bluuebunny/crossref_metadata_2025 huggingface dataset:
Respiratory plasticity induced by intermittent hypoxia following mid‐cervical contusion in unanesthetized rats
Authors: Chiang Shu‐Chi , Lee Kun‐Ze
Published: November 2023

Intermittent hypoxia has been considered as a potential approach to induce respiratory neuroplasticity. The purpose of present study was to investigate the impact of intermittent hypoxia on respiratory function following cervical contusion in unanesthetized rats. Adult male rats were received laminectomy or unilateral contusion at C3–C4 spinal cord using the MASCIS Impactor (height: 6.25 or 12.5 mm). At 4 weeks post‐injury, the breathing pattern of unanesthetized rats was measured by the whole body plethymography before, during and after 10 episodes of 5‐min hypoxia (10 % O2, 4 % CO2, balance N2) with 5‐min normoxia intervals. The result demonstrated that animals received severe cervical contusion had a lower tidal volume than uninjured animals. Hypoxia induced a significant increase in frequency and tidal volume in uninjured and contused animals. However, there is a progressive decline in the magnitude of hypoxic ventilatory response during intermittent hypoxia. Specifically, the frequency response is gradually reduced in uninjured animals, and the tidal volume response is decreased in contused animals with successive hypoxic stimuli. In addition, the tidal volume was significantly enhanced in contused rats but not uninjured rats at 30 min post‐hypoxia, suggesting intermittent hypoxia can induce long‐term facilitation of tidal volume following cervical contusion. These results demonstrated that intermittent hypoxia can induce differential forms of respiratory plasticity in uninjured vs. contused animals, and it may be a promising neurorehabilitation approach to improve respiratory function after cervical spinal cord injury.

Support or Funding Information

National Health Research Institutes (NHRI‐EX104‐10223NC), Ministry of Science and Technology (Most 102‐2320‐B‐110‐004‐MY3)
Hyperbaric Oxygen Treatment Following Mid‐Cervical Spinal Contusion Injury
Authors: Turner Sara , Hoyt Aaron , Gregory Nichole , Tester Nicole , Reier Paul , Fuller David
Published: November 2023

Mounting evidence suggests daily hyperbaric oxygen treatment (HBOT) can enhance functional motor recovery after thoracic spinal cord injury (SCI), possibly through alterations in inflammatory signaling and reduced apoptosis at or near the lesion epicenter. The purpose of this initial study was to explore using HBOT as a rehabilitative tool after cervical spinal contusion injury. Rats were randomly divided into the following groups: 1) naïve (no prior treatment), 2) sham operated (laminectomy only), 3) lateral contusion at C3/4 (cSCI) and, 4) cSCI + daily HBOT. The HBOT consisted of a 1 hr exposure to 2.0 atmospheres of 100% O2, beginning on the day of SCI and continued for 10 days. Forelimb function was analyzed on day 10 post‐surgery using open field forelimb locomotor scoring (FLS) and a “water grooming” test. Compared to naïve and sham rats, ipsilateral forelimb function was severely disrupted following cSCI in both the untreated and HBOT‐treated groups (p<0.05). However, HBOT‐treated rats showed a 17% increase in ipsilateral forelimb joint mobility (FLS score = 4.1 ± 1.6) when compared to the untreated cSCI group (FLS score = 1.25 ± 0.25; p = 0.074). In addition, a greater proportion of HBOT‐treated cSCI animals regained the ability to groom with the ipsilateral forelimb when compared to untreated cSCI animals (Chi‐Square Test, p = 0.016). After the 10th treatment day, cervical spinal tissues (C2–C5) were harvested for analyses of mRNA gene expression. Compared to non‐treated rats, the HBOT cohort showed increased mRNA expression (p<0.05) of genes involved in axon growth/guidance, including: SLIT2, CNTN1, ABLIM1, PLXNB1, and ARHGEF12. In addition HBOT treated rats showed decreased mRNA expression (p<0.05) of genes involved in the induction of apoptosis via DR3 and DR4/5 death receptors, such as: RELA, CASP9, NFKB1A, FADD, TNFSF12, and APAF1. Our data are consistent with prior reports of improved motor function following HBOT in SCI animal models, and the mRNA data raise the possibility that HBOT may be influencing axonal growth and regeneration, as well as attenuating cell death. Thus, HBOT following spinal cord injury may have exciting rehabilitative potential.

Support or Funding Information

CTSA U54‐TR001012, Craig H. Neilsen Foundation 313369 (SMFT), 1 R01 NS080180‐01A1 (DDF), State of Florida Brain and Spinal Cord Injury Research Program (DDF and PJR)
3D Bioprinting Nanocomposite Scaffolds for Complex Tissue Regeneration
Authors: Zhang Grace
Published: November 2023

Cells within the human body are in intimate contact with a 3D extracellular matrix composed of numerous organic and inorganic components. As a result, one of the revolutionary changes in the tissue and organ fields is to develop 3D bioprinting techniques to create complex tissue construct mimicking native tissue. In particular, 3D bioprinting offers great precision and control of the internal architecture and outer shape of a scaffold, and prints complicated patient specific structures. Furthermore, human tissue is nano in the smallest dimension and many current 3D bioprinting techniques still exhibit difficulty in achieving a nano resolution. Therefore, the main objective of our research is to develop novel biologically inspired nanomaterials as the key component in our 3D bioprinting “ink” and advanced 3D printing techniques to fabricate the next generation of nanocomposite scaffolds for complex tissue regeneration. For this purpose, we designed and synthesized innovative biologically inspired nanomaterials (i.e., nano hydroxyapatites, carbon nanomaterials, DNA based self‐assembly nanotubes, and core‐shell nanospheres with bioactive factors). Through 3D printing in our lab, a series of biomimetic vascularized bone, osteochondral and neural tissue scaffolds with nano and micro features were fabricated. Our results show that these 3D printed nanocomposite scaffolds have not only improved mechanical properties but also excellent cytocompatibility properties for enhancing various cell growth and differentiation, thus promising for complex tissue regeneration.
Novel Method for the 3D Printing of Bioactive Biomedical Devices
Authors: Weisman Jeff A. , Murthy UdayaBhanu , Tappa Karthik , Mills David K
Published: November 2023

