bluuebunny/crossref_metadata_2025 huggingface dataset:
Type III Transforming Growth Factor‐ β Receptor regulates proliferation and apoptosis in epicardial cells
Authors: Sanchez Nora Sylvia , Austin Anita F , Compton Leigh A , Love Joey D , Barnett Joey V
Published: June 2021

Epicardial cells transform into the myocardium and form coronary vessels. The Type III Transforming Growth Factor‐ β Receptor (Tgfbr3) is required for coronary vessel development. Tgfbr3‐/‐ embryos have an irregular epicardium, few coronary vessels, and die at E14.5. To explore TGF βR3 signaling we made immortalized epicardial cell lines from E11.5 embryos. Tgfbr3+/+, +/‐, and ‐/‐ cells form a tight epithelium. TGF β1 or TGF β2 (250 pM) caused transformation and induced the smooth muscle marker, SM22α. Circular wounds were made in cell monolayers and the percent wound closure calculated. Tgfbr3 +/+ and +/‐ cells closed the wound by 48h. Tgfbr3‐/‐ cells required 72h (p<0.05). Tgfbr3 +/+ and +/‐ cells gave similar proliferation rates when measured by BrdU, peaking at 48h (33.1% and 33.93%) and returning to basal levels (19.9% and 20.85%) by 72h. Tgfbr3‐/‐ cells sustained a basal proliferation rate 13% lower at 48h (p=0.001). As a second measure of proliferation, we used the in vivo reduction of MTS tetrazolium. Tgfbr3‐/‐ cells had a rate of proliferation 2‐fold lower at 48h and 2.6 fold lower at 72h (p<0.05). Apoptosis was determined by Apo‐One Caspase 3/7 Homogenous assay. The apoptosis rate in Tgfbr3‐/‐ cells was 1.8, 3.3 and 5.9‐ fold higher than Tgfbr3+/+ cells at 24, 48, 72h, respectively (p<0.05). In summary, TGFβR3 is not required for transformation in epicardial cells, but does regulate proliferation and apoptosis.

Source: HL085708, HL076133 (AFA), AHA0655129 (JVB), GM007628 (NSS), & GM07347 (LAC).
Theoretical simulation of microvascular network growth and adaptation
Authors: Secomb Timothy W. , Alberding Jonathan P. , Pries Axel R.
Published: June 2021

In the microcirculation, vessel formation (angiogenesis), structural adaptation (remodeling), and regression (pruning) redistribute flow in response to varying demands. Theoretical simulations were used to explore the roles of these processes in the formation of efficient and functionally adequate network structures. A model for structural adaptation, including responses to pressure, shear stress and local oxygen level (PO2), was combined with a model for sprouting angiogenesis, interconnection and pruning. A diffusible growth factor (GF) was assumed to be produced in hypoxic tissue. For a given network, the blood flow distribution and the PO2 and GF fields are computed. Probability of sprout formation and direction of growth depend on GF concentrations and gradients. Pruning occurs when diameter drops below 3 μm. In simulations starting with a sparse network and tissue hypoxia, vascular density first increases, with many non‐flowing segments. Redundant vessels drop out, yielding stable network structures with little or no hypoxia, resembling structures observed in the rat mesentery. Both remodeling and pruning are needed for formation of functional and efficient networks. The model was extended to include vessel migration due to axial tension in vessels, yielding a distribution of branching angles similar to that observed. Supported by NIH HL034555.
Chase‐ing regeneration: engineering chondroitinase for mammalian expression
Authors: Curinga Gabrielle Mercedes , Jin Ying , Zhang Chen , Mashburn Charles , Snow Diane M , Smith George M
Published: June 2021

Neurons fail to regenerate following spinal cord injury partly due to the up‐regulation of chondroitin sulfate proteoglycans (CSPGs). Animal models of neuronal injury show enhanced regeneration when treated with chondroitinase, a bacterial enzyme that degrades inhibitory CSPG glycosaminoglycan (GAG) side chains. Unfortunately, this treatment paradigm is limited by the lack of a non‐invasive, sustained delivery method. This study aims to increase the therapeutic potential of chondroitinase by "engineering" the bacterial chondroitinase gene for maximum sustained expression in mammals and by using a lentiviral gene therapy approach for delivery. Injection of "engineered" chondroitinase lentivirus resulted in sustained activity as detected by digestion of GAG chains two weeks post‐injection. Transplantation of primary astrocytes and Schwann cells that were previously transduced with chondroitinase lentivirus also resulted in GAG chain degradation at the two week time point. Lack of macrophage immunoreactivity following both methods of chondroitinase delivery suggests minimal host immune response and supports the non‐invasive nature of our approach. Finally, in contrast to control neurons, preliminary data suggests that chondroitinase transduced neurons extend axons through regions of CSPG in vitro. These data significantly advance the clinical therapeutic potential for chondroitinase. Supported by NIH‐NIDA 1T32 DA022738, NIH‐NINDS 1 P30 NS051220‐01, and KSCHIRT #6‐13.

