bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ablation of the Carotenoid Cleavage Enzyme β‐Carotene‐15,15′‐Dioxygenase (BCO1) Reduces Serum Testosterone and Prostatic Androgen Receptor Signaling in Mice
Authors: Smith Joshua W. , Ford Nikki A. , Thomas‐Ahner Jennifer M. , Moran Nancy E. , Bolton Eric C. , Clinton Steven K. , Erdman John W. 
Published: November 2023

β‐carotene‐15,15′‐dioxygenase (BCO1) cleaves dietary carotenoids at the central 15,15′ double bond, most notably acting on β‐carotene to yield retinal. Ablation of this gene in mice fed carotenoid‐rich diets has been shown to affect tissue carotenoid accumulation. However, Bco1 ablation also impacts diverse physiologic endpoints independent of dietary carotenoid feeding, including expression of genes controlling androgen metabolism. Using the Bco1−/−mouse model, we sought to probe the effects of Bco1 ablation on testicular steroidogenesis, serum testosterone levels, and prostatic androgen signaling. Male wild‐type (WT) and Bco1−/− mice were weaned and raised on carotenoid‐free, semi‐purified AIN‐93G diets before sacrifice at 10–14 weeks of age. In three independent trials, Bco1 ablation significantly and consistently decreased serum testosterone and altered prostatic homeostasis. Organ weights of the prostate (all lobes) and seminal vesicles were significantly lower in Bco1−/−than WT mice (−18% and −29%, respectively; p<0.001). Bco1−/− mice had significantly reduced levels of serum testosterone (−79%; p<0.05). Additionally, Bco1 ablation decreased testicular mRNA expression of Hsd17b3 (−32% p<0.01), while increasing Hsd17b2 expression (+34%, p<0.05), suggesting a possible inhibition of testosterone synthesis. Furthermore, prostatic NanoString gene expression analysis revealed that Bco1 loss significantly (p<0.005) disrupted androgen receptor signaling, cell cycle progression, and proliferation. Reduced mRNA expression (by qPCR) of the androgen‐induced gene Msmb (−30%, p<0.05) and reduced protein expression of the proliferation marker Ki67 (by immunofluorescence microscopy; −47%, p<0.001) support these findings. While BCO1 has been suggested to function in physiologic processes more diverse than carotenoid cleavage and production of vitamin A, this is the first demonstration that Bco1 ablation impacts murine prostatic proliferation and androgen receptor signaling.Support or Funding InformationFunded by NIH grant PHS‐1‐R01 CA125384
Acute excess caloric intake negatively affects the rhythmic expression of the human growth hormone gene and its association with the circadian‐linked transcription factor Bmal1
Authors: Vakili Hana , Jin Yan , Cattini Peter A. 
Published: November 2023

Rhythmicity of biological functions is fundamental for optimal adaptations to environmental cues. Growth hormone (GH) is a major metabolic homeostatic factor that is secreted with a circadian pattern, but whether it is produced rhythmically is unknown. We used transgenic (TG) mice containing the intact human GH gene locus (human GH/CS‐TG) to investigate the rhythmicity of human GH gene (hGH‐N) expression. Human GH RNA expression oscillates over a 24‐hour cycle. Moreover, we have previously reported that excess caloric intake is associated with a decrease in human GH synthesis with reduced hGH‐N promoter function. Sequence analysis of the hGH‐N promoter revealed the presence of an enhancer motif (E‐box) and as such a putative binding site for circadian transcriptional machinery (Bmal1 and Clock). Results from electrophoretic mobility shift assay support participation of Bmal1/Clock and the hGH‐N E‐box DNA element in a common complex. Transfection studies combined with site‐directed mutagenesis revealed that this E‐box DNA element contributes to basal hGH‐N promoter activity. Furthermore, Bmal1/Clock were able to transactivate the hGH‐N promoter. The ability of Bmal1 to associate