bluuebunny/crossref_metadata_2025_split huggingface dataset:
TopoCell – An image analysis tool to study intracellular topography
Authors: Fachada Vasco Rui Carvalho Pereira Fernandes , Fachada Nuno , Turpeinen Tuomas , Rahkila Paavo , Rosa Agostinho , Kainulainen Heikki 
Published: June 2021

We aim to build an open source bioimage informatics tool capable of analyzing 2D and 3D topography (absolute and relative localization, shape, size and intensity) of distinct types of intracellular particles (such as organelles, proteins, lipids and other detectable biomolecules) in different types of cells in a high‐throughput fashion.To achieve this, TopoCell is primarily designed to run fluorescence microscope images, where all possible channels imaged are used to semi‐automatically detect different particles, define cell type and segment cells. To each detected particle, a coordinate, intensity, volume, and sphericity value are attributed. Cells are detected through a plasmalemma marker, where volume and coordinate values are generated after the segmentation step. Numerous cell types can be defined by the user accordingly to the density of given intracellular particles. By controlling a dedicated filter, the user can select which cells and particles to consider in the analysis according to their detected values, including relative distance between two or more types of particles. Visualization of results and statistical analysis are also available.TopoCell allows us to comprehensively measure in still microscope images, the distribution of intracellular phenomena which otherwise would be negligible to the naked eye, as for instance translocation or particle association.
New Cell Biology Assays Using a Stabilized Luciferase Exhibiting Red Luminescence
Authors: Jiang Ying , Naleway John Joseph 
Published: June 2021

Bioluminescence is the light produced in certain organisms as a result of luciferase enzyme mediated oxidation of a luciferin substrate. This oxidation is linearly dependent on the presence of adenosine tri‐phosphate (ATP). We have recently developed a codon‐optimized luciferase based upon the gene sequence of the natural luciferase isolated from Luciola cruciata (Japanese firefly). This new gene codes for a modified amino acid sequence which exhibits improved expression levels in mammalian cells, red‐shifted emission wavelengths (560 to 619 nm) as well as improved thermal stability, while still utilizing D‐luciferin and ATP as substrates.This new recombinant enzyme was purified to 98% homogeneity from both transfected E. coli bacteria and HEK 293 mammalian cells using Ni+2‐affinity chromatography as the His6‐tagged protein. Enzymes isolated from either source were found to have equivalent activity in ATP dependent assays. The purified recombinant luciferase was used to develop a number of ultrasensitive biological assays useful in measuring cell number, cell viability as well as the cytotoxicity effects of several chemotherapeutic drugs (Etoposide and Doxorubicin) when added to murine NIH3T3 cells in culture. We also utilized the purified enzyme to sensitively detect bacterial cell number down to 1 × 102 cells and adapted the methods for use in high‐throughput antibiotic screening protocols. By cloning the luciferase downstream of TFkB responsive elements, the activation of TF‐κB (with stimulation by TNFα) in human HEK 293 cells could be quantitated by directly monitoring the increase in luminescence from activation of luciferase expression. Finally, since this new luciferase has a red‐shift in light emission, it was utilized in multiplexed assay conditions with other green (firefly) or blue‐emitting (Renilla) luciferases in a number of coupled biological assays. Moreover, application of this new luciferase for use in in vivo imaging was explored.
Galactooligosaccharides: effects on calcium absorption and gut microflora in young premenarcheal girls
Authors: Whisner Corrie Marie , Martin Berdine R , Clavijo Andrea , Nakatsu Cindy H , McCabe George P , McCabe Linda D , van den Heuvel Ellen GHM , Schoterman Margriet HC , Weaver Connie M 
Published: June 2021

