bluuebunny/crossref_metadata_2025 huggingface dataset:
Mechanisms of 1α, 25‐Dihydroxyvitamin D regulation of hypoxia‐inducible factor‐1α in breast epithelial cells
Authors: Jiang Yan , Harris Jessica , Teegarden Dorothy
Published: June 2021

The purpose of this study was to investigate 1α, 25‐dihydroxyvitamin (1,25(OH)2D) regulation of hypoxia inducible factor‐1α (HIF‐1α), an angiogenic transcriptional regulator, in untransformed MCF10A (10A) and H‐ras transfected MCF10A (10A‐ras) breast epithelial cells, a model of cancer progression. In 10A cells, 1,25(OH)2D increased HIF‐1α mRNA abundance (64% ± 19 greater than vehicle control, p=0.01) and protein level (137% ± 30, p<0.001) at 12 hr. In contrast, 1,25(OH)2D treatment did not alter HIF‐1α mRNA abundance in 10A‐ras although, similar to 10A cells, the protein level was increased at 12 hrs (108% ± 38, p=0.05). A transcriptional inhibitor prevented the 1,25(OH)2D‐mediated increase in HIF‐1α protein expression in 10A but not in 10A‐ras cells. In contrast, inhibition of proteasomal degradation prevented the 1,25(OH)2D‐induced HIF‐1α protein level in 10A‐ras but not in 10A cells. Knock‐down of VDR (siRNA) prevented 1,25(OH)2D‐mediated induction of HIF‐1α protein in 10A but not in 10A‐ras cells and VDR was associated with a putative VDR response element on HIF‐1α gene promoter in 10A cells (chromatin IP assay). These results support that 1,25(OH)2D regulates HIF‐1α protein level in 10A cells via transcriptional regulation in contrast to through proteosomal degradation in 10A‐ras cells, suggesting a differential mechanism in the two cell types.

Grant Funding Source: Supported by NIH CA128770
Redox modulation as a mechanistic feature of biological effects of cysteine and glutathione mixed disulfide conjugates of Allium thiosulfinates
Authors: ZHANG GUODONG , PARKIN KIRK L
Published: June 2021

The objective of this work was to determine if phase II enzyme induction and inhibition of nitric oxide (NO) evolution by cysteine and glutathione mixed disulfide conjugates of Allium thiosulfinates (CySSR/GSSR) were linked to redox homeostasis in cultured cells. The CySSR/GSSR species induced quinone reductase (QR) through the Nrf2 pathway in Hepa 1c1c7 cells, and inhibited iNOS expression and NO evolution in LPS‐activated macrophages. CySSR/GSSR also modulated intracellular redox status by boosting intracellular GSH levels. The effects of the CySSR and GSSR species were progressively attenuated by the addition of exogenous thiol agents that influenced redox status in a dose‐dependent manner in both intracellular and extracellular spaces. Thus, the biological activities of CySSR/GSSR appear to be dependent on both extracellular and intracellular redox status. The effects of CySSR, but not GSSR, treatments resulted in an increase in extracellular thiol levels. Evidence was obtained for a mechanism involving cellular uptake of CySSR by L transporter, disulfide bond reduction by thioredoxin reductase (TRxR) and the export of cysteine and RSH into the medium by multiple drug resistance protein. This study adds to the evidence that Allium organosulfur compounds and metabolic derivatives exert a range of biological effects through a redox‐modulatory mechansism.

Supported by USDA‐NRI‐CGP.
Identification of selenoproteins exhibiting resistance to nonsense‐mediated decay under selenium deficiency
Authors: Seyedali Ali , Gilman Christy L , Seale Lucia A , Berry Marla J
Published: June 2021

Selenoproteins, defined by the presence of the 21st amino acid selenocysteine (Sec), exhibit functions vital to cellular well‐being. Sec translation into protein occurs via the stop codon UGA and requires recoding by means of secondary structures in the 3′UTR associating with trans‐acting factors. This Sec‐specific UGA creates competition between selenoprotein biosynthesis and the nonsense‐mediated decay pathway (NMD); a pathway functioning to protect the cell from bulk translation of potentially deleterious truncated proteins. A recent study reported that eIF4a3, a protein component of the exon junction complexes deposited near exon junctions during splicing, binds the SECIS elements of select selenoprotein mRNAs under selenium deficiency and prevents Sec incorporation into protein. This translational inhibition does not necessarily increase NMD. Characterization of selenoprotein mRNAs reveal insights into NMD resistance. Several studies have identified variability in selenoprotein mRNA and protein levels under selenium deficiency. Our aim was to characterize expression of selenoproteins and their mRNAs under selenium deficiency and to elucidate underlying mechanisms. We have found that selenoproteins exhibit a wide variety of tissue specific dependence on selenium supply and that overexpression of eIF4a3 in HEK293 cells resulted in a decrease in certain selenoprotein mRNAs.
A novel compound isoangustone A dampens chronic high glucose‐induced mesangial matrix deposition and fibrosis
Authors: Li Jing , Lee Eun Sook , Lim Soon Sung , Kang Young‐Hee
Published: June 2021

Isoangustone A, a phenolic compound found in licorice, has been shown to exhibit various pharmacological effects including anti‐virus, anti‐allergic and anti‐tumor activities. Glomerulosclerosis is a prominent feature of diabetic nephropathy (DN), which leads to renal failure and end‐stage renal disease. Mesangial matrix mainly comprised of type IV collagen is expanded in DN, where connective tissue growth factor (CTGF) is excessively secreted. The present study was to test whether isoangustone A can inhibit production of collagen and CTGF promoted by high ambient glucose (HG) in human mesangial cells (HRMC). Serum starved HRMC were cultured in media containing 5.5 mM glucose plus 27.5 mM mannitol as an osmotic control or 33 mM glucose for 3 days in the presence of 1–20 μM isoangustone A. Exposure of HRMC to HG caused marked increases in collagen secretion and CTGF expression, which was dose‐dependently suppressed by isoangustone A. Furthermore, isoangustone A appeared to retard HG‐triggered matrix expansion, repealing MT1‐MMP attenuation and TIMP‐2 elevation accompanied by HG. Such effects of isoangustone A was mediated most likely through disturbing TGF‐β/Smad signaling pathways. The results demonstrate that isoangustone A may be a potential therapeutic agent for the prevention and treatment of glomerulosclerosis.
Men and not women showed cholesterol lowering after 12 weeks of supplementation with secoisolariciresinol diglucoside
Authors: Ye Zhong , Kong Kai Ling , Wu Xianai , Lee Sun Ok , Williamson‐Hughes Patricia , Hendrich Suzanne
Published: June 2021