Grant Funding Source

NIH
Polarity and Morphogenesis‐‐‐ the Planar Cell Polarity Pathway in Mammalian Embryogenesis
Authors: Wang Jianbo
Published: June 2021

Originally identified in flies, the planar cell polarity (PCP) pathway is crucial in establishing the cellular polarity in the plane of the epithelium, perpendicular to the apical‐basal axis of the cell. In frogs and zebrafish, homologous PCP genes regulate directional cell behavior to mediate polarized tissue morphogenesis. In the mouse, we and others have found that a highly conserved mammalian PCP pathway regulates both cell polarity and morphogenetic movement to assemble cells into tissues and organs with distinct anatomical forms and structures. For example, auditory sensory hair cells develop uniformly oriented stereocilia on the apical surface and PCP proteins localize asymmetrically on the apical membrane during this process. Mutations in PCP genes disrupt polarized membrane distribution of other PCP proteins and cause randomized orientation of stereocilia. We have also found that the PCP pathway is essential for coordinated lengthening and narrowing of the neural plate during neural tube closure. The PCP pathway is also required for outflow tract formation in the heart and our genetic studies pinpointed a specific role of this pathway in cardiomyocytes derived from the secondary heart field. While the detailed cellular mechanisms are yet to be defined, the mammalian PCP pathway is undoubtedly a critical force in shaping various tissues and organs and may underlie a number of birth defects in humans.
Rotation of organizer tissue contributes to left‐right asymmetry in avian embryos
Authors: Rongish Brenda J , Cui Cheng , Little Charles D.
Published: June 2021

Establishment of left‐right sidedness is a crucial early event in vertebrate morphogenesis. It is currently believed that left‐right asymmetry patterns result from genetic regulatory networks via extracellular morphogens and/or ciliary activity in all vertebrates. However the role of biomechanical motion near the organizer region, has been overlooked. Electroporation of histone 2B‐YFP nuclear marker into avian nodal epiblastic cells and time‐lapse imaging (HH 4‐6) revealed the movements of labeled cells during the interval when morphological and gene expression asymmetries are first observed. The recordings indicated that relative motion occurs between the organizer (Hensen's node) and the surrounding epiblastic epithelium in avians. Specifically, an asymmetric counterclockwise rotation of epiblastic nodal tissue was observed following electroporation on the embryonic left side of the node. However, on the embryonic right, a two‐pronged migratory path was observed. The rotational motion occurs during the interval when initial expression of Shh and Fgf8 is symmetrical, and continues until establishment of Shh and Fgf8 left‐right asymmetry. Ongoing work is aimed at influencing the biomechanical tissue motion and analysis of resulting gene expression patterns. Supported by an AHA Fellowship 0620105Z, the NIH P20 RR016475 from the INBRE Program, NCRR; NIHLB R01 068855; and the Mathers Foundation.
Microfibril‐associated glycoprotein‐1 binds multiple active TGF‐beta family members, affecting homeostasis in multiple organ systems
Authors: Pierce Richard A , Craft Clarissa S , Weinbaum Justin S , Broekelmann Thomas , Mecham Robert P
Published: June 2021

Microfibril‐associated glycoprotein‐1 (MAGP‐1) is a small molecular weight extracellular matrix component of the fibrillin‐rich microfibril that forms the scaffold for elastic fibers. Surprisingly, mice null for MAGP‐1 have normal elastic fiber morphology and their elastic tissues function normally. The complex phenotype of mice null for MAGP‐1 includes increased body weight and size due to excess body fat, an altered wound healing response in bone and skin, and a bleeding diathesis. The obesity and bone phenotype is intriguing given the localization of obesity and osteoporosis candidate genes to human chromosome 1, which also houses the MAGP‐1 gene (mfap2). These phenotypic traits are consistent with transforming growth factor‐beta (TGF‐beta) dysregulation, and we have determined that MAGP‐1 binds TGF‐beta and multiple BMPs with high affinity. Further, these traits are opposite to those associated with mutations in MAGP‐1's binding partner fibrillin‐1, and many fibrillin‐1 mutations result in enhanced TGF‐beta signaling. Together, our data implicates MAGP‐1 as an important modulator of microfibril‐mediated growth factor signaling.
A protein chip approach to unraveling networks and pathways
Authors: Zhu Heng
Published: June 2021

The protein chip technology is emerging as a versatile tool in the field of proteomics. It provides an unbiased, high‐throughput, and comprehensive platform to investigate various aspects of protein properties; the results from which greatly facilitate construction of networks and pathways on a global scale. Our group has established new assays and applied them to globally investigate protein posttranslational modifications, with focus on lysine residues. We have also built new protein chip platform to decode the DNA‐protein interactome in humans. In this presentation, exciting new discoveries will be shared with the audience.
Comparison of bioavailability of the vitamin A‐forming carotinoids (beta‐cryptoxanthin. beta‐carotene and alpha‐carotene).
Authors: Burri Betty Jane , Chang Jasmine , Neidlinger Terry Ray
Published: June 2021

Beta‐carotene (BC), beta‐cryptoxanthin (CX) and alpha‐carotene (AC) are common vitamin A (VA)‐forming carotenoids in human diets and blood. Recently conversion ratios for VA formation were set at 12:1, 24:1, and 24:1 for BC‐, CX‐ and AC‐rich foods, respectively. The conversion ratio for BC is based on scientific data of its bioavailability, but ratios for CX‐ and AC‐rich foods are based primarily on the chemical structure of these carotenoids without regard for their bioavailability from their primary food sources. However, in our studies CX serum concentrations have been unexpectedly high compared to dietary intakes. We hypothesized that CX serum concentrations might also be elevated compared to dietary intake in other studies. We calculated the ratio of dietary intake to serum concentration for AC, BC, and CX from our studies and several large US and European surveys. We then compared the ratios of AC/BC and CX/BC. The ratio of AC/BC approached 1 (AC/BC = 0.74 to 1.1), suggesting that eating similar amounts of AC‐ and BC‐rich foods result in similar blood concentrations, and thus that the primary food sources of AC and BC have comparable bioavailability. However, the ratio of CX/BC was 5.6 to 35, suggesting that eating comparable amounts of CX and BC resulted in much higher serum CX concentrations. This suggests that the bioavailability of CX from its main food sources is higher than BC or AC, and that CX‐rich foods might be better sources of VA than expected from their abundance in the diet.
Lycopene‐accumulating E. coli transformed with human CMO II as a lycopene metabolism model
Authors: Lu Chi‐Hua , Gong Xiaoming , Marisiddaiah Raju , Rubin Lewis P. , Erdman John W.
Published: June 2021

