bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of mineralized water consumption on postprandial lipemia
Authors: Zaïr Yassine , Housez Béatrice , Pichelin Matthieu , Raoux François , Cazaubiel Murielle , Ouguerram Khadija
Published: June 2021

Dietary factors may affect lipid metabolism and thus impact cardiovascular disease development. The objective of this study was to analyze the effects of 8 wk ingestion of mineralized water compared with control on fasting and postprandial lipoproteins levels in hyperlipidemic men. Subjects (N=12) consumed mineralized water (Saint Yorre) or low mineralized Water (Ogeu) in random order for 8‐wk periods, separated by a 1‐wk washout. Lipoprotein lipids were assessed prior to (fasting) and following a standard meal for 8 h (postprandial). Fasting and postprandial plasma triglycerides (TG) were decreased following mineralized water compared to control (p<0.01 and <0.05, respectively). This decrease was related to a significant decrease in fasting and postprandial levels of very low‐density lipoprotein (VLDL) TG (respectively p<0.01 and p<0.05) and a slight decrease in fasting total cholesterol (TC) (p<0.1). High‐density lipoprotein cholesterol (HDL‐C) increased postprandially (p<0.05) following the mineralized water compared to the low mineralized water. The reduction in postprandial concentrations of VLDL TG and the increase in HDL‐C suggest improvements in liver lipid metabolism and postprandial lipemia, which may favorably affect cardiovascular health.
Effect of ProAlgaZyme on plasma lipid profile in diet induced hypercholesterolemic hamsters
Authors: Geamanu Andreea , Saadat Nadia , Goja Arvind , Wadehra Monika , Gupta Smiti
Published: June 2021

Increased plasma LDLc and low HDLc are associated with risk of cardiovascular disease (CVD), the leading cause of death worldwide. This study investigated the effect of an algae infusion, ProAlgaZyme(P) and its sub‐fractions on plasma lipids in hamster model. Sixty (n=10/group) male Golden Syrian hamsters (8 w old) fed a high fat diet (30% calories from fat) were randomized into 6 groups: control, P and its subfractions P1, P2, P3, and P4. The infusion (20% v/v) was added to the drinking water of all experimental groups, while the control animals received water. Total cholesterol (TC), HDLc, phospholipids, and cholesterol ester levels were assayed enzymatically after 4 weeks. LDLc was significantly reduced in P (p<0.05), P3 and P4 (p<0.001) as compared to the control, while HDL was considerably increased in P, P3 and P4 groups (p<0.001). Moreover, the TC/HDLc ratio was significantly lower in all experimental animals (P, P1, P2 p<0.05; P3, P4 p<0.001). The shift in plasma cholesterol towards the higher density fractions was validated by density gradient ultracentifugation. Also, the HDL diameter was significantly increased in P3 and P4, as compared with control (p<0.05). This has been associated with protection against CVD. Thus, intake of ProAlgaZyme and its sub‐fractions significantly improved plasma lipid profile by lowering LDLc and increasing HDLc. This may potentially provide benefits in cardiovascular health.

Grant Funding Source : Health Enhancement Products Inc.
Oxidation of U‐13C‐α‐linolenic acid is independent of body weight in pigs fed flaxseed diets
Authors: Martinez-Ramirez Hector Ramon , Kramer J. K.G. , de Lange C. F.M.
Published: June 2021

Twelve pigs (14.6±0.41 kg initial body weight, BW; 4 pigs per treatment) were used to assess the effect of BW and cumulative intake of flaxseed (FS) on the oxidation of α‐linolenic acid (ALA). Pigs were fed FS free (CON) or FS diets according to three feeding regimes: R1, 10% FS diet from 15 to 37 kg BW; and then 5% FS diet; R2, CON from 15 to 37 kg BW; and then 5% FS diet; and R3, 10% FS diet form 15 to 37 kg BW; and then CON. At 25 and 55 kg BW, a single bolus dose of U‐13C‐ALA (2.75 mg/kg BW) was fed and recovery of 13C in expired CO2 was quantified during 26 h to estimate oxidation. There were no interactive effects (P>0.10) of feeding regime and BW on ALA oxidation. There was no effect of previous FS intake on ALA oxidation at 55 kg BW (P=0.20; 8.0% for R1 and 4.9% for R2). Across BW, R1 tended to have higher mean ALA oxidation than R2 (P=0.06; 9.8 vs 5.1%), whereas mean oxidation was similar for R2 and R3 (P=0.94; 5.1 vs 5.7%). BW did not affect the oxidation of ALA (P=0.42; 7.0 at 25 kg and 5.6% at 55 kg BW); pigs on FS tended to have higher ALA oxidation than pigs on CON (P=0.07; 8.0 vs 4.7%). ALA oxidation was generally less than 10% of intake, not affected by BW and previous nutrition, and reduced when feeding low ALA diets.

