorsal root ganglia; DRG) injury sites of a high-fat diet (HFD)-induced mouse model of obesity/prediabetes at early and late disease stages. Self-organizing map and differentially expressed gene analyses followed by pathway enrichment analysis identified genes and pathways altered across disease stage and injury site. Pathways related to immune response, inflammation, and glucose and lipid metabolism were consistently dysregulated with HFD-induced PN, irrespective of injury site. However, regulation of oxidative stress was unique to the SCN while dysregulated Hippo and Notch signaling were only observed in the DRG. The role of the immune system and inflammation in disease progression was supported by an increase in the percentage of immune cells in the SCN with PN progression. Finally, when comparing these data to transcriptomic signatures from human patients with PN, we observed conserved pathways related to metabolic dysregulation across species, highlighting the translational relevance of our mouse data. Our findings demonstrate that PN is associated with distinct site-specific molecular re-programming in the peripheral nervous system, identifying novel, clinically relevant therapeutic targets.

Evaluating Barriers to Opioid Use Disorder Treatment From Patients' Perspectives.

Introduction: Utilizing medications to treat opioid use disorder (MOUD) is both highly effective and unfortunately underutilized in the US health care system. Stigma surrounding substance use disorders, insufficient provider knowledge about substance use disorders and MOUD, and historical lack of physicians with X-waivers to prescribe buprenorphine contribute to this underutilization. Our study aimed to elucidate barriers to accessing MOUD in Milwaukee, Wisconsin. Methods: We conducted semistructured interviews with patients receiving MOUD at a family medicine residency program in Milwaukee, Wisconsin. Interviews were audio-recorded, transcribed verbatim, and analyzed using the qualitative analysis Framework Method. Researchers in our team reviewed transcripts, coding for specific topics of discussion. Coded transcript data were then sorted into a matrix to identify common themes. Results: Interviews with 30 participants showed that motivations to seek treatment appeared self-driven and/or for loved ones. Eighteen patients noted concerns with treatment including treatment denial and efficacy of treatment. Housing instability, experiences with incarceration, insurance, and transportation were common structural barriers to treatment. Conclusions: Primary drivers to seek treatment were patients themselves and/or loved ones. Barriers to care include lack of effective transportation, previous experience with the carceral system, and relative scarcity of clinicians offering MOUD. Future studies may further explore effects of structural inadequacies and biases on MOUD access and quality.

Sugar reduction in chocolate compound by replacement with flours containing small insoluble starch granules.

Consumer concern about intake of added sugars has increased commercial demand for sugar-reduced chocolates. However, substitution with high-potency sweeteners is not possible as sugar serves as an important bulking agent. Here, we investigated replacement of sucrose in chocolate compound with oat or rice flours, with starch granules smaller than 10 µm, focusing on texture, sweetness, and acceptability. In Study 1, six chocolates were made: a control with 54% sucrose, four sucrose-reduced versions (reductions of 25% or 50%, using either oat or sweet rice flour), and one 54% sucrose chocolate with reduced refining time. These chocolates were compared in a Difference from Control (DFC) test in a within-subject design, with and without nose clips. Particle size distribution, yield stress, and plastic viscosity were measured. Chocolates with 25% sucrose reduction by either sweet rice or oat flour (or reduced refining time) were not significantly different from the blind control (p > 0.05), regardless of nose clip use. In open-ended comments, participants reported differences in rice-flour-containing chocolates were due to a chalkier texture, while oat-flour-containing chocolates were described as smoother, softer, and creamier. DFC scores from the chocolates were positively correlated with plastic viscosity and negatively correlated with yield stress. In Study 2, 25% reduced sugar chocolates made with rice flour were liked significantly less than control, but the oat flour sample did not differ from control. Collectively, these results suggest up to 25% of sucrose in chocolate can be replaced with oat flour without negatively affecting texture or consumer acceptance.

Using Herbs/Spices to Enhance the Flavor of Commonly Consumed Foods Reformulated to Be Lower in Overconsumed Dietary Components Is an Acceptable Strategy and Has the Potential to Lower Intake of Saturated Fat and Sodium: A National Health and Nutrition Examination Survey Analysis and Blind Tasting.

