bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Prenatal DHA Supplementation on Child Development at age 5 years in Mexico
Authors: Ramakrishnan Usha , Pallo Beth C , Schnaas Lourdes , DiGirolamo Ann , Feregrino Raquel Garcia , Quezada David Amado , Hao Wei , Neufeld Lynnette , Stein Aryeh D , Rivera Juan A , Martorell Reynaldo
Published: June 2021

Docosahexanoic acid (DHA) is an important constituent of the brain that accretes during the first 1000 days of life. Evidence of long‐term benefits of increasing DHA intakes during pregnancy is sparse. We have completed the age 5 y follow‐up of offspring of women who participated in a double‐blind randomized controlled trial of prenatal DHA supplementation in Cuernavaca, Mexico. Pregnant women (n=1094) were randomized to receive a daily supplement of 400 mg of DHA or placebo from 18–22 weeks of pregnancy until delivery. We assessed child development at age 5 y for 798 children (82% of live births) using the Spanish language version of the McCarthy Test for Global Development. Loss to follow‐up did not differ by treatment group and the groups remained balanced for maternal baseline and infant birth characteristics. Intent to treat analysis showed no significant differences by treatment group (p>;0.05) for measures of global development. We however find evidence of selective effects by quality of home environment (measured at age 12 mo; p<0.05 for interaction). Among children from poor home environments, those exposed to DHA in utero had better verbal, memory and overall scores compared to those in the placebo group. No difference was observed for children from better home environments. Children lacking an enriched learning environment may be able to “catch‐up” to their peers by DHA supplementation in pregnancy. (Support: NIH – HD043099).
Anti‐atherogenic effect of blue‐green algae in male apolipoprotein E knockout mice fed an atherogenic diet
Authors: Ku Chai Siah , Kim Bohkyung , Pham Tho , Wegner Casey J , Yang Yue , Park Youngki , Lee Jiyoung
Published: June 2021

Development of atherosclerotic plaque in the arteries involves hyperlipidemia and inflammation. We previously reported hypolipidemic and anti‐inflammatory properties of blue‐green algae (BGA) or their organic extracts in vivo and in vitro, respectively. Therefore, BGA may be protective against atherogenesis. To evaluate an anti‐atherogenic effect of Nostoc commune var. sphaeroides Kützing (NO) and Spirulina platensis (SP), 45 male apolipoprotein E knockout mice were fed a high fat/high cholesterol control diet (HF/HC, 15% fat and 0.2% cholesterol by wt) or a HF/HC diet supplemented with 5% of dry, powdered NO or SP by wt for 12 wk. NO supplementation significantly decreased plasma total cholesterol concentrations at 6 and 12 wk than other groups. Plasma triglyceride levels were lower in NO‐fed mice than SP mice. BGA supplementation did not alter liver weight and the expression of hepatic genes for cholesterol synthesis, LDL uptake and lipogenesis compared with control. Plasma tumor necrosis factor α levels were not significantly different between groups. However, when splenocytes isolated from NO and SP‐fed mice were stimulated by lipopolysaccharide ex vivo, interleukin 6 secretion was significantly decreased compared with control splenocytes. In conclusion, different types of BGA may be athero‐protective by exerting hypolipidemic and/or anti‐inflammatory functions. (Funded by NIH R21AT005152)

Grant Funding Source: NIH R21AT005152
Central adiposity predicts hippocampal‐dependent relational memory in prepubertal children
Authors: Khan Naiman A , Baym Carol L , Raine Lauren B , Drollette Eric , Scudder Mark , Kramer Arthur F , Cohen Neal J , Hillman Charles H
Published: June 2021

Central adiposity is a major risk factor for metabolic syndrome and emerging evidence suggests that excess fat mass may negatively affect brain and cognitive health as well. However, the influence of fat distribution on memory has not been examined in children. We investigated the relationship between adiposity and memory in prepubertal children (8–9 years). Participants (N=50) completed tasks assessing working memory and item and relational long‐term memory, which are known to have different brain substrates. Percent fat mass (%FM) and central adiposity (FM‐abd) were measured by DXA. Item and relational memory accuracy were negatively correlated (p<0.05, one‐tailed) with BMI (r=−0.27 and r=−0.36), %FM (r=−0.27 and r=−0.34), and FM‐abd (r=−0.29 and r=−0.40). Working memory was not related to adiposity. Adjusting for demographics (age, sex, SES), IQ, and VO2max, regression analyses showed that FM‐abd (β= −0.37, p=0.02) and BMI (β=−0.33, p=0.03) were negatively associated with relational memory. Neither item nor working memory was related to adiposity, and no memory outcomes were related to %FM. Our findings suggest that distinct forms of memory may be differentially affected by weight status and fat distribution. Given the known dependence of relational memory on the hippocampus, central adiposity may selectively affect hippocampal function in prepubertal children. (Funding source: NIH HD069381)

Grant Funding Source: NIH HD069381
Association of food intake with prostate cancer aggressiveness of African and Caucasian Americans in a population‐based study
Authors: Vance Terrence M , Wang Ying , Su L. Joseph , Fontham Elizabeth T. H. , Bensen Jeannette T. , Mohler James L. , Chen Ming‐Hui , Chun Ock K.
Published: June 2021

