bluuebunny/crossref_metadata_2025 huggingface dataset:
Structure‐Function Analysis of Ursolic Acid and its Analogs to Identify Their Anti‐inflammatory Mechanism of Action
Authors: Nguyen Huynh Nga , Kim Hong Seok , Ullevig Sarah , Asmis Reto
Published: June 2021

We showed that ursolic acid (UA), a pentacyclic triterpenoid with anti‐inflammatory properties, reduces atherosclerotic lesion size and monocyte dysfunction in diabetic mice. We proposed that UA protects monocytes against metabolic stress‐induced monocyte dysfunction or “priming,” by preventing the induction of Nox4, the rate‐limiting step promoting hyper‐responsiveness to chemoattractants and accelerated chemotaxis induced by metabolic stress. We have also demonstrated that UA rescues MKP‐1 protein expression and activity in metabolically primed cells. The aim of this study was to investigate nine of UA's naturally occurring analogs and identify the structural features of UA required to inhibit monocyte priming and prevent Nox4 induction. We determined the doses (0.03 ‐ 3 μM) at which UA and its analogs blocked metabolic stress‐induced acceleration of chemotaxis in response to MCP‐1 in metabolically primed THP‐1 monocytes. Only the UA analogs hederagenin (IC50 = 0.64 μM), oleanolic acid (IC50 = 1.9 μM), and uvaol (IC50 = 2.46 μM), protected monocytes against metabolic priming. UA was the most potent analog (IC50 = 0.1 μM). All protective analogs also prevented Nox4 induction and rescued MKP‐1 from degradation. Our data suggests that hederagenin, oleanolic acid and uvaol prevent monocyte priming and dysfunction through the same mechanism as UA and thus may all be atheroprotective. Identifying the structural features responsible for UA's beneficial effects will enable us to develop more potent, dietary supplements for the prevention of atherosclerosis and other monocyte‐mediated diabetic complications.​

Funding Source: Supported by a grant to RA from the NIH (R01‐AT006885)
How Does Homestead Food Production Improve Child Nutrition? Path Analysis of the AAMA Project in Nepal
Authors: Self Julie , Haardörfer Regine , Stein Aryeh , Pandey Pooja , Martorell Reynaldo , Webb Girard Amy
Published: June 2021

The Action Against Malnutrition through Agriculture (AAMA) Project is an Enhanced Homestead Food Production Project implemented by Helen Keller International and local partners in Far Western Nepal that combined a nutrition‐sensitive agriculture intervention with behavior change communication to improve nutrition and health. Data from a cross‐sectional endline survey were used to measure variables along HKI's hypothesized Program Impact Pathway (PIP), from inputs through key outcomes of interest, including child growth and maternal and child hemoglobin. Survey respondents included mothers with children aged 12‐48 months at the time of the interview (n=2614). We used path analysis to assess the relative contribution of specific mechanisms of effect on nutrition and health as outlined in the PIP. The hypothesized model for height‐for‐age z‐score fit the data well (χ²(19)=55.8, p<0.001; RMSEA=0.027; CFI=0.945), while the model for child hemoglobin (χ²(20)=297.7, p<0.001; RMSEA=0.073, CFI=0.913) was a moderate fit, and some models did not achieve adequate fit. Agricultural inputs is the program component with the strongest path in the model (β=0.022, p<0.001). Areas of weak association and poor model fit may be attributed to weaknesses in measurement, intervention fidelity, and contextual factors unaccounted for in the PIP. These findings guide program improvements by highlighting areas for enhancements in program design, implementation or evaluation.
Differential Effects of Carbohydrates on Changes in Acute Childhood Cognitive Control
Authors: Khan Naiman , Raine Lauren , Winter Adam , Cohen Neal , Kramer Arthur , Hillman Charles
Published: June 2021

Understanding how diet affects cognition has important public health implications. However, the effects of specific nutrients on acute childhood cognitive function are unclear. This double‐blind, placebo controlled trial assessed the effects of formulae containing carbohydrates with different glycemic properties, on cognitive control among 9‐10‐year‐olds. All participants (N=103) consumed a non‐caloric placebo and were randomized to 1 of 3 treatments containing carbohydrates with varying absorption rates (FAST [n=36], MODERATE [n=34], or SLOW [n=33]). A modified Eriksen flanker task was used to assess cognitive control during the fasted state and 10 min and 70 min postprandial. 70 min postprandial, the FAST carbohydrate increased reaction time during the congruent trials and response variability during congruent and incongruent trials. The SLOW carbohydrate decreased response accuracy interference (congruent – incongruent) compared to the MODERATE carbohydrate. These results demonstrate that ingestion of carbohydrates with varying glycemic properties elicits differential changes in acute cognitive control whereby rapidly absorbed carbohydrates result in longer reaction times and greater intraindividual response time variability. Conversely, moderating the glycemic response may improve children's ability to modulate cognitive control when faced with higher task demands. (Supported by a grant from Abbott Nutrition through the Center for Nutrition, Learning, and Memory at the University of Illinois)
Pro‐inflammatory Cytokine, IL‐1B, Diminishes Butyrate Oxidation in Colorectal Cancer Cells
Authors: Johnstone Megan , Donohoe Dallas
Published: June 2021