With the advancement of 3D printing new and creative filament materials are being quickly introduced into the marketplace. These filaments are commonly manufactured with a large batch manufacturing method used to ensure that large product volumes can be rapidly manufactured with cost efficiencies. Small batch manufacturing is a more promising approach offering the potential for developing new and novel patient treatment modalities. These can be designed, prepared, and applied locally rather than having specialty implants fabricated offsite, with a significant savings in time and cost and the ability to provide customized treatments. In this study, commercially available 3D equipment (extruders and printers) and the role they may play in onsite small batch manufacturing was examined. Characterization of 3D printed constructs was performed to study the effects of using bioactive compounds in these fabrication processes and to ensure that bioactive reactivity of the dopant compounds was maintained throughout the manufacturing process. Preliminary results have shown that commercial 3D equipment can be used successfully for both antibiotic and chemotherapeutic doping of biodegradable polymers already in use with commercial 3D printers, and that these manufactured implants show greater elution properties than commercial‐grade PMMA/MMA bone cements. This novel method permits the fabrication of heterogeneous physical objects or structures of high complexity, without loss of resolution, and permits control over many design features including stack orientation, dopant and polymer composition. Doped constructs were also shown to be effective in inhibiting bacterial growth and reducing osteosarcoma cell proliferation.
3D Printing of Customized Implants for Treatment of Orofacial Deformities and Defects
Authors: Tayebi Lobat
Published: November 2023

Acute or refractory orofacial deformities and defects present surgeons with numerous reconstructive challenges.

Such defects are currently reconstructed using microvascular free flaps, non‐vascular autogenous bone and/or skin grafts, or reconstruction with xenografts or synthetic materials. Using these methods, surgeons are constrained by the size and complexity of the defect; defects larger than 5 cm are difficult to reconstruct with these techniques, so are those defects that involve the anterior mandibular region. Lack of predictability of graft success, as well as the lack of predictability of the esthetic and functional outcome of a non‐vascular grafting makes it necessary to use a vascularized free flap in many of these cases. Such procedures are lengthy and challenging despite the improvements in the technology and the techniques. Patients experience long recovery periods and are exposed to related perioperative morbidities that increase health care costs, increase time lost from work, and negatively affect the patients’ quality of life.

3D printing is currently being intensely studied by a diverse set of applications, including the development of bioengineered tissues, as well as the production of functional biomedical materials and devices for dental and orthopedic applications. Producing vascularized 3D‐printed customized implants/scaffolds that are infused with stem cells, and designed to replace many of the traditional surgical procedures can be considered as a new method to treat orofacial deformities and defects. These implants/scaffolds will be tailored to meet the patient's current condition, age, and prior medical history and designed for tension free soft tissue closure, enhanced vascularity and tissue formation, and the prevention of micromobility

We anticipate that this method will significantly improve the quality of the life of patients affected by numerous oral and craniofacial pathologic conditions.
Assessing the Role of Saturated Fats in Adipose Tissue Inflammation
Authors: Morgan‐Bathke Maria , Faubion William , Jensen Michael
Published: November 2023

Background

A multitude of studies have suggested that there is a link between obesity and inflammation. Various in vitro and animal studies have shown that saturated fatty acids are largely responsible for the induction of various pro‐inflammatory pathways. As vegetarians consume about half the amount of saturated fat as omnivores, we hypothesized that they would have less inflammation for any given amount of obesity.

Objective

To quantitate the inflammatory status of adipose tissue from obese vegetarians and omnivores.

Methods

The study included 8 obese vegetarians (1 male) and 8 obese omnivores matched for age, sex, and BMI. Body composition was measured by DXA. A blood sample to measure plasma free fatty acids (FFA) and abdominal and femoral subcutaneous adipose tissue biopsies to measure macrophage content, and the fatty acid composition were collected. Each participant also completed a Food Frequency Questionnaire (FFQ) to determine their average dietary intake.

Results

By design there were no differences in age, weight, BMI, percentage body fat, fat free mass or adipocyte size between the omnivore and vegetarian group. The plasma FFA of vegetarians revealed that they had greater proportions of the polyunsaturated fatty acids linoleic, linolenic and DHA (p = 0.04, 0.05, 0.05 respectively). Surprisingly, there was no difference in the fatty acid composition of the adipose tissue samples (abdominal or femoral) between the two groups. The FFQ data showed no significant difference in total fat, protein, or carbohydrate intake between the groups. However, the omnivore group had significantly greater intake of saturated fat and cholesterol than the vegetarian group (p = 0.05, 0.04, respectively). The inflammatory status of adipose tissue was quantified by measuring macrophage content (total, pro‐inflammatory M1, and anti‐inflammatory M2 macrophages). There was no difference in total, M1 or M2 macrophage content between the vegetarian and omnivore groups. We also quantified the senescent cell population within the adipose tissue and again found no difference between the groups.