This study aimed to determine the association between major food groups that contribute to antioxidant intake and prostate cancer aggressiveness (PCA). Data are from the North Carolina‐Louisiana Prostate Cancer Project, a study of incident prostate cancer in African Americans (AA) (n=925) and Caucasian Americans (CA) (n=1,059). Dietary data were collected by a modified NCI DHQ. Prostate cancer cases were classified as low aggressive (LA) (Gleason score <7, stage ≤T2, PSA <10 ng/ml), high aggressive (HA) (Gleason score ≥8; PSA ≥20 ng/ml; or Gleason score = 7 if stage ≥T3), or intermediate aggressive (IA) (others). AA consuming 1 to 2 cups of fruit drink a week were found to have an odds ratio (OR) for HA vs. LA of 0.43 (95% CI: 0.20, 0.96) and 1 to 2 glasses of wine a week were found to have an OR for IA vs. LA of 0.30 (95% CI: 0.09, 1.00). CA drinking >;2 cups a day of tea were found to have an OR for HA vs. LA of OR 0.44 (95% CI: 0.20, 0.94) and 1 to 2 cups a week of orange juice were found to have an OR for IA vs. LA of OR 0.52 (95% CI: 0.27, 0.98). These results imply that antioxidant rich foods may contribute differently to PCA in CA and AA.

Grant Funding Source: NIH Cancer Epidemiology Small Grant #1R03CA159421–01A1.
Reduction in average child consumption of sugar‐sweetened beverages without decrease in home inventory
Authors: Ferris Ann M. , Wakefield Dorothy , Quesada Catalina , Lora Karina
Published: June 2021

Longitudinal study tested hypothesis that an educational intervention would decrease both home beverage inventory (HBI) of sugar‐sweetened beverages (flavored milk, juice drinks, iced tea, energy drinks, soda) (SSBs) and reported daily consumption the 3–5 y old child in the home. Study included 238 low‐income caregivers of 3–5 y. old children whose children consumed SSBs at baseline. Caregivers provided demographics and completed an HBI at baseline and 6 months. Intervention group received SSB education and the control group received a sham food safety curriculum. Non‐parametric tests tested for differences in total ounces available in home and consumed by child. At baseline, there was no difference in total ounces available in the home (median (min, max) (control:349 [6, 2442], intervention: 282 [6, 8524]) or ounces of consumed by the child each day (control: 10 [1, 106], intervention:12 [1, 62]). At 6 months, both groups showed a significant (p<0.001) decrease in ounces the child consumed per day (control: 6.2 [0, 48], intervention: 2.9 [0, 53]). The difference between the groups neared significance (p=0.06). Among SNAP recipients, the intervention group consumed less than the control group (p=0.02) at 6 months. In contrast, there was no decrease in total available SSBs. Preliminary results show a decrease in daily consumption of SSBs without a corresponding decrease in HBI.
Role of Caveolae in the development of abdominal aortic aneurysms
Authors: Crawford Kevin , Takayanagi Takehiko , Eguchi Satoru , Rizzo Victor
Published: June 2021

Abdominal aortic aneurysm (AAA) is a major cardiovascular disease and involves enhancement of rennin‐angiotensin system and recruitment/activation of inflammatory factors such as matrix metalloproteinases (MMP's). Caveolae has been shown to play a role in a number of different cardiovascular diseases through different mechanisms including regulation of oxidative stress, inflammation and degradation of extracellular matrix components through MMP's. In addition, endothelial cell caveolae are known to localize the Ang‐II (AT1) receptor and regulate rennin‐angiotensin signaling. Based on these findings, we evaluated the role of caveolae in a murine model of AAA formation. Here, eight week old mice were co‐infused with AngII and BAPN, a lysyl oxidase inhibitor, to induce AAA. We found that mice lacking the main structural protein of caveolae, caveolin‐1, did not develop AAA compared to WT animals in spite of hypertensive blood pressures measured by telemetry in both groups. This finding suggests that intact Ang‐II signaling remains in place in caveolin‐1 knockout mice. To begin to address the underlying mechanism by which caveolae contributes to AAA, we measured the level of oxidative stress and MMP's in aneurysms. We found an increased expression of MMP‐2 and MMP‐9 in vessels of WT mice displaying aneurysms. This increase in expression was not observed in Cav‐1 knockout mice. Furthermore, KO mice showed less oxidative stress then their WT counterparts as assessed by anti‐nitrotyrosine staining. In conclusion, caveolae appears to play a critical role in the formation of AAA in mice via oxidative stress, and recruitment and/or activation of MMPs, specifically MMP‐2 and MMP‐9.
Nano‐metal oxide aspiration exposure induces endothelial dysfunction in mice.
Authors: Frame Mary (Molly) , Dewar Anthony M , Brenner Sara
Published: June 2021

Metal oxide nanoparticles (nMO) are widely used in many diverse industries. We evaluated the effect of aspiration exposure to three nMO: SiO2 (length 51.9 ± 16.2 nm, aspect ratio 1.1 ± 0.1), Al2O3 (84.6 ± 20 nm, 1.4 ± 0.2) and CeO2 (52.5 ± 19.1 nm, 1.5 ± 0.4). Anesthetized (isoflurane) mice (25g) aspirated 20ug of SiO2, Al2O3 or CeO2 in 100ul of 1g% albumin in saline. 24 h later bronchial‐alveolar lavage showed and increase in total cells of 80% SiO2, 85% Al2O3, and 60% CeO2. Only Al2O3 caused weight loss 3.3% overnight. Using intravital microscopy, vasoactive responses were obtained in the cremaster m. model. Compared to controls, baseline diameters were unaltered for the same class of arteriole. Dilation to adenosine (10–4M) was attenuated for each nMO. Constriction to phenylephrine (10–6M) was attenuated for only Al2O3. Exposure to SiO2 (N=4) did not alter the dilation to acetylcholine (ACH 10–7 – 10–4M), but attenuated dilation to bradykinin (BK 10–8 – 10–5M) and isoproterenol (ISO 10–8 – 10–4M). Exposure to Al2O3 (N=4) or CeO2 (N=4) eliminated dilation to BK and attenuated dilation to ACH or ISO. Further, for SiO2 and Al2O3, a significant dilation to ISO was uncovered in the pM range (10–14 – 10–10M) that is mimicked by blocking clathrin endosome formation with dynasore (80uM) in control mice (N=6). Thus, aspiration of 20ug nMO induced moderate to severe endothelial dysfunction that was MO‐type specific. AHA 0655908T
Drug‐induced glutamine depletion hinders the growth of β‐ catenin mutated human liver cancer xenografts
Authors: Chiu Martina , Tardito Saverio , Pillozzi Serena , Arcangeli Annarosa , Campanini Nicoletta , Silini Enrico M. , Bussolati Ovidio
Published: June 2021