Grant Funding Source : Ontario Pork, OMAFRA, AAFC
EPA and DHA increase insulin receptor in proliferating but reduce GLUT4 in differentiated C2C12
Authors: Kim Jeffrey , Li Yong , Watkins Bruce A
Published: June 2021

Endocannabinoids (EC) are messengers derived from polyunsaturated fatty acids (PUFA) that play a role in disrupting glucose regulation in skeletal muscle. The objective of our research is to study the mechanistic aspects of glucose regulation and the EC system in the mouse myoblast cell line, C2C12, when supplemented with and without n‐3 and n‐6 PUFA. C2C12 myoblasts were cultured until a confluency of 50%. Cells were assigned to two groups: one being maintained in the proliferating state while the other was induced to differentiate. Both groups were then treated with 100μM of either eicosapenaenoic acid (EPA), docosahexaenoic acid (DHA), arachidonic acid (AA), or a bovine serum albumin (BSA) control for 24 hours. RNA was isolated and analyzed using real time PCR revealing an increase in the mRNA of the insulin receptor in both the EPA and DHA treated proliferating cells compared to BSA and AA groups, but decreased in differentiated cells. In addition, glucose uptake transporter 4 mRNA was reduced in the differentiated EPA and DHA groups while Akt1 expression was reduced in only the differentiated DHA group. The mRNA for the primary EC receptor in skeletal muscle, CB1, was found to be less than in the n‐3 PUFA treated proliferating cells. Changes in EC synthesizing and degrading enzymes complement this observed reduction in structural and key mechanistic components of the EC system when treated with n‐3 PUFA.

Grant Funding Source : Diet and Health Initiative (UConn)
Twice‐weekly consumption of farmed Atlantic salmon increases plasma content of phospholipid n‐3 fatty acids
Authors: Picklo Matthew J , Rosenberger Thad A , Burr Gary S , Wolters William R , Raatz Susan K
Published: June 2021

Elevated intake of the n‐3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), is related to risk reduction of cardiovascular and other diseases. Consumption of seafood such as farmed Atlantic salmon is an effective way to consume n‐3 but there is a paucity of data as to how much to eat. The goal of this work was to evaluate the effect of salmon consumption on increasing n‐3 in the plasma phospholipids fatty acids (PLFAs), a surrogate marker for lipid membrane fatty acids. In this randomized, cross‐over designed study, 17 healthy volunteers were fed portions of baked salmon (90 g, 180 g, 270 g raw weight) twice weekly for 4 weeks with washouts of 4–6 weeks. PLFA content was determined at the beginning and end of each treatment. EPA increased significantly in a portion‐dependent manner with no effect observed with the 90 g portion, a doubling of EPA content in the 180 g consumption and a nearly three‐fold increase at the 270 g level. DHA content in PLFA was elevated similarly (approximately 50%) without respect to portion size. Arachidonic acid was reduced (~10%) without respect to portion size with no changes in linoleic acid. Further, the n6/n3 ratio decreased and total n‐3 content increased in a portion dependent manner. These results indicate that the PLFA content of n‐3 fatty acids are highly responsive to the consumption of farmed Atlantic salmon. USDA 54505100004800D, 19153100000300D NIH 2P20RR01769909.
Exogenous modification of platelet membranes with the omega‐3 fatty acids DHA and EPA impairs thrombogenesis
Authors: Ensz Nicole M , Hjelmen Carl E , McComas Leah S , Tormoen Garth W , Patel Ishan A , McCarty Owen J.T. , Larson Mark K
Published: June 2021

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are associated with lower risk for adverse cardiovascular events. EPA and DHA‐enhanced platelets exhibit relatively normal platelet aggregation characteristics, suggesting that cardioprotective mechanisms are not inherent to platelet reactivity. We hypothesize that EPA and DHA incorporation hinders platelet‐mediated thrombin production. To this end, platelet membranes were modified by exogenous addition of EPA and DHA to whole blood or platelet‐rich plasma followed by thrombin generation assays. Whole blood treatment inhibited occlusive thrombus formation when perfused over collagen‐coated surfaces compared to matched vehicle controls. EPA and DHA‐treated platelets also showed a kinetic impairment of thrombin generation as measured by both thrombin‐specific substrate conversion and thrombin precursor protein levels in the presence of Factor Xa and prothrombin. A functional deficiency in thrombin production was observed as EPA and DHA‐treated blood formed more diffuse thrombi and less fibrin versus matched controls after perfusion across collagen‐coated surfaces. These results strongly suggest that EPA and DHA impair platelet‐mediated thrombin generation, providing a unique cellular mechanism behind the cardioprotective phenotype seen after increased omega‐3 fatty acid intake. Funded by NIH NCRR 2 P20 RR016479.
Single molecule investigation of Rad51 and Srs2
Authors: Myong Sua , Qiu Yupeng , Antony Edwin , lohman Timothy
Published: June 2021