Most colorectal cancers undergo the Warburg effect. This shifts cell energetics toward increased glycolytic metabolism and away from oxidative metabolism and butyrate oxidation. Butyrate, derived from the fermentation of dietary fiber, is the preferred energetic substrate of the non‐cancerous colonocyte. Butyrate, at physiologically relevant levels in the colon, inhibits histone deacetylases to regulate gene expression and subsequently increase apoptosis in colorectal cancer cells. Accordingly, we sought to determine the effects of the pro‐inflammatory cytokine interleukin‐1 β (IL‐1β) on butyrate oxidation in real‐time cellular respiration using a Seahorse Analyzer. We utilized 2 colorectal cancer cell lines (HCT116 and HT‐29) in these studies. Cells were placed in fatty acid oxidation buffer supplemented with or without IL‐1B (1ng/mL) treatment for one hour prior to Seahorse assay. Butyrate and 2‐deoxyglucose (2DG) were injected into wells.. Baseline measurements reveal injected butyrate (final concentration 5mM) to significantly increase oxidative consumption rate (OCR) as compared to control (p<0.05, n=5). Concurrently, cells injected with butyrate and treated with IL‐1B showed significantly increased OCR as compared to control (p<0.05, n=5); yet significantly decreased butyrate oxidation as compared to butyrate injected wells (p <0.05, n=5). Upon injection of 2DG (final concentration 5mM), which blocks glucose oxidation leaving only butyrate as the sole energetic substrate, only butyrate‐treated cells showed significant increases in OCR (p<0.0, n=5). Thus, IL‐1β appears to diminish butyrate oxidation in colorectal cancer cells, which may affect subsequent genomic acetylation and all downstream targets including proliferation and apoptotic pathways.
Dried Plums Modify Colon Microbiota Composition And Spatial Distribution, And Protect Against Chemically‐Induced Carcinogenesis
Authors: Seidel D. , Hicks K. , Taddeo S. , Azcarate‐Peril M. , Carroll R. , Turner N.
Published: June 2021

Differences in microbial populations in the proximal and distal colon may impact apparent site‐specific differences in pathology. Diet is known to alter metabolism and composition of colon microbiota, which has major implications for disease prevention and treatment. The hypothesis tested by this experiment was that consumption of dried plums would promote retention of beneficial microbiota and patterns of microbial metabolism throughout the colon, and that by doing so would reduce colon cancer incidence. Male Sprague‐Dawley rats were provided either a control (CD, n=25) or plum diet (PD, 5% of calories, n=26) 3 wk before being given two injections of AOM (15 mg/kg BW) or saline, and sacrificed 8 wk later. Tissues were resected and fecal contents isolated separately from the proximal and distal colon. Irrespective of treatment, the PD increased Bacteroidetes (p<0.0001) and reduced Firmicutes (p<0.0001) in the distal colon without affecting their proximal proportions, compared to the CD, which suppressed Bacteroidetes and increased Firmicutes in the distal colon. Additionally, rats consuming PD had significantly reduced numbers of aberrant crypts (p=0.0025), aberrant crypt foci (p=0.0060), and high multiplicity aberrant crypt foci (p=0.0008) compared to CD rats. These data support our hypothesis that dried plums protect against colon cancer, and this may be due in part to their ability to establish putatively beneficial colon microbiota compositions in the distal colon. Supported by California Dried Plum Board PN 12‐20.
Anthocyanin‐rich Black Elderberry Extract improves Inflammatory Markers and Insulin Resistance in a High‐Fat Diet‐Induced Obesity Model of Mice
Authors: Farrell Nicholas , Norris Gregory , Ryan Julia , Blesso Christopher
Published: June 2021

Despite increased knowledge of healthy eating and exercise habits, obesity continues to be a public health concern in the U.S., resulting in many preventable comorbidities such as type 2 diabetes mellitus (T2DM) cardiovascular disease (CVD), and premature death. Identifying new prevention and treatment strategies for obesity‐related conditions are a primary interest. Black elderberry has one of the highest concentrations of polyphenols per gram of dry weight of all commercially available foods and may be useful in mitigating the inflammatory effects of obesity. The objective of this study was to determine if anthocyanin‐rich black elderberry extract (Sambucus nigra) (BEE) could protect against low‐grade chronic inflammation and insulin resistance in a high‐fat diet induced obesity model using C57BL/6J mice. Mice were fed either a low fat diet (LFD), high‐fat diet (HFD), HFD + 0.25% (w/w) BEE (LBEE), or HFD + 1.25% (w/w) BEE (HBEE) for 16 weeks. After 16 weeks, there were significant reductions in liver weight, fasting triglycerides, fasting insulin, serum monocyte chemoattractant protein‐1 (MCP‐1), and serum tumor necrosis factor‐α (TNFα) in the LBEE and HBEE groups compared to the HFD group. No differences were observed in body weight or food intake between high‐fat diet groups. The homeostatic model assessment method (HOMA) was used to quantify insulin resistance and the LBEE and HBEE groups were found to have significantly lower values, indicating higher insulin sensitivity. These results suggest that black elderberry may have potential as a natural product to help reduce obesity‐associated chronic disease. [Supported by the UConn Research Foundation].
Complementary Feeding Pattern Reduces the Impact of Diarrhea on Relative Weight: A Longitudinal Analysis of the Interaction of Diet and Diarrhea on Weight‐for‐length in Cebu Infants
Authors: Wright Melecia , Adair Linda
Published: June 2021

Undernutrition and diarrheal disease are major contributors to infant morbidity and mortality worldwide. We investigated the combined roles of infant feeding and diarrhea on relative weight (weight‐for‐length z scores or WFL) in 2814 infants from the Cebu Longitudinal Health and Nutrition Survey. Indicator coding was used to create eight primary predictors based on combinations of current breastfeeding status (breastfed/BF vs non‐breastfed/NBF), high or low diversity score (DDS) in the complementary feeding diet (consumption of 4+ or <4 of 7 possible food groups) and presence of diarrheal disease (sick vs healthy with respect to diarrheal disease). WFL was predicted using sex‐stratified, fixed‐effects longitudinal models that also adjusted for age and energy from complementary foods.