Human liver cancer HepG2 cell line is β‐catenin mutated and exhibits features of glutamine addiction. HepG2 cells in culture are sensitive to glutamine starvation induced with L‐asparaginase from Erwinia chrysanthemi (ASNase) and the inhibitor of Glutamine Synthetase (GS) methionine‐L‐sulfoximine (MSO). To evaluate HepG2 behaviour in vivo, 8‐day subcutaneous HepG2 xenografts were treated for three weeks with ASNase, MSO, or both. The administration of ASNase and MSO produced a marked serum depletion of both asparagine and glutamine, a decrease in liver and tumor glutamine content, and the inhibition of liver GS activity without any evidence of systemic toxicity. Xenograft growth was significantly hindered by either ASNase or MSO and completely prevented by the combined treatment. Histopathological evaluation confirmed high GS expression and nuclear positivity for β‐catenin in HepG2 xenografts, as well as the inhibition of tumor growth in mice treated with both ASNase and MSO. These results strengthen the hypothesis that liver cancers carrying β‐catenin mutations are glutamine‐addicted and represent the first evidence for antitumor activity of GS inhibitors.
Higher‐order chromatin folding and gene regulation
Authors: Bickmore Wendy
Published: June 2021

It is widely appreciated that histone modifications correlate to gene expression states and can, in some cases, directly impact on gene expression and repression. But, our understanding of chromatin states beyond the level of the nucleosome itself is much more rudimentary. Until quite recently there has been little exploration of how these levels of higher‐order chromatin structure might contribute to the regulation of gene expression.

Using specific developmentally regulated loci in the mouse, I will discuss how chromatin compaction can act to regulate the spatial and temporal activation of genes during mouse development. I will compare different methods that can probe higher order chromatin structure and will demonstrate their use in both cell culture systems that reflect developmental stages, and in embryos.
Transforming growth factor‐beta‐induced protein ig‐h3 is essential for the formation of human colorectal carcinoma liver metastasis in vivo
Authors: Castronovo Vincent , Blomme Arnaud , Delvaux David , Delvenne Philippe , Detry Olivier , Turtoi Andrei
Published: June 2021

Transforming growth factor‐beta‐induced protein ig‐h3 (TGFBI) is extracellular matrix component important for cell‐collagen interaction. We and others have reported elevated expression of TGFBI in several human cancers, where its role remains controversial. Current study reports for the first time on the function of TGFBI in colorectal carcinoma liver metastasis (CRC‐LM), a frequently encountered malignancy with no satisfactory treatment to date.

We show that TGFBI is highly expressed in human CRC‐LM and in LM originating from breast, lung and pancreatic tumors. From functional perspective, silencing TGFBI in SW1222 human colorectal carcinoma cells lead to a decrease in migration (−70%) and proliferation (−20%). To understand how this effects translate in vivo we have developed a novel animal model of colorectal carcinoma based on chicken chorioallantoic membrane (CAM) that mimics human CRC‐LM. TGFBI silencing resulted in 50% reduction of tumor volume in vivo. Finally, we have demonstrated that TGFBI is systemically reachable target for homing fluorescently‐labeled antibodies injected in tumor bearing animals. Collectively the data show that TGFBI is essential for promoting the development of CRC‐LM and therefore a promising target for designing novel therapeutic approaches.

This work is supported by FNRS, Belgium and FP7 EU program.
Active learning: a small group histology laboratory exercise in a whole class setting utilizing virtual slides and peer education
Authors: Bloodgood Robert Alan
Published: June 2021

Histology laboratory instruction is embracing virtual microscopy, facing reduced curricular time and being ask to focus on small group active learning. The challenge lies in developing meaningful active learning experiences that take advantage of virtual microscopy while engaging and challenging the students, maintaining the involvement of experienced medical educators and reaping the benefits of learning in a small group environment, even when the exercise is given within a large learning space. We developed an histology laboratory exercise, occurring in a Learning Studio housing the entire medical class, that utilizes virtual slides in the context of small group active learning and peer instruction. Students sit at round tables and work in groups of three. Even and odd‐numbered tables access different sets of virtual slides of lymphoid tissues and organs using laptop computers. Each group of three is given a worksheet to complete in which they identify the unique characteristics of each lymphoid tissue or organ. The 2nd part of the exercise has selected pairs of medical students present a virtual slide to the entire class (using large screens mounted around the circular room), describe the unique characteristics allowing identification and fielding questions from their classmates. The last part of the exercise involves the class being presented with unknown Virtual Slides that they identify and discuss.
Effects of subcutaneous daidzein on recovery of function in the rat after ischemic stroke
Authors: Knapp Austen N , Stout Jessica M , Grisley Elizabeth D , Wallace Doug G , Cheatwood Joseph L
Published: June 2021