Mouse and ferret CMO II have been reported to asymmetrically cleave beta‐carotene or lycopene at the 9', 10' double bond and form apo‐10′‐carotenal or apo‐10′‐lycopenal. To investigate the ability of human CMO II to cleave lycopene, four E. coli strains were prepared: control, lycopene‐accumulating (LYC), lycopene‐accumulating transformed with human CMO II (Lyc‐CMO II), and lycopene‐accumulating transformed with an enzymatic site mutated human CMO II (Lyc‐mtCMO II). E coli were extracted with acetone by sonication, and lycopene metabolic products (lycopenoids) were determined by HPLC. PCR and western blot confirmed that the CMO II gene was successfully expressed in E. coli cells. Among the strains tested, Lyc‐mtCMO II accumulated the highest lycopene content, followed by LYC, Lyc‐CMO II, and control, while the lycopenoids, apo‐8′‐lycopenal, apo‐10′‐lycopenal, apo‐12′‐lycopenal, apo‐10′‐lycopenoic acid, and apo‐15′‐lycopenoic acid, were below limits of detection. (Supported by CA125384A, HD42174, and RR18728)

Grant Funding Source

CA125384A
Overweight, Obesity and television viewing in adolescents of Public School from Mexico
Authors: Morales‐Ruán Ma. del Carmen , Gómez Luz María , Shamah‐Levy Teresa
Published: June 2021

Objective

To determine whether time spent in television (TV) viewing is related to overweight and obesity in adolescents from Public Schools.

Materials and Methods

Cross sectional design. Adolescents aged 10 to 19 were included in the National Health Survey in Scholar Population (ENSE 2008). Weight and Height were measured and BMI was calculated. TV watching time was also measured and finally the time dedicated to moderated and vigorous physical activity was recorded. A logistic regression model was created to estimate the relationship between the overweight and obesity and time spent watching TV, physical activity, age, gender, and school level (elementary or junior high school).

Results

A total sample of 39,653 adolescents was studied. Overweight and obesity prevalence were of 33.3% and 30.9% in men and women respectively. A positive relation between hours spent viewing TV and overweight and obesity was found. Adolescents who spent watching TV from 7 to 13 hr/week increase the chance of suffer overweight or obesity (OR 1.20, p < 0.05), watch TV from 14 to 20 hr/week increase 29% the probability of to develop these conditions and watching TV more than 21 h/week increases the same risk in 32% (OR 1.32, p<0.05) in comparison with those adolescents who dedicate less than 7 h/week to watch TV.

Conclusion

A positive association exists between the spent time in front of TV and the overweight and obesity.
Hepatic synthesis of palmitate and oleate during the postprandial period is suppressed in response to low fat/high carbohydrate intake in subjects with type 2 diabetes
Authors: Wilke Michaelann , French Margaret A , Goh Yeow K , Ryan Edmond A , Clandinin Michael T
Published: June 2021

The objective of this research was to determine whether diabetes and/or low fat intake would increase postprandial de novo lipogenesis of saturated fatty acids (FAs), negatively affecting VLDL‐triglyceride composition. It was hypothesized that low fat/high carbohydrate intake, particularly in diabetes, would result in higher hepatic synthesis of total and saturated FAs. A short‐term low glycemic/low fat diet was compared to a high oleate/high fat diet in seven subjects with type 2 diabetes mellitus and 7 matched non‐diabetic control subjects in a blinded randomized crossover design. For the first time in humans, de novo synthesis of myristate, palmitate, palmitoleate and oleate was estimated using deuterium incorporation techniques. The synthesis of all FAs increased significantly during the postprandial period, but there was large variation in synthetic rates. Myristate exhibited the highest rate of synthesis, whereas palmitate was the major net de novo FA synthesized. The lower fat diet acutely stimulated FA synthesis in control but not diabetic subjects. These results indicate a lack of short term lipogenic response to dietary carbohydrate in diabetes subjects. Funded by the CDA and CIHR.

Nutrition & Metabolism

Biochemistry of Vitamins and Minerals (400‐ASN)

Energy and Nutrient Metabolism (401‐ASN)

Human and Clinical Nutrition (402‐ASN)

Metabolic and Disease Processes (403‐ASN)

Late Breaking Category ‐ Nutrition & Metabolism

400‐ASN Biochemistry of Vitamins and Minerals

401‐ASN Energy and Nutrient Metabolism

402‐ASN Human and Clinical Nutrition

403‐ASN Metabolic and Disease Processes

Abstract 870

Grant Funding Source

Canadian Diabetes Association
Caffeine free polyphenolic extracts from Yaupon holly (Ilex vomitoria) have chemopreventive potential and reduce the expression of inflammatory genes in non‐cancer human myofibroblast (CCD‐18) cells
Authors: Noratto Giuliana Doris , Kim Youngmok , Talcott Steve Terel , Mertens‐Talcott Susanne U.
Published: June 2021

The objective of this study was to evaluate chemopreventive potential of different fractions of phenolics isolated from green tea (GT) and yaupon (YU) on human HT‐29 colon cancer cells and the anti‐inflammatory effects on colonic myofibroblasts CCD‐18 cells. Infusions from GT and YU leaves were fractionated into phenolics acids (Pa) and flavonoids (Fv) followed by caffeine removal (noC). Proliferation was determined by MTT assay, inhibition of hydrogenperoxide‐induced reactive oxygen species (ROS) by dichlorofluorescein (DCF) assay, and modulation of gene expression by RT‐PCR. Among extracts the order of antiproliferative

potency in HT‐29 cells was YU‐Fv‐noC > YU‐Fv > GT‐noC ~ GT. The IC50 values of YU‐Fv‐noC for HT‐29 and CCD‐18 were 15 and 68 mg gallic acid equiv./L, respectively.