with the hGH‐N promoter region containing the E‐box element was confirmed in the human GH/CS‐TG mouse pituitary in situ by chromatin immunoprecipitation assay. Occupancy was reduced in mice fed a high fat diet during the light cycle. This decrease was not seen, however, during the dark cycle. Initial assessment of the mechanism suggests an impact of excess caloric intake on the chromatin and/or DNA structure. In summary, the presence of an E‐box DNA element as well as association of Bmal1 with the hGH‐N promoter region in situ supports the notion that hGH‐N is a cyclic gene. Acute excess caloric intake for three days was sufficient to reprogram the circadian rhythm mediated by chromatin remodeling of the hGH‐N promoter region and interfer with recruitment of Bmal1 to the E‐box element. Further investigation of the rhythmic expression of human GH may provide insight into the changes in GH availability under pathophysiological conditions such as metabolic changes associated with overeating and obesity.Support or Funding InformationFunded by the Canadian Institutes of Health Research
Soy protein in the prevention of hepatic steatosis in obese OLETF rats
Authors: Rector R Scott , Schuster Colin M , Phillips Kathryn E , Eck Kevin J , Meers Grace M , Butteiger Dustie N , Krul Elaine S , Thyfault John P 
Published: November 2023

Soy protein has shown beneficial effects on hepatic lipid metabolism and prevention of hepatic steatosis. Here we sought to test our hypothesis that a soy protein versus dairy protein‐based diet would significantly improve hepatic mitochondrial function and attenuate hepatic steatosis development in hyperphagic Otsuka Long‐Evans Tokushima Fatty (OLETF) rats. Five week old male OLETF rats were randomized to isocaloric diets containing 21% kcal as protein from milk protein isolate (MPI), soy protein isolate (SPI), or 50:50 MPI/SPI (M/S) (N=9–10/group) for 16 weeks. SPI significantly (p<0.05) attenuated fat mass and percent body fat by ~10% compared with M/S, but did not differ from MPI. Cumulative food intake was not different between SPI and other groups, but was significantly less in MPI rats compared with M/S (p<0.05), which corresponded with lowered serum insulin (p<0.05 vs SPI and M/S); serum glucose did not differ among groups. SPI reduced serum cholesterol and serum thiobarbituric acid reactive substances (TBARS) content (p<0.05) and tended to lower serum free fatty acids and increase serum antioxidant capacity compared with the other groups (p=0.1). Histological examination of liver revealed significant attenuation (p<0.05) in hepatic steatosis in the SPI group compared with MPI and M/S and biochemical assessment of liver triglyceride content showed a similar trend. Lipidomics analyses also indicated significant reductions (p<0.05) in hepatic diacylglycerols and phosphatidylcholine in SPI vs MPI. Intriguingly, reduced hepatic steatosis with SPI was associated with dramatic reductions in markers of hepatic de novo lipogenesis (acetyl‐coA carboxylase and fatty acid synthase protein content) but not with improvements in hepatic mitochondrial content or function. In fact, MPI livers displayed higher hepatic mitochondrial respiration, β‐hydroxy‐acyl‐CoA dehydrogenase activity and citrate synthase activity (p<0.05) compared with SPI and M/S groups. Furthermore, milk protein and soy protein appear to impact different pathways in hepatic lipid metabolism, with milk protein upregulating hepatic mitochondrial content/function and soy protein downregulating hepatic de novo lipogenesis. In summary, soy protein may be more beneficial than milk protein in the attenuation of hepatic steatosis development in hyperphagic obese OLETF rats.Support or Funding InformationSupport: MU School of Medicine Summer Research Fellowship (CMS), DuPont Nutrition & Health, NIH R01DK088940 (JPT) and VHA‐CDA2 IK2BX001299 (RSR).