Stroke is a leading cause of lasting disability in humans and often results in deficits in sensory and motor functions. There is still a need for a treatment that will enhance recovery and/or reduce behavioral deficits following stroke. In a previous study, our laboratory demonstrated a decrease in post‐stroke behavioral deficits when provided a soy protein‐based diet prior to stroke. Daidzein, a bioactive isoflavone found in soy, is a potential contributor to the observed effects. In the current study, we aimed to examine the effectiveness of daidzein as a post‐stroke treatment by assessing forelimb function to determine post‐stroke behavioral effects and gauge functional recovery. Young adult male Long Evans Hooded rats underwent a middle cerebral artery occlusion (MCAO), which induced a stroke lesion. Following MCAO, rats were treated with vehicle or daidzein by a subcutaneous osmotic minipump. The skilled ladder rung walking task was used to measure forelimb function following stroke. The number of total, slight, and deep slips were compared between groups. No significant differences were found in total slips (slight plus deep) or slight slips(p>;0.05). However, the daidzein group had significantly less deep slips than the vehicle group (p<0.05). These results suggest daidzein mediates neuroprotection and provides enhanced behavioral recovery post‐stroke.

Supported by: SIUSOM, SIU REACH, SIU CIRCNS

Grant Funding Source: SIU CIRCNS
Single Molecular Analysis of the Encounter of Replication Forks with DNA Interstrand Crosslinks
Authors: Huang Jing , Liu Shuo , Wang Yinsheng , Thazhathveetil Arun Kalliat , Wang Weidong , Seidman Michael M.
Published: June 2021

DNA interstrand crosslinks (ICLs) are absolute blocks to replication. Progress of ICL repair study, particularly at replication forks, has been limited by the lack of approaches for the direct analysis of replication in the vicinity of ICLs. Additionally, most crosslinking agents generate primarily monoadducts, with only a small fraction of actual ICLs, complicating interpretation of experiments with these compounds.

We have developed a novel approach for single‐molecule analysis of replication to study the encounter of individual replication forks with ICLs. We employed antigen tagged psoralen (dig‐pso) which forms a high frequency of ICLs following photoactivation by ultraviolet light (UVA). Cells are treated with dig‐pso/UVA and then pulsed with halogenated thymidine derivatives. The location of replication tracts in the vicinity of ICLs can be determined by immunofluorescent visualization of the thymidine analogues and dig‐pso on DNA fibers. Single fork collisions with the ICLs do occur, but replication on either side of the ICLs is observed more frequently. Similar replication patterns were obtained in both repair proficient and deficient cells, suggesting that replication decisions are made independently of repair competence. Our results argue that cells frequently follow a replication‐replicationrepair model in response to replication blocks.
RECQL5 plays both co‐operative and complementary roles with WRN syndrome helicase.
Authors: Popuri Venkateswarlu , Huang Jing , Mahesh Ramamoorthy , Tadokoro Takashi , Deborah Croteau L , Bohr Vilhelm A.
Published: June 2021

Humans have five RecQ helicases whereas lower organisms like bacteria and yeast have only one member per species. Little is known about whether and how these RecQ helicases cooperate and/or complement each other in response to cellular stress. Here we show that RECQL5 associates longer at laser‐induced DNA double strand breaks (DSBs) in the absence of Werner syndrome (WRN) protein and that it interacts physically and functionally with WRN both in vivo and in vitro. RECQL5 co‐operates with WRN on synthetic stalled replication fork‐like structures and stimulates its helicase activity on DNA fork duplexes. Both RECQL5 and WRN re‐localize from the nucleolus into the nucleus after replicative stress and significantly associate with each other during S‐phase. Further, we show that RECQL5 is essential for cell survival in the absence of WRN. Loss of both RECQL5 and WRN severely compromises DNA replication, accumulates genomic instability and ultimately leads to cell death. Collectively, our results indicate that RECQL5 plays both co‐operative and complementary roles with WRN. This is an early demonstration of a significant functional interplay and a novel synthetic lethal interaction among the human RecQ helicases.
Argonaute 2‐loaded promoter‐targeted double stranded RNA (Ago2‐ptRNA) mediates gene activation by interacting with the core transcriptional machinery
Authors: Portnoy Victoria , Li Long‐Cheng
Published: June 2021

Promoter‐targeted double‐stranded RNA (ptRNA) has been shown to mediate potent activation of gene expression in a phenomenon known as RNA activation (RNAa). Previous studies revealed requirement of RNAa on Argonaute 2 (Ago2) protein as well as RNAa association with epigenetic changes at the target promoter. However, its precise molecular nature remains elusive.

Using human p21 promoter as a model system, we further characterized RNAa mechanism. ptRNA designed to target p21 promoter is loaded by Ago2 protein, which uses one of the RNA strands to guide it binding to promoter DNA. ptRNA‐Ago2 binding to the promoter target site activate p21 gene expression on transcriptional level. Our biochemical studies revealed the identity of the RNAa effector complex, which contains ptRNA‐loaded Ago2 protein and at least two additional protein components: RNA helicase A (RHA) and RNA polymerase II (RNAP II)‐associated factor CTR9, both capable of recruiting histone modifying enzymes to facilitate transcription initiation and elongation. We show evidence that RNAa effector complex physically associates with RNAPII to facilitate re‐initiation of transcription.

Together, our results provide important new insights into RNAa mechanism and allow us to build a working model for RNAa.