YU‐Fv‐noC showed low toxicity exerted in non‐cancer colon cells, though the protective effect against the H2O2 induced ROS was not significant. However, YU‐Fv‐noC significantly decreased the expression of the pro‐inflammatory genes IL‐6 and COX‐2, up to 99% and NF‐κB from 8 to 35% compared to untreated cells. The expression of cell‐protective catalase was increased by 1.1 to 1.4 fold in a dose‐dependent manner at 10‐40mg/L. In summary, our results indicate that flavonoids from YU seem to have anti‐cancer effects and also may have potential for use in chemoprevention based on their anti‐inflammatory effects.
Both black and green tea consumption associated with reduced risk of stroke
Authors: Arab Lenore , Liu Weiqing , Elashoff David
Published: June 2021

Animal experiments show tea extracts to reduce the infarct volume in a dose dependent fashion. To examine whether tea consumption in man is associated with a reduced risk of cerebral infarction in man we conducted a research librarian supported literature search and a meta analysis of all published human clinical and epidemiologic research on tea and stroke. Risk estimates in each study were calibrated to estimate the effect on stroke associated with consumption of 3 cups of tea. Of 59 citations, 10 studies were identified with original data on either fatal or non‐fatal ischemic stroke in relationship to tea consumption. All of these studies provided risk point estimates below 1.0. The meta analysis demonstrated no significant heterogeneity across studies. The pooled, fixed effect relative risk was 0.79 with a 95% confidence interval of 0.73‐0.85. These effects were quite consistent for black tea (RR=0.76, 0.67‐0.86) and green tea (RR=0.79, 0.72‐0.86). We found that, regardless of country of origin or type of tea consumed, the additional consumption of 3 cups of tea per day is associated with an average 21% lower risk of ischemic stroke, suggesting that the onset of ischemic stroke in man may be prevented through daily tea consumption.
Identification of bacterial species with phenolic‐degrading activity in the human oral cavity
Authors: Ye Zhong , Rieser Erinn , Li Li , Hendrich Suzanne
Published: June 2021

We hypothesized that human oral bacteria had significant but highly variable phenolic degradation capability, which may influence ability of these compounds to prevent periodontal disease. Caucasian (2 men, 7 women) and Asian (3 men, 8 women)subjects who had no dental disease and were not taking supplements provided 5‐7mL saliva before toothbrushing. Rutin, caffeic acid, daidzein, quercetin, naringenin, luteolin and myricetin (each 100μM)were incubated with saliva in anaerobic BHI media at 37°C. Phenolics were measured at 0, 3, 6, 9, 12 and 24h by HPLC. Saliva bacterial 16S rRNA gene sequences were amplified and separated. Saliva degradation rates were caffeic acid/rutin > quercetin/ myricetin/naringenin > luteolin >daidzein (p < 0.05). High & low degradation rate clusters were found for rutin, caffeic acid and naringenin; degradation rates of these compounds were significantly correlated with each other. DNA bands from high degraders of caffeic acid were concordant with Streptomyces coelicolor and S. avermitilis, and Lactobacillus brevis/L. reuteri for high degraders of rutin. These species may affect human oral degradation of these phenolics and prevention of gum disease. (Funded by Nutrition & Wellness Res. Ctr, Iowa State University).
Estrogenic compounds alter cell growth in non‐cancerous colonocytes
Authors: Weige Charles Conan , Allred Clinton D
Published: June 2021

Studies have shown that hormone replacement therapy can reduce the risk of colon cancer in post‐menopausal women, and dietary phytoestrogens are also associated with lower risk of the disease. We have demonstrated that estradiol (E2) and genistein (Gen), a model soy phytoestrogen, do not inhibit colon cancer cell growth regardless of whether the cell expresses estrogen receptor beta (ERβ). These data demonstrate the need to determine the role of estrogenic compounds in non‐cancerous colonocytes, but data has been limited due to lack of appropriate in vitro models. We have employed young adult mouse colonocyte (YAMC) cells which were shown to express ERβ. Treatment of YAMCs with E2 or Gen caused a reduction in cell number in a dose responsive manner. This effect was significantly inhibited by treatment with ICI, an ER antagonist. Apoptosis was induced in response to estrogenic treatments, but preliminary data suggests these treatments do not significantly alter the cell cycle. When the oncogene Ras is over‐expressed in YAMC cells, the effects of the compounds are lost. This data is critical because it is the first to show estrogenic compounds alter the cellular activity of non‐cancerous colonocytes, which is the first step towards determining how these compounds reduce colon cancer risk.

Grant Funding Source

The American Institute for Cancer Research
Selenium‐regulated expression of non‐selenoprotein genes
Authors: Raines Anna M. , Sunde Roger A.
Published: June 2021

Recent studies have linked super‐nutritional Se intake to both beneficial and adverse outcomes such as lowered risk for cancer and increased risk for type II diabetes. These paradoxical effects of high Se intake highlight the need for biomarkers of high Se status. While a subset of selenoprotein mRNAs are useful as biomarkers of Se deficiency, no selenoprotein mRNAs further increase at super‐nutritional intakes, precluding their use as biomarkers of high Se. To identify non‐selenoprotein genes regulated by Se, we used microarrays to analyze gene expression in liver and kidney from mice fed < 0.007 (‐Se), 0.05 (~Se), or 0.2 μg Se/g diet (+Se). Comparison analysis of expression in +Se vs. ~Se and +Se vs. ‐Se yielded 103 unique candidate biomarker genes changed = 1.4 fold. Detected selenoproteins show the expected pattern of Se‐regulation. The Se‐dependent expression of non‐selenoprotein genes is being confirmed by qRT‐PCR and tested in rats fed 0‐0.8 μg Se/g diet. So far, we have identified 6 genes changed > 3 fold only in +Se, and 1 transporter gene upregulated only in ‐Se in both mice and rats. Interestingly, four distinct patterns of Se‐regulation have emerged among the candidates: high in +Se; low in +Se; high in ‐Se; low in ‐Se (selenoprotein pattern). Candidate genes may be useful as high Se biomarkers and could be involved in Se homeostasis in Se deficiency or Se excess. Support: DK74184, DK07665, WISC04909