Adolescence presents an opportunity to influence peak bone mass with prebiotic agents like galactooligosaccharides (GOS) that increase calcium absorption in the large intestine in animal models and adults. The objectives of this study were 1) to investigate the dose response of GOS supplementation on calcium absorption during growth and 2) to assess total colonic microbiota and bifidobacteria to better understand the mechanism of action. Using a randomized, double‐blind, crossover design, thirty‐one 10–13 year old girls consumed smoothie drinks enriched with 0, 2.5 or 5 g GOS twice a day for 3 weeks. The 3‐week periods were separated by 2‐week washout periods. Fractional calcium absorption was determined, using dual stable isotope methods, as the ratio of oral and intravenous tracers in urine over 48 hours. DNA was extracted from fecal samples to quantify total bacteria and bifidobacteria using qPCR. Preliminary results show that GOS increased fractional calcium absorption. Bifidobacteria as a proportion of total bacteria was also significantly greater after consumption of GOS. Results suggest that GOS consumption increases fractional calcium absorption in young girls through possible mechanisms in the large intestine.Grant Funding Source : FrieslandCampina Domo
Individualized counseling for a Healthy People 2010 diet results in surpassed fruit and vegetable intakes
Authors: Sidahmed Elkhansa , Cornellier Maria , Askew Leah , McCoy Alexandra , Rapai Mary , Sen Ananda , Djuric Zora 
Published: June 2021

The objective of the study was to evaluate adherence to two different diets that were implemented in a colon cancer prevention study. A total of 120 subjects were randomized to 6 months of individualized counseling for a Healthy Eating or a Mediterranean diet. The counseling was implemented starting with one face to face meeting with a registered dietitian followed by telephone counseling at the same intervals in both arms. A checklist was developed for assistance in meeting goals in both arms. The percentage of goals met in the two groups was similar with 87% of goals met in the Healthy Arm and 82% in the Mediterranean arm. Although the fruit and vegetable goals were lower in the Healthy arm, these goals were surpassed resulting in statistically similar fruit and vegetable intakes in the two groups (7.8 vs 8.1 servings/day). Carotenoid intakes and blood carotenoid levels also increased to similar extents in the two arms. These results indicate that individualized counseling for meeting Healthy People 2010 goals can result in better than expected compliance, resulting in carotenoid intakes similar to what can be achieved with a Mediterranean diet. Supported by grant NIH CA120381Grant Funding Source : NCI
Dietary Behaviors and Diet Quality of Low‐Income Children in the Supplemental Nutrition Assistance Program
Authors: Leung Cindy W , Hoffnagle Elena E , Jensen Helen H , Foerster Susan B , Murphy Suzanne P , Nestle Marion , Cheung Lilian , Turrell Sophia , Mozaffarian Dariush , Willett Walter C , Blumenthal Susan J 
Published: June 2021

The Supplemental Nutrition Assistance Program (SNAP) is the national nutrition safety net for low‐income families. Few studies have thoroughly examined dietary differences by children's SNAP participation status. We examined if SNAP participation was associated with weight and dietary outcomes in low‐income children. Using 1999–2008 NHANES, the study included 5,730 children aged 4–19 with household incomes <130% of federal poverty level. Diet was measured using 1–2 24‐hour recalls. No significant associations were observed between SNAP participation and weight status among all children (OR, obesity, 0.94, 95% CI 0.67, 1.32), or within age, sex and food security groups. For SNAP and non‐SNAP children, mean intakes of whole grains, fruits, vegetables, fiber, and potassium were ≤50% of the recommendations. Mean intakes of sugary beverages, saturated fat, sodium and iron were ≥150% of the recommendations. After multivariate adjustment, SNAP participants consumed 30% more vegetables (95% CI 4%, 62%), 34% more processed meats (95% CI 6%, 69%), 36% more high‐fat dairy (95% CI 13% 63%), and 63% more water (95% CI 14%, 31%) than nonparticipants. Additional dietary differences were observed among food‐insecure children only. SNAP participation was associated with a 5% lower Alternate Healthy Eating Index (95% CI 1%, 9%). Regardless of SNAP participation, the diets of low‐income children need improvement.Grant Funding Source : Robert Wood Johnson Foundation; Aetna Foundation; National Institutes of Health
Novel secreted maltase activity enables suckling mouse pup starch digestion
Authors: Nichols Buford Lee , Diaz-Sotomayor Maricela , Avery Stephen , Chacko Shaji , Hadsell Darrel , Baker Susan , Yan Like , Lin Amy (Hui-Mei) , Ao Zi-Hua , Quezada-Calvillo Roberto , Hamaker Bruce 
Published: June 2021