Conclusions

While vegetarians have significantly greater proportions of polyunsaturated fatty acids in plasma FFA than omnivores, this was not true for adipose tissue fatty acid composition. As expected, omnivores consume significantly greater amounts of saturated fatty acids, but there was no difference in adipose tissue inflammatory status between the two groups when measured using flow cytometry and senescent cell population. It is possible that the burden of obesity is more important than the consumption of saturated fatty acids in terms of human adipose tissue inflammation.
Mitochondrial Respiration Regulates Proliferation, Micromotion, and Wound Healing Migration of Human Mesenchymal Stem Cells
Authors: Lee Yu Wei , Tung Tse Hua , Gomez Sofia , Lo Chun Min
Published: November 2023

Mitochondria plays an important role in human mesenchymal stem cell (hMSC) proliferation and differentiation. Carbonyl cyanide 4‐(trifluoromethoxy) phenylhydrazone (FCCP), which uncouples mitochondrial oxidative phosphorylation from the electron transport chain, is known to increase oxygen consumption rates. In this study, hMSCs derived from umbilical cord were chronically exposed to different concentrations of FCCP. Cell proliferation and mitochondrial activities were measure by CyQUANT® NF cell proliferation assay and Seahorse XF‐24 extracellular flux analyzer respectively. Compared with control group, chronic exposure of hMSCs to 1 μM and 3 μM FCCP for two days led to approximately 10% and 60% reduction of proliferation. Furthermore, chronic exposure of hMSCs to 3 μM FCCP for 20 hours significantly reduced their mitochondrial function. Electric cell‐substrate impedance sensing (ECIS) was also used to detect the alternation of hMSC micromotion and wound healing migration in response to different concentrations of FCCP treatment for 20 hours. The variance and energy of level 1 DWT (discrete wavelet transform) detail coefficients of the measured resistance time series at 4 kHz were calculated and the results demonstrated a dose‐dependent decrease from 1 μM to 3 μM FCCP, implying the dose‐dependent reduction of cell micromotion. For wound healing migration, the recovery resistance curves were fitted by sigmoid curve and the hill slope showed a dose‐dependent decline from 0.3 μM to 3 μM FCCP, indicating the decrease in migration rate. Moreover, dose dependent incline of the inflection points from 0.3 μM to 3 μM FCCP implied the increase of the half time for wound recovery migration. If the measured capacitance time series at 40 kHz were analyzed and compared with the previous resistance results, the variance and energy of level 1 DWT detail coefficients dose‐dependently decreased from 0.1 μM to 3 μM FCCP. Together, these results demonstrate that partial uncoupling of mitochondrial oxidative phosphorylation reduces proliferation, micromotion, and wound healing migration of hMSCs.

Support or Funding Information

This work was supported by the Ministry of Science and Technology, Taiwan. (Grant number: NSC 102‐2628‐B‐010‐010‐MY3)
Effects of Dexamethasone on Tissue Engineered Skeletal Muscle Units
Authors: Larson Alexie
Published: November 2023

Volumetric muscle loss (VML), muscular trauma that overwhelms the native repair mechanism, necessitates surgical intervention. Limitations to current VML treatments present a need for exogenous graft muscle sources. To date, engineered muscle produces forces substantially lower than native muscle, limiting its potential for repair. In this study, we examined the trophic effects of dexamethasone (DEX), a glucocorticoid that stimulates myoblast differentiation and fusion into myotubes, on our tissue engineered skeletal muscle units (SMUs). Using our established SMU fabrication protocol, muscle isolates were exposed to two DEX concentrations (10 and 25 nM). Following seeding onto a laminin‐coated Sylgard substrate, DEX addition was initiated on day 0 until the first feeding of growth media (day 4), or sustained until SMU formation as a control (days 0–20). Cell proliferation was measured with a BrdU assay (day 4), and myogenesis was observed through immunostaining for MyoD (day 4), and α‐actinin (day 11). After SMU formation (day 20), isometric tetanic force production was recorded to quantify functional improvement. The sustained addition of 10 nM DEX yielded optimal SMUs with advanced structural maturity and functionality. Though 25 nM DEX administered days 0–4 showed a 12% increase in myogenic proliferation determined by a BrdU and MyoD co‐stain at day 4, by day 11 the SMUs with sustained addition of 10 nM DEX demonstrated the greatest advancement in sarcomeric structure observed by α‐actinin immunostaining and a 2‐fold rise in myotube density determined by light microscopy analysis on day 14, compared to both 10 nM and 25 nM DEX days 0–4 groups. Additionally, the SMUs with sustained 10 nM DEX exhibited a 16% increase in force production compared to both 10 nM and 25 nM DEX days 0–4 groups. Future research will focus on further investigation of the timing of DEX addition, understanding the mechanism behind these effects and evaluating the regenerative potential of these SMUs in vivo.