Table 1 Results of longitudinal regression of WFL on dietary and diarrhea patterns during infancy

Boys' Beta
95% CI
p
Girls' Beta
95% CI
p

Diet x diarrhea pattern

Healthy + NBF + Low DDS
‐0.141*
(‐0.221, ‐0.062)
0.001
‐0.114*
(‐0.208, ‐0.020)
0.018

Healthy + BF + Low DDS
0.009
(‐0.062,0.080)
0.801
0.048
(‐0.035,0.132)
0.257

Healthy + NBF + High DDS
‐0.127*
(‐0.216, ‐0.037)
0.006
‐0.093+
(‐0.195,0.009)
0.073

Healthy + BF + High DDS
Referent
n/a
n/a
Referent
n/a
n/a

Sick + NBF + Low DDS
‐0.319*
(‐0.404, ‐0.235)
<0.001
‐0.224*
(‐0.324, ‐0.125)
<0.001

Sick + BF + Low DDS
‐0.148*
(‐0.228, ‐0.069)
<0.001
‐0.064
(‐0.155,0.026)
0.164

Sick + NBF + High DDS
‐0.224*
(‐0.332, ‐0.116)
<0.001
‐0.152*
(‐0.276, ‐0.029)
0.016

Sick + BF + High DDS
‐0.101
(‐0.241,0.038)
0.154
‐0.091
(‐0.237,0.056)
0.224

Age
‐2.232*
(‐2.402, ‐2.062)
<0.001
‐2.090*
(‐2.263, ‐1.917)
<0.001

Age‐squared
0.770*
(0.700,0.840)
<0.001
0.708*
(0.638,0.778)
<0.001

Energy intake
‐0.286*
(‐0.428, ‐0.143)
<0.001
‐0.427*
(‐0.573, ‐0.281)
<0.001

Age x energy intake
0.375*
(0.275,0.475)
<0.001
0.481*
(0.378,0.584)
<0.001

Intercept
0.719*
(0.593,0.845)
<0.001
0.627*
(0.495,0.759)
<0.001

Within strata of diarrhea status or diet diversity, non‐breastfed babies were thinner than their breastfed counterparts. Interestingly, regardless of breastfeeding status, diarrhea was consistently related to lower WFL in babies with low diet diversity but diarrhea did not significantly reduce the WFL of babies with high diet diversity. Furthermore, amongst non‐breastfed boys with diarrheal disease, infants with high diet diversity had significantly higher relative weight than those with low diet diversity.

image

These findings suggest that high diversity in the complementary diet may mitigate the detrimental impact of diarrhea on infant nutritional status.
Pulsatile Delivery of Cortisol Regulates Lipid Storage in Adipocytes in Vitro and Human Adipose Gene Expression in Vivo
Authors: Karastergiou Kalypso , Pramyothin Pornpoj , Port Ava , Apovian Caroline , Lee Mi‐Jeong , Fried Susan
Published: June 2021

Glucocorticoids (GC) regulate the distribution and function of human adipose tissue, and their pathophysiological effects are attributed to changes in their concentration. We hypothesized that the diurnal cortisol rhythm contributes to its effects.

Microarray analysis and qPCR were used to identify genes with expression changes as circulating cortisol declines between 9:00 and 15:00, in human abdominal subcutaneous adipose tissue of healthy volunteers (n=5). Placebo or a single, physiological dose of cortisol were administered out of phase (11:30) to identify genes with expression changes dependent on the cortisol rhythm. A novel in vitro model was established to study the long‐term effects of pulsatile versus constant GC in human adipocytes.

We identified 280 genes significantly upregulated and 292 downregulated from 9:00 to 15:00. Cortisol changed the expression pattern of 191 (33.4%) genes. We verified changes in the clock gene PER1, the GC receptor co‐chaperone FKBP5, and KLF15, a regulator of circadian nitrogen homeostasis. In vitro, pulsatile GC established rhythmic clock gene expression in human adipocytes, led to enlarged lipid droplets and lower basal lipolysis and de novo fatty acid synthesis compared to constant GC.

These findings suggest that higher turnover of triglycerides may occur in patients with disrupted glucocorticoid rhythm and contribute to increased cardiometabolic risk.

Research support: NIH DK080448, P30 DK046200 and UL1‐TR000157.
Effect of a new dietary SIRT1 activator on high fat diet‐induced obesity and the involvement of microRNAs
Authors: Ahn Jiyun , Lee Hyunjung , Jung Chang Hwa , Ha Tae Youl , Jang Young Jin
Published: June 2021

SIRT1 plays a key role in regulating metabolism, and SIRT1 activation may be a promising strategy to treat metabolic syndrome. This study investigated whether zerumbone ameliorated diet‐induced obesity through SIRT1 activation. We used differentiated 3T3‐L1 fibroblasts to examine the effect of zerumbone on adipogenesis through the miR‐146b‐SIRT1 cascade. To investigate the anti‐obesity effect of zerumbone in vivo, we fed zerumbone to high‐fat diet‐induced obese C57BL/6J mice for 8 weeks and measured body weight, adipose tissue size, and blood lipid profiles. We also measured the effect of zerumbone supplementation on SIRT1 and AMPK signaling pathways in the white adipose tissue of these mice. Zerumbone inhibited adipogenesis through the miR146b‐SIRT1 pathway and significantly reduced diet‐induced obesity in mice. Zerumbone supplementation was associated with miR‐146b downregulation, followed by SIRT1 activation and the deacetylation of FOXO1 and PGC1α, respectively, in the white adipose tissue of these mice. Zerumbone also activated AMPK and modulated lipid metabolism in adipose tissue by increasing fatty acid oxidation and reducing adipogenesis gene expression, respectively. Zerumbone inhibited adipogenesis via the miR‐146b/SIRT1 pathway and ameliorated diet‐induced obesity in mice via SIRT1 activation, suggesting that zerumbone could target obesity‐related metabolic disorders through its stimulation of SIRT1 activity.
The Association Between Serum Prostate‐Specific Antigen and Glycemic Index, Glycemic Load, and Metformin in Individuals with Diabetes: a Cross‐sectional Analysis
Authors: Jayalath Viranda , Ireland Christopher , Augustin Livia , Nishi Stephanie , Mirrahimi Arash , De Souza Russell , Kendall Cyril , Jenkins David
Published: June 2021

Background

Improving glycemic control with Metformin therapy, glycemic index (GI), and glycemic load (GL) have been protectively associated with prostate cancer risk. No studies have yet assessed the relationship between GI, GL, and Metformin on serum prostate‐specific antigen (PSA), a controversial‐yet‐widely used marker of prostate health.