We hypothesized that secoisolariciresinol diglucoside (SDG, Beneflax™) derived from flaxseed, would lower serum cholesterol in hypercholesterolemic humans and that the effect would be correlated with serum lignan metabolites. 90 subjects ingested 0 mg (18 male, 12 female), 150 mg (19 male, 11 female), or 300 mg SDG (, 20 male, 10 female) total daily dose for 12 w in a randomized, placebo‐controlled double‐blind trial. At 0 and 12 weeks, serum secoisolariciresinol (SECO), enterodiol (ED), and enterolactone (EL) were measured by LC/MS. Fasting serum cholesterol, glucose, and HbA1c were measured at −2, 0, 4, 8 and 12 wks. Decrease of total cholesterol (TC) and LDL‐cholesterol (LDL‐C) from baseline were significant in men ingesting either dose of SDG. Plasma glucose was not changed from baseline by any treatment. The decrease of TC and LDL‐C from baseline was significantly greater in men than in women ingesting 300 mg SDG/d. Serum SECO, EL and ED were significantly increased in subjects ingesting either dose of lignans. LDL‐C was negatively correlated with serum ED (r = − 0.50, P = 0.03); and LDL decrease from baseline was positively correlated with serum SECO (r = 0.59, P = 0.01) in subjects ingesting 150 mg SDG/d. In conclusion, SDG decreased serum TC and LDL‐C in men but not in women and this function was related to the bioavailability of lignan metabolites. Research funded by Archer Daniels Midland.

Grant Funding Source: Archer Daniels Midland
Differential Regulation of Protein Synthesis and mTOR Signaling in Skeletal Muscle and Visceral Tissues of Neonatal Pigs after a Meal
Authors: Gazzaneo Maria C , Suryawan Agus , Orellana Renán A. , Torrazza Roberto Murgas , Nguyen Hanh V. , Wilson Fiona A. , Fiorotto Marta L. , Davis Teresa
Published: June 2021

Neonates have a high rate of growth driven by elevated rates of skeletal muscle protein synthesis. Feeding stimulates protein synthesis but the time course of the changes in different tissues has not been determined. The aim of this study was to evaluate the time course of the changes in protein synthesis rates and translation initiation factor activation in skeletal muscle and visceral tissues after a meal. Neonatal pigs (n=36; 5–7 day old) received a meal providing one‐sixth of daily requirements and were sacrificed at 0, 30, 60, 90, 120, or 240 min after feeding. Protein synthesis increased 30 to 120 min in glycolytic muscles, 30 to 90 min in mixed fiber type muscle and did not increase in oxidative muscle post‐feeding. Protein synthesis increased in liver 30 to 120 min, jejunum 30 to 60 min and pancreas 30 min post‐feeding and did not increase in kidney and heart. Eukaryotic initiation factor (eIF) 4E binding protein (4EBP1) and rp S6 kinase 1 (S6K1) phosphorylation increased 30 to 90 min in all tissues. These results suggest that feeding stimulates mTOR signaling in skeletal muscle and visceral tissues. However, mTOR activation alone was not sufficient to stimulate protein synthesis in all tissues. (Supported by NIH grants AR‐44474 and AR‐51563 and USDA/ARS 6250510000‐33).

Grant Funding Source: NIH grants AR‐44474 and AR‐51563 and USDA/ARS 6250510000‐33
Folic acid source, usual intake, and serum folate concentrations in US children, the National Health and Nutrition Examination Survey (NHANES) 2003–2006
Authors: Yeung Lorraine , Cogswell Mary , Carriquiry Alicia , Bailey Lynn , Berry Robert
Published: June 2021

Our objective was to estimate among US children usual folic acid intake above the tolerable upper intake levels (UL) established for that age group and folate concentrations by three major folic acid sources: enriched cereal grain products (ECGP), ready‐to‐eat cereals (RTE), and supplements (SUP). We analyzed data from 7397 children aged 1–18 years (y) in NHANES 2003–2006. Among children aged 1–3 y, 4–8 y, 9–13 y, and 14–18 y, 21%, 18%, 34%, and 47%, respectively, reported consumption of folic acid from ECGP only. Fifty‐two percent, 45%, 44% and 36%, respectively, reported consumption from ECGP+RTE. Eight to nine percent reported consumption from ECGP+SUP. Eighteen percent, 28%, 13%, and 9%, respectively, reported consumption from ECGP+RTE+SUP. Among all age groups, 0% of the children who consumed folic acid from ECGP only exceeded the UL for their age group. The proportion above the UL for their age group was greater with consumption of RTE and SUP. Compared with consumption of ECGP only, the adjusted geometric mean folate concentrations were progressively higher among those who consumed ECGP + RTE, ECGP + SUP, and ECGP+SUP+RTE in all age groups except children aged 4–8 y. Post‐fortification, US children who consume ECGP only are unlikely to exceed the UL for their age group and the majority of children consume folic acid from either RTE or SUP. Research funded by CDC.
Plasma folate's association with depressive symptoms among US adults: the role of dietary intake and diet quality
Authors: Beydoun May A , Fanelli‐Kuczmarski Marie T , Beydoun Hind A , Shroff Monal R , Mason Marc , Evans Michele K , Zonderman Alan B
Published: June 2021