Support or Funding Information

Supported by NIH/NIAMS 1R01AR067744‐01
Reseeding Decellularized Porcine Ventricular Extracellular Matrix with Isolated Primary Human Atrial Cells
Authors: Muller Alison L , Haromy Alois , Freed Darren H
Published: November 2023

As a result of pioneering research unveiling the influence of extracellular environment in determining stem/progenitor cell differentiation, a myriad of cellular scaffolds have been developed in order to optimize the delivery of these cells in a diseased environment as a technique to improve their efficacy in stem cell therapy. Although a variety of scaffolds are being studied in different contexts, the most physiologically relevant ones are those that most closely resemble the environment to be treated. Our lab has a keen interest in cardiovascular disease and has recently optimized a technique that decellularizes and reseeds isolated sections of healthy porcine ventricular tissue with primary human atrial cells. Decellularization involved a multi‐step, multi‐reagent washing process over a period of four days, after which confirmation of a lack of cells was determined by DAPI staining and subsequent western blotting looking for the absence of β‐tubulin. After decellularization, our objective was to successfully seed the decellularized tissue slices via co‐incubation of isolated primary human atrial fibroblasts (hAFs) or human c‐kit(+) cardiac progenitor cells (hCCs). These two cell types were chosen to compare how hAFs and hCCs respond differently to healthy extracellular matrix (ECM). Cells were isolated from the atrial appendage of patients undergoing open heart surgery by mincing followed by collagenase digestion and subsequent isolation of hCCs was performed using a c‐kit(+) magnetic bead isolation procedure. HAFs and hCCs were incubated separately on sections of decellularized porcine ventricular ECM over a period of 24 hours and then visualized using immunofluoresence. Collagen‐1, fibronectin, and versican antibodies were used to image the ECM structure and DAPI was used to identify cells embedded in the recellularized matrix. Successful seeding of cells was also evaluated by western blot detection of β‐tubulin in samples not prepared for immunofluorescence. In conclusion, our technique was successful in reseeding decellularized porcine ventricular extracellular matrix and can be extrapolated to detect other proteins in these tissues without having to reseed an entire decellularized heart. This can be used to compare differences among cell types and their responses between healthy and damaged or diseased cardiac ECM among different hearts or even within the same heart.

Support or Funding Information

Mazankowski Alberta Heart Institute University of Alberta Hospital Foundation Alberta Innovates Health Solutions
Application of a new miniature bioreactor system to generate and test artificial tumor and normal breast ductal tissues using MCF‐7/MCF10A cells
Authors: Lyden Timothy , Cole Kris , Goodman Steve
Published: November 2023

During the past decade, the UWRF Tissue and Cellular Innovation Center has been focused on application of natural extra‐cellular matrix materials as biomimetic scaffolds for small‐scale 3D artificial tissue (AT) and artificial tumor tissue (ATT) modeling. These constructs represent much more physiologically relevant in‐vitro models than most standard 2D cultures and are now the focus of a series of studies aimed at developing new models of cancer progression and metastasis. Of particular interest is the modeling of breast adenocarcinoma and monitoring/testing of the progression of these constructs into metastasis‐related processes. Although we have been successful in generating ATT's from several cells lines and primary patient tumors, our approach has always been limited by relatively non‐standardized culture conditions. These conditions were consistent enough for basic construct characterization and general “proof‐of‐concept” validation but next‐step direct experimentation has been limited by these culture conditions. In late 2013, we partnered with Microscopy Innovations, LLC to begin testing their mPrep capsules as miniature bioreactors in combination with our standard natural matrix/scaffold materials. In these studies, we are working to standardize culture conditions and develop a prototype reactor system approach to study MCF‐7 breast adenocarcinoma cell‐derived ATT's and their progression toward metastasis. Early loading studies of mPrep capsules with MCF‐7, MCF10A non‐cancerous ductal cells as well as stromal fibroblast and pre‐adipocyte (3T3‐Swiss and L1) cell lines have generated substantial tumor and control artificial tissue constructs. Ongoing characterization studies using flow cytometry and Western blot analysis are examining the relative population dimensions of cancer stem cells as well as EMT/MET markers and cell cycle status of both AT/ATT cells and the shed cells appearing in the flow‐through effluent of the capsule chambers. To date, this system has shown very significant promise for in‐vitro modeling of the complex relationship between tumors or control tissues and the surrounding fluid compartments in which they develop and function in vivo.
Synergistic targets between ASC‐differentiated endothelial and neural lineage cells to prevent hypoxic‐ischemic brain injury
Authors: Huang Chia‐Wei , Chang Ya‐Ju , Chien Shu , Wu Chia‐Ching
Published: November 2023

Neonatal hypoxic‐ischemic (HI) brain injury causes disruption of neurovascular integrity and leads to life‐long functional deficit in human development. These devastating consequences can be prevented by protecting the architecture of the neurovascular unit. Our lab has demonstrated the possibility to differentiate the adipose‐derived stem cells (ASCs) into endothelial lineage cells (ELCs) or neuronal lineage cells (NLCs) via microenvironmental induction and studied the therapeutic effect of the cells by transplanting ASCs, ELCs, NLCs, or combination of ELCs and NLCs (E+N) into HI brain injured neonatal rats. The combined E+N treatment showed a significantly greater decrease of infarction and apoptotic areas, and better preservation of the neurovascular structure. The E+N combinations increased the cell migration under in vitro hypoxia microenvironment. Moreover, the transplanted ELCs and NLCs were able to engraft into host tissue and promote endogenous angiogenesis and neurogenesis. We performed the RNAseq to fish out the potential targets for the differentiated ELCs and NLCs. The ELCs, induced by endothelial growth factors and laminar shear stress, migrated and contributed to the vascular structure by activating the Akt signaling through vascular endothelial growth factor receptor 2 (VEGFR2) and neuropilin 1 (NRP1). NLCs showed better ability in preserving the neural structure. The synergistic benefits in E+N combination were revealed via the cell‐cell interactions for NRP1 signaling in ELCs and C‐X‐C chemokine receptor 4 (CXCR4) and fibroblast growth factor receptor 1 (FGFR1) signaling in NLCs. Blockage of target signals in either ELCs or NLCs diminished the synergistic interaction in cell migration, homing, and protection of neurovasculature in the HI injured brain. The results in current study pointed out the potential molecular targets for the synergistic effect between ELCs and NLCs to improve the therapeutic outcome for preserving brain integrity and function after HI injury.