The mechanosensitive ion channel Piezo2 is selectively expressed in enterochromaffin (EC) cells of the human and mouse gastrointestinal mucosa
Authors: Knutson Kaitlyn , Wang Fan , Alcaino Constanza , Bernard Cheryl , Farrugia Gianrico , Beyder Arthur 
Published: November 2023

BACKGROUNDEnterochromaffin (EC) cells are enteroendocrine cells in the gastrointestinal (GI) epithelium that release serotonin (5‐HT) in response to a variety of stimuli including mechanical force. In turn, EC cell 5‐HT affects GI secretion, motility and sensation. The molecular mechanism of EC cell mechanotransduction is unknown. The recently discovered mechanosensitive ion channel Piezo2 is important in several mechanosensory cell types.AIMto determine if the mechanosensitive ion channel Piezo2 is expressed and functional in EC cells.METHODSMurine small bowel was collected, fixed in 4% paraformaldehyde, and frozen. Human and mouse RNA from human jejunum and mouse duodenum, jejunum and ileum was analyzed by quantitative RT‐PCR for Piezo2 mRNA. Piezo2 and 5‐HT were also identified by fluorescence immunolabeling. QGP‐1, a human enteroendocrine cell model, was analyzed by RT‐PCR for the presence of Piezo2 and immunolabeling was done to detect Piezo2 and 5‐HT. Piezo2 mechanically activated currents were recorded from QGP‐1 cells in whole‐cell configuration. Voltage‐clamped QGP‐1 cells were subjected to a series of mechanical perturbations using a piezo‐electrically driven glass probe at steps of 0.3 μm for stimulation.RESULTSHuman jejunum and all segments of mouse small bowel mucosa (duodenum, jejunum, ileum) expressed Piezo2 mRNA. In human jejunum and all segments of mouse small bowel Piezo2 co‐localized with 5‐HT in >90% of 5‐HT containing cells. No Piezo2 immunoreactivity was seen outside of 5‐HT positive enterochromaffin cells. The enteroendocrine cell model QGP‐1 expressed both Piezo2 and 5‐HT by immunohistochemistry and RT‐PCR showed message for Piezo2. Application of force to the surface of voltage‐clamped QGP‐1 cells elicited whole cell fast inward non‐selective cation currents of ~200 pA (at −60 mV) that were blocked by Gd3+ with kinetic properties compatible with Piezo2 channels.CONCLUSIONSPiezo2 is specifically expressed in human and murine small bowel EC cells and in the enteroendocrine cell model QGP‐1. QGP‐1 cells have a mechanosensitive inward cation current with kinetic and pharmacological properties similar to Piezo2. Our data suggest that Piezo2 is involved in EC cell mechanotransduction.Support or Funding InformationSupported by NIH DK106456, DK84567, DK052766
Effect of Pediococcus pentosaceus at different doses on in vivo model of acute ulcerative colitis.Effect of Pediococcus pentosaceus at different doses on in vivo model of acute ulcerative colitis
Authors: AGUILAR LORENA TORRES , FRAGOSO LOURDES RODRIGUEZ , REYES‐ESPARZA JORGE 
Published: November 2023

Ulcerative colitis (UC) is a chronic inflammatory disease of the colon, whose characteristic symptoms include rectal bleeding or bloody diarrhea, abdominal pain and weight loss. Factors associated with the development of CU is the change of intestinal microbiota, for example declining species of Pediococcus. Therefore a new alternatives for the treatment of UC is the use of probiotics. Probiotics are live microorganisms which when administered in adequate amounts exert a beneficial effect on human health. There are several strains of Pediococcus pentosaceus with probiotic properties. The aim of this study is to evaluate the efficacy of different doses of Pediococcus pentosaceus on in vivo model of acute ulcerative colitis induced with dextran sodium sulfate (DSS). Thirty‐six male BALB/c mice weighing 20–22 g were housed