BACKGROUND: Foods lower in saturated fat, sodium, and added sugars (ie, overconsumed dietary components) must have an acceptable flavor profile to promote intake. OBJECTIVE: The aim of this research was to model the influence of using herbs/spices as flavor-enhancers when reducing overconsumed dietary components in commonly consumed foods and evaluate acceptance of these flavor-enhanced reformulations. DESIGN: Ten leading sources of overconsumed dietary components were identified using the National Health and Nutrition Examination Survey 2015-2018 dietary data. These foods were reformulated to reduce overconsumed dietary components and herbs/spices were used to preserve acceptability. The influence of consumer adoption of the reformulated foods on intake of overconsumed dietary components was modeled using National Health and Nutrition Examination Survey data. Consumer acceptability of the reformulated recipes was assessed with blind taste testing. PARTICIPANTS/SETTING: Dietary data from adults aged 19 years and older (n = 9,812) included in the National Health and Nutrition Examination Survey 2015-2018 were used to identify foods for reformulation and model the potential influence of reformulation. The blind taste testing included 85 to 107 consumers per panel. MAIN OUTCOME MEASURES: Estimated daily change in total intake of saturated fat, sodium, added sugars, and energy with intake of the reformulated foods instead of the original foods. Consumer ratings of overall liking of the reformulated recipes vs the original recipes were assessed using standard 9-point hedonic scales. STATISTICAL ANALYSES PERFORMED: Descriptive statistics with use of survey procedures were used to model the influence of reformulated food adoption. Mixed effect models were used for analysis of the blind tasting data. RESULTS: With intake of the reformulated foods, instead of the original versions, by 25% to 100% of current consumers, estimates suggest lowering of saturated fat (25% consumer adoption to 100% consumer adoption -2.9% to -11.4%, respectively), sodium (-3.2 to -11.5%, respectively), and added sugars (-0.5 to -2.7%, respectively) intake. The overall liking ratings for seven of the 10 reformulated foods were superior or at parity with the original foods. CONCLUSIONS: This proof-of-concept research suggests that using herbs/spices to create flavor-enhanced recipes lower in overconsumed dietary components has the potential to reduce intake and is acceptable to consumers.

Inducible desensitization to capsaicin with repeated low-dose exposure in human volunteers.

Responses to capsaicin are reduced following repeated exposure, a phenomenon known as capsaicin desensitization. Heavy consumers of chilies consistently report reduced oral burn relative to infrequent consumers, presumably due to chronic desensitization. However, the mechanism(s) underlying capsaicin desensitization remain poorly understood. We hypothesized that reduced response to capsaicin due to repeated oral exposure may result from a change in the expression of the capsaicin receptor (TRPV1) gene. To test this, we conducted two longitudinal desensitization studies in healthy human volunteers. In Study 1, 51 adults completed a 17-day capsaicin desensitization protocol. The study consisted of three in-person visits where they were asked to sample stimuli, including 3, 6, and 9 ppm capsaicin, and rate intensity on a general labeled magnitude scale (gLMS). Between days 3 & 17, participants rinsed at home with 6 ppm capsaicin (n = 31) or a control (n = 20) solution (20 uM sucrose octaccetate; SOA) twice a day. Before and after the oral exposure protocol, a clinician collected fungiform papillae. Participants randomized to the capsaicin rinse showed a statistically significant reduction in oral burn ratings that was not observed in controls, indicating repeated low-dose exposure can systematically induce desensitization. TRPV1 expression was not associated with reported capsaicin burn, and there was no evidence of a decrease in TRPV1 expression following capsaicin exposure. In Study 2, participants (n = 45) rinsed with 6 ppm capsaicin in a similar protocol, rating capsaicin, vanillyl butyl ether (VBE), cinnamaldehyde, ethanol, menthol, and sucrose on days 1, 3, & 17. Burn from capsaicin, VBE, cinnamaldehyde, and ethanol all showed a statistically significant change - capsaicin, VBE and cinnamaldehyde burn all dropped ∼20 %, and a larger reduction was seen for ethanol - while menthol cooling and sucrose sweetness did not change. Collectively, this suggests reductions in oral burn following chronic capsaicin exposure generalizes to other stimuli (i.e., cross desensitization) and this cannot be explained by a change in TRPV1 mRNA expression. More work is needed to elucidate the underlying mechanism for capsaicin desensitization in the oral cavity.