The research was supported by grants from NIH (1R01GM090293–0109 to L.C.L.) and Department of Defense (W81XWH‐08–1‐0260 to L.C.L.)
Binding Characteristics of TAR RNA and Tat‐Cyc peptide from the Human Immunodeficiency Virus
Authors: Sun Josh , Najib Mohammad
Published: June 2021

The trans‐activation of transcription (Tat) peptide and its binding to the trans‐activating response element (TAR) RNA are observable in the human immunodeficiency virus (HIV). The interaction between the TAR RNA and the Tat peptide immensely increases the transcription efficiency of HIV by preventing cells from inducing apoptosis. Optimal binding of Tat to TAR requires the presence of a host protein, cyclin T1 to allow phosphorylation of the stalled RNA polymerase II. To study the interaction between trans‐activation of transcription (Tat) peptide and the trans‐activating response element (TAR) RNA of HIV, the cDNA for a fusion of the Tat peptide with a cyclin T1 peptide (Tat‐Cyc) and TAR RNA were both cloned separately by PCR‐assisted gene assembly using overlapping primers. The PCR products were initially ligated into the pGEM‐T vector (Promega). The cDNA for the TAR RNA was then subcloned into the BamHI and EcoRI restriction sites of vector pUC18 and will be transcribed via T7 RNA polymerase. The Tat‐cyc peptide was subcloned into the expression vector pSUMO‐Star (LifeSensors) at the BsaI and Xho I restriction sites. Once both the TAR RNA and Tat‐cyc peptides have been purified, the binding characteristics of the two will be analyzed and observed using Isothermal Titration Calorimetry (ITC) and gel shift binding assays. This TAR RNA and TAT peptide recognition study could promote the development of therapeutic agents to block or inhibit the viral activation and transcription of HIV.
Preparation and characterization of guar gum microparticles loaded with Cisplatin
Authors: Ribeiro Camyla Mara da Silva , Gomes Anderson J
Published: June 2021

Systems for controlled release of drugs have been extensively studied and they are very attractive alternative in cancer treatment, in vivo, to achieve clinical benefit. Among the biopolymers used in the characterization of microparticles, the highlights are the polysaccharides, including guar gum. This study produced and characterized guar gum microparticles to encapsulate anticancer drugs, such as Cisplatin, in order to reduce adverse effects and enhance bioavailability with sustained delivery. The microparticles were prepared using the method of emulsification/ionic crosslinking. The microparticles of guar gum obtained 60% efficiency of encapsulation, the mean particles size was 1.7 ± 0.6 μm, and zeta potential of −16.5 mV. The study of release profile, in vitro, appeared to be sustained and bimodal during 216 hours. The study of stability profile showed low tendency of aggregation and good physicochemical stability during 264 hours. Therefore, guar gum microparticles appear, technologically, a promising anticancer drugs delivery system. Supported by FAPDF, CAPES, CNPq, FINATEC and DPP‐UnB.
Plasma Membrane Ca2+‐ATPase 4 Splice Variants are Acquired by Murine Sperm During Epididymal Transit via Epididymosomes
Authors: Patel Ramkrishna , Stabley Deborah , Barone Carol , Galileo Deni , Strehler Emanuel , Martin‐DeLeon Patricia
Published: June 2021

Plasma Membrane Ca2+‐ATPase isoform 4 (PMCA4) is the primary Ca2+ efflux pump in murine sperm where it regulates motility. In Pmca4 null sperm hyperactivated motility is lost, resulting in infertility. We have shown that PMCA4b – CASK interaction in sperm is key in regulating Ca2+ homeostasis. However, recent work showed that in bulls the PMCA4a splice variant (not in testis) is epididymally‐expressed, along with 4b, and may be transferred to sperm. Here we show, via conventional and in situ RT‐PCR, that both splice variants of Pmca4 mRNA are expressed in the murine testis and along the epididymis. Immunohistochemistry localized PMCA4a in the epididymal epithelium and Western analysis showed, for the first time, that PMCA4a is secreted in the epididymal luminal fluid (ELF). Flow cytometry indicated the presence of PMCA4a on mature caudal sperm where it was increased ~5‐fold compared to caput sperm (detected by Western) and~2‐fold and ~2.75‐fold after incubation in ELF and epididymosomes, respectively, revealing in vitro uptake. Using pan‐PMCA4 antibodies, CO‐IP assays revealed that both variants associate with CASK, indicating possible function as a complex. Our study suggests a mechanism for combining the different functional attributes of both variants, leading to greater efficiency of the pump in maintenance of Ca2+ homeostasis, which is crucial for sperm motility and male fertility.
Defining the role of two sulfur dioxygenases of Azotobacter vinelandii in sulfur trafficking
Authors: Nguyen Diep Minh Ngoc , Piel Robert B. , Laughlin Caitlin T. , Larson Timothy J.
Published: June 2021

The human ETHE1 gene encodes sulfur dioxygenase (SD) which, together with likely participation of sulfurtransferase (rhodanese), is essential for sulfide detoxification in mitochondria. The current work describes a similar pathway in Azotobacter vinelandii. This Gram‐negative bacterium has two ETHE1‐like genes (Avin_46910 & Avin_01930) potentially involved in sulfide clearance. Biochemical and genetic approaches were used to test if these genes encode SD. The corresponding proteins (SD1 and SD2) were purified and had strong SD activity (sulfur‐ and glutathione‐dependent consumption of oxygen; ~25 U/mg). The SD1 gene is found in the same operon with genes for a SulP family transporter (sulP) and a rhodanese (rhdB). Glutathione persulfide, the substrate for SD, could be generated by RhdB using thiosulfate as sulfur donor and glutathione as sulfur acceptor, implying tight coupling of the rhodanese and SD reactions. Mutant strains deficient in SD1 and/or SD2 were constructed. Higher levels of sulfide accumulated in the double mutant compared to the wild type and single mutants during late log and stationary phase of growth with sulfate as sulfur source. During growth with thiosulfate, the double mutant achieved lower cell density and accumulated more sulfide relative to the wild‐type strain. The results indicate that these SDs participate in oxidation of sulfide. Supported by Fralin Life Sciences Institute.
The green tea polyphenol EGCG induces Mesenchymal to Epithelial Transition (MET) and tumor regression in Triple Negative Breast Cancer (TNBC) cells and mouse‐xenograft model: Involvement of CCN5
Authors: Das Amlan , Banerjee Snigdha , De Archana , Haque Inamul , Maity Gargi , McEwen Matt , Banerjee Sushanta K
Published: June 2021