Methods

A cross‐section analysis of baseline PSA data from men with type 2 diabetes mellitus (T2DM) who previously completed a trial at this research center was conducted. Multivariate linear regression quantified associations between PSA and log‐PSA with GI, GL, and Metformin dose.

Results

Data were available from 317 men, with a mean PSA of 1.41±0.09ng/mL. Medians exposures (ranges) were 80 (56 – 96) for GI, 164 (62 – 386) for GL, and 1500mg (0 ‐ 3000 mg) for Metformin; 90% of men were under Metformin therapy. Neither PSA nor log‐PSA were significantly related to GI or GL. A negative trend was seen with PSA and metformin dose (p=0.09) in multivariate models. PSA was 24% lower among individuals taking 蠅2000mg of Metformin (n=125) compared to individuals taking <1000mg (n=62) (p=0.06). Log‐PSA significantly decreased by 0.15±0.09 ng/mL for every 1000mg increase in Metformin dose (r=‐0.38, p=0.02).

Conclusions

Metformin, but not GI or GL, inversely related with log‐PSA in a sample of men with T2DM. Modification of PSA‐test sensitivity as a marker of prostate cancer with diet and Metformin warrants attention.

Funding CIHR, Barilla, Loblaw Brands ltd
Incidence of Pulmonary Fibrosis in Lungs of CUX‐1 Transgenic Mice
Authors: Helgeson Scott , Harris Ethan , Herndon Betty , Vanden Heuvel Gregory , Baybutt Richard , Molteni Agostino
Published: June 2021

CUX‐1 is a transcriptional repressor gene and part of the network controlling G1‐S phase transition by repressing the cyclin‐kinase inhibitors p21 and p27. CUX‐1 transgenic mice develop polycystic kidneys, reduced spermatogenesis, and liver fibrosis as a consequence of aberrant repression of p27. We reported that these mice also show severe pulmonary inflammation with vasculitis, bronchitis, bronchiectasis, and diffuse presence of fibroblasts.

This study reports the presence of collagen using Image J software (NIH program) on slides of lungs of CUX‐1 mice stained by trichrome. Wild type mice were used as controls. An average of seven photographs at 400x magnification were taken on the lungs of each mouse.

Collagen presence and intensity was 17.06% in lungs of CUX‐1 mice versus 8.73% the wild type mice (p<0.05). Fibrosis was mostly evident in the basal membrane and musculature of the ectasic bronchi and the peribronchial arteries. CUX‐1 mice also showed medial thickening, reduced lumen patency (64% versus 76% in wild type, p 0.015), and reduced media/adventitia ratio (42% versus 64% in wild type, p 0.018). Fibroblasts were particularly evident in the adventitia. Data using a specific rat stain for SMA‐1 confirm those ones observed with trichrome.

Our data quantitatively confirms the presence of severe pulmonary fibrosis in these mice. Would some patients with polycystic kidney disease also have aberrant repression of p21 and p27 and develop pulmonary fibrosis?
Survivin is a Therapeutic Target in Ewing Sarcoma
Authors: Dioufa Nikolina , Tsokos Maria
Published: June 2021

Ewing sarcoma, a bone and soft tissue sarcoma of young adults, is genetically defined by the EWS/FLI1 fusion. Current treatment is not effective in patients with macro‐ (25%) and micro‐metastatic (30%) disease, which is fatal in 80% of them. Our goal is to study the metastatic phenotype of Ewing sarcoma in order to identify molecules that can predict and be targeted to treat metastatic disease. In order to survive in the circulation and metastasize tumor cells escape anoikis, a form of cell death that occurs when they detach from the extracellular matrix. Therefore, three dimensional spheroid cultures based on anchorage independent survival of tumor cells have been used to study metastatic tumor phenotypes. We established three dimensional spheroid cultures of four Ewing sarcoma cell lines and showed that when compared to their respective adherent cells they 1) show increased chemoresistance and migration in vitro, 2) grow faster and form larger tumors in mice 3) show increased levels of EWS/FLI1 in the blood of mice consistent with circulating tumor cells and 4) have higher levels of survivin, an inhibitor of apoptosis protein with preferential expression in cancer cells. We also showed that survivin has a role in the formation of the spheres and drug‐resistant phenotype, because its downregulation with siRNA and YM155, a small molecule inhibitor currently tested in clinical trials, reversed the sphere formation and chemoresistance. These data support that survivin is a biologic marker of aggressiveness and can be targeted to treat patients with metastatic Ewing sarcoma.
A Multi‐omics Approach to Identify Biomarkers of Clinically Advanced Retinoblastoma for Diagnostics and Therapeutic Applications
Authors: Guha Nilanjan , S A Deepak , Lateef Syed , Padmanabhan Arunkumar , Gundimeda Seetaraman , Ghosh Arkasubhra , Mallipatna Ashwin , Suresh Babu Vishnu
Published: June 2021