Folate deficiency has been implicated in the etiology of unipolar depression. Using a population‐based study of 1,681 subjects aged 30–64 years (Healthy Aging in Neighborhoods of Diversity across the Lifespan Study) the association of folate with depressive symptoms, measured with the Center for Epidemiologic Studies Depression scale (CES‐D), was examined. Folate was measured by plasma levels and dietary intakes based on two 24‐hr recalls. The 2005‐Healthy Eating Index (HEI) was computed. Multivariate logistic regression and structural equation modeling (SM) were conducted. Compared to the lowest tertile, the middle and uppermost tertiles of plasma folate were associated with a 39–40% reduced odds of elevated CES‐D (≥16) among women [adjusted OR (T3 vs. T1)=0.60(95% CI=0.42–0.86), P=0.006]. Confounding of this association by HEItotal was noted, though dietary folate did not confound this association appreciably. In SM, plasma folate completely mediated the inverse HEItotal‐CES‐D association among men only, particularly in six HEI components. Among women, HEItotal and four components had an inverse direct effect on CES‐D score suggesting an independent mechanism from plasma folate. Depressive symptoms may be alleviated by improving dietary quality with plasma folate playing a potential mediating role only among men.

Plasma folate's association with depressive symptoms among US adults: the role of dietary intake and diet quality
Effect of prenatal supplementation with docosahexanoic acid on infant development: a randomized placebo‐controlled trial in Mexico.
Authors: Ramakrishnan Usha , DiGirolamo Ann , Schnaas Lourdes , Stein Aryeh D , Wang Meng , Martorell Reynaldo , Neufeld Lynnette , Rivera Juan
Published: June 2021

The omega‐3 fatty acid docosahexanoic acid (DHA) is deposited in the brain early in life and plays an important role in neural and visual function. Little is known however about the benefits of ingesting DHA supplements during pregnancy for infant development. We conducted a randomized double‐blind placebo‐controlled trial of 400 mg/day algal DHA (Martek) from week 20 of gestation through delivery (n=1094 pregnant women) and followed the offspring (n=973 live births). Child development was measured in 730 children (75% of birth cohort) using the Spanish version of the Bayley Scales of Infant Development (BSID)‐II at 18 mo of age; we calculated standardized psychomotor development (PDI) and mental development indices (MDI). PDI and MDI scores (mean ± S.D.) were 94.3 ± 10.7 and 93.0 ± 8.9 in the DHA group (n=365) and 95.2 ± 9.3 and 93.3 ± 9.8 in the placebo group (n=365), respectively. Intent to treat differences in PDI and MDI scores at 18 mo were 0.90 (95% CI: −0.55, 2.35; p=0.23) and 0.26 (95% CI: −1.10, 1.63; p=0.70), respectively. Prenatal supplementation with 400 mg algal DHA did not result in improved global development scores at 18 mo of age in a study population (Mexico) with low intakes of DHA. Follow‐up of the cohort is ongoing. Funding: NIH (HD043099) and March of Dimes Foundation.
A surprising level of unmetabolized folic acid in serum from the Tsimane Amerindians of Bolivia
Authors: Bailey Steven W. , Gurven Michael , Kaplan Hillard , Alverson Patricia B , Ayling June E.
Published: June 2021

We have previously analyzed plasma from fasting subjects in the U.S., who were neither consumers of supplements nor breakfast cereals, and found persistent levels of unmetabolized folic acid (FA). We have also previously detected low levels of unmetabolized FA in samples from Europe, despite the lack of national FA fortification in these countries. We hypothesized that the presence of FA, even in European samples, could be due to any of the following:

Subjects in countries without FA fortification were, none the less, consuming some FA in their purchased foods, or

FA is generated by oxidation of natural food tetrahydrofolate during cooking or storage, or

FA is generated by oxidation of endogenous tetrahydrofolate within the body.

We have now determined the folate status of the Tsimane, a remote population of forager‐horticulturalists in the lowland Bolivian Amazon. The total serum folate averaged 13 nM, consistent with a relatively modest folate intake. Despite this and the lack of exposure to manufactured FA, unmetabolized FA was detected in all samples. Although lower on average than the above group in the U.S., this was significantly higher than those from Europe. Thus, it appears that humans may have been subject to persistent low FA throughout most of their recent evolution. Efforts are now underway to resolve whether this FA is due to cooking and/or oxidative stress.
DMT1 is not essential for Cu uptake from blood by internal organs
Authors: Linder Maria C. , Kidane Theodros Z. , Sauble Eric N.
Published: June 2021

Divalent metal transporter 1 (DMT1) is the main mechanism by which Fe(II) is imported into enterocytes from the intestinal lumen. This uptake is inhibited by Cu(I) and some divalent metal ions; knockdown of expression is accompanied by a reduction in Cu as well as Fe uptake. In contrast, cells of internal organs mainly obtain their Fe from blood plasma transferrin by receptor mediated endocytosis, and DMT1 is mainly localized in endosomes, implying little would be at the cell surface to mediate Cu uptake. To verify this, we compared initial (1h) rates of uptake of tracer 64Cu(II) by organs of virgin and lactating Belgrade rats with those in the background (Fisher) strain. Lactation greatly increased 64Cu uptake by the mammary gland and reduced uptake by liver; but no significant differences ascribable to lack of DMT1 were detected in any of the organs tested. HepG2 cells were used to further explore potential involvement of DMT1 by measuring initial (30 min) rates of 64Cu uptake from purified plasma proteins. No inhibition by Fe(II) was observed. Conversely, uptake of 59Fe(II) (± ascorbate) was not inhibited by excess Cu(I) or Cu(II). Pretreatment of cells with Fe for 24h enhanced rates of 59Fe(II) uptake but did not alter those for Cu. Fe pretreatment did not alter DMT1 mRNA or protein expression. We conclude that DMT1 is unlikely to be important for uptake of Cu by cells of internal organs. Supported in part by USPHS Grant HD 46949.
Ganoderma lucidum suppresses colon carcinogenesis and inflammation
Authors: Sliva Daniel , Jedinak Andrej , Jiang Jiahua , Adamec Jiri , Sandusky George , Dudhgaonkar Shailesh
Published: June 2021