Support or Funding Information

This study was supported in part by grants from the Ministry of Science and Technology (NSC102‐2320‐B‐006‐009‐MY3) and National Health Research Institute (NHRI‐EX101‐10115EC) in Taiwan.
Are Foods of Higher Nutritional Quality More Expensive Than Their Less Healthy Counterparts? An Analysis of Canadian Packaged Foods
Authors: Labonté Marie‐Eve , Noorhosseini Sheida , Bernstein Jodi , Ahmed Mavra , L'Abbé Mary
Published: November 2023

Price‐related differences in the nutritional quality of food products may create unintended disparities in healthy food availability between different socioeconomic segments of the population. This study assessed whether healthier products are more expensive than less healthy products within specific food categories in the Canadian food supply. Data on nutritional composition and price were obtained from the University of Toronto Food Label Information Program (FLIP) 2013 database, which contains information on n=15,401 packaged products from the four largest national retailers by sales. Ready‐to‐eat breakfast cereals (n=250) and yogurts (n=236) were included in the present analyses. The Food Standards Australia New Zealand Nutrient Profiling Scoring Criterion (FSANZ‐NPSC) was used to calculate a summary score of the healthfulness of each product based on its content of nutrients to limit (e.g. sodium) and nutrients to encourage (e.g. fibre). Additionally, food products were classified either as “healthier” or “less healthy” based on the established cutoff score in the FSANZ‐NPSC model. The Student's t‐test was used to compare different price measures (price per 100 g, per serving, and per 100 Kcal) between “healthier” and “less healthy” breakfast cereals and yogurts. “Healthier” breakfast cereals had a higher price per serving than breakfast cereals of lower nutritional quality (geometric mean [95% confidence interval] = CAD $0.53 [0.49–0.57] vs. 0.44 [0.41–0.47], respectively, P=0.0004). Price per 100 g and price per 100 Kcal did not differ according to the degree of healthfulness of breakfast cereals (both P≥0.60). “Healthier” yogurts had a lower price per serving and price per 100 g than yogurts of lower nutritional quality (per serving: CAD $0.82 [0.77–0.87] vs. 1.13 [0.91–1.40], respectively, P=0.006; per 100g: CAD $0.67 [0.64–0.70] vs. 0.81 [0.72–0.92], P=0.03), but a higher price per 100 Kcal (CAD $0.93 [0.88–0.98] vs. 0.61 [0.57–0.65], P<0.0001). These data suggest that healthier food products are not necessarily more expensive than less healthy products. Importantly, the relationship between the healthfulness of foods and their price varies depending on the type of food products evaluated and the selected price measures. These factors must be taken into consideration when communicating research results to a general audience.

Support or Funding Information

Canadian Institutes of Health Research (CIHR) Fellowship (MEL); CIHR Open Operating Grant (ML); and Burroughs Wellcome Fund – Innovation in Regulatory Science (ML)
Mapping inflammation and cancer‐modifying regions in broccoli genome
Authors: Bussler Weston , Alley Jessica , Chandler Holli , Thetford Claire , Yousef Gad , Brown Allan , Esposito Debora , Lila Mary Ann , Komarnytsky Slavko
Published: November 2023

Consuming a healthy diet is one of the controllable factors that influences the development or severity of multiple chronic diseases, including metabolic, inflammatory, and neoplastic disorders. Diets high in cruciferous vegetables consistently show a decreased incidence of colorectal cancer and cardiovascular disease. Understanding the dynamic interrelationships that exist between bioactive food components in broccoli and human disease prevention is recognized as a high priority. A collection of 125 near‐isogenic broccoli lines (Brassica oleracea subsp. italica) previously developed by our group, represents a unique tool for studying effects of multiple dietary bioactive components by utilizing physiologically relevant concentrations of the agents and eliminating experimental noise associated with food matrix effects. By using broccoli near‐isogenic lines, we successfully observed direct nutrient‐nutrient or nutrient‐disease interactions that vary significantly (order of magnitude) in their ability to suppress inflammation in RAW 264.7 macrophages or inhibit proliferation of HT‐29 colorectal cancer cells. When combined with quantitative trait loci (QTL) or genome‐wide association (GWAS) studies, this approach allowed for direct mapping of human health‐modifying regions (HMRs) in the broccoli genome – even if the interacting bioactive metabolites, or their respective biosynthetic and genetic regulatory pathways, were unknown. Our findings suggested that 3 health‐modifying regions (HMRs) on 2 chromosomes were sufficient to explain over 52% of the variation in health‐promoting activity observed among broccoli varieties tested. Within these regions, known gene sequences were assessed for relevance to synthesis or regulation of bioactive metabolites to identify candidate genes and advance breeding programs aimed at targeted improvement of broccoli nutritional value.