Grant Funding Source

NIH
Kidney Gpx3 is bound to tubular basement membranes
Authors: Olson Gary E , Winfrey Virginia P , Hill Kristina E , Burk Raymond F
Published: June 2021

Glutathione peroxidase 3 (Gpx3) is a plasma and extracellular selenoprotein postulated to protect against H2O2 and lipid hydroperoxides. In this study we show that kidney Gpx3 is insoluble, unlike plasma Gpx3. Immunohistochemistry localized Gpx3 to the perimeter of kidney tubules in the cortex and outer medulla of Gpx3+/+ mice. Absence of staining in Gpx3−/− kidneys verified antibody specificity. Tissue overlay experiments using kidney sections from Gpx3−/− mice demonstrated specific Gpx3 binding at the perimeter of the cortical tubules. Basement membranes isolated from Gpx3+/+ kidneys contained Gpx3 and, while basement membranes from Gpx3−/− mice did not contain Gpx3, they displayed Gpx3‐binding activity. Reducing conditions did not disrupt Gpx3 binding to basement membranes, indicating that binding was not mediated by disulfide or selenenylsulfide formation. Feeding selenium‐deficient diet resulted in depletion of the basement membrane‐bound Gpx3 pool. The reducing substrate glutathione is secreted by the tubular epithelium and that may allow the basement membrane‐associated Gpx3 pool to protect plasma membranes of kidney tubule cells. Supported by HD44863 and ES02497.
Involvement of Sep15 family members in immune response
Authors: Kasaikina Marina V , Fomenko Dmitry E , Jones Clinton J , Reddy Jay , Hatfield Dolph L. , Gladyshev Vadim N
Published: June 2021

Selenium and selenoproteins are known to have immunomodulatory properties. 15 kDa selenoprotein (Sep15) was previously shown to interact with UDP‐glucose:glycoprotein glucosyltransferase, a protein involved in the calnexin quality control cycle for many secreted glycoproteins, including proteins with the immune protection function (e.g., MHC class I receptor). In this study, we examined the resistance of Sep15 knockout mice (Sep15 KO) to viral infection and autoimmune disease. In both cases the development of the disease in the Sep15 KO group was different from that in wild type mice, and resulted in higher severity of symptoms and higher percentage of death cases. Examination of immunological parameters in mouse spleenocytes, including MHC class I and II, and CD4/CD8 population of T cells, did not show significant differences between WT and Sep15 KO groups. However, cell culture experiments with Sep15 KO MEFs demonstrated a decrease in â2‐microglobulin subunit when Sep15 KO was combined with deficiency in a distant Sep15 homolog, Selenoprotein M. Altogether, these data show the role of Sep15 family members in the immune function.
Indices of insulin sensitivity and secretion from a standard liquid meal test in subjects with type 2 diabetes, impaired and normal fasting glucose
Authors: Maki Kevin C. , McKenney James M. , Farmer Mildred V. , Reeves Matthew S. , Dicklin Mary R.
Published: June 2021

This study examined insulin sensitivity and secretion indices from a standard liquid meal tolerance test protocol in subjects with normal (NFG, n = 19), impaired fasting glucose (IFG, n = 19) and type 2 diabetes mellitus (n = 35). Areas under the curve (AUC) for glucose and insulin from pre‐meal to 120 min after a liquid meal were calculated, as were homeostasis model assessments of insulin resistance (HOMA2‐IR) and the Matsuda index of insulin sensitivity. Subjects with NFG, IFG and diabetes had mean fasting glucose levels of 5.22, 5.87 and 7.53 mmol/L and insulin of 52.8, 82.0 and 124.8 pmol/L, respectively. Subjects with NFG, IFG and diabetes had mean HOMA2‐IR values of 1.00, 1.57 and 2.48; glucose AUC of 11.1, 14.1 and 23.1 h*mmol/L; insulin AUC of 894, 1346 and 922 h*pmol/L; insulin/glucose AUC ratio of 78.6, 95.5 and 40.7 pmol/mmol; and Matsuda Index values of 15.6, 8.8 and 6.0, respectively. The product of the insulin/glucose AUC ratio and the Matsuda index, calculated as an indicator of the ability of insulin secretion to match the level of insulin resistance, was 995.6, 684.0, 188.3 for NFG, IFG and diabetes, and discriminated between fasting glucose categories (p < 0.001 for each comparison). These results provide initial evidence of the usefulness of a standard liquid meal tolerance test for evaluation of insulin secretion and sensitivity in clinical and population studies. Funded by Cargill Corporation (Wayzata, MN).
Poverty reduction program for extreme poor in Bangladesh improves nutritional status of preschool children
Authors: Jalal Chowdhury Sufi Bhakta , Frongillo Edward A
Published: June 2021

Poverty alleviation programs for the extreme poor improve participants' economic status and may impact other important outcomes that are seldom evaluated. A program targeted to the extreme poor by BRAC, a development organization in Bangladesh, has been successful in significantly alleviating extreme poverty. We hypothesized that the program also improved nutritional status (i.e., weight‐for‐height) of preschool children. A non‐equivalent control, pre‐ and post‐test quasi‐experimental design that was longitudinal at village level was used to test the hypotheses. Data were collected from a random sample of 4131 (1934 program and 2197 control) children from 3409 households across 169 villages of 3 northern districts of Bangladesh in 2002 and 2006. Linear mixed random‐intercept models accounted for clustering effects and potential confounders. Weight‐for‐height of children between ages 24‐35 months of program households was significantly higher (p<0.05) than that of control households. We found no significant program‐control difference in other age categories of preschool children. These results are highly important as this is a large‐scale program already extended to half of the country. Findings will contribute in judging the cost‐benefit and cost‐effectiveness of the program, and in garnering support for the expansion of such programs.
Impact of supplementary feeding on intra‐household food distribution in rural Kenya
Authors: Gewa Constance , Murphy Suzanne P , Weiss Robert E , Neumann Charlotte G
Published: June 2021