Background

Epithelial to Mesenchymal transition (EMT) is an important and coordinated series of events associated with tumor metastasis and invasionRecent studies had shown the importance of CCN5 in the regulation of various carcinomas including the breast cancer. EGCG, the most abundant green tea polyphenol, has been extensively studied as a bioactive dietary component against various types of carcinomas. However, the mechanism of action of EGCG in breast carcinoma is uncertain.

Objective

The objective of the present study is to determine whether CCN5 plays any significant role in EGCG‐mediated cytotoxicity in triple negative breast cancer cells.

Exposure of triple negative human breast cancer (TNBC) cells, MDA‐MB‐231 and HCC70 to EGCG resulted in a dose‐dependent inhibition of proliferation after 72h and the IC50 was observed around 75μM for MDA‐MB‐231 cells and 50μM for HCC70 respectively. We found EGCG‐treatment effectively induces MET and inhibits the in vitro migration parallel with the induction of CCN5 expression in TNBC cells in a dose‐dependent fashion. Furthermore, consistent with in vitro findings, tumor progression was drastically inhibited in EGCG‐treated MDA‐MB‐ 231‐tumor xenograft in nude mouse model.

Conclusion

EGCG imparts its anti‐cancer activity via induction of CCN5.

(Support: VA Merit Grants, S. Banerjee and S.K. Banerjee)
Investigation of RGS16 mediated inhibition of pancreatic cancer metastasis
Authors: Carper Miranda Bernel , Boskovic Goran , Denvir James , Primerano Donald , Hardman W. Elaine , Claudio Pier Paolo
Published: June 2021

Pancreatic cancer is ranked as the fourth cause of cancer‐related deaths in the United States with a five‐year survival rate of <5%. This is due in part, to early systemic dissemination. Due to the aggressive nature of this disease, new therapies are needed to inhibit and treat metastatic pancreatic cancer. We have previously performed a microarray analysis to determine proteins regulated by the tumor suppressors p53 and pRb to identify proteins involved in the p53 and pRb cross‐talk pathway. We have found that Regulator of G‐protein signaling (RGS16) is regulated by both p53 and pRb in WI38 normal lung fibroblast cells. RGS16 turns off signaling of several oncogene pathways that are involved in proliferation, chemoresistance, and metastasis. Recently it has also been found that RGS16 is downregulated in metastasized pancreatic cancer. The objective is to investigate the ability of RGS16 to inhibit metastasis by examining pancreatic cancer cell invasion and migration.

RGS16 was overexpressed in BxPC‐3 and PANC‐1 pancreatic cancer cell lines. Migration and invasion were investigated using wound‐healing assays and Boyden chambers, respectively. The results suggest that RGS16 is able to inhibit migration and invasion of pancreatic cancer cells.

Our data indicates that RGS16 is regulated by both p53 and pRb and overexpression of RGS16 is able to inhibit pancreatic cancer metastasis.
Effects of substituting whole milk with skim milk on the distribution of diet macronutrients and serum triglycerides in indigenous children and teenagers of the state of Hidalgo, Mexico
Authors: Zamora Lilia Susana Pedraza , Villalpando Salvador , Figueroa Héctor Lamadrid , Silva Claudia Ivonne Ramirez
Published: June 2021

The cardiovascular health effects of high intake of saturated fatty acids are considerable since childhood. A significant source of this type of fat is whole milk. Substitution with skim milk could cause energy compensation and the nutrients used for energy replacement might affect serum triglycerides. Data from a randomized double blinded study with children and teenagers in indigenous shelters of Hidalgo was used. Shelters were randomized to receive whole milk, low fat milk or skim milk for four months. Shelters assigned to receive whole milk were the control group, and the other two were the treatment groups. Cluster adjusted random effect models were used for analysis. There was a 62% increase in the carbohydrate intake of the skim milk group, in comparison to the control group. Total fat percentage of energy was 24% lower in the low fat milk group. Serum triglycerides were higher in the skim milk group comparing with the control group. There was an increase in the amount of tortilla consumed by the skim milk group of 116 grams. The substitution of whole milk with skim milk produced a compensation of energy with carbohydrates in our study population. The food source from where the carbohydrates were obtained is mainly tortilla and its derivatives. Such an increase in the intake of carbohydrates also generates an important increase in serum triglycerides, which induce cardiovascular risk even at a young age.
A social franchising approach to strengthening infant and young child feeding (IYCF) counseling improves the quality of services at health facilities in Vietnam
Authors: Nguyen Phuong H. , Menon Purnima , Kim Sunny , Hajeebhoy Nemat , Tran Lan M. , Ruel Marie T. , Rawat Rahul
Published: June 2021