Support or Funding Information

This work was supported in part by NCSU faculty start‐up funds (SK); Plant Pathways Elucidation Project; and Cell Culture and Phenotyping Core of the Plants for Human Health Institute, NC State University.
Evo‐Devo Anatomy Dissected: An Approach Using Soft Embalming that Transforms the Regional‐Versus‐Systemic Debate
Authors: Boaz Noel T. , Bernor R. L , Meshida K.
Published: November 2023

Gross anatomists have been at odds with curricular initiatives in medical education that emphasize a systems approach, primarily because of the impracticality of dissecting widespread bodily systems rather than regions. An evolutionary and developmental (“evo‐devo”) anatomical approach, however, is systemic and is eminently compatible with a systems‐based integrated curriculum. The flexibility, resilience, and mobility of soft‐embalmed structures allow dissection of body segments, such as pharyngeal arches and myodermatomal derivatives, thus elucidating important neurovascular and clinical aspects of the anatomy. A sequence of dissection following embryological appearance of systems provides a new logic to dissection that effectively correlates to other courses and modules in a systems‐based curriculum. A soft‐embalmed human male has been completely dissected with such an approach and is presented. Representative clinical correlations that utilize the unique attributes of soft‐embalmed cadavers, such as ultrasound imaging, physical examination practica, and invasive procedure practica, are illustrated.
The Metabolic Fate of Isotopically‐Labeled Trimethylamine‐N‐oxide (TMAO) in Humans
Authors: Taesuwan Siraphat , Cho Clara E , Malysheva Olga V , Bender Erica , Yan Jian , Thalacker‐Mercer Anna , Caudill Marie A
Published: November 2023

Although trimethylamine‐N‐oxide (TMAO) is associated with chronic disease risk, little is known about the metabolic fate of orally consumed TMAO. As part of a randomized, controlled cross‐over study, healthy young men (n=40) consumed 50 mg deuterium‐labeled methyl‐d9‐TMAO (d9‐TMAO) and TMAO metabolites were measured in plasma, urine, stool and muscle. Plasma d9‐TMAO comprised ~75% of total TMAO at 2 h and remained elevated after 6 h. d9‐trimethylamine (d9‐TMA) and d6‐dimethylamine (d6‐DMA) became enriched in plasma with averages of 52% and 3% of total TMA and DMA, respectively, at 6 h. d9‐TMAO was excreted more rapidly during 0–6 h than 6–24 h, comprising 67% and 52%, respectively, of urinary TMAO. No d9‐TMAO was detected in feces. Interestingly, d9‐TMAO and d9‐TMA were detected in skeletal muscle (n=6) at 6 h, and the enrichment ratio of d9‐TMAO:d9‐TMA was influenced by a genetic variant in flavin‐containing monooxygenase (FMO3 G472A). The heterozygous GA genotype had lower muscle d9‐TMAO:d9‐TMA than the homozygous wildtype GG genotype. In conclusion, orally consumed TMAO is rapidly absorbed, is metabolized in part to TMA and DMA, and is taken up by skeletal muscle, where evidence of impaired conversion of TMA to TMAO among carriers of the FMO3 472A variant allele is observed.

Support or Funding Information

Egg Nutrition Center, Beef Checkoff through the National Cattlemen's Beef Association, Canadian Institutes of Health Research Postdoctoral Fellowship.
Phenolic recovery and bioaccessibility from milled and finished whole grain oat products
Authors: Li Min , Koecher Katie , Hansen Laura , Ferruzzi Mario G
Published: November 2023

Whole grain oats and oat products are a rich source of nutrients, fiber and dietary phenolics. While much is known regarding the benefits of oat fiber, limited information is available on the relevance of dietary phenolics from oat products. This study focused on defining the profile and stability of phenolics in native oats during common milling and processing and to determine the bioaccessibility of oat phenolics from common food preparations. Phenolic content from 26 oat cultivars as well as milled oat ingredients and ready to eat (RTE) products (puffed cereals and snack bars) were profiled. Free phenolic content ranged from 17.2~228.1 μg/g DW with avenanthramides accounting for 41~91% of free phenolics. Bound phenolic content ranged from 100.5~680.9 μg/g DW with ferulic acid accounting for 62~94 % of bound phenolics. Selected oat groats were ground to flour and prepared as wet cooked porridges (~20 % oat flour in boiling water) or introduced as RTE products to a three stage in vitro digestion to determine phenolic bioaccessibility. Relative bioaccessibility for wet cooked porridges ranged 0.3~1.6% and 3.4~37.0% for individual phenolic acids and avenanthramides, respectively. Puffed oat cereal had significantly higher bioaccessibility compared to matching wet‐cook porridge made from the same oat flour (e.g. 83.8% versus 19.1% for Ave A; P<0.05). Intestinal uptake of bioaccessible oat phenolics from digesta was confirmed by experiments with Caco‐2 human intestinal cells. Overall intestinal uptake of oat phenolics was low ranging between 0.16 % and 2.7 % across digesta from all oat products tested. In conclusion, these experiments demonstrate that phenolics naturally present in whole grain oats are well recovered through traditional grain processing, milling and food processing. Processing oat cultivars to a RTE cereal may have positive impact on digestive release and bioaccessibility of oat phenolics in the upper GI tract.

Support or Funding Information

Supported by General Mills, Inc and the Purdue University Ingestive Behavior Research Center.
Through Metabolic Reprogramming Walnut May Mitigate the Stemness of Colon Cancer Stem Cells
Authors: Choi Sang Woon , Choi Jina , Kim Junil , Friso Simonetta , Kim Yuri
Published: November 2023

