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Article: CRIPTO promotes extracellular vesicle uptake and activation of cancer associated fibroblasts.

Freeman, David W / Gates, Brooke L / Spendlove, Mauri D / Gulbahce, H Evin / Spike, Benjamin T

bioRxiv : the preprint server for biology

2024

Abstract: Expression of CRIPTO, a factor involved in embryonic stem cells, fetal development, and wound healing, is tied to poor prognosis in multiple cancers. Prior studies in triple negative breast cancer (TNBC) models showed CRIPTO blockade inhibits tumor ... ...

Abstract	Expression of CRIPTO, a factor involved in embryonic stem cells, fetal development, and wound healing, is tied to poor prognosis in multiple cancers. Prior studies in triple negative breast cancer (TNBC) models showed CRIPTO blockade inhibits tumor growth and dissemination. Here, we uncover a previously unidentified role for CRIPTO in orchestrating tumor-derived extracellular vesicle (TEV) uptake and fibroblast activation through discrete mechanisms. We found a novel mechanism by which CRIPTO drives aggressive TNBC phenotypes, involving CRIPTO-laden TEVs that program stromal fibroblasts, toward cancer associated fibroblast cell states, which in turn prompt tumor cell invasion. CRIPTO-bearing TEVs exhibited markedly elevated uptake in target fibroblasts and activated SMAD2/3 through NODAL-independent and - dependent mechanisms, respectively. Engineered expression of CRIPTO on EVs enhanced the delivery of bioactive molecules.
Language	English
Publishing date	2024-04-02
Publishing country	United States
Document type	Preprint
DOI	10.1101/2024.03.01.583059
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Article: Mcam stabilizes luminal progenitor breast cancer phenotypes via Ck2 control and Src/Akt/Stat3 attenuation.

Balcioglu, Ozlen / Gates, Brooke L / Freeman, David W / Hagos, Berhane M / Mehrabad, Elnaz Mirzaei / Ayala-Talavera, David / Spike, Benjamin T

bioRxiv : the preprint server for biology

2024

Abstract: Breast cancers are categorized into subtypes with distinctive therapeutic vulnerabilities and prognoses based on their expression of clinically targetable receptors and gene expression patterns mimicking different cell types of the normal gland. Here, we ...

Abstract	Breast cancers are categorized into subtypes with distinctive therapeutic vulnerabilities and prognoses based on their expression of clinically targetable receptors and gene expression patterns mimicking different cell types of the normal gland. Here, we tested the role of Mcam in breast cancer cell state control and tumorigenicity in a luminal progenitor-like murine tumor cell line (Py230) that exhibits lineage and tumor subtype plasticity. Mcam knockdown Py230 cells show augmented Stat3 and Pi3K/Akt activation associated with a lineage state switch away from a hormone-sensing/luminal progenitor state toward alveolar and basal cell related phenotypes that were refractory to growth inhibition by the anti-estrogen therapeutic, tamoxifen. Inhibition of Stat3, or the upstream activator Ck2, reversed these cell state changes. Mcam binds Ck2 and acts as a regulator of Ck2 substrate utilization across multiple mammary tumor cell lines. In Py230 cells this activity manifests as increased mesenchymal morphology, migration, and Src/Fak/Mapk/Paxillin adhesion complex signaling
Language	English
Publishing date	2024-03-20
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.05.10.540211
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Article ; Online: Growth Differentiation Factor 15 and Diet Quality Trajectory Interact to Determine Frailty Incidence among Middle-Aged Urban Adults.

Beydoun, May A / Noren Hooten, Nicole / Fanelli-Kuczmaski, Marie T / Maino Vieytes, Christian A / Georgescu, Michael F / Beydoun, Hind A / Freeman, David W / Evans, Michele K / Zonderman, Alan B

The Journal of nutrition

2024 Volume 154, Issue 5, Page(s) 1652–1664

Abstract: Background: Elevated plasma growth differentiation factor 15 (GDF15) and poor diet quality may be associated with increased frailty incidence, although their interactive associations have not been assessed in urban middle-aged adults.: Objectives: We ...