Medicinal mushroom Ganoderma lucidum, well recognized natural product that have been used in the traditional Chinese medicine, is currently used as a popular dietary supplement. Here, we show that an oral application of G. lucidum triterpene extract (GLT) suppressed colon carcinogenesis induced by the food‐borne carcinogen (2‐Amino‐1‐methyl–6‐phenylimidazol[4,5‐b]pyridine [PhIP]) and inflammation (dextran sodium sulfate [DSS]) in mice. PhIP and DSS induced formation of colonic tumors, whereas GLT treatment significantly decreased the number of tumors. Furthermore, GLT suppressed focal hyperplasia and aberrant crypt foci (ACF) formation. These anti‐proliferative effects of GLT were confirmed by the decreased staining with Ki‐67 in colon tissues. PhIP/DSS‐induced colon inflammation was demonstrated by the significant shortening of the large intestine, whereas GLT treatment reversed the shortening of colon lengths to the levels comparable to the control group. Moreover, PhIP/DSS‐induced infiltration of macrophages was significantly reduced in colon tissue in animals treated with GLT. Our data suggest that GLT could be considered as an alternative dietary approach for the suppression of the food‐borne carcinogen‐ and inflammation‐induced colon carcinogenesis.

Supported by P50 AT00477 from NCCAM and the NIH Office of Dietary Supplements.
Role of Grape‐Derived Polyphenols in the Prevention of Alzheimer's disease and the Promotion of Healthy Aging
Authors: Pasinetti Giulio Maria , Chen Tzu‐Ying , Janle Elsa M. , Cheng Alice
Published: June 2021

A high consumer demand exists for foods and dietary supplements with the ability to help preserve cognitive functions by preventing processes involved in age‐related cognitive decline. Bioactive dietary botanicals in food and supplements are complex mixtures of numerous chemical entities, and the identity of these compounds is often not well established. Ongoing studies on our laboratory are underway to characterize compounds derived from grape seed polyphenolic extract (GSPE) to investigate their potential role in healthy aging. Recent evidence indicates that age‐related cognitive decline may be associated with impaired energy metabolism and increased oxidative stress damage in the brain. Based on this evidence, ongoing preclinical studies in our laboratory were designed to identify and characterize the bioavailable and bioactive GSPE which may help to preserve cognitive functions through mechanisms involving improvement of mitochondrial function and maintenance of energy metabolism. Collectively, these ongoing studies will help to clarify the role of GSPE in the preservation of cognitive functions and healthy aging. Supported by PO1AT004511 to GMP.
Activation of the Wnt/β‐catenin pathway leads to enhanced proliferation and liver regeneration in mice
Authors: Nejak‐Bowen Kari Nichole , Thompson Michael , Monga Satdarshan
Published: June 2021

An important role for β‐catenin in liver regeneration is observed in mice, rats and more recently in zebrafish and humans. In order to elucidate any stimulatory impact of β‐catenin on liver regeneration, we utilized transgenic (TG) mice that overexpress β‐catenin under an albumin promoter/enhancer in FVB strain. Using the partial hepatectomy (PHx) model of liver regeneration, we found a dramatic increase in cell proliferation in TG mice as compared to WT at 40H after PHx, before the onset of proliferation in WT FVB mice that occurs at 72H. Coincident with the earlier onset of proliferation, β‐catenin translocates to the nucleus at 40H post‐PHx in TGs, whereas this translocation does not occur until 72H in WT. Analysis of cyclin D1 expression by Western blotting showed that levels of this cell‐cycle regulator are several‐fold higher in TG than in WT at 40H. We tested if hydrodynamic delivery of Wnt‐1 naked DNA could also impact regeneration in WT mice. We observed a significant increase in total Wnt‐1, β‐catenin, glutamine synthetase, cyclin‐D1 and PCNA in the Wnt‐1 and not the control plasmid‐injected mice, similar to that seen in the β‐catenin overexpressing TG mice. In conclusion, stimulation of Wnt/β‐catenin signaling induces liver regeneration through induction of such downstream targets as cyclin‐D1, and thus might have implications in regenerative medicine.
Thymosin‐β4 (Tβ4) Inhibits Transdifferentiation of Human Hepatic Stellate Cells by an Akt‐dependent, PI3K‐independent Mechanism
Authors: Reyes‐Gordillo Karina , Shah Ruchi , Fu Sidney , Hindle Anna , Brody Frederick , Rojkind Marcos
Published: June 2021

Hepatocyte growth factor (HGF) is used in the treatment of liver fibrosis. It is known to up‐regulate Thymosin‐β4 (Tβ4), a chemokine expressed by hepatic stellate cells (HSC), the main collagen producing cells of the liver. Thus, we investigated whether Tβ4 could prevent the expression of platelet‐derived growth factor receptor (PDGF‐βR), a hallmark of HSC activation, in cells treated with PDGF‐BB.

METHODS

Human HSCs (passage 4–6) were either treated with PDGF‐BB (50 ng/ml), Tβ4 (10 ng/ml), or both. Cell proliferation was determined by the MTT assay and cell migration analyzed by wounding confluent cultures of HSCs. Expression of PDGF‐βR and several protein kinases was determined by Western analysis. PDGF‐βR, α1(I) collagen and α‐SMA mRNAs were determined by qPCR.