A network of social franchises was established in 780 government health facilities in Vietnam under the Alive & Thrive initiative to provide a standardized package of IYCF counseling services with upgraded physical spaces, educational materials, and specialized training of health staff. We studied the quality of care at franchise facilities (FF) compared to standard facilities (SF) across 3 dimensions: structure, process and outcome. A mixed methods approach was used to study quality of care in the context of a process evaluation. Data collection included facility inventories, staff surveys, observations of counseling sessions and client exit interviews. Compared to SF, FF had better physical infrastructure and materials (structure), service delivery (process), and client satisfaction (outcome). FF counselors had greater interpersonal communication skills than SF counselors (score 14.2 vs. 9.0, range 0–22), and better knowledge of optimal breastfeeding practices (score 11.5 vs. 9.8 range 0–13). Smaller differences were observed for counseling on complementary feeding between FF and SF (score 5.5 vs. 4.3, range 0–9). In conclusion, a social franchise approach enables the provision of higher quality nutrition counseling services within government health facilities and has potential to improve IYCF practices. Impact will depend upon high utilization and adherence to counseling recommendations.

Funded by Bill & Melinda Gates Foundation, through Alive & Thrive, managed by FHI360
Trends in dietary carbohydrate consumption from 1991–2008 in the Framingham Heart Study offspring cohort
Authors: Makarem Nour , Scott Marc , Quatromoni Paula , Jacques Paul , Parekh Niyati
Published: June 2021

Longitudinal studies evaluating trends in dietary carbohydrate consumption among U.S. adults are limited. More recent studies reflective of the changes in food supply are needed to evaluate long‐term intakes and to compare them to dietary recommendations. We examined trends in dietary carbohydrate intake in the Framingham Heart Study (FHS) Offspring Cohort over a 17‐year period. The FHS offspring generation was recruited in 1971–75. Follow‐up exams were conducted every 4 years. Dietary data collection began at Exam 5 in 1991 using a semi‐quantitative food frequency questionnaire. This analysis included 3265 adults ages ≥25y with complete dietary data in at least three exams from 1991–2008. Mixed effects linear regression was used to examine trends in macronutrient and food intake using SASv9.3. Over 17 years of follow‐up, % energy from total carbohydrates (50.95% to 46.84%; p‐trend<0.0005) and from total sugars (18.19 % to 16.62%; p‐trend<0.0005) decreased. There was a decrease in % energy from fructose (5.41 to 4.76 %; p‐trend <0.0005) and % energy from sugar in beverages (1.43% to 0.9%, p‐trend<0.0005). Dietary fiber intake increased (18.01 to 19.21g, p‐trend<0.0005). Similar trends were observed when the analyses were stratified by sex. These results suggest some improvement in dietary carbohydrate quality over time in this cohort.

Grant Funding Source: American Cancer Society Research Scholar Grant (#RSG‐12–005–01‐CNE)
Prevalence of metabolic syndrome (MetS) and hyperglycemia in US adults: NHANES 2003–06 and 2007–10
Authors: Gee David L. , Bailey Regan L.
Published: June 2021

MetS consists of a cluster of risk factors that strongly predict developing diabetes and is associated with increased mortality. The purpose of this analysis was to characterize MetS and hyperglycemia in US adults using NHANES 2003–6 (n=3532) & 2007–10 (n=4717). The prevalence of MetS remained constant at 33% between the two time periods. The prevalence of MetS was unchanged within gender, age group, weight categories, or race/ethnic group except among male Mexican‐Americans which significantly increased (32% to 38%). However, the prevalence of hyperglycemia (FBG >; 100 mg/dl) increased from 36% to 41% and was greatest in young adults (20–39y, 25% increase) and for those with a normal BMI (28% increase). The increase in the prevalence of pre‐diabetes & diabetes was observed regardless if the criteria were fasting blood glucose or fasting glycohemoglobin. The prevalence of pre‐diabetes and diabetes significantly increased in normal, overweight, and obese weight categories. Fasting plasma insulin concentrations, homeostasis model assessment of insulin resistance (HOMA‐IR), and glycohemoglobin concentrations significantly increased by 17%, 19%, & 2% respectively between these two time periods. These findings indicate that while the prevalence of MetS remained constant, the significant increase in diabetes and pre‐diabetes reflects an additional 22 million cases in adult Americans.
Exploring Nutrition Education Opportunities in Alternative School Settings
Authors: Phelps Josh Andrew , Norquest Michele , Hermann Janice , Kennedy Tay
Published: June 2021

Participants included alternative school students participating in a Supplemental Nutrition Assistance Program‐Education (SNAP‐Ed) program. Data collection involved focus groups for collecting student perspectives regarding preferred teaching and learning styles, and nutrition topics of interest. Twenty‐four students (12 females and 12 males) ranging in age from 15–18 years participated in the project. Data revealed experiential learning as the preferred learning style facilitated by hands‐on teaching approaches. Disconnected learning was revealed as their least favorite learning style, which students associated with teaching styles that left them having to learn on their own without guidance for how to apply what they are learning. General nutrition, personalized nutrition, and empowering nutrition were three areas of nutrition education identified as important by students. Topics of interest included understanding what they were putting in their bodies, staying fit and healthy, and shopping, cooking, and planning meals. Based on student responses, alternative school students are interested in receiving nutrition education if it involves an engaging process utilizing applied approaches. Opportunities exist to develop participant‐centered nutrition topics in alternative school settings with materials and delivery methods tailored to the identified needs of the audience.
Fortification of Donor Human Milk for Premature Infants
Authors: Bissett Emily , Fogleman April , Cohen Ronald , Sakamoto Pauline , Allen Jonathan
Published: June 2021

Premature infants at risk for metabolic bone disease may need increased minerals for normal bone mineralization, in which case supplementation of donor human milk (DHM) with calcium (Ca), phosphorus (P), and post‐discharge formula (PDF) is common. The bioavailability of these additives has not been proven. Objectives were to study the effect of Ca, P, and PDF supplementation on the bioaccessibility of calcium and phosphate. A novel in‐vitro model of the premature infant's gastrointestinal tract was developed to simulate effects of digestion, absorption and transport. Calcium and phosphate were measured before and after in‐vitro digestion in DHM with and without added supplements. Bioaccessible calcium was greater in DHM with supplemental calcium and calcium and phosphate together than in DHM alone or with supplemental PDF (p < 0.0001). Bioaccessible phosphate was not significantly different between treatment groups. The addition of minerals rather than PDF provides the most bioaccessible calcium.