Abstract	Background: Elevated plasma growth differentiation factor 15 (GDF15) and poor diet quality may be associated with increased frailty incidence, although their interactive associations have not been assessed in urban middle-aged adults. Objectives: We aimed to examine GDF15 and its interactive association with diet quality in relation to frailty incidence among a sample of middle-aged urban adults. Methods: The relationship between GDF15 and diet quality trajectories in relation to incident frailty was examined in a longitudinal study of participants in the Healthy Aging in Neighborhoods of Diversity across the Life Span (2004-2017). Serum GDF15 concentration and frailty incidence were primary exposure and outcome, respectively. Group-based trajectory models were used to assess diet quality trajectories (≤3 visits/participant, N = 945, N' = 2247 observations) using the Healthy Eating Index 2010 version (HEI-2010), Dietary Inflammatory Index, and mean adequacy ratio (MAR). Cox proportional hazards models were used, testing interactive associations of GDF15 and diet quality trajectories with frail/prefrail incidence (N = 400 frailty-free at first visit, N' = 604 observations, n = 168 incident frail/prefrail). Results: Both elevated GDF15 and lower diet quality trajectories were associated with a lower probability of remaining nonfrail (≤13 y follow-up). Among females, the "high diet quality" HEI-2010 trajectory had a hazard ratio (HR) of 0.15 [95% confidence interval (CI): 0.04, 0.54; P = 0.004; fully adjusted model] when compared with the "low diet quality" trajectory group. Among males only, there was an antagonistic interaction between lower HEI-2010 trajectory and elevated GDF15. Specifically, the HR for GDF15-frailty in the higher diet quality trajectory group (high/medium combined), and among males, was 2.69 (95% CI: 1.06, 6.62; P = 0.032), whereas among the lower diet quality trajectory group, the HR was 0.94 (95% CI: 0.49, 1.80; P = 0.86). Elevated GDF15 was independently associated with frailty among African American adults. Conclusions: Pending replication, we found an antagonistic interaction between GDF15 and HEI-2010 trajectory in relation to frailty incidence among males.
MeSH term(s)	Humans ; Male ; Growth Differentiation Factor 15/blood ; Female ; Frailty/epidemiology ; Frailty/blood ; Middle Aged ; Incidence ; Diet ; Longitudinal Studies ; Urban Population ; Aged
Language	English
Publishing date	2024-03-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	218373-0
ISSN	1541-6100 ; 0022-3166
ISSN (online)	1541-6100
ISSN	0022-3166
DOI	10.1016/j.tjnut.2024.03.006
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Article ; Online: GDF15 and its association with cognitive performance over time in a longitudinal study of middle-aged urban adults.

Beydoun, May A / Noren Hooten, Nicole / Weiss, Jordan / Beydoun, Hind A / Georgescu, Michael / Freeman, David W / Evans, Michele K / Zonderman, Alan B

Brain, behavior, and immunity

2022 Volume 108, Page(s) 340–349

Abstract: Serum GDF15 levels are correlated with multiple neurodegenerative diseases. Few studies have tested this marker's association with middle-aged cognitive performance over time, and whether race affects this association is unknown. We examined associations ...

Abstract	Serum GDF15 levels are correlated with multiple neurodegenerative diseases. Few studies have tested this marker's association with middle-aged cognitive performance over time, and whether race affects this association is unknown. We examined associations of initial serum GDF15 concentrations with longitudinal cognitive performance, spanning domains of global mental status, visual and verbal memory, attention, fluency, and executive function in a sub-sample of adults participating in the Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) study (n = 776, Age
MeSH term(s)	Female ; Humans ; Male ; Cognition ; Cognitive Dysfunction ; Cross-Sectional Studies ; Executive Function ; Growth Differentiation Factor 15 ; Longitudinal Studies ; Memory ; Middle Aged
Chemical Substances	GDF15 protein, human ; Growth Differentiation Factor 15
Language	English
Publishing date	2022-12-19
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	639219-2
ISSN	1090-2139 ; 0889-1591
ISSN (online)	1090-2139
ISSN	0889-1591
DOI	10.1016/j.bbi.2022.12.015
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Article ; Online: Safety, feasibility, and impact on the gut microbiome of kefir administration in critically ill adults.