Reasons for meal termination, eating frequency, and typical meal context differ between persons with and without a spinal cord injury.

Overeating associated with neurogenic obesity after spinal cord injury (SCI) may be related to how persons with SCI experience satiation (processes leading to meal termination), their eating frequency, and the context in which they eat their meals. In an online, cross-sectional study, adults with (n = 688) and without (Controls; n = 420) SCI completed the Reasons Individuals Stop Eating Questionnaire-15 (RISE-Q-15), which measures individual differences in the experience of factors contributing to meal termination on five scales: Physical Satisfaction, Planned Amount, Decreased Food Appeal, Self-Consciousness, and Decreased Priority of Eating. Participants also reported weekly meal and snack frequency and who prepares, serves, and eats dinner with them at a typical dinner meal. Analysis revealed that while Physical Satisfaction, Planned Amount, and Decreased Food Appeal were reported as the most frequent drivers of meal termination in both groups, scores for the RISE-Q-15 scales differed across the groups. Compared to Controls, persons with SCI reported Physical Satisfaction and Planned Amount as drivers of meal termination less frequently, and Decreased Food Appeal and Decreased Priority of Eating more frequently (all p < 0.001). This suggests that persons with SCI rely less on physiological satiation cues for meal termination than Controls and instead rely more on hedonic cues. Compared to Controls, persons with SCI less frequently reported preparing and serving dinner meals and less frequently reported eating alone (all p < 0.001), indicating differences in meal contexts between groups. Individuals with SCI reported consuming fewer meals than Controls but reported a higher overall eating frequency due to increased snacking (p ≤ 0.015). A decrease in the experience of physical fullness, along with a dependence on a communal meal context and frequent snacking, likely contribute to overeating associated with neurogenic obesity after SCI.

Human neural stem cells restore spatial memory in a transgenic Alzheimer's disease mouse model by an immunomodulating mechanism.

Introduction: Stem cells are a promising therapeutic in Alzheimer's disease (AD) given the complex pathophysiologic pathways involved. However, the therapeutic mechanisms of stem cells remain unclear. Here, we used spatial transcriptomics to elucidate therapeutic mechanisms of human neural stem cells (hNSCs) in an animal model of AD. Methods: hNSCs were transplanted into the fimbria fornix of the hippocampus using the 5XFAD mouse model. Spatial memory was assessed by Morris water maze. Amyloid plaque burden was quantified. Spatial transcriptomics was performed and differentially expressed genes (DEGs) identified both globally and within the hippocampus. Subsequent pathway enrichment and ligand-receptor network analysis was performed. Results: hNSC transplantation restored learning curves of 5XFAD mice. However, there were no changes in amyloid plaque burden. Spatial transcriptomics showed 1,061 DEGs normalized in hippocampal subregions. Plaque induced genes in microglia, along with populations of stage 1 and stage 2 disease associated microglia (DAM), were normalized upon hNSC transplantation. Pathologic signaling between hippocampus and DAM was also restored. Discussion: hNSCs normalized many dysregulated genes, although this was not mediated by a change in amyloid plaque levels. Rather, hNSCs appear to exert beneficial effects in part by modulating microglia-mediated neuroinflammation and signaling in AD.

Human neural stem cells restore spatial memory in a transgenic Alzheimer's disease mouse model by an immunomodulating mechanism.

INTRODUCTION: Stem cells are a promising therapeutic in Alzheimer's disease (AD) given the complex pathophysiologic pathways involved. However, the therapeutic mechanisms of stem cells remain unclear. Here, we used spatial transcriptomics to elucidate therapeutic mechanisms of human neural stem cells (hNSCs) in an animal model of AD. METHODS: hNSCs were transplanted into the fimbria fornix of the hippocampus using the 5XFAD mouse model. Spatial memory was assessed by Morris water maze. Amyloid plaque burden was quantified. Spatial transcriptomics was performed and differentially expressed genes (DEGs) identified both globally and within the hippocampus. Subsequent pathway enrichment and ligand-receptor network analysis was performed. RESULTS: hNSC transplantation restored learning curves of 5XFAD mice. However, there were no changes in amyloid plaque burden. Spatial transcriptomics showed 1061 DEGs normalized in hippocampal subregions. Plaque induced genes in microglia, along with populations of stage 1 and stage 2 disease associated microglia (DAM), were normalized upon hNSC transplantation. Pathologic signaling between hippocampus and DAM was also restored. DISCUSSION: hNSCs normalized many dysregulated genes, although this was not mediated by a change in amyloid plaque levels. Rather, hNSCs appear to exert beneficial effects in part by modulating microglia-mediated neuroinflammation and signaling in AD.