Starch requires 6 enzymes for digestion to free glucose: 2 amylases (salivary and pancreatic) and 4 mucosal maltase activities; Sucrase‐isomaltase (Si) and Maltase‐glucoamylase complexes (Mgam). All 6 activities are deficient in suckling rodents.Objective: Test 13C‐starch digestion before weaning (at 21d) from mother's milk by measuring enrichment of blood 13C‐glucose in Mgam Null and Wild Type mice.MethodsMgam gene was ablated at the N‐terminal expressing the membrane binding domain. Dams were fed low 13C‐diet and litters kept on low 13C‐diet. Pups were weaned at 21d. Digestion was tested at 13d and 25d by intragastric feeding of amylase predigested UL 13C‐α‐limit dextrins (LDx). Blood 13C‐glucose enrichment was measured by GCRMS using pentaacetate derivatives.Results4 h after feeding, blood 13C‐glucose MPE% was enriched by 26×103 in null and 18×103 foldat 13d and 0.3×103 and 0.2×103 fold at 25d (vs. fasting p = 0.045 and p = 0.045). By jejunal enzyme assay, imunohistochemistry or Western blots there was no maltase activity or brush border staining with anti‐Si or Mgam antibodies at 13d but these were fully developed in WT by 25d. In 13d null mice lumenal contents were stained by both specific antibodies.Conclusion1. 13C‐LDx was rapidly digested to 13C‐glucose in 13d mice independent of age, Mgam genotype or mucosal maltase activity. 2. This novel observation demonstrates that a maltase activity is secreted which enables suckling pup starch digestion well before brush border enzyme development. 3. The origin of this secreted starch digesting activity is presently uncertain.
The metabolic effects of a high fructose versus a high glucose diet in healthy overweight men
Authors: Johnston Richard David , Stephenson Mary , Crossland Hannah , Cordon Sally , Taylor Moira , Aithal Guruprasad P , Macdonald Ian 
Published: June 2021

This study assessed the effects of high glucose and fructose intakes, both at energy balance and with overfeeding, on liver and muscle lipid contents and other metabolic parameters.32 healthy, centrally overweight men (age 34 yrs and BMI 29.4kg/m2) were randomised to 2 periods each of 2 weeks of either a high fructose or high glucose intake (25% energy). Groups were well matched at study entry. In the first 2 weeks, isoenergetic status was maintained by providing all foodstuffs, followed by a 6 week washout. The second 2 week period was an ad libitum overfeeding period.

Period
Fructose
Glucose
Sig between groups

Mean
SD
Mean
SD

Weight (kg)
Isoenergetic
−0.26
0.9
−0.12
0.7
0.62

Overfeeding
1.03*
1.4
0.57 *
1.0
0.29

Hepatic lipid (%)
Isoenergetic
0.30
2.2
−0.05
2.1
0.65

Overfeeding
1.70*
2.6
2.05*
2.9
0.73

ALT (U/L)
Isoenergetic
−4.0
7.9
−2.9
6.5
0.67

Overfeeding
*5.8
8.7
4.1
9.8
0.62

Triglyceride (mg/dL)
Isoenergetic
−6.1
35.1
11.4
61.4
0.35

Overfeeding
31.6
70.2
28.9**
35.1
0.91

Absolute changes during each 2 week period
=p<0.05,
=p<0.01 compared to that group's baseline