Gupta, Vinod K / Rajendraprasad, Sanu / Ozkan, Mahmut / Ramachandran, Dhanya / Ahmad, Sumera / Bakken, Johan S / Laudanski, Krzysztof / Gajic, Ognjen / Bauer, Brent / Zec, Simon / Freeman, David W / Khanna, Sahil / Shah, Aditya / Skalski, Joseph H / Sung, Jaeyun / Karnatovskaia, Lioudmila V

BMC medicine

2024 Volume 22, Issue 1, Page(s) 80

Abstract: Background: Dysbiosis of the gut microbiome is frequent in the intensive care unit (ICU), potentially leading to a heightened risk of nosocomial infections. Enhancing the gut microbiome has been proposed as a strategic approach to mitigate potential ... ...

Abstract	Background: Dysbiosis of the gut microbiome is frequent in the intensive care unit (ICU), potentially leading to a heightened risk of nosocomial infections. Enhancing the gut microbiome has been proposed as a strategic approach to mitigate potential adverse outcomes. While prior research on select probiotic supplements has not successfully shown to improve gut microbial diversity, fermented foods offer a promising alternative. In this open-label phase I safety and feasibility study, we examined the safety and feasibility of kefir as an initial step towards utilizing fermented foods to mitigate gut dysbiosis in critically ill patients. Methods: We administered kefir in escalating doses (60 mL, followed by 120 mL after 12 h, then 240 mL daily) to 54 critically ill patients with an intact gastrointestinal tract. To evaluate kefir's safety, we monitored for gastrointestinal symptoms. Feasibility was determined by whether patients received a minimum of 75% of their assigned kefir doses. To assess changes in the gut microbiome composition following kefir administration, we collected two stool samples from 13 patients: one within 72 h of admission to the ICU and another at least 72 h after the first stool sample. Results: After administering kefir, none of the 54 critically ill patients exhibited signs of kefir-related bacteremia. No side effects like bloating, vomiting, or aspiration were noted, except for diarrhea in two patients concurrently on laxatives. Out of the 393 kefir doses prescribed for all participants, 359 (91%) were successfully administered. We were able to collect an initial stool sample from 29 (54%) patients and a follow-up sample from 13 (24%) patients. Analysis of the 26 paired samples revealed no increase in gut microbial α-diversity between the two timepoints. However, there was a significant improvement in the Gut Microbiome Wellness Index (GMWI) by the second timepoint (P = 0.034, one-sided Wilcoxon signed-rank test); this finding supports our hypothesis that kefir administration can improve gut health in critically ill patients. Additionally, the known microbial species in kefir were found to exhibit varying levels of engraftment in patients' guts. Conclusions: Providing kefir to critically ill individuals is safe and feasible. Our findings warrant a larger evaluation of kefir's safety, tolerability, and impact on gut microbiome dysbiosis in patients admitted to the ICU. Trial registration: NCT05416814; trial registered on June 13, 2022.
MeSH term(s)	Adult ; Humans ; Critical Illness/therapy ; Dysbiosis ; Feasibility Studies ; Gastrointestinal Microbiome ; Kefir/analysis
Language	English
Publishing date	2024-02-20
Publishing country	England
Document type	Clinical Trial, Phase I ; Journal Article
ZDB-ID	2131669-7
ISSN	1741-7015 ; 1741-7015
ISSN (online)	1741-7015
ISSN	1741-7015
DOI	10.1186/s12916-024-03299-x
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Article ; Online: Association between GDF15, poverty and mortality in urban middle-aged African American and white adults.

Freeman, David W / Noren Hooten, Nicole / Kim, Yoonseo / Mode, Nicolle A / Ejiogu, Ngozi / Zonderman, Alan B / Evans, Michele K

PloS one

2020 Volume 15, Issue 8, Page(s) e0237059

Abstract: Mortality disparities are influenced by race and poverty. There is limited information about whether poverty influences biologic markers of mortality risk. Emerging data suggests that growth differentiation factor 15 (GDF15) is associated with mortality; ...