in groups of 6 mice/cage. Acute colitis was induced by administration of DSS (molecular weight 40 kDa) 3.5% wt/vol in drinking water ad libitum for 7 days. At day 2 during induction of colitis, four groups were administered via gastric tube once daily at different doses of Pediococcus pentosaceus (1×107, 1×108, 1×109, and 1×1010 Colony Forming Units(CFU)). The clinical course of colitis was monitored by a daily disease activity index consisting of the three parameters weight loss, stool consistency, and perianal bleeding. The animals were sacrificed on the seventh day, the colon length was measured and histology were performed using H & E staining. P. pentosaceus at dose of 1×107, 1×108, 1×109, 1×1010 CFU didn't change the weight in animals with ulcerative colitis. However, the animals treated with P. pentosaceus at dose of 1×1010 delay the onset of diarrhea and bleeding during the induction of damage and showed a disease activity index by 41% less than the UC group. Furthermore, this group has a 21.3% less colon shortening than the CU group (p <0.05). The groups treated with P. pentosaceus at different doses (1×107, 1×108, 1×109 and 1×1010 UFC) didn't show damage to the submucosa of the colon, because the damage is only present in the area of the mucosa and muscularis mucosa. In conclusion Pediococcus pentosaceus treatment at the dose of 1×1010 CFU delayed the onset of some of the symptoms of acute ulcerative colitis in animals.Support or Funding InformationLorena Torres Aguilar es fellowship from CONACYT
Effects of probiotic bacteria on cytokine expression in cultured intestinal epithelial cells
Authors: Lyle Sarah , Erickson Tony , Walton Kristen LW 
Published: November 2023

The benefits of probiotic bacteria in intestinal physiology and disease may be mediated through multiple mechanisms, such as affecting the composition of the gut microbiome, having direct action on host intestinal cells, or modulating mucosal immune responses. The commercially available probiotic VSL#3 includes a mixture of eight species, mostly lactobacilli and bifidobacteria. VSL#3 has been shown to have clinical benefits in ulcerative colitis patients, and has also been demonstrated to alter expression of mucins and other genes in cultured intestinal epithelial cells. This study aimed to assess effects of Lactobacillus acidophilus or VSL#3 on pro‐ and anti‐inflammatory cytokine and tight junction protein expression in CMT‐93 and Caco‐2 colonic epithelial cells. Cells were treated for 4 hours with L. acidophilus or VSL#3 (1×108 CFU/ml) and expression of the anti‐inflammatory cytokine transforming growth factor‐beta‐1 (TGFβ‐1), pro‐inflammatory interleukin (IL)‐6 and the housekeeping gene GAPDH was analyzed by semi‐quantitative PCR. The number of viable cells was analyzed after 24 hours of exposure to probiotic bacteria using a cell survival assay. Expression of claudin‐2 and localization of nuclear factor kappa‐B (NFkB) were detected by immunofluorescent staining. Probiotic‐treated CMT‐93 cells showed less induction of IL‐6 than LPS‐treated cells. TGFβ expression was not altered by probiotic treatment compared to untreated controls. Immunofluorescent staining showed increased claudin‐2 staining in cells treated with VSL#3 for 24 hours compared to untreated controls. In conclusion, probiotic bacteria stimulate reduced IL‐6 production compared to a strong inflammatory stimulus in murine intestinal epithelial cells. In addition, VSL#3 stimulated claudin‐2 expression in CMT‐93 cells, providing a potential mechanism for protective effects of VSL#3 on the intestinal epithelium.Support or Funding InformationThis project was supported by the Missouri Western State University PORTAL program.