Funded by San Jose Mothers’ Milk Bank
Association between abdominal adipose tissue mass with anthropometric and cardiometabolic variables in a subgroup of males and females from the Canola Oil Multicentre Intervention Trial (COMIT)
Authors: Liu Xiaoran , Kris‐Etherton Penny M. , West Sheila G. , Lamarche Benoit , Jenkins David J. , Fleming Jennifer A. , Jones Peter J.
Published: June 2021

Subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) differ metabolically. Excessive VAT is linked to obesity‐related disease. We evaluated the association between baseline abdominal SAT, VAT and cardiometabolic (glucose) and anthropometric measurements in a subgroup of participants from COMIT (F=7, M=7) with increased waist circumference (F: >;80 cm; M: >; 94 cm). MRI images were analyzed to separate the SAT and VAT; 3 slices centered between lumbar vertebrae L4–5 were calculated. Abdominal and total fat mass were measured by DXA. In females, SAT was positively correlated with abdominal (r=0.86, p=0.013) and total (r=0.82, p=0.02) fat mass and with waist circumference (r=0.84, p=0.02). In males, VAT was positively associated with abdominal fat mass (r=0.93, p=0.002) and demonstrated a strong tendency with waist circumference (r=0.75, p=0.051). SAT also correlated with total fat mass (r=0.73, p=0.06). Glucose levels were positively associated with SAT and VAT in females (r=0.84, p=0.02; r=0.86, p=0.04). In summary, in females, SAT is positively correlated with fat mass, waist circumference; SAT and VAT may contribute to elevated glucose levels. In males, VAT alone was positively associated with abdominal fat mass and increased waist circumference. Gender differences were observed for SAT and VAT depots; further research on differing metabolic responses between males and females is warranted.

Grant Funding Source: Canola Council of Canada
Preventive Effect of Artemisia capillaris extract on Bone Loss in Ovariectomized Rat Model
Authors: Jang Hae‐Dong , Lee Sang‐Hyun , Kim Young‐Ho
Published: June 2021

Artemisia capillaris Thunberg has been used as a folk medicine for treating various diseases including inflammatory and immune‐related diseases. The aim of the present study was to investigate the protective effect of Artemisia capillaris extract (ACE) on bone loss in ovariectomized rats. A total of sixty‐8‐week‐old female Spraque‐Dawley rats were divided randomly into sham‐operated group and four ovariectomized (OVA) groups: OVA, OVA + 17β‐ estradiol (E2, 50 μg/kg/day) and OVA + ACE (250 or 500 mg/kg/day). Daily oral administration of E2 or ACE began 3 weeks after surgery and lasted for 6 or 10 weeks. ACE prevented total BMD and BMC decrease in the femur induced by OVA, which was accompanied by a significant decrease in bone remodeling, as it was evidenced by the regulated levels of the bone turnover markers such as osteocalcin (OC), alkaline phosphatase (ALP), procollagen type1 C‐terminal peptide (PICP), osteoprotegerin (OPG), receptor activator of NF‐κB ligand (RANKL), tartrate resistant acid phosphatase (TRAP), and cross‐linked C‐terminal telopeptide of type 1 collagen (ICTP). These results indicate that Artemisia capillaris extract may be utilized as a therapeutic agent for the prevention of bone metabolism‐related diseases such as postmenopausal osteoporosis.
One‐carbon Metabolism Related B‐vitamins Alter the Expression of microRNAs Associated with the Wnt Pathway in Mouse Colonic Epithelium
Authors: Mohn Emily , Ma Xianyong , Lee Brianna , Choi Sang‐Woon , Crott Jimmy W. , Lai Chao‐qiang , Liu Zhenhua
Published: June 2021

MicroRNAs regulate numerous biological processes and thereby contribute to a variety of diseases including cancer. One‐carbon metabolism related B‐vitamins (folate, B2, B6, B12) have been indicated to influence the expression of a variety of genes, likely via their role in biological methylation. This study investigated whether these vitamins alter the expression of Wnt‐signaling related microRNAs and thereby influence intestinal tumorigenesis. MicroRNA expression profiles were measured using microRNA arrays on colonic epithelial cells from Apc1638N mice fed with diets deplete or sufficient in those vitamins. 38 Wnt pathway‐related microRNAs were also examined by real‐time PCR. 6 microRNAs in a total of 609 were identified to be significantly altered by those vitamin depletion (p<0.05) with additional 11 microRNAs having a mild degree of alterations (p<0.10). Among the 38 Wnt pathway‐related microRNAs, the expressions of 4 microRNAs were altered (p<0.05). The microRNA array data with the validation of Wnt pathway‐related microRNAs collectively indicate that the expressions of miR‐21 and miR‐122, which are biologically linked, miR‐29 family and miR‐34 family are dysregulated by those B‐vitamin depletion. In summary, these findings indicate that microRNAs may constitue a mechanism by which one‐carbon B‐vitamins regulate the Wnt‐signaling.