Inequitable food distribution practices between different household members: males and females, adults and children, household heads and non‐household heads and high and low earners have been associated with the differential distribution of malnutrition and poor health status among populations. Feeding programs that provide foods to selected household members may lead to changes in intrahousehold food allocation patterns. Our objective is to evaluate the effect of school snack provision on intra‐household nutrient distribution in rural Kenya and to assess if the effects differ by type of snack received. There was significant increase in school children's total daily intake between baseline and follow‐up period which approximately matched the energy content of the snack when snack and non‐snack days were compared. There was a general decline in the quantity and quality of non‐supplementary foods received by school children and in family member's intakes. However, there was no evidence that other family members received more energy on days that the schoolchild received a snack at school. There was no evidence that supplementary snack provision was associated with the changes noted in school children's intake of home foods and family members' intakes. Neither were there any consistent between‐group differences in the changes that were reported.
Caffeine biomonitoring: using LC‐MS/MS to estimate caffeine exposure in the U.S. population
Authors: Rybak Michael E , Pao Ching‐I , Pfeiffer Christine M
Published: June 2021

Caffeine is a psycho‐stimulatory plant alkaloid commonly encountered in the diet. Food frequency reporting is most often used for the epidemiological assessment of caffeine consumption. Biomarker measurement has been proposed as a means of assessing caffeine exposure but is not widely used at present. Our laboratory has developed an isotope‐dilution LC‐MS/MS analysis method that will be used to develop reference ranges for urinary caffeine and its metabolites for the 2009‐2010 National Health and Nutritional Examination Survey (NHANES). These ranges will provide biomarker estimates of caffeine consumption and may also be used to estimate the prevalence of several detoxification enzyme activity classifications (i.e., phenotypes) in the U.S. population. The method is capable of measuring caffeine and all its anticipated metabolites in urine at sub‐micromolar detection limits. Metabolites proposed for assessing caffeine exposure or enzyme activity phenotyping could be quantified in >99% of samples in a random subset. Between‐run coefficients of variation (CVs) were generally <10% for these compounds at concentrations at or above the subset 10th percentile, and within‐run repeatability CVs were <3%. In this paper we present and discuss the analytical performance of our method in the perspective of population biomonitoring studies such as NHANES.
Effects of signaling through the retinoic acid receptors in motor neurons: implications for amyotrophic lateral sclerosis
Authors: Kolarcik Christi , Bowser Robert
Published: June 2021

Amyotrophic lateral sclerosis (ALS), the most common form of motor neuron disease, is a progressive, fatal neurodegenerative disorder which is poorly understood and for which effective therapeutic interventions are lacking. Our laboratory has identified potential biomarkers for ALS and has characterized these proteins and related signaling pathways using a primary motor neuron‐enriched cell culture system. We focused on retinoid signaling as we have observed alterations in this pathway in sporadic ALS patients. Using all‐trans retinoic acid (ATRA) to stimulate the retinoic acid receptors (RARs), we further investigated the impact of signaling through the RARs on motor neuron survival. In the presence of ATRA, the number of branch points/cell was decreased in motor neurons. RAR β was localized to the nucleus irrespective of the presence of ATRA. RARα localized to axonal processes without ATRA, and this decreased with increasing ATRA concentrations. As similar phenomena have been observed in sporadic ALS patients, we believe that motor neurons may attempt to recapitulate a developmental process in an effort to maintain axonal connections as ALS progresses.

Research Support: NIH grant ES013469 (RB); NIH T32 EB001026 (CK); NIH T32 05 TL1 RR024155‐02 (CK)
Saturated fatty acid, palmitate, promotes smooth muscle phenotypic modulation and exacerbates neointima formation.
Authors: Shen Hua , Manabe Ichiro , Eguchi Kosei , Fujiu Katsuhito , Nagai Ryozo
Published: June 2021

Metabolic syndrome is a major risk factor for atherosclerotic cardiovascular disease. Circulating free fatty acid (FFA) levels are known to be increased in obese individuals.Increased levels of FFAs have been shown to be associated with higher risks for cardiovascular disease. However, little is known regarding effects of FFAs on vascular cells.We found that palmitate, the major circulating free fatty acid, induced expression of KLF5 and inflammatory genes, such as MMPs, and elicited smooth muscle cell (SMC) phenotypic modulation. When palmitate was administered to the mice whose carotid arteries were ligated, palmitate dramatically exacerbated neointima formation. Palmitate induced ROS formation, partly by induction of NADPH oxidase, NOX1 in cultured SMCs. NOX1 expression levels were also induced by palmitate infusion in mouse aortas and carotid arteries in vivo. Inhibition of NOX1 attenuated palmitate‐induced phenotypic modulation. Furthermore, we found that toll‐like receptor (TLR) signaling mediated the response to palmitate. Knockdown of Myd88, adaptor protein for TLRs, significantly reduced NOX1 induction and ROS formation. Taken together, palmitate promotes neointima formation via TLR‐signaling mediated ROS production and inflammatory gene expression. Palmitate appears to be a key common obesity‐related factor involved in pathogenesis of both cardiovascular and metabolic diseases.
Beta‐catenin/TCF complex regulates HNF4‐alpha expression in the liver
Authors: Zeng Gang , Monga Satdarshan S
Published: June 2021