Retinoblastoma is a pediatric eye cancer affecting children usually less than five years of age. Inactivation of both the copies of the RB1 gene in a child's retina initiates cancer formation. While a lot of literature exists regarding pathogenesis and genetic changes in retinoblastoma, there is still a lack of revelation of specific progression markers and correlation of expression markers with disease mechanism. In this study, we aim to determine transcriptomic, metabolomic and proteomic profile of this cancer from different sample types viz. tissue, aqueous humor, vitreous humor and tear from enucleated eyes of 9 patients and 2 deceased controls, whose cause of death is not due to any eye related disease. There are 5 male and 4 female patients and comprises of both unilateral and bilateral cases. In the first stage of the study, we performed pathway analysis of mRNA and miRNA microarray data looking for gene enrichment that would enable functional characterization of tumors. We were able to identify many of the key pathways that are known to be involved in the progression of retinoblastoma including cell cycle pathway. The study also revealed 18 novel miRNAs which has not been previously implicated in the disease. Metabolomics and proteomics studies are underway to identify linkages and data concordance that will contribute to a more comprehensive understanding of the underlying mechanism of retinoblastoma. Overall, the study shows the utility of applying an integrative approach to study molecular mechanisms in retinoblastoma by co‐analyzing multi‐omics data that will also help identify signatures that are unique to aggressive retinoblastoma.
Immunohistochemical Analysis of LGR5 Expression in Pediatric Liver Disease
Authors: Khan Zahida , Orr Anne , Michalopoulos George , Ranganathan Sarangarajan
Published: June 2021

BACKGROUND

In regenerating liver, hepatic progenitor cells (HPCs) are recruited in response to liver injury; however, there is a lack of highly specific human HPC markers for the hepatocyte lineage. LGR5, a Wnt‐associated adult stem cell marker, has been extensively studied in intestinal stem cells. Although LGR5 appears to be exclusively expressed by HPCs in mice, little is known about its expression in human liver. We hypothesized that LGR5+ cells are induced in the regenerative response to liver injury to potentially rescue dysfunctional hepatocytes.

METHODS

Using immunohistochemistry, we characterized LGR5 expression in pediatric liver diseases, using mainly liver explants (n=36 cases total).

RESULTS

We found cytoplasmic LGR5 over‐expression in all 36 pediatric cases, including acute hepatic necrosis and cholestatic liver diseases. In 44% of cases, >50% of hepatocytes were LGR5+, signifying a robust regenerative response. In contrast, there was only weak staining in bile ducts, suggesting hepatocyte‐specific expression. Marked peri‐portal expression was prominent in cholestatic diseases, consistent with known patterns of regeneration. Comparison to other regeneration markers will further characterize this hepatocyte progenitor cell phenotype.

CONCLUSIONS

Although we observed some degree of regenerative response in all cases examined, LGR5 was highly expressed in cholestatic liver diseases, possibly due to alternate regeneration pathways. These studies are the first to support a unique role for LGR5 in human hepatocyte regeneration and as a potential predictive marker for recovery of liver function in children.

LGR5 INTENSITY
TOTAL CASES
WEAK +
1+
2+
3+

A1AT Deficiency (ATD)
15
0
2
5
8

Biliary Atresia (BA)
5
0
0
2
3

PFIC
10
1
1
5
3

Acute Hepatic Necrosis (ALF)
6
1
4
1
0

TOTAL CASES
36
2
7
13
14

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Differential And Cooperative Roles Of β‐Catenin And Yap During Hepatoblastoma Pathogenesis
Authors: Tao Junyan , Li Xiaolei , Singh Sucha , Chen Xin , Monga Satdarshan
Published: June 2021

Hepatoblastoma (HB) is the most frequent pediatric liver tumor. In our previous study we demonstrated that coordinated activation of Yap and β‐catenin is a major mechanism in promoting HB development. However, the individual contribution of Yap and β‐catenin and how they may interact to contribute to induce HB remains unknown. Here, instead of co‐delivering yap and β‐catenin sleeping beauty plasmids with transposase, we hydrodynamically delivered either Yap first followed 3 days or 7 days later by β‐catenin or vice versa. The mouse livers were harvested at 6 weeks, 7 weeks, and 9 weeks after the second injection for comparing tumor size and phenotype to the Yap‐b‐catenin co‐injected group. There was no difference in the liver phenotype when Yap injection preceded β‐catenin compared to Yap‐β‐catenin co‐injection. However, the tumors were significantly smaller and fewer when β‐catenin injection preceded Yap as compared to the co‐delivery group at the early time‐points (6 & 7 weeks). Intriguingly, there was no difference in tumors between all 3 groups at late stage (9 weeks). Lastly, when Yap with defective TEAD binding and b‐catenin or b‐catenin with defective TCF4 binding and Yap were co‐delivered, there was alleviation of tumorigenesis. In summary, Yap is important early in reprograming hepatocyte to hepatoblasts, while both Yap and β‐catenin with preserved ability to bind their canonical transcription factors are required for tumor growth and progression through activation of downstream targets.
Crosstalk In DNA‐Alkyltransferase Functions
Authors: Fried Michael , Tessmer Ingrid
Published: June 2021

DNA alkyltransferases protect genomic integrity, but under some conditions can also defend tumor cells against chemotherapeutic alkylating agents. Here we examine mechanisms that regulate the DNA‐binding and repair activities of human O6‐alkylguanine‐DNA alkyltransferase (AGT). AGT binds O6‐methylG‐C and O6‐methylG‐T lesions with specificity far too low for efficient lesion search. This suggests that other factors are needed to direct AGT to lesion sites. Recently we have found that AGT binds the human MutSβ homologue and PCNA proteins. In vitro, MutSβ inhibits alkyltransferase activity on short duplex DNAs, while PCNA increases repair rates on single‐stranded DNAs and long duplexes. The compositions and binding activities of MutSβ‐AGT and PCNA‐AGT complexes suggest roles for these interactions in lesion search. In addition to lesion‐specific binding, the surveillance process includes interactions with lesion‐free DNAs. The non‐specific binding of AGT is homo‐cooperative, allowing high densities to be attained in the absence of other proteins. Protein‐protein contacts that contribute to cooperativity are predicted to be optimal over a narrow range of DNA twist values. Outside of those values, torsional free energy will limit cooperative cluster size. Assays using superhelical DNAs show that binding densities and alkyltransferase rates decrease with the linking difference, ΔLk. These results predict that AGT and its DNA‐repair activities will partition in favor of torsionally‐relaxed, relatively protein‐free DNA structures like those found near sites of chromatin remodeling. This binding preference may play an important role in genome surveillance. Supported by NIH grant GM070662 to MGF.
Crosstalk between Bone and Muscle: Deletion of Mbtps1 in Bone Leads to Age‐Dependent Increase in Muscle Size and Contractile Function
Authors: Gorski Jeffrey , Huffman Nichole , Seidah Nabil , Bonewald Lynda , Brotto Marco
Published: June 2021