Abstract	Mortality disparities are influenced by race and poverty. There is limited information about whether poverty influences biologic markers of mortality risk. Emerging data suggests that growth differentiation factor 15 (GDF15) is associated with mortality; however, the interplay between GDF15, sociodemographic factors and mortality is not known. We sought to evaluate the interactions between GDF15 and sex, race and poverty status on mortality. Serum GDF15 was measured in 1036 African American and white middle-aged men and women above and below 125% of the Federal poverty status from the Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) study. Multivariable adjusted Cox regression models were used to assess the association between log-transformed GDF15 (logGDF15) and 12-year mortality outcomes (all-cause, cardiovascular- and cancer-specific outcomes) and interactions with sex, race and poverty status. Likelihood ratio tests were used to assess significance of the interaction terms. Median GDF15 was 655.2 pg/mL (IQR = 575.1). During 12.2 years of follow-up, 331 died of which 94 cardiovascular- and 87 were cancer-specific deaths. One unit of increase in logGDF15 was associated with a hazard ratio for all-cause mortality, cardiovascular- and cancer-specific mortality of 2.26 (95% confidence interval [CI], 1.94-2.64), 2.74 (95%CI, 2.06-3.63) and 1.41 (95%CI, 1.00-2.00), respectively. There was an interaction between logGDF15 and poverty status on all-cause mortality (p<0.05). The GDF15×poverty status interaction term improved model calibration for all-cause mortality. Our study provides the first evidence that the effect of elevated GDF15 on all-cause mortality is modified by poverty status.
MeSH term(s)	Adult ; African Americans ; Biomarkers/blood ; European Continental Ancestry Group ; Female ; Growth Differentiation Factor 15/blood ; Healthy Aging/blood ; Humans ; Male ; Middle Aged ; Mortality ; Poverty ; Proportional Hazards Models ; United States/epidemiology ; Urban Health ; Urban Population
Chemical Substances	Biomarkers ; GDF15 protein, human ; Growth Differentiation Factor 15
Keywords	covid19
Language	English
Publishing date	2020-08-07
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Intramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0237059
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Article ; Online: Extracellular vesicles in diabetes mellitus induce alterations in endothelial cell morphology and migration.

Wu, Sharon F / Noren Hooten, Nicole / Freeman, David W / Mode, Nicolle A / Zonderman, Alan B / Evans, Michele K

Journal of translational medicine

2020 Volume 18, Issue 1, Page(s) 230

Abstract: Background: Inflammation-related atherosclerotic peripheral vascular disease is a major end organ complication of diabetes mellitus that results in devastating morbidity and mortality. Extracellular vesicles (EVs) are nano-sized particles that contain ... ...

Abstract	Background: Inflammation-related atherosclerotic peripheral vascular disease is a major end organ complication of diabetes mellitus that results in devastating morbidity and mortality. Extracellular vesicles (EVs) are nano-sized particles that contain molecular cargo and circulate in the blood. Here, we examined EV protein cargo from diabetic individuals and whether these EVs cause functional changes in endothelial cells. Methods: We quantified inflammatory protein levels in plasma-derived EVs from a longitudinal cohort of euglycemic and diabetic individuals and used in vitro endothelial cell biological assays to assess the functional effects of these EVs with samples from a cross-sectional cohort. Results: We found several significant associations between EV inflammatory protein levels and diabetes status. The angiogenic factor, vascular endothelial growth factor A (VEGF-A), was associated with diabetes status in our longitudinal cohort. Those with diabetes mellitus had higher EV VEGF-A levels compared to euglycemic individuals. Additionally, EV levels of VEGF-A were significantly associated with homeostatic model assessment of insulin resistance (HOMA-IR) and β-cell function (HOMA-B). To test whether EVs with different inflammatory cargo can demonstrate different effects on endothelial cells, we performed cell migration and immunofluorescence assays. We observed that EVs from diabetic individuals increased cell lamellipodia formation and migration when compared to EVs from euglycemic individuals. Conclusions: Higher levels of inflammatory proteins were found in EVs from diabetic individuals. Our data implicate EVs as playing important roles in peripheral vascular disease that occur in individuals with diabetes mellitus and suggest that EVs may serve as an informative diagnostic tool for the disease.
MeSH term(s)	Cross-Sectional Studies ; Diabetes Mellitus ; Endothelial Cells ; Extracellular Vesicles ; Humans ; Vascular Endothelial Growth Factor A
Chemical Substances	Vascular Endothelial Growth Factor A
Language	English
Publishing date	2020-06-09
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Intramural
ISSN	1479-5876
ISSN (online)	1479-5876
DOI	10.1186/s12967-020-02398-6
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Article: Using the NRC to manage horse nutrition (an over view of management issues in the Nutrient Requirements of Horses, sixth revised edition)

Freeman, David W

Proceedings of the ... Mid-Atlantic Nutrition Conference. 2007, no. 5

2007

Keywords	horses ; animal nutrition ; nutrient requirements ; animal feeding ; guidelines ; publications
Language	English
Size	p. 171-177.
Document type	Article
Note	Conference held March 28-29, 2007, Timonium, Maryland.
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Association of Extracellular Vesicle Protein Cargo with Race and Clinical Markers of Mortality.