Effect of Salba‐Chia (Salvia Hispanica L), an Ancient Seed, in the Treatment of Overweight and Obese Patients with Type 2 Diabetes: A Double‐blind, Parallel, Randomized Controlled Trial
Authors: Jenkins Alexandra L , Brissette Christy , Jovanovski Elena , Au‐Yeung Fei , Ho Hoang Vi Thanh , Zurbau Andreea , Sievenpiper John , Vuksan Vladimir 
Published: November 2023

Preliminary studies indicate that consumption of Salba‐chia (Salvia hispanica L) improves management of type 2 diabetes and acutely suppresses appetite. In this study, we assessed the effect of Salba‐chia on body weight and related cardiovascular disease risk factors in overweight or obese adults with type 2 diabetes over 6 months.Using a double blind, parallel, controlled trial design, 77 overweight or obese patients with type 2 diabetes (HbA1c 6.8–8.5%; BMI 25–40kg/m2) were recruited. Both groups followed a 6‐month calorie‐restricted diet; one group consumed 30g/1000kcal of Salba‐chia, the other 36g/1000kcal oat bran‐based control, per day. Primary endpoint was change in body weight over 6‐months. Secondary endpoints included changes in waist circumference, body fat %, HbA1c, blood pressure, lipids, C‐reactive protein, and obesity‐related gut hormones.Fifty four participants completed the full 6‐month protocol and 58 (31 Control, 27 Salba‐Chia) were included in the final analysis. At 6‐months, participants on Salba‐chia lost significantly more weight than those on oat bran‐based control (1.9±0.5kg and 0.3±0.4kg, respectively; p=0.020), accompanied by a greater reduction in waist circumference (3.5±0.7cm and 1.1±0.7cm, respectively; p=0.027). C‐reactive protein was reduced by 1.1±0.5mg/L (39±17%) on Salba‐chia, compared to 0.2±0.4mg/L (7±20%) on control (P=0.045) and plasma adiponectin was increased on Salba‐chia by 6.5±0.7% compared to 0.0±0.6% on control (P=0.022). There were no significant differences between treatments on blood pressure, lipids and HbA1c, and fat distribution, however, a within intervention reduction of android (3.7±2.8%, P=0.031) and gynoid fat (6.9±3.9%, P=0.047) was observed in the Salba‐chia group, but not in control.The current trial demonstrates the potential of Salba‐chia to promote weight loss, and improve obesity‐related risk factors, while maintaining good glycemic control. The results suggest Salba‐chia may represent a promising supplementation to conventional therapy in the treatment of overweight and obesity in diabetes.Support or Funding InformationCanadian Diabetes Association Operating Grant, Salba‐chia was provided by Salba Smart Natural Products, LLC
Thinking Positive: The Effect of Emotional Context on Respiratory Sensory Gating
Authors: Chan Pei‐Ying Sarah , Ju Ya‐Jhih 
Published: November 2023

Respiratory sensory gating function has been found decreased by induced negative emotional stimulation in healthy adults. The increased ratio of the respiratory‐related evoked potentials (RREP) N1 peak amplitude for the second occlusion (S2) versus the first occlusion (S1), S2/S1, is indicative of decreased central neural gating mechanism. In this study, we tested the hypothesis that positive emotional context would enhance respiratory sensory gating function in healthy individuals.We tested respiratory sensory gating in a group of healthy individuals by the paired inspiratory occlusion paradigm of two transient obstructions of 150 milliseconds (msec) with 500 msec interval under both pleasant and neutral emotional contexts. The S2/S1 amplitude in the pleasant context and the neutral context were 0.57 and 0.45, respectively. There was a significant decrease in the S2/S1 ratio under the pleasant context (p < .05). After post‐hoc analysis, there was a significant decrease in S2 N1 peak amplitude in the pleasant compared to neutral context (p < .05).Our results demonstrated that positive emotional context enhances brain inhibitory mechanisms by filtering out repetitive respiratory stimuli. Further investigation on how positive emotional context modulates respiratory sensation in clinical patients is necessary.Support or Funding InformationMOST 103‐2420‐H‐182 ‐003 ‐MY2 CMRPD1B0332
Ventral Medullary Inspiratory Neurons Do Not Contribute to Increased Inspiratory Drive during Swallow
Authors: Pitts Teresa , King Suzanne , Poliacek Ivan 
Published: November 2023

Schluckatmung (swallow‐breath) is believed to be an evolutionarily preserved strategy from deglutition to eupnea. The schluckatmung is composed of both alterations in respiratory phase timing as well as a transient increase in inspiratory motor drive. This increased inspiratory motor drive creates a negative trans‐diaphragmatic pressure that moves the bolus from the pharynx into the esophagus. Our previous work showed that inspiratory neurons in the region of the dorsal respiratory column (DRC) increase instantaneous firing rate during the schluckatmung, consistent with an important role for this population of neurons in swallowing. We hypothesized that this role of DRC neurons in the generation of the schluckatmung is unique, and not shared by other inspiratory neurons in the ventrolateral medulla. A multi‐channel electrode array was used to record ventral respiratory column (VRC) neurons in anesthetized, spontaneously breathing cats. Swallowing was induced by injection of water into the oropharynx. Electromyograms (EMG) were recorded from pharyngeal, laryngeal, and chest wall muscles, and intra‐esophageal pressure was recorded from the mid‐thoracic esophagus. Unlike previously reported dorsal respiration column neurons, VRC inspiratory neurons significantly decreased their firing rate during swallow compared to eupnea. Suppression of VRC inspiratory neurons could play an important role in phase timing for swallow and phase‐lag of esubsequent inspirations for breathing and cough. Supported by HL 111215, P30‐GM103507.Support or Funding InformationSupported by HL 111215, P30‐GM103507.