Hepatocyte nuclear factor‐alpha (HNF4α) is a liver‐enriched nuclear receptor of the steroid and thyroid hormone receptor superfamily. It is expressed at the highest levels in the liver, kidney, intestine, and pancreas, and plays an essential role in development, organogenesis, and maintenance of organ function. Previously, we have shown that loss of β‐catenin during prenatal liver development compromises hepatocyte maturation. We identified low HNF4α expression in the β‐catenin‐deficient hepatocytes. Also, livers from transgenic mice overexpressing stable β‐catenin and hepatocyte‐specific β‐catenin knockout mice show high and low HNFα expression, respectively. Here, we investigate the regulation of HNF4α expression by β‐catenin. Transfection of HepG2 cells, a human hepatoma cell line, which harbors constitutively active β‐catenin, with HNF4α‐ reporter plasmids demonstrates high luciferase activity, which was significantly reduced in response to β‐catenin siRNA transfection. Co‐transfection of Hep3B cells (contain wild‐type β‐catenin) with stable β‐catenin plasmid and HNF4α‐promoter luciferase expression plasmid also exhibited high reporter activity. ChIP assays utilizing β‐catenin and TCF antibodies clearly show binding to HNF4α promoter. These data indicate that β‐catenin can directly regulate HNF4α, a nuclear transcription receptor and plays a critical role hepatocyte homeostasis.
Dickkopf‐1 regulates epithelial cell migration by inhibiting directional orientation
Authors: Koch Stefan , Capaldo Christopher T , Nava Porfirio , Skerra Arne , Sacks David B , Parkos Charles A , Nusrat Asma
Published: June 2021

Wnt signaling pathways regulate proliferation, motility and survival in a variety of human cell types. Dickkopf‐1 (Dkk‐1) is a secreted Wnt antagonist that has been shown to regulate tissue homeostasis in the intestine. In this report, we show that Dkk‐1 is secreted by migrating intestinal epithelial cells after wounding and during intestinal inflammation in vivo, and inhibits cell migration by attenuating directional orientation. We found that physiological concentrations of recombinant Dkk‐1 protein are sufficient to inhibit polarization of the microtubule organizing center and Golgi apparatus towards the leading edge of migrating epithelial cells. Cells exposed to Dkk‐1 exhibit a strong, mis‐localized activation of Cdc42, which coincides with an irregular localization of the polarity protein Par6 and increased phosphorylation of atypical PKC. In contrast, classical PKC phosphorylation is attenuated in the presence of Dkk‐1, suggesting that the observed effects on polarized cell migration might be mediated by inhibition of non‐canonical Wnt pathways. Taken together, these data demonstrate a previously undescribed role of Dkk‐1 in the establishment of directional polarity in epithelial cells, which contributes to the detrimental effects of Dkk‐1 in vivo.

Supported by grants from the National Institutes of Health (DK72564, DK61379 and DK79392 to C.A.P., DK53202, DK55679, DK59888 to A.N.).
Talin mediates monocyte α4β1 integrin‐mediated migration, but not chemokine‐triggered α4β1 affinity up‐regulation or adhesion
Authors: Hyduk Sharon , Rullo Jacob , Puig Cano Adrianet , Chen Mian , Cybulsky Myron I.
Published: June 2021

α4β1 integrin plays an important role in monocyte adhesion and emigration. Activation by chemokines and chemoattractants is required to increase α4β1 integrin affinity and mediate adhesion to VCAM‐1 expressed by cytokine‐activated endothelium. We investigated the role of talin, a cytoskeletal protein that interacts with integrin cytoplasmic tails, in α4β1 integrin‐dependent functions using formyl peptide receptor‐transfected U937 monocytic cells. fMLP‐induced affinity up‐regulation, measured by binding of soluble LDV‐FITC peptide, was unaffected by siRNA knockdown of talin‐1 or sequestration of talin through expression of a dominant inhibitory talin‐binding fragment of phosphatidylinositol phosphate kinase type I‐90. Similarly, talin‐1 knockdown did not inhibit fMLP or SDF‐1‐stimulated adhesion to VCAM‐1. Talin was however, essential for PMA‐induced adhesion to VCAM‐1, but not PMA‐induced α4β1 integrin affinity up‐regulation. In addition, talin knockdown reduced fMLP‐driven, α4β1 integrin‐dependent chemotactic migration across VCAM‐1 coated filters.

These results suggest that talin association with the cytoplasmic tail of α4β1 integrin is not involved in chemokine‐induced affinity changes or adhesion. However, talin plays an important role in α4β1 integrin‐mediated chemotactic migration and PMA‐induced adhesion to VCAM‐1, steps that follow affinity modulation. Funded by CIHR.
An insight into the Inhibition of HIV‐1 replication by co‐infecting HHV‐6, HHV‐7 or GVB‐C viruses: Role of Mutually Homologous miRNAs in Downregulation of Viral Replication
Authors: Bagasra Omar , Aggarwal Mayank , Addanki krishna chaitanya , Alsayari Mohammad , Noorali samina , Haynes Naiomi , Panasik Nicholas , Mahalingham Kuha
Published: June 2021