Proprotein convertase Mbtps1 is required for transcription of bone matrix proteins including fibronectin, collagen XI, and PHEX by osteoblastic/osteocytic cells. Since complete loss of Mbtps1 is lethal at E4, we asked if conditional deletion of Mbtps1 using early osteocyte DMP1 cre would disrupt bone structure. Mbtps1 cKO male mice were slightly larger than controls (43.6 g vs. 40.2 g, p=0.057) but displayed no significant changes in bone dimensions, BMD, mineral content, or cortical/trabecular bone parameters at 12 and 40 weeks of age. However, 40 week old soleus muscles from cKO mice weighed 20% more and contracted with 50% more force ex vivo even when normalized for size despite the fact that Mptps1 expression was unchanged (0.94 cKO/control +/‐ 0.42 STD). Immunostaining revealed cKO soleus muscles contained about 35% type I and 60% type 2 myosin heavy chain positive cells, were devoid of myosin 2b fiber+ cells which comprised 5% of control cells, and displayed a higher percentage of central nuclei than controls. Central nuclei, which can reflect muscle repair or regeneration, were restricted to type I myosin heavy chain positive cells. The osteocyte specific deletion of Mbtps1 along with genome arrays that demonstrate increased expression of muscle performance markers such as α‐actinin3 support a cross‐talk signaling mechanism by which Mbtps1‐deficient bone cells increase muscle contractile function in adult mice. Revealing the molecular mechanism responsible could lead to new therapies for a host of muscle disorders.
Signal‐dependent Regulation of Transcription Factor Activity
Authors: Gill Grace , Saia Greg , Lalonde Jasmin
Published: June 2021

Signal dependent regulation of transcription factor activity in the nervous system controls gene expression programs that underlie learning and memory. Transcription factor Sp4 is zinc finger DNA binding protein that regulates neuronal connectivity, brain development and function. In humans, altered Sp4 has been linked to psychiatric disorders such as schizophrenia and bipolar disorder. We are investigating the function and regulation of Sp4 in neurons. Our recent studies have revealed two distinct calcium‐dependent signaling pathways that regulate Sp4 via altered patterns of post‐translational modification. We have found that stimulation of the NMDA receptor activates a PP1/PP2A protein phosphatase signaling pathway that dephosphorylates Sp4. Our data suggests that this phosphorylation reduces Sp4 function independent of an effect on Sp4 levels. We have also found that Sp4 protein, but not mRNA, abundance is highly regulated in response to changes in membrane potential associated with neuronal activity. In resting neurons, Sp4 is ubiquitinated and rapidly degraded by the proteasome. We have found that calcium influx via store operated calcium entry (SOCE) and the ER calcium sensor Stim1 regulates Sp4 ubiquitination. Our studies reveal new aspects of the complex mechanisms that determine signal‐dependent regulation of transcription factor activity important for neuronal plasticity, brain development and disease.
15‐PGDH Prevents LPS‐induced Acute Liver Injury through Activation PPAR‐γ
Authors: yao lu , han Chang , wu tong
Published: June 2021

The NAD+‐dependent 15‐hydroxyprostaglandin dehydrogenase (15‐PGDH) catalyzes the oxidation of Prostaglandin E2 (PGE2), converts PGE2 to 15‐keto‐PGE2 (an endogenous ligand for peroxisome proliferator‐activated receptor‐gamma [PPAR‐γ]). PPAR‐γ is a ligand‐dependent transcriptional factor, which controls genes relevant to the regulation of liver cytokines production and hepatocyte apoptosis. To evaluate the significance of 15‐PGDH/PPAR‐γ cascade in hepatocyte apoptosis and liver injury, we generated transgenic mice with targeted expression of 15‐PGDH in the liver by using an albumin promoter‐enhancer–driven vector and the animals were subjected to lipopolysaccharide (LPS)‐induced acute liver injury. Specifically, the 15‐PGDH transgenic (Tg) and wild type mice were administered intraperitoneally with LPS (60ng/g) in combination with D‐galactosamine (D‐GalN) (800µg/g) for 5 hours and the extent of liver injury was assessed by histopathology, serum aminotransferases, caspase‐3 staining, and caspase activation. Comparing to wild type mice, the 15‐PGDH Tg mice exhibited lower alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, less liver damage, and less hepatocyte apoptosis, and the 15‐PGDH Tg livers showed reduced production of inflammatory cytokines and reactive oxygen species (ROS), higher PPAR‐γ downstream gene expression, and weaker activation of TNF‐α/JNK cascade compared. Pretreatment 15‐PGDH Tg mice with inhibitor/antagonist of 15‐PGDH/PPAR‐γ restored the susceptibility of 15‐PGDH Tg mice to LPS‐induced acute liver injury. Our findings suggest that 15‐PGDH ameliorates LPS‐induced liver injury through its enzymatic product, 15‐keto‐PGE2, which activates PPAR‐γ and thus inhibits ROS and TNF‐α/JNK cascades.
Regulation of Kupffer Cell Activation by Hepatic Stellate Cells
Authors: Roth Katherine , Albee Ryan , Copple Bryan
Published: June 2021