There were no changes during the isoenergetic period and similar changes in each group during overfeeding. There were no inter‐group differences in intramyocellular lipid, systemic or hepatic insulin resistance or haemodynamic parameters. Changes during the overfeeding period were strongly associated with weight changes. Hence these were an energy, as opposed to a nutrient, specific effect. There was no evidence for specific, potentially deleterious effects of fructose compared to glucose.Clinical trials registry: NCT01050140Financial support: CORE charity UK
Advertising for food and dietary supplements in the print media in South Africa
Authors: Temple Norman Joseph , Mchiza Zandile , Abrahams Zulfa , Steyn Nelia P 
Published: June 2021

ObjectiveTo survey advertisements for food and beverages, slimming products, and dietary supplements in South African magazines.MethodsWe examined the 5 most popular magazines over one year. They are in 3 major languages (English, Afrikaans, and Xhosa).ResultsWe recorded 959 advertisements; 776 (80.9%) were for food and beverages, 86 (9.0%) for slimming products, and 97 (10.1%) for dietary supplements. The most common advertisements for food and beverages were for restaurants (n=126), supermarkets (119), alcoholic beverages (79), cereal products (59), confectionery items (59), vegetables (40), and fruit or fruit juices (29). Thirty‐nine of them (4.1%) carried health claims; these were most common for dairy products (14), followed by non‐alcoholic beverages (7), and meat and poultry (6). The most commonly mentioned health benefits were for healthy growth (18), performance claims (13), therapeutic claims (9), and non‐serious risk reduction claims (6), and for and energy (6). The majority of health claims referred to iron (17), B vitamins (12), vitamins and minerals (11), essential fatty acids (11), and calcium (9).ConclusionA fraction of advertisements were for healthy products (such as vegetables, fruit, and low‐fat milk) but the large majority were for unhealthy foods. Misleading advertisements for slimming products and dietary supplements are becoming common.
Allergen‐derived proteinases: Isolation, characterization and signaling via proteinase‐activated receptors (PARs)
Authors: Polley Danny , Mihara Koichiro , Saifeddine Mahmoud , Renaux Bernard , Vliagoftis Harrisios , Boitano Scott , Daines Michael , Hollenberg Morley D. 
Published: June 2021

ObjectiveTo identify proteolytic enzymes in insect and mould asthma allergens and assess their ability to signal via the PARs.MethodsAllergen‐derived trypsin‐like enzymes known to contribute to lung inflammation were visualized (western blot) with serine proteinase activity‐based probes (ABP). Enzymes were purified using ion‐exchange chromatography and analyzed for their ABP reactivity & substrate‐inhibitor kinetic profiles. Each enzyme was tested for its activation of PAR–dependent calcium signaling in PAR1 & PAR2‐expressing KNRK cells. ABP‐labeled enzymes were isolated by avidin affinity chromatography and their peptide sequences determined (mass spectrometry).ResultsBoth allergens contained ABP‐labeled proteinases with MWs of 20–26 kDa. Each enzyme had distinct kinetic substrate (Km) and inhibitor (Ki) profiles. The enzymes all caused PAR2 calcium signaling, and differentially activated PAR1. Sequencing revealed that the allergen proteinases were homologous with previously described trypsin‐like enzymes.ConclusionsDiverse allergens contain multiple biochemically distinct trypsin‐like proteinases that are able to target PARs 1 and 2. Thus, PARs and their activating allergen proteinases may be attractive therapeutic targets for the treatment of allergen‐induced asthma.Support: Canadian Institutes of Health Research, NIH USA and Lung Association of Alberta & NWT
β‐arrestin2 and ARRDC proteins have distinct roles in β2AR trafficking and signaling
Authors: Han Sang-oh , Kommaddi Reddy Peera , Venkataramanan Vidya , Shenoy Sudha K 
Published: June 2021