RESULTS

Tβ4 downregulated the expression of PDGF‐βR and abolished PDGF‐BB‐induced HSC proliferation and migration, as well as expression of PDGFβR, α1(I) collagen and α‐SMA mRNAs. Tβ4 blunted PDGF‐BB‐dependent phosphorylation of AKT (Thr 308) and PRAS40 (Thr 246) but had no effect on PTEN, 85PI3K, and PDK1 phosphorylation.

CONCLUSION

Tβ4 prevents expression of key parameters of HSC activation by an AKT‐dependent, PI3K‐independent mechanism. Because Tβ4 is the major actin sequestering protein in mammalian cells we speculate that Tβ4 may prevent binding of AKT to actin and thus its phosphorylation by PDK1. Supported by NIAAA.
Hypoxia Stimulates Hepatocyte Epithelial to Mesenchymal Transition
Authors: Copple Bryan L , Sullivan Bradley , Bai Shan
Published: June 2021

Studies have shown that hepatocytes differentiate into myofibroblasts during fibrosis by a process called epithelial to mesenchymal transition (EMT). In kidney fibrosis, hypoxia, through activation of the transcription factor hypoxia‐inducible factors‐1α (HIF‐1α) is an important stimulus of EMT. Accordingly, we tested the hypothesis that hypoxia stimulates hepatocyte EMT by a HIF‐dependent mechanism. Exposure of primary mouse hepatocytes to hypoxia increased fibroblast‐specific protein‐1 (FSP‐1) and SNAIL, and decreased E‐cadherin and zona occludins‐1 consistent with EMT. Upregulation of FSP‐1 and SNAIL was prevented in hepatocytes isolated from HIF‐1β‐deficient mice. In addition, upregulation of FSP‐1 and SNAIL was partially prevented by pretreatment with the activin inhibitor, follistatin, and by pretreatment with a transforming growth factor‐β1‐neutralizing antibody. This suggested that activins and TGF‐β1 were required for hypoxia‐induced EMT. Consistent with these in vitro results, mRNA and protein levels of FSP‐1 were increased to a greater extent in the livers of wild‐type mice after bile duct ligation when compared to HIF‐1α‐deficient mice. Results from these studies demonstrate that hypoxia stimulates hepatocyte EMT in vitro and in vivo by a HIF‐dependent mechanism. Supported by NIH grant DK073566.
So Close Yet So Far: Who Were the Neanderthals?
Authors: Tattersall Ian
Published: June 2021

The relatively recently extinct Neanderthals have long been known to differ from living Homo sapiens in a host of osteological characteristics. Yet many paleoanthropologists continue to view them as a bizarre variant of Homo sapiens, if only because they possessed brains that are of modern human size. Still, many congeneric primates of similar brain size belong to different species. And a recent reconstruction of an entire Neanderthal skeleton shows a significantly different postcranial aspect from Homo sapiens. Most strikingly expressed in the rib cage and pelvis, these differences would certainly have expressed themselves in gait, and these two hominids would have presented a very different affect as living entities on the landscape. Multiple lines of evidence, including molecular data, thus now converge on the conclusion that the lineages leading to Homo neanderthalensis and Homo sapiens diverged well over half a million years ago, and that the former was entirely displaced following the invasion of Europe by Homo sapiens some 40 thousand years ago, with no significant biological intermixing between the two. Clearly, large brains are not synonymous with the unique symbolic way in which modern Homo sapiens process information about the world around them.
Mitochondrial Basis of Collateral Growth
Authors: Chilian William M
Published: June 2021

Patients with obesity, insulin resistance and type II diabetes have more major complications following cardiac events than those without these risk factors. There is strong evidence that ROS from mitochondria activate pathological pathways that induce complications in insulin resistance. The prediabetic metabolic syndrome (MS) with insulin resistance leads to many pathological phenotypes including impaired arteriogenesis. Mitochondrial (mt) oxidative stress (mtOxStr) is postulated to contribute to the development of such phenotypes, but whether reduction of mtOxStr would restore coronary collateral growth (CCG) has not been determined. We tested the hypothesis that in the MS, mtOxStr plays a key role in inhibition of coronary arteriogenesis. Lean and obese Zucker obese fatty rats (model of MS) were subjected to a protocol of repetitive ischemia (RI) for 10 days to stimulate CCG, which was determined from the ratio of flows to collateral‐dependent and normal zones (CZ/NZ). Prior to RI, CZ/NZ was 0.2–0.3. After RI, CZ/NZ was 0.8 in LN, which was markedly higher than that in the OB (0.2) indicating impaired CA in the OB rats. To reduce mtOxStr, OB animals were given mito‐Quinone (mt‐directed free radical scavenger) in drinking water; this treatment restored CA (CZ/NZ = 0.8) comparable to that in LN. We conclude that mitochondrial oxidative stress is pivotal in the abrogation of CA in the MS.
Anatomy ‐ Assuring a firm foundation in a fluid veterinary curriculum
Authors: Mizer Linda Allen
Published: June 2021

Veterinary anatomy is the foundation of veterinary medicine and surgery, yet it is a foundation that must be laid on a very uncertain terrain, the veterinary curriculum. A graduating veterinarian must be capable of constructing a mental image of the animal body whether for physical examination, imaging, surgery or pathology. In no other profession is the individual expected to be competent in so many potential specialties, all of which require a firm foundation in anatomy from gross to microscopic to developmental.

Gross anatomy is usually taught as one companion animal course and a second equine or production animal course. Histology and embryology are usually offered as separate courses. Horizontal integration of multiple disciplines within a single course is not generally achieved.

Vertical integration over the four years may include required core components such as applied anatomy or surgical anatomy and elective components. Reinforcement in the clinical years of the curriculum is highly variable. The total hours of anatomy in a curriculum footprint vary greatly among veterinary colleges, as does the mode of delivery and the range of resources is used.