The advent of the cancer stem cell (CSC) theory has accelerated our understanding of cancer. CSCs are defined as a small population of cells within a tumor that can self‐renew and drive tumorigenesis. The major reason for the failure of treatment is ascribed to the presence of CSCs in the tumor mass. Strategies that reduce CSC number or expansion represent a rational approach for colorectal cancer treatment. It is also well known that CSCs have demonstrated impaired mitochondrial functions and cancer cells exhibit altered metabolism to meet energy need, such as the Warburg effect. In our previous study, walnut phenolic extracts (WPEs) decreased the stemness of colon CSCs by suppressing their self‐renewing capacity and enhancing the differentiation ability. To investigate the mechanism behind the effect of walnuts on CSCs, using the Illumina HiSeq2500 we performed RNA‐sequencing on CSCs isolated from the HCT116 colon cancer cell line by the fluorescence‐activated cell sorting method and treated with or without 40ug/ml WPE (n=3). Among the total of 28305 transcripts, 1983 genes were found to be differentially expressed by WPE treatment (1168 genes were up‐regulated and 815 genes were down‐regulated)(fold change >1.5, p<0.05). Differentially expressed genes were categorized into functional clusters using the bioinformatic analysis tools, such as Database for annotation, visualization and integrity discovery (DAVID), Gene regulatory network (GRN) and Ingenuity pathway analysis (IPA). We found a novel phenomenon that WPE alters the expression of CSC genes associated with metabolic pathways, such as oxidative phosphorylation (NDUFSs, CYC1, COX8A, ATP5D), pentose phosphate pathway (TKT, TALDO1, ALDOA), glycolysis (GPI, PFKL, ENO1), TCA cycle (MDH2, IDHs), purine metabolism (APRT, GUK1), and pyrimidine metabolism (TK1, NME1, POLD2). The expression patterns of genes encoded for mitochondrial enzymes, such as NDUFB10, ATP5D, CYC1, and COX8A, as well as genes encoding mitochondrial ribosomal‐structure proteins, such as MRPLs, were also up‐regulated, indicating that WPE may affect the mitochondrial function. It appears that the WPE reduces the stemness of colon CSCs by restoring altered metabolic pathways and mitochondrial function.

Support or Funding Information

This work has been supported by the California Walnut Commission and the Korean Food Research Institute.
The Role of Intra‐abdominal Adipose Tissue in Cancer
Authors: Bernard Jamie , Bullard Blair
Published: November 2023

The obesity pandemic is causing worldwide morbidity and mortality. Recent research suggests that a high‐fat diet (HFD) and/or obesity (increased adiposity) are key players in the pathogenesis and prognosis of many common cancers. However the molecular changes induced by adipose tissue (AT) that actually enhance cancer development are poorly understood. Our preliminary studies directly implicate secreted factors from parametrial (around the uterus) AT (the largest visceral fat pad in female mice) in promoting tumor formation. We demonstrated an increase in pro‐inflammatory adipocytokines in parametrial AT of mice fed a diet high in saturated animal fats (HFD). Removing the parametrial AT of these HFD‐fed mice by lipectomy surgery reduced the number of UVB‐induced skin cancers by 80% compared to sham‐operated mice. Similarly, in an obesity‐resistant mouse model of mammary tumor growth, we showed that a HFD increased parametrial adiposity and pro‐inflammatory adipocytokines in parametrial AT suggesting that diet‐induced parametrial adiposity and adipocytokine increases are not unique to diet‐induced obesity models. We hypothesize that a diet high in saturated animal fat promotes tumor formation by increasing pro‐inflammatory adipocytokines/growth factors from an enlarged parametrial fat pad. To understand the role that parametrial AT and saturated HFDs play in cancer, we developed a system where molecules released and filtered from AT (fat tissue filtrate) stimulate JB6 P+ cell proliferation and transformation. The mouse BALB/c skin epithelial JB6 P+ cell line is a well‐characterized model for neoplastic transformation that is responsive to tumor promoter‐induced transformation growth in soft agar and is uniquely suited for studying tumor promotion and promotion‐relevant molecular events in vitro. Our data demonstrate that fat tissue filtrate made from the parametrial AT of obese mice fed a HFD had 160% more transforming activity than that from mice fed a LFD and formed malignant mesenchymal tumors in vivo. These differences between parametrial AT from mice fed a saturated HFD and a LFD were associated with their density of crown‐like structures (CLS; surrogate markers for dead adipocytes) and adipocytokine secretomes. Moreover, we have found specific pro‐inflammatory adipocytokines and growth factors that are preferentially induced in the parametrial AT in both obese and obesity‐resistant mice with HFD‐feeding that stimulate the transformation of JB6 P+ cells. These studies provide an in vitro model and the basis for investigations on the mechanisms of parametrial AT‐induced cell transformation and tumor growth. Findings from these studies may lead to the discovery of new therapeutic targets for saturated HFD‐promoted cancer and will have implications for the prevention and reversal of disease.

Support or Funding Information

CA177868, ES007148
Microenvironment folate stress promotes Warburg‐metabolic switched‐cancer stem cell signaling network and stemness features to mediate invasion and metastasis of lung cancers
Authors: Chen Wan‐Jing , Huang Rwei‐Fen S.
Published: November 2023