ObjectiveTo investigate the roles of the 7TMR adaptor β‐arrestin2 (βarr2) and the related arrestin domain containing α‐arrestins (ARRDC 2, 3, and 4) in β2 adrenergic receptor (β2AR) trafficking and signaling.Resultsβarr2 binding to activated β2ARs is rapid and detected at the plasma‐membrane, whereas GFP‐tagged ARRDC binding is delayed and detected on endosomes. Gene‐silencing of βarr2, but not of ARRDC3 blocks internalization, MAP Kinase activation, receptor ubiquitination and lysosomal trafficking of activated β2AR. βarr2 binds clathrin, AP‐2 and pERK robustly upon agonist‐activation of the β2AR whereas ARRDC3 does not. ARRDC3 has canonical poly‐proline domains, which are binding regions for WW‐domain containing E3 ubiquitin ligase Nedd4 that ubiquitinates agonist‐activated β2AR. Mutations within the poly‐proline domains in ARRDC3 not only ablate its binding with Nedd4, but also prevent its localization on endosomes and eliminate the interaction with internalized β2AR.ConclusionsThe role of βarr2 in β2AR internalization, ubiquitination, lysosomal trafficking and ERK signaling cannot be substituted by ARRDC3. The apparent association of ARRDC3 with the β2AR results from ARRDC3/Nedd4 primarily and Nedd4/β2AR association secondarily. ARRDC2 or ARRDC4 can substitute for ARRDC3 in associating with Nedd4 during β2AR post‐endocytic trafficking. (Supported by HL 080525 to SKS).
ROS Sensitive Calcium Channel TRPM2 Regulates VEGF Induced Angiogenesis
Authors: Mittal Manish , Malik Asrar B 
Published: June 2021

TRPM2 is a cation channel involved in calcium influx in response to cellular oxidative stress. We hypothesized that TRPM2 regulates VEGF induced vascular angiogenesis secondary to ROS production. We show that TRPM2 knockdown suppressed VEGF induced human lung endothelial cell (hLEC) hyper‐permeability, which is essential for angiogenesis, as measured by trans‐endothelial resistance and albumin influx (EBA). VEGF induced phosphorylation of VE cadherin at Tyr658 residue was inhibited by TRPM2 knockdown. Additionally, VEGF stimulation increased ROS production in hLEC, and application of apocynin (ROS scavenger) inhibited VEGF induced phosphorylation of Tyr658 of VE cadherin. TRPM2 knockdown reduced VEGF induced calcium entry in hLEC, and delayed migration of HUVEC. VEGF induced angiogenesis measured by in vivo matrigel plug assay was markedly reduced in TRPM2 KO mice compared to wild type. We conclude that calcium influx from ROS activated TRPM2 channels on VEGF stimulation is a key mechanism signaling angiogenesis.Research support: NIH grants RO1 HL45638‐23 and P01 HL077806
Long‐term effects of indocyanine green on lymphatic pump function
Authors: Weiler Michael , Dixon J. Brandon 
Published: June 2021

IntroductionNear‐infrared (NIR) imaging with indocyanine green (ICG) has emerged as a potential technique for diagnosing lymphatic diseases, however, the long‐term effects of ICG on lymphatic function have not been studied in vivo. Therefore, we quantified the retention rate of ICG in lymphatic vessels and the effect of repeated ICG injections on lymphatic pump function.MethodsRats were given intradermal ICG injections and subsequently imaged using NIR to quantify lymphatic function as members of one of four treatment groups: (i) initial ICG injection with no follow‐up injection, (2) follow‐up ICG injection after 1 week, (3) 2 week follow‐up ICG injection, and (4) 1 month follow‐up ICG injection.Results and ConclusionsICG remained visible in rat collecting lymphatics for approximately two weeks, and NIR lymphatic function measurements after follow‐up ICG injections of 1 and 2 weeks showed significant reductions in fluid velocity, pulsatile flow frequency and clearance rate while no significant differences were observed after 1 month follow‐up ICG injections. These results indicate that ICG is retained in the local tissue space for an extended period and may suggest that lymphatic function is reduced during this time. ICG‐based NIR imaging diagnostics for lymphatic function, therefore, may necessitate a minimum of 1 month between subsequent ICG injections to avoid erroneous measurements of function.
Resolving inflammation in stroke through FPR2/ALX
Authors: smith helen k , gavins felicity 
Published: June 2021