My goal is to present some strategies that have been used in selected veterinary curricula and to open discussion of strategies for the vertical integration of veterinary anatomy training.
Diagnostic imaging of Mummified Soft tissue: Pathologies, Pitfalls and Perspectives
Authors: Ruhli Frank J
Published: June 2021

Ancient mummified remains are a unique source to learn not only about the past but also for the present. Such tissue‐based historic information can contribute substantially to the understanding of ongoing interactions e.g. between mankind and environment as echoed e.g. by alterations in prevalence and morbidity of disease. Since the very initial attempts of diagnostic imaging of such unique samples in the late 1890's, technological evolutions were tremendous and nowadays particularly radiological techniques allow more than ever non‐ and minimal invasive diagnostic analyses of ancient soft and hard tissue. Based on our own experience by using state‐of‐the‐art imaging modalities such as MSCT, Micro‐CT, or MRI, major technological pitfalls such as restrictions in sample size or low specifity of spatial tissue discrimination in ancient mummies will be addressed. Future imaging modalities such as DECT or THz will been addressed briefly too. Examples of disease we found in the soft‐tissue compartments of ancient mummified remains include e.g. severe atherosclerosis in ancient Egyptian mummies or the laceration of a major vessel with subsequent lethal bleeding in the Neolithic South Tyrolean Iceman (“Ötzi”).
Imaging the dynamics of embryonic development
Authors: Fraser Scott E.
Published: June 2021

The explosion of progress in cell biology, biochemistry, and molecular biology has offered unprecedented knowledge of the components involved complex events such as embryonic development. This poses the challenge of integrating this knowledge of the building blocks into an understanding of the functioning system. Advanced imaging techniques offer important stepping‐stones between disparate approaches, permitting us to spy on the assembly process and to pose questions about cellular and molecular events in the most relevant setting of the intact system. Intravital imaging presents major tradeoffs between the rapidity of data collection and the limited photon budget. We are attempting to advance this tradeoff by constructing more efficient microscopes and creating labeling reagents with greater photostability and brightness such as second harmonic generating nanoparticles. These nanoparticles give sufficient signal for clear imaging down to the single molecule level, even in complex optical environments, and avoid the limitations of quantum dots (blinking and bleaching). Such new labels, combined with new imaging tools, are giving unique insights into the processes of embryonic development.
Discreteness, heterogeneity, and stochasticity: confronting realities in modeling embryogenesis
Authors: Newman Timothy
Published: June 2021

The past few years have seen a dramatic increase in modeling efforts in the field of embryo development, with interdisciplinary teams arising connecting developmental biologists with engineers, physicists, computer scientists, and mathematicians. Each field brings valuable modeling expertise, e.g. finite element modeling from engineering and non‐linear differential equations from mathematics. In this talk I will bring a statistical physics perspective and try to connect modeling with realities of the developing embryo ‐ namely, that order and form arise robustly despite an environment that is stochastic and heterogeneous. I will discuss some modeling efforts in my own group, using a cutting‐edge discrete computational algorithm ‐ the Subcellular Element Model. I will focus on an important early stage of amniote development, namely, primitive streak formation, and describe our attempts to use the SEM to distinguish between conflicting experimental findings.
Astrocytes in the Amygdala
Authors: Johnson Ryan Taylor , Breedlove Marc , Jordan Cynthia L
Published: June 2021

The amygdala is connected to numerous brain regions and is involved in complex behaviors such as survival and reproduction. This highly conserved brain region is sexually dimorphic in several species with the most dramatic differences found in the medial subregion. Neuroendocrine research has repeatedly demonstrated the plasticity of the medial amygdala across the lifespan. Astrocytes are a type of glial cell involved in synapse formation and functioning, and in several brain nuclei, astrocytes change morphology in response to gonadal hormones, leading to alterations in surrounding neurons. We have begun investigating astrocytes in the medial postero‐dorsal amygdala and our results reveal interesting sex and hemisphere differences in this dynamic brain region. Furthermore, our use of testicular feminized mutant rats demonstrates that functional androgen receptors may be necessary for formation of these differences.

Grant Funding Source: NIH
How enzymes work
Authors: Herschlag Daniel
Published: June 2021

Decades of research in enzymology has remarkably advanced our understanding of enzyme function. Indeed, it is now possible to draw convincing reaction mechanisms for the vast majority of enzymes. As we move to confront challenges in understanding enzyme evolution and the design of enzymes that catalyze novel reactions, a still‐deeper understanding of the structural and energetic underpinnings of catalysis will be required. We have chosen particularly tractable enzymes to ask specific questions about the nature of complementary interactions with transition states and the electrostatic environment within the active site, and we have approached these questions with a battery of structural, biophysical, chemical and biochemical approaches.
Transcriptional Control of Brown Adipogenesis and Energy Homeostasis
Authors: Spiegelman Bruce M.
Published: June 2021

Obesity is a disorder of energy balance, and can occur when food intake exceeds energy expenditure. We have been particularly interested in the transcriptional pathways controlling energy expenditure through regulation of mitochondrial function, using brown adipose cells as a rich source from which to “mine” molecules with broad roles in these processes. Most recently, we have found that the large, nuclear zinc‐finger protein PRDM16 is a “master” regulator of the determination of brown fat cell fate. This molecule can alter the differentiation pathway of fat cell precursors and hence, might represent a new pathway for the treatment of obesity. PRDM16 can alter the gene expression pattern of white fat precursor cells, so that they differentiate into brown fat cells. Expression of sh‐RNA against PRDM16 in immortalized brown fat cells shows that PRDM16 is required for essentially the entire brown fat program. Interestingly, when the same sh‐RNA against PRDM16 is expressed in primary brown preadipose cells, these cells differentiate into muscle cells, strongly suggesting a relationship between brown fat and skeletal muscle. Conversely, expression of PRDM16 alone is sufficient to convert primary brown fat precursor cells into skeletal muscle cells, replete with expression of myofibrillar proteins. Fate mapping experiments indicate that skeletal muscle and brown adipose arise from myf‐5‐expressing cells, while white fat does not come from this lineage. Finally, purification of the PRDM16 complex in brown fat cells indicates that PRDM16 binds to PPAR‐γ and C/EBPβ. The coactivation of these proteins indicates how this protein can be adipogenic in a muscle cell. These data, taken together, show that PRDM16 is a “master” regulator of the brown fat lineage and shows that brown fat and skeletal muscle arise from a similar or identical cell lineage. Thus brown and white fat represent a cellular and biochemical convergence through expression and function of PPAR‐γ but they arise from very different developmental pathways. The potential uses of the PRDM16 pathway in novel approaches to the therapy of human obesity will be discussed.
Binge drinking exacerbates asthmatic‐like pulmonary inflammation in cockroach‐allergen (CRA) induced allergic mice
Authors: Bouchard Jacqueline , Kim Jiyoun , Vaickus Louis , Remick Daniel
Published: June 2021