When cells in the liver are damaged or killed by trauma, macrophages become activated to release cytokines and phagocytose dead cell debris, which facilitates liver repair. The mechanisms that regulate macrophage activation after liver damage remain poorly understood. We found previously that the transcription factor hypoxia‐inducible factor‐1a (HIF‐1a) is activated in hepatic stellate cells (HSCs) after injury. Deletion of HIF‐1a in HSCs prevented hepatic macrophages from polarizing to an M1 phenotype after liver injury, which reduced production of proinflammatory cytokines and prevented removal of dead cell debris. In the present study, we tested the hypothesis that necrotic hepatocytes activate HIF‐1a in HSCs, which regulates release of a mediator that in turn activates Kupffer cells. In our current studies, we show that HIF‐1a is activated and HIF‐1a target genes are upregulated in HSCs exposed to necrotic hepatocytes. Our data reveal further that exposure of HSCs to necrotic hepatocytes also activates several MAPK pathways, including ERK1/2 and p38, and upregulates the chemokines CCL2, CCL7, and CCL9. Finally, we demonstrate that treatment of Kupffer cells with conditioned medium from HSCs treated with necrotic hepatocytes upregulates the proinflammatory genes CXCL1 and CXCL2 and down‐regulates the anti‐inflammatory gene IL‐10. Together, these data support the mechanism whereby necrotic cells activate HIF‐1a in HSCs, which upregulates a mediator that is released. This mediator then stimulates Kupffer cells to produce proinflammatory cytokines and to phagocytose dead cell debris. These studies could help identify novel targets for reducing liver injury and improving liver repair by modulating the hepatic inflammatory response.
Investigating Multimodal Classroom Experiences across a Biology Curriculum
Authors: Petzold Andrew , Metzger Kelsey , Dunbar Robert
Published: June 2021

The Center of Learning Innovation at the University of Minnesota Rochester aims to deliver an undergraduate curriculum that advances learner‐centered, technology‐enhanced, competency‐based, assessment‐driven education in the health sciences. To accomplish this, the faculty within the life sciences have created multimodal classrooms that are based on recent pedagogical innovations including backward course design, flipped classroom engagement, and highly collaborative learning spaces to create a student‐centered approach to learning. By combining these approaches with a variety of active‐learning implementation strategies and multiple instructors in the classroom space guiding and supporting student activities, we are able to facilitate an environment for learning that is most suited for utilizing cognitive research driven instructional practices. This multimodal approach to learning, entitled the “confluent classroom,” provides faculty the opportunity to focus, at the course level and the series level, on furthering the specific aims of the Center. All course data (e.g. artifacts of student learning, metrics of student performance, course metadata including Learning and Development Outcomes) is collected in an in‐house database and preliminary analysis can be performed with a faculty‐developed Browser of Student and Course Objects (BoSCO). Effectiveness of the confluent classroom model is explored by tracking and comparing individual student performance across a three‐semester core life sciences course series that culminates in a sophomore‐level Anatomy and Physiology II course.
Program Assessment in a Large Undergraduate Physiology Curriculum
Authors: Stanescu Claudia , Henriksen Erik , Delamere Nicholas
Published: June 2021

Program assessment is a challenging task, especially in the Physiology Undergraduate major at the University of Arizona, which must accommodate over 2000 students in large classrooms. We describe an assessment plan that includes six student learning outcomes (SLOs). Students are expected to demonstrate knowledge of cellular (SLO 1) and organ systems function (SLO 2); show the ability to integrate knowledge from the cellular and molecular to the organ system level of organization (SLO 3); effectively read, evaluate and communicate scientific information (SLO 4); conduct and/or evaluate physiology laboratory experiments (SLO 5); and demonstrate knowledge of current physiology topics (SLO 6). Faculty have developed a sustainable assessment program that includes direct and indirect measures of assessment and is being implemented over a four‐year period. The first year included assessment of SLOs 1, 2, and 3 that will be measured annually. SLOs 4, 5, and 6 will be evaluated in subsequent years on a rotating basis. Assessment activities include: 1) pre‐post clicker questions in large core courses that determine cellular and organ systems knowledge (SLOs 1, 2); 2) use of a grading rubric to assess the ability to integrate physiology in a subsample of graduating seniors participating in a focus group (SLO 3); 3) use of a grading rubric on a subsample of students in a large advanced elective course to evaluate the ability to effectively communicate scientific information (SLO 4); and 4) graduating senior surveys to obtain student ratings for program SLOs, major courses, advising and overall satisfaction with the Physiology major. The overall goal is to collect and analyze data, discuss findings with the faculty, refine the curriculum as needed, and regularly improve the assessment plan.
“Dialogical Story‐Telling” Approach to Enhance Analytical Thinking and Student Engagement During Lecture‐Based Classes
Authors: ElSayed Suzan , Loftus Stephen
Published: June 2021

The “Dialogical Story‐Telling” approach is an interactive method to teach physiology for health professional students during didactic lectures. This interactive method integrates an interactive conversational style within a story‐telling method. The story telling incorporates dialogue with the students, which is in a question‐answer format that not only creates a motivating learning context but also builds an educationally safe and supportive environment.

A week before the lecture, students are provided with copies of the PowerPoint slides to be used. The students are required to review all the slides. During the lecture, the instructor guides the students through the material using story‐telling and question‐answer styles. The instructor also uses questions related to the material that encourage analytical thinking.