Recent advances suggest that cancer stem cells in minute percentages of tumour bulk mediate change of epithelial‐to‐mesenchymal transition (EMT), invasion and metastasis, and drug‐resistance of malignancy tumours. Whether the molecular mechanisms by which metabolic folate stress may promote Warburg‐metabolic switched‐cancer stem cell signaling network and stemness features to mediate invasion and metastasis of lung cancer cells are unknown. Lung embryonic fibroblasts (MRC5), non‐small cell lung cancer cells (NSCLC) including adenocarcinoma (A549, H23) and squamacarcinoma cells (H520) were used as the experiment models to test the hypothesis. As compared with the controls, cultivation of NSCLC cells with clinically low serum folate levels (LF) for 4–8 days gradually and significantly promoted (1) protein expressions of cancer stem cell signature molecules (Shh, Sox2, Oct4, ALDH1A1, HIF‐1 alpha and Foxo3a, (2) expressions of EMT markers (E‐cadherin and vimentin), and (3) morphological EMT alteration. Energetic metabolic switch in the LF‐NSCLC cells, but not the controls, was evident by lactate release, glycolytic enzymes activity and redux status of NADP/NADPH ratios. When replacing the NSCLC cells into the oncogenic suspension medium to mimic human metastatic microenvironment, the LF‐NSCLC, but not the controls, massively transformed to tumour spheroids, developing stemness features. Blockage of mTOR, sonic hedgehog signaling pathways, epigenetic acetylation (Sirt1) and DNA methylation (5AZA) abolished LF‐promoted stem cell signatures, reversed Warburg metabolic switch and eliminated tumour spheroid formations. Antifolate drugs treatment did not completely prevent from LF‐induced tumour spheroid formation. Injection of LF‐NSCLC and control‐NSCLC cells into the NNK‐mutagenized C57BL/6 mice fed with various folate diets exhibited differentially modulations on metastasis tumours. Collectively, metabolic folate stress promotes Warburg‐metabolic switched‐cancer stem cell signaling network and stemness features to mediate invasion and metastasis of lung cancers.
Cranberry extract initiates intrinsic apoptosis in HL‐60 cells by increasing Bad activity through the inhibition of Akt phosphorylation
Authors: Mansouri Rasha , Percival Susan S
Published: November 2023

Cranberry's role as a potential chemopreventive agent has been examined by various studies since 1996. One of the proposed mechanisms is to eliminate cancerous cells by inducing apoptosis. A limited number of studies have shown that cranberry can activate apoptosis in cancer cells. Therefore, we sought to determine the mechanism by which cranberry extract initiates apoptosis. In our study, HL‐60 cells were incubated with 25 μg/ml of cranberry extract for 24 hours. Cranberry treatment caused a significant increase in caspase 3/7 activity. In addition, the data showed a significant increase in the concentration of caspase‐9, but no change in the level of caspase‐8. Moreover, cranberry treatment caused a loss of mitochondrial transmembrane permeability, leading to a significant increase in release of cytochrome C and Smac. These results indicate that cranberry treatment initiates a cascade of events associated with intrinsic apoptosis. These apoptotic events were correlated with a significant decrease in Akt phosphorylation at both Ser473 and Thr308 sites, followed by a significant increase of Bad (proapoptotic regulator) dephosphorylation at Ser136. Together, these findings provide an important molecular framework for the apoptosis initiated by cranberry extract in HL‐60 cells. The overall mechanism includes the suppression of Akt phosphorylation, causing an increase in dephosphorylated Bad. Active (dephosphorylated) Bad then promotes the permeability of the mitochondrial outer membrane and the subsequent release of pro‐apoptotic proteins, including cytochrome C and Smac, into the cytosol. These pro‐apoptotic proteins activate caspase 9, which causes a caspase cascade and leads to intrinsic apoptosis. This sequence of events describes a novel mechanism to explain how cranberry extract can trigger intrinsic apoptosis in HL‐60 cells. This may give direction to future studies investigating the effect of cranberry extract on cancer in vivo. The cranberry extract utilized was provided by Ocean Spray Cranberries, Inc. (Lakeville, MA).
Novel Role of Glutathione Peroxidase‐1 in Liver and Muscle Protein Metabolism of Mice
Authors: Tao Ling , Zhao Ze Ping , Lei Xin Gen
Published: November 2023

Our laboratory has revealed novel roles and mechanisms of glutathione peroxidase‐1 (GPX1) in glucose and lipid metabolism, whereas little is known about its function in protein metabolism. This study was conducted to elucidate the physiological impacts of the GPX1 overexpression and knockout on gene expression and activity of key factors related to protein/amino acid synthesis, transport, and catabolism in mice. Three genotypes of Se‐adequate young adult (2‐mo old) mice: wild type (WT), GPX1 knockout (GKO), and GPX1 overexpressing (OE), were used to collect tissue samples (n = 5 to10). Although the GPX1 genotype showed no effects on total protein of liver and muscle, and plasma activities of alanine transaminase and aspartate aminotransferase, the concentrations of plasma urea nitrogen were approximately 33% lower (P < 0.05) in both GKO and OE mice than WT mice. Meanwhile, a number of protein degradation‐related genes including cathepsin K, calpain 2, ubiquitin‐like modifier activating enzyme 1, and ubiquitin protein ligase E3A were down‐regulated (P < 0.05) in the liver of OE (29%–49% of decrease) and GKO (13–38% of decrease) mice. In contrast, amino acid transporter and protein synthesis‐related genes such as solute carrier family 3 member 2, mechanistic target of rapamycin (serine/threonine kinase), ribosomal protein S6 and eukaryotic translation initiation factor 4E binding protein 1 were down‐regulated (P < 0.05) in the liver of only GKO but not OE mice. The muscle mRNA levels of genes involved in protein degradation were decreased (P < 0.05) and elevated (P < 0.05), respectively, by the overexpression and knockout of GPX1, compared with the wild‐type controls. Additionally, the muscle glutamate deamination activities of glutamate dehydrogenase (GDH), an important enzyme involved in oxidative deamination, were elevated (P < 0.05) in the GKO mice. In conclusion, both GPX1 overexpression and knockout exerted effects on the gene expressions and biochemical functions of multiple pathways of protein metabolism in the liver and muscle of mice. The net metabolic outcome and human health implication of those effects warrant future research.

Support or Funding Information

Supported in part by a NIH grant DK53018