Noren Hooten, Nicole / McFarland, Minna H / Freeman, David W / Mode, Nicolle A / Ezike, Ngozi / Zonderman, Alan B / Evans, Michele K

Scientific reports

2019 Volume 9, Issue 1, Page(s) 17582

Abstract: Differential mortality rates remain a significant health disparity in the United States, suggesting the need to investigate novel potential molecular markers associated with mortality. Extracellular vesicles (EVs), including exosomes, microvesicles and ... ...

Abstract	Differential mortality rates remain a significant health disparity in the United States, suggesting the need to investigate novel potential molecular markers associated with mortality. Extracellular vesicles (EVs), including exosomes, microvesicles and apoptotic bodies, are lipid-bound vesicles secreted by cells into the circulation. EVs mediate intercellular communication by shuttling functional signaling molecules as cargo. EV characteristics by race in the context of mortality risk factors have not been described. We isolated plasma EVs from a cross-sectional cohort of African Americans (AA) and whites and found no significant differences in EV size, distribution or concentration between race or by sex. However, EV cargo showed increased levels of phospho-p53, total p53, cleaved caspase 3, ERK1/2 and phospho-AKT in white individuals compared to AAs. phospho-IGF-1R levels were significantly higher in females compared to males. EV concentration was significantly associated with several clinical mortality risk factors: high-sensitivity C-reactive protein (hsCRP), homeostatic model assessment of insulin resistance (HOMA-IR), alkaline phosphatase, body mass index, waist circumference and pulse pressure. The association of EV proteins with mortality markers were dependent on race. These data suggest that EV cargo can differ by race and sex and is associated with mortality risk factors.
MeSH term(s)	African Americans/statistics & numerical data ; Biomarkers/analysis ; Blood Proteins/analysis ; Cell-Derived Microparticles/chemistry ; Continental Population Groups/statistics & numerical data ; Cross-Sectional Studies ; European Continental Ancestry Group/statistics & numerical data ; Exosomes/chemistry ; Extracellular Vesicles/chemistry ; Female ; Health Status Disparities ; Humans ; Male ; Middle Aged ; Mortality ; Risk Factors ; Sex Factors ; United States/epidemiology
Chemical Substances	Biomarkers ; Blood Proteins
Language	English
Publishing date	2019-11-26
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-019-53640-1
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Article ; Online: Whence CRIPTO: The Reemergence of an Oncofetal Factor in 'Wounds' That Fail to Heal.

Freeman, David W / Rodrigues Sousa, Elisa / Karkampouna, Sofia / Zoni, Eugenio / Gray, Peter C / Salomon, David S / Kruithof-de Julio, Marianna / Spike, Benjamin T

International journal of molecular sciences

2021 Volume 22, Issue 18

Abstract: There exists a set of factors termed oncofetal proteins that play key roles in ontogeny before they decline or disappear as the organism's tissues achieve homeostasis, only to then re-emerge in cancer. Although the unique therapeutic potential presented ... ...

Abstract	There exists a set of factors termed oncofetal proteins that play key roles in ontogeny before they decline or disappear as the organism's tissues achieve homeostasis, only to then re-emerge in cancer. Although the unique therapeutic potential presented by such factors has been recognized for more than a century, their clinical utility has yet to be fully realized1. This review highlights the small signaling protein CRIPTO encoded by the tumor derived growth factor 1 (
MeSH term(s)	Animals ; Humans ; Signal Transduction/genetics ; Signal Transduction/physiology ; Stem Cells/metabolism ; Stem Cells/physiology ; Transforming Growth Factor beta/metabolism
Chemical Substances	Transforming Growth Factor beta
Language	English
Publishing date	2021-09-21
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms221810164
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