There is an emphasis on building an educational relationship between students and teachers. Relationship‐centered learning goes beyond student‐centered learning, in recognizing and rehabilitating the role of the teacher. There are strong theoretical foundations for relationship‐centered learning. There is the dialogism of Bakhtin, with the recognition of the importance of interactivity. There is also social constructionism where it is accepted that meaning, knowledge and understanding are jointly constructed with others.
Changes of Chemical Elements in the Special Embalming Fluid after Immersed in Human Fresh Amputated Remains
Authors: Luo Jia , Liu Tianxiong , Liu Lian , Hu Tu , Wang Jianlong , Chen Yanliang , Zeng Leping , Huang Jufang
Published: June 2021

Objectives

To evaluate the effect of the special embalming fluid preserved on the human fresh amputated remains at a micro level, and to find out the temporal and spatial changes of chemical elements leaching from tissues.

Methods

The whole remains and tissues (skin, muscle, nerve, artery) cut off from the remains were immersed into the special embalming fluid for 6, 12 and 24 months. After paraffin embedding, H&E staining was performed, and the concentrations of chemical elements in embalming fluid were determined by ICP‐AES.

Results

The morphological structure of tissues was preserved well in 6 and 12 months, and the chemical elements, especially the trace elements leached few into the embalming fluid. The common elements leached earlier than the trace elements. As the time passing, the type and the concentrations of chemical elements were increased. The trace elements in the whole remains were preserved better than the removed tissues, so were that in muscle and artery better than in skin and nerve.

Conclusion

The embalming fluid preserved the fresh amputated remains well in short time. Specific embalming fluid should be developed to preserve different tissues better and longer.

This work was supported by the National Key Technology Support Program (No.2012BAK14B03), Chinese postdoctoral Science Foundation (No. 2014M562136) and Hunan Provincial Innovation Foundation For Postgraduate(No.1145‐71380100001)
3D Rendering of Porcine Recurrent Laryngeal Neuroanatomy
Authors: Mason Nena , Wisco Jonathan
Published: June 2021

Recurrent laryngeal nerve (RLN) palsy is a common post‐operative complication of many head and neck surgeries. The best treatment to restore partial function to a damaged RLN is reinnervation of the posterior cricoarytenoid (PCA) muscle via anastomosis of the RLN and phrenic nerves. The pig is an excellent model of human laryngeal anatomy and physiology. A thorough knowledge of porcine laryngeal anatomy is necessary before the pig can be used to improve existing surgical strategies, and develop new ones. This study provides state of the art 3D renderings of the porcine larynx and the location of the RLN and its branches. Anatomical variation of primary RLN branching was analyzed in 25 specimens to reveal 3 major branching patterns. Pattern A (44%) consisted of a medial primary branch that bifurcated into two secondary branches. Pattern B (32%) consisted of three primary branches. Pattern C (24%), consisted of two primary branches. Several heat maps depict the relative spatial variability of 32 bilateral specimens of RLN trunks and primary branches. We show a high degree of variability in whole nerve medial/lateral location within the larynx in left RLN specimens as compared to right RLNs. However, the location of RLN primary branches was more highly conserved among left RLNs. We intend this study to be useful to groups using a porcine model to study PCA reinnervation techniques. This research is funded by the BYU College of Life Sciences Start‐Up Mentoring Grant.
Seriated C.T‐Scan Of The Osseous Head In Pantera Pardus
Authors: LE FLOCH‐PRIGENT PATRICE , VERDEILLE STEPHANE , GILLOT JEAN BERNARD
Published: June 2021

The osseous head in a panther (Pantera pardus) was CT‐scanned (Siemens somatom definition AS apparatus) was systematically seriated CT‐scanned, every 0,8 mm with a 1 mm thickness : 252 cross‐sections were obtained in the frontal plane with the head laid horizontally on the radiological table. In the para‐sagittal plane and in the horizontal (orbito‐meatal or Francfort plane), respectively 151 and 252 cross‐sections were reconstructed from the native ones. The morphological results were numerous and often tremendous ; the osseous structure was perfectly defined on every level of cross‐section. The exact description of the roots of the dentition was possible. Two metallic wires which solidarized the cranium and the mandible did not alter the examination. The absence of the central structures in the nasal cavities was only a missing set of data without commitment for the interpretation of the peripheral walls which were unaltered. The external reconstructions under several views were a great help in the localization of the structures.
Codistribution of Collagens V and VI with Collagens I and III in Hepatic Fibrosis of Elderly Cadavers
Authors: Mak Ki
Published: June 2021

Collagen V is a fibrillar collagen of the extracellular matrix (ECM). It forms heterotypic fibrils with collagen I and regulates the growth of collagen I fibrils. Collagen VI is a microfilamentous collagen that forms a network in the ECM, linking matrix components and cells. In the liver, collagens V and VI are minor ECM proteins, but their levels are significantly elevated in cirrhosis. Fibrosis is often seen in aged livers and is prevalent in elderly cadavers. Using immunoperoxidase staining in paraffin sections, this study assessed collagens V and VI expression and their association with fibrillar collagens I and III in progressive stages of liver fibrosis of elderly embalmed cadavers. In liver with minimal fibrosis, staining for collagens V and VI was uniform in the perisinusoidal space of the lobules, stroma and vessel walls of portal tracts, and rims of central veins. Perisinusoidal hepatic stellate cells (HSCs) expressed collagens V and VI, revealing the cellular source of these collagens. In foci of severe perisinusoidal fibrosis, collagens V and VI staining was enhanced, codistributing with the increased presence of collagens I and III. Collagens V, VI, I, and III were abundant in the ECM of fibrotic portal tracts, fibrous septa of sepal fibrosis and cirrhosis. In sections post‐stained with Sirius red for collagen fibers—mainly collagens I and III—the immunoreactivity of collagens V and VI was seen localized to the fiber bundles.

Conclusion

The association of collagen V with collagen I in fibrotic foci is in accord with its regulatory role for fibrillogenesis of collagen I, while collagen VI—a connecting protein—links fibrillar collagens to the surrounding ECM. Thus, collagens V and VI could serve as biomarkers of liver fibrogenesis.