Ischaemia/reperfusion (I/R) injury in stroke can worsen patient outcome through excess inflammation. We investigated the pharmacological potential of an endogenous anti‐inflammatory mechanism, formyl peptide receptor 2/lipoxin receptor (FPR2/ALX; Fpr2/Fpr3 in mouse), in reducing I/R‐associated inflammation in a mouse model of stroke.Mice underwent 5 min global cerebral ischaemia, followed by 40 min or 2 h reperfusion. FPR agonists—Ac2‐26 (Annexin A1 pharmacophore) and 15‐epi‐lipoxin A4—were administered at the start of reperfusion. Subsequent leukocyte‐endothelial (L‐E) interactions (characteristic of an inflammatory response) were quantified through a cranial window using intravital video microscopy.15‐epi‐lipoxin A4 administration dramatically reduced L‐E interactions at 40 min reperfusion but less so at 2 h. Ac2‐26 treatment decreased leukocyte adhesion at 40 min and all L‐E interactions at 2 h. Combined treatment with Ac2‐26 plus FPR antagonists Boc2 or WRW4 (FPR2/ALX‐selective) abrogated the effects of Ac2‐26.We show that activity through Fpr2/Fpr3 reduces L‐E interactions in a model of global cerebral I/R. These data provide evidence that FPR2/ALX could be a therapeutic target in stroke.This was funded by the BBSRC.
Forty five minutes of ischemia and 24 h of reperfusion in the Wistar rat induce an acute pre‐renal injury which is not improved by 4 days of pre‐ischemia treatment with Spironolactone
Authors: Acuña Dulce Mariela Montes , López-Rodríguez Juan Francisco , Calvo-Turrubiartes Miriam Zaharí , Varela Norma Leticia Dávila , LLamazares-Azuara Lilia , Rodríguez-Martínez Manuel 
Published: June 2021

To characterize what kind of renal injury is induced by 45 min of ischemia and 24 h of reperfusion (I/R) and assess the effect of 4 days of pre‐ischemia treatment with Spironolactone (Sp = 20 mg/kg/day/o.g.gavage) or Vehicle (V = 560μl/day/o.g. gavage) on this I/R model, Wistar male rats (~250g of bw) were random assigned to three groups: V+Sham‐I/R, V+Real‐I/R & Sp+Real‐I/R and the following parameters were measured: glomerular filtration rate (GFR = CCr = ml/min/100g), urinary protein excretion (UPrV = mg/24h/100g), fractional excretion of sodium (FENa = %), free water clearance (CH20 = ml/min/100g).

n
GFR
UPrV
FENa
CH2O

V+Sham‐I/R
8
0.51±0.04
7.2±0.6
2.1±0.8
−2.0±3.3

V+Real‐I/R
8
0.28±0.03#
16.2±3.0°
2.7±1.2
−3.4±3.0

Sp+Real‐I/R
8
0.12±0.03&
17.3±3.0°
15.7±5.5°*
−0.7±4.0

Values are mean ± SEM;
°p< 0.05,
p=0.01,
p=0.0001 vs. V+Sham‐I/R;
p<0.05,
p<0.005 vs. V+Real‐I/R

There were no differences among groups in 24h urinary nitrogen oxides excretion, renal malondialdehyde content or baseline renal superoxide production either.Conclusions45 min of ischemia and 24h of reperfusion in the Wistar rat induce an acute pre‐renal injury, which unexpectedly was not improved by 4 days of pre‐ischemia treatment with spironolactone. Supported by CO9‐FAI‐03‐20.20.