In recent years, several reports have documented a better prognosis for HIV‐1‐infected patients co‐infected with HHV‐6, 7 or GBV‐C who may live three times longer as compared to those without these co‐infections. Recent studies have also identified anti‐viral roles of host miRNAs in plants, worms, mammals and possibly humans. To evaluate whether the apparent beneficial effects of the co‐infecting viruses are due to mutually homologous miRNAs, we computationally analyzed miRNAs that have significant homologies to both HIV‐1 and the co‐infecting viruses. We discovered 50 miRNAs that showed >80% homology to HIV‐1 and among these a significant number also mutually targeted co‐infecting viral genomes. We selected four miRNAs based on a tool that detects C+·GC and T.AT triplets which imparts greater stability between miRNA and HIV‐1 provirus. We synthesized four double‐stranded 20‐21mer, fluorophore‐conjugated oligonucleotides, representing HIV‐1 proviral DNA motifs and four homologous, quencher‐conjugated oligonucleotides representing HIV‐specific TF‐miRNA. Each was confirmed for their ability to form highly stable TF as indicated by melting curve data. Next, we developed stably transfected Hela‐CD4+cell lines expressing each of the miRNAs. Two miRNAs exhibited a significant inhibition of HIV‐1 as measured by HIV‐1p24 ELISA and Real Time PCR (>90%; p>0.001). In this study we provide a possible molecular mechanism by which co‐infecting non‐pathogenic viruses impart resistance to HIV‐1, suggest a new miRNA‐based therapeutic strategy for HIV‐1, and demonstrate the anti HIV‐1 protective effects of certain miRNAs that are mutually homologous to HIV‐1 and human herpes virus 6, HHV‐7 or GBV‐C co‐infecting viruses.
Effect of melatonin as HDAC inhibitor on diurnal HDAC expression in isolated neonatal cardiac cells
Authors: Hong Yunkyung , Lee Sang‐kil , Park Sookyoung , Kim Joo‐heon , Hong Yonggeun
Published: June 2021

Many biological systems are regulated by an intricate set of feedback loops that oscillate with a circadian rhythmicity of roughly 24 hr. For example, increases in body temperature, heart rate, blood pressure, and cortisol secretion occur early in the day. Histone proteins exist in a delicate balance between acetylated and deacetylated states. Acetylation leads to nucleosomal relaxation and increased expression of numerous genes. Recent work implicates this chromatin remodeling response in hypertrophic growth of the myocardium. trichostatin A(TSA), a potent inhibitor of histone deactylase(HDAC) activity, imposes a dose‐dependent blockade to hypertrophy and fetal gene activation in neonatal cardiac cells. To explore the role of HDAC activation in an in vitro, we studied the effects of TSA, NaB and melatonin in neonatal and H9c2 cells. Chemicals such as TSA and NaB treated neonatal cardiac cells had an increase of global histone H3 and H4(P<0.05), but decrease of these proteins by melatonin(P<0.01). To determine whether these chemicals act as trigger on the expression of H3 and H4 in cells, we studied cardiac cells subjected to RT‐PCR and immunoblot analysis. At 8 hours after treatment of TSA and NaB, the amplitude of expression of H3 and H4 was significantly decreased, and that of melatonin was significantly damped. Strikingly, diurnal variation of H3 and H4 expression were altered by treatment of TSA and NaB, suggesting a role in anticipating and responding to nuclear processes. However, melatonin induced a 4‐8 hour phase shift of H4 protein expression, and rhythmic expression of H3 was amplified. Key words : melatonin, HDAC, cardiac cells, trichostatin A(TSA), NaB. Supported by KRF‐E00344, BioGreen21 Program (#20070401034006) of Rural Development Administration, South Korea.
Gender modulation of cardiac gene expression in rats with chronic volume overload
Authors: Walton Karen L. , McLarty Jennifer L. , Bennett Shante' , Melendez Giselle C. , Sabo‐Attwood Tara , Brower Gregory L.
Published: June 2021

We have previously described gender differences in cardiac remodeling secondary to chronic volume overload in rats. However, the mechanisms underlying ovarian hormone mediated cardioprotection have not yet been elucidated. Accordingly, this study sought to identify altered cardiac gene expression contributing to the protection from adverse cardiac remodeling seen in female rats. To that end, left ventricular (LV) gene expression in male and intact and ovariectomized adult female rats (N=3 per group) with an aortocaval fistula was compared to sham‐operated controls. At three days post‐surgery, the hearts were excised, separated and the LV prepared for RNA isolation. An Applied Biosystems Low Density Inflammation Array allowed us to simultaneously examine 96 genes of interest. There were significant gender differences observed in the expression of genes kallikrein‐15, cyclooxgenase‐1 and ‐2, thromboxane synthases and nerve growth factor which were subsequently confirmed by RT‐PCR. Cytokine gene expression, including the TNF‐alpha superfamily, IL‐6 and stem cell factor were also similarly affected. Thus, ovarian hormone regulation of prostaglandin, cytokine and serine protease inflammatory gene expression in the heart are implicated in the attenuation of pathogenic remodeling induced by a chronic volume overload.

Funding Sources: NIH grants RO1‐HL‐62228 and RO1‐HL‐073990
Blastocyst Injection of Wild Type Embryonic Stem Cells Induces Global Corrections in Mdx Mice
Authors: Vitale Joseph M , Stillwell Elizabeth , Khadim Farah , Elson Genie , Altaf Aneela , Schneider Joel , Yehia Ghassan , Fraidenraich Diego
Published: June 2021

Duchenne muscular dystrophy (DMD) is an incurable neuromuscular degenerative disease, caused by a mutation in the dystrophin gene. Mdx mice recapitulate DMD features. Here we show that injection of wild‐type (WT) embryonic stem cells (ESCs) into mdx blastocysts produces mice with improved pathology. A small fraction of WT ESCs incorporates into the mdx mouse nonuniformly to upregulate protein levels of dystrophin in the skeletal muscle. The chimeric muscle shows reduced regeneration and restores dystrobrevin, a dystrophin‐related protein, in areas with high and with low dystrophin content. WT ESC injection also normalizes the amount of fat, a tissue that does not express dystrophin. ESC injection without dystrophin does not prevent the appearance of phenotypes in the skeletal muscle or in the fat. Thus, dystrophin supplied by the ESCs reverses disease in mdx mice globally.