This study investigates the effects of binge drinking on pulmonary inflammation in CRA‐induced murine asthma. Outbred mice were immunized intratracheally with CRA and challenged 2 times (Day 14 and 21). On Day 21 the Ethanol and Water groups were given 300μl of 32% ethanol or water by oral gavage 30 minutes prior to final challenge (FC). Blood ethanol levels peaked 1 hour post‐gavage (0.13%) and were undetectable by 5 hours. At 1.5 hours post‐FC, acute alcohol consumption led to an increase in bronchoalveolar lavage absolute cells, eosinophils, neutrophils, eotaxin‐1, RANTES, KC, and MIP‐2, and an increase in plasma IgE levels. At 5.5 hours post‐FC AHR exacerbation was measured by increases in enhanced pause (PenH) and time of inspiration (Ti), and decreases in minute ventilation (MV) and peak inspiratory flow (PIF). N = 3–4 mice per group; *pg/ml, **ng/ml; values ± SEM. Supported by 5R01ES013538‐04.

Water
Ethanol
P‐value

Absolute cells x 103
140 ± 21
329 ± 17
<0.001

Eosinophils x 103
42 ± 6
160 ± 21
0.001

Neutrophils x 103
1.4 ± 0.5
12.3 ± 5.0
0.037

Eotaxin‐1*
12.4 ± 4.5
34.3 ± 4.9
0.009

RANTES*
2.3 ± 1.1
6.3 ± 1.2
0.025

KC*
1810 ± 210
3884 ± 757
0.019

MIP‐2*
353 ± 58
1022 ± 172
0.005

IgE**
20 ± 4
53 ± 12
0.022

PenH (% above base)
38 ± 25
463 ± 246
0.048

Ti (milliseconds)
72 ± 6
93 ± 8
0.042

PIF (ml air/second)
5.1 ± 0.1
3.9 ± 0.4
0.014

MV (ml air/minute)
65 ± 5
46 ± 4
0.018
Swap 70 promotes allergic airway inflammation
Authors: Bahaie Nooshin S , Hosseinkhani Reza M , Kang Bit Na , Ge Xiao Na , Ha Sung Gil , Jessberger Rolf , Rao Savita P , Sriramarao P
Published: June 2021

Swap 70 is a signaling molecule with a known role in B cell and mast cell activation and function. In this study, the involvement of Swap 70 in airway inflammation was investigated in a murine model of ovalbumin (OVA)‐induced acute (28 days) allergic inflammation. Bronchoalveolar lavage fluid (BALF) and lung tissue collected from wild type (WT) and Swap 70 knock‐out (KO) mice after the last challenge were evaluated. Expression of Swap 70 was induced in the lungs of WT allergen‐challenged mice. Compared to WT mice, OVA‐challenged Swap 70 KO mice demonstrated a significant reduction in the recruitment of inflammatory cells, especially eosinophils, in the BALF by differential cell counts and in lung tissue by immunohistology for eosinophil‐expressed major basic protein. Reduced eosinophil recruitment in Swap 70 KO mice was associated with diminished Th2 cytokine (IL‐4, IL‐5, IL‐13) and inflammatory chemokine (RANTES, MCP‐1 and eotaxin) levels in the BALF compared to allergen‐challenged WT mice. Mucus secretion and expression of found in inflammatory zone 1 (FIZZ1), a pro‐angiogenic protein expressed during hypoxia and allergic inflammation, was also substantially reduced in lung tissue of Swap 70 KO mice compared to WT mice. Overall, these findings suggest that Swap 70 signaling is important in the development of allergic airway inflammation. 1Supported by National Institutes of Health grants HL0793041 and AI35796.
Adenosine A2B Receptor Expression by Myeloid Cells Mediates Airway Inflammation in Asthma
Authors: Belikoff Bryan , Viackus Louis , Sitkovsky Michail , Remick Daniel G
Published: June 2021

Allergic‐type airway inflammation characteristic of human asthma is modeled in mice by direct inhalation of German cockroach allergens (CRA). The role of adenosine A2B receptors (A2B) in asthma was studied in mice lacking A2B (A2B KO) and myeloid specific A2B KO (Lys‐M). CRA was given intratracheally on day 0 with two pulmonary challenges on day 14 and 21. 24 hours after final challenge lungs and brochoalveolar lavage fluid (BAL) were collected.

Virtually all measures of pulmonary inflammation were reduced in the A2B KO and Lys‐M compared to wild type mice. Lung homogenate (LH) eosinophil peroxidase (WT, 1.0 ± 0.2; A2B KO, 0.2 ± 0.04*; Lys‐M, 0.3 ± 0.04*; *p < 0.01), but not myeloperoxidase (WT, 0.4 ± 0.1; A2B KO, 0.4 ± 0.1; Lys‐M 0.6 ± 0.1) levels were decreased. Our results demonstrate A2B expression by myeloid cells is an important contributor to the pulmonary inflammation in asthma.