Regional interneuron transcriptional changes reveal pathologic markers of disease progression in a mouse model of Alzheimer's disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder and leading cause of dementia, characterized by neuronal and synapse loss, amyloid-ß and tau protein aggregates, and a multifactorial pathology involving neuroinflammation, vascular dysfunction, and disrupted metabolism. Additionally, there is growing evidence of imbalance between neuronal excitation and inhibition in the AD brain secondary to dysfunction of parvalbumin (PV)- and somatostatin (SST)-positive interneurons, which differentially modulate neuronal activity. Importantly, impaired interneuron activity in AD may occur upstream of amyloid-ß pathology rendering it a potential therapeutic target. To determine the underlying pathologic processes involved in interneuron dysfunction, we spatially profiled the brain transcriptome of the 5XFAD AD mouse model versus controls, across four brain regions, dentate gyrus, hippocampal CA1 and CA3, and cortex, at early-stage (12 weeks-of-age) and late-stage (30 weeks-of-age) disease. Global comparison of differentially expressed genes (DEGs) followed by enrichment analysis of 5XFAD versus control highlighted various biological pathways related to RNA and protein processing, transport, and clearance in early-stage disease and neurodegeneration pathways at late-stage disease. Early-stage DEGs examination found shared, e.g ., RNA and protein biology, and distinct, e.g ., N-glycan biosynthesis, pathways enriched in PV-versus somatostatin SST-positive interneurons and in excitatory neurons, which expressed neurodegenerative and axon- and synapse-related pathways. At late-stage disease, PV-positive interneurons featured cancer and cancer signaling pathways along with neuronal and synapse pathways, whereas SST-positive interneurons showcased glycan biosynthesis and various infection pathways. Late-state excitatory neurons were primarily characterized by neurodegenerative pathways. These fine-grained transcriptomic profiles for PV- and SST-positive interneurons in a time- and spatial-dependent manner offer new insight into potential AD pathophysiology and therapeutic targets.

Palmitate and glucose increase amyloid precursor protein in extracellular vesicles: Missing link between metabolic syndrome and Alzheimer's disease.

The metabolic syndrome (MetS) and Alzheimer's disease share several pathological features, including insulin resistance, abnormal protein processing, mitochondrial dysfunction and elevated inflammation and oxidative stress. The MetS constitutes elevated fasting glucose, obesity, dyslipidaemia and hypertension and increases the risk of developing Alzheimer's disease, but the precise mechanism remains elusive. Insulin resistance, which develops from a diet rich in sugars and saturated fatty acids, such as palmitate, is shared by the MetS and Alzheimer's disease. Extracellular vesicles (EVs) are also a point of convergence, with altered dynamics in both the MetS and Alzheimer's disease. However, the role of palmitate- and glucose-induced insulin resistance in the brain and its potential link through EVs to Alzheimer's disease is unknown. We demonstrate that palmitate and high glucose induce insulin resistance and amyloid precursor protein phosphorylation in primary rat embryonic cortical neurons and human cortical stem cells. Palmitate also triggers insulin resistance in oligodendrocytes, the supportive glia of the brain. Palmitate and glucose enhance amyloid precursor protein secretion from cortical neurons via EVs, which induce tau phosphorylation when added to naïve neurons. Additionally, EVs from palmitate-treated oligodendrocytes enhance insulin resistance in recipient neurons. Overall, our findings suggest a novel theory underlying the increased risk of Alzheimer's disease in MetS mediated by EVs, which spread Alzheimer's pathology and insulin resistance.