Expiratory Diaphragm Activity Reduces Atelectasis Formation
Authors: Hedenstierna Goran , Pellegrini Mariangela , Roneus Agneta , Segelsjo Monica , Larsson Anders , Perchiazzi Gaetano 
Published: November 2023

ObjectiveWe hypothesized that the major inspiratory muscle, the diaphragm, also acts during expiration as a brake to maintain lung aeration and prevent collapse. We tested this hypothesis in a porcine model of surfactant‐depleted lung.MethodsAcute lung injury was induced in 8 anesthetized, tracheostomized pigs by repeated lung lavages, targeting a PaO2/FiO2 of 250 mmHg. After stabilization, the animals were switched to spontaneous breathing (SB) and underwent a decremental continuous positive end‐expiratory pressure (PEEP) trial of 15, 12, 9, 6, 3 and 0 cmH2O. Diaphragmatic electric activity during ongoing expiration (EAdiexp) and at no‐flow end‐expiration (EAdimin) (six electrodes esophageal‐gastric catheter) and the trans‐diaphragmatic pressure (gastric minus esophageal pressure, Pdi) were measured. Para‐diaphragmatic dynamic‐CT scans (dCTs) were obtained. The dCT scans were collected at end‐expiration and the amount of collapse (atelectasis) in that cut was calculated. The atelectatic tissue was defined as the sum of voxels with a density between −100 and +100 Hounsfield Units. In 4 pigs, the same protocol was repeated during mechanical pressure control ventilation (PCV) after complete muscle paralysis.ResultsIn spontaneously breathing pigs, a linear correlation was found between expiratory EAdi and expiratory Pdi (R2 > 0.82, p < 0.01). During mechanical ventilation (PCV) with muscle paralysis, no electrical activity of the diaphragm was seen.When PEEP was lowered, with concomitant decrease in lung volume, the EAdiexp increased significantly throughout the expiration. The EAdimin increased when PEEP was reduced from 12 to 0 cmH2O. Gas flow was lower at any given time point of expiration during SB without muscle paralysis than during PCV with muscle paralysis. Atelectasis increased in size with the lowering of PEEP but significantly more so during PCV than during SB so that the difference between SB and PCV increased with decreasing PEEP (Figure 1).ConclusionsOur findings show that the diaphragm electrical activity is well related to the diaphragmatic muscle force and that the diaphragm acts as an expiratory brake to limit the fall in end‐expiratory lung volume and to prevent lung collapse. Thus, the diaphragm is an active muscle also during expiration and loss of its function, as during mechanical ventilation and muscle paralysis, may be a contributing factor to unsuccessful respiratory support. We also speculate that expiratory work of breathing may be an additional component of work, added to the inspiratory work that contributes to respiratory muscle fatigue.Support or Funding InformationSupported by grants from the Swedish Research Council, the Swedish Heart and Lung Fund, and institutional funds at Uppsala and Bari Universities