*LH and ^BAL cytokines ng/mL. *^p < 0.05 compared to WT

Parameter
WT
KO
Lys‐M

Eotaxin 1
2.6 ± 0.2
0.9 ± 0.1*
1.0 ± 0.2*

Eotaxin 2
14.7 ± 1.6
2.3 ± 0.3*^
4.8 ± 0.1*^

IFN‐γ
8.6 ± 0.9
4.1 ± 0.2*
5.6 ± 0.6*

IL‐4
3.3 ± 0.3
1.8 ± 0.1*
1.8 ± 0.2*

IL‐5
3.2 ± 0.3
1.3 ± 0.1*
1.8 ± 0.3*

IL‐13
7.8 ± 0.9
4.1 ± 0.3*
4.3 ± 0.6*

TNF‐α
5.0 ± 0.6
1.9 ± 0.4*
2.7 ± 0.2*

BAL cells *p < 0.05 compared to WT

Parameter (x 104)
WT
KO
Lys‐M

Eosinophils
26.2 ± 5.8
1.7 ± 0.3*
1.5 ± 0.6*

Macrophages
14.9 ± 1.8
9.5 ± 0.4*
9.0 ± 0.9*

Lymphocytes
7.9 ± 1.3
2.0 ± 0.3*
3.4 ± 0.5*

Neutrophils
55.4 ± 5.4
43.8 ± 2.4
35.4 ± 3.8*

This work was supported by NIH Grant R01 ES0135283‐03
Pivotal role of Protease‐activated receptor‐2 in bleomycin‐induced pulmonary fibrosis
Authors: Borensztajn Keren Sarah , Bresser Paul , von der Thusen Jan , van der Loos Chris , van der Blink Bernt , den Bakker Michael , Daalhuisen Joost , Groot Angelique , Peppelenbosch Maikel , Spek Arnold
Published: June 2021

Idiopathic pulmonary fibrosis (IPF) constitutes the most devastating form of fibrotic lung disorders, and remains refractory to current therapies. The coagulation cascade is frequently activated during pulmonary fibrosis, but this observation has so far resisted a mechanistic explanation. Recent data suggest that protease‐activated receptor (PAR)‐2, a receptor activated by coagulation factor(F) Xa, plays a key role in fibrotic disease. Here, we assessed the role of PAR‐2 in pulmonary fibrosis. We show that PAR‐2 is upregulated in the lungs of IPF patients. The bronchoalveolar lavage fluid of these patients displays increased procoagulant‐dependant mitogenic activity. In a murine model of pulmonary fibrosis, we show that bleomycin induces PAR‐2 expression, myofibroblast differentiation and collagen synthesis. In PAR‐2−/− mice, the extent and severity of fibrotic lesions is reduced whereas myofibroblast differentiation is abolished and collagen expression is decreased. Moreover, fibrin deposition in lungs of fibrotic PAR‐2−/− mice is reduced compared to wild type mice due to differential tissue factor expression in response to bleomycin. Finally, anticoagulant targeting FXa significantly reduced fibrosis development. Taken together, PAR‐2 is instrumental in pulmonary fibrosis, and inhibition of the PAR‐2‐coagulation axis may provide a novel therapeutic approach for this devastating disease.
Potential of Denufosol as an Early Intervention Therapy for CF Lung Disease
Authors: Navratil Tomas , Schaberg Amy , Tian Wei , Durham Todd , Evans Carole , Lindroos Christina , Barrus Susan , Mathews Dave , Barnes Matthew , Ratjen Felix A. , Moss Richard B. , Accurso Frank J.
Published: June 2021

CF lung disease arises from ion transport defect. Disease origins in small airways before 1 yr of age necessitate early intervention. Denufosol (DEN), a novel chloride channel activator, stimulates Cl− transport, inhibits Na+ absorption and increases ciliary beat frequency. A Phase 3, double‐blind, placebo (PL)‐controlled trial (TIGER‐1) was conducted in 352 CF pts to compare inhaled DEN 60 mg TID to PL for 24 wks. Pts were ≥ 5 yrs (mean 14.6 yrs) and had FEV1 ≥ 75% predicted (mean 92%). Pulmonary function tests and exacerbations, QOL measures and systemic exposure of DEN were evaluated. This work examines endpoints relevant to DEN use as an early intervention agent. The ability to reach small airways was confirmed by significant improvement in FEF25%–75% in pts ≤ 110% predicted FEV1 (p=0.025). There was little to no systemic exposure to denufosol, no accumulation with chronic dosing, and no evidence of systemic AEs including liver enzyme levels, blood biochemistry and differential cell counts. DEN's mechanism of action addressing the basic defect, the lack of systemic exposure and ability to target small airways makes this a potentially promising therapy for early intervention in CF lung disease.
Nucleosome thermodynamics, histone modifications, and histone chaperone function
Authors: Luger Karolin , Stargell Laurie A , Andrews Andrew J , Chen Xu , Zevin Alex
Published: June 2021

Genome accessibility is regulated to a large part by the posttranslational modification of histones (PTM). Surprisingly, and despite the fact that nucleosomes were first characterized over 35 years ago, very little is known about the free energy required to assemble and maintain nucleosomes under physiological conditions, and even less is known about the effect of PTMs on nucleosome structure and stability. We have determined the structure of nucleosome bearing several PTMs, and investigated their effect on higher order structure. We also present a thermodynamic framework to describe nucleosome stability and how it is affected by histone modifications. A picture emerges in which various histone modifications have very differentiated effects on nucleosome and chromatin structure and stability, thereby fine‐tuning chromatin structure. Histone chaperones are acidic proteins that interact directly with histones and are implicated in nucleosome (dis)assembly through unknown mechanisms. We show that nucleosome assembly requires elimination of non‐nucleosomal histone‐DNA interactions through the action of the histone chaperone Nap1. Predictions from this unexpected model concur perfectly with our in vivo findings, where the absence of Nap1 results in an excess of non‐nucleosomal histone‐DNA complexes in cells. The accumulation of these atypical complexes has a dramatic effect on transcription activation and repression in vivo.