DNA double strand break (DSB) is the principal cytotoxic damage from ionizing radiation, chemical reagents and the cellular metabolic processes such as DNA replication. Unrepaired DSBs give rise to harmful mutations and chromosome rearrangements which lead to carcinogenesis. The primary repair pathway for DSB is homologous recombination, which is mediated by the Rad51 recombinase. This recombinase activity is counter‐balanced by anti‐recombinases, which are DNA helicases that remove the recombinase from DNA. We investigated the molecular mechanism involved in the beginning steps of recombination and anti‐recombination using highly sensitive single molecule fluorescence assays. We present our result on the (1) detection of the initiation steps involved in Rad51 filament formation on single stranded DNA, (2) mechanism of the ATP fueled translocation activity of anti‐recombinase, Srs2 by which it disrupts the Rad51 filament and (3) the role of Rad51 in anti‐recombinase mediated unwinding activity.
Chemical characteristics and antioxidant activity of the laver (Porphyra tenera) grown in southwestern seashore of Korea
Authors: Lee Kyongae , Shin Malshik , Oh Soojung , Choe Eunok
Published: June 2021

This study determined the chemical characteristics (proximate composition, lipid and fatty acid composition, pigments and antioxidants) of the laver, and antioxidant activity of methanol‐, water‐, ethanol‐ and methanol‐water (1:1, v/v) extracts of the laver. The laver consisted of moisture, protein, lipid, carbohydrates, and ash at 11.6, 38.4, 0.31, 42.7, and 7.95%, respectively. The laver lipids were consisted of triacylglycerols, 1,2‐diacylglycerols, 1,3‐ diacylglycerols, monoacylglycerols and free fatty acids at 36.1,9.1, 8.7, 20.3, and 25.8%, respectively. Major fatty acid was eicosapentaenoic acid (EPA, 54.4%), with omega‐3 fatty acids totally at 56.6%. Contents of chlorophyll, phycocyanin, phycoerythrin, polyphenols and α‐tocopherol were 392.7, 2,100, 6,930, 8,670, and 0.2 mg/kg, respectively. DPPH radical scavenging activity and reducing power were the highest in the methanol‐water extract of the laver, 93–94 and 75–78%, respectively, of control α‐tocopherol and L‐ascorbic acid. No nitrite scavenging ability was observed in the laver extracts. The results clearly showed the dried laver was a good source of dietary fiber and polyphenols, which could contribute to its high antioxidant activity.
Oleanolic acid retards resistin induction in differentiating adipocytes: Role of Tyk2‐Stat signaling
Authors: Kim Hyun-Sung , Kang Young-Hee
Published: June 2021

Oleanolic acid (OA), a naturally occurring triterpenoid widely distributed in foods and medicinal plants, has anti‐cancer, antioxidant, and anti‐aging properties. This study elucidated anti‐obesity actions of OA in terms of production of inflammatory adipokines of resistin during adipogenic differentiation. 3T3‐L1 adipocytes were cultured in adipogenic DMEM containing MDI and insulin for 7 d in the absence and presence of 1–25 μM OA. Production of the inflammatory adipokine resistin was markedly enhanced during differentiation, which was dampened by treating OA. Stat1 and Stat3 were activated during differentiation with disparate temporal fashions. Stat1 was phosphorylated 5 d after initiating differentiation, whereas Stat3 was rapidly activated within 1 d. When 25 μM OA was supplied to differentiating adipocytes, Both Stat1 and Stat3 phosphorylation was substantially suppressed. The upstream Tyk2 was rapidly activated in a similar manner to Stat3, which was blocked by incubating adipocytes with 25 μM OA. Taken together, OA suppressed adipocyte differentiation‐associated production of resistin through disturbing Tyk2‐Stat1/3 signaling. Therefore, OA can be a potential agent blunting obesity and related inflammatory diseases.
GLUT‐1 transporter expression is up‐regulated on human CD4+ and CD8+ T lymphocytes following T cell activation
Authors: MENG HUICUI , TURBITT WILLIAM J , ROGERS CONNIE J
Published: June 2021

The availability and uptake of nutrients play a critical role in the generation of adequate immune responses. In particular, the metabolic demands of T lymphocytes increase following activation to provide energy for proliferation and other important effector functions. Glucose, one of the major energy sources of T cells, can be transported into cells via several glucose transporters, including GLUT‐1. However, the distribution of GLUT‐1 on T cell subsets remains unclear. Therefore, the goal of the current study was to evaluate the effect of T cell activation on GLUT‐1 expression in CD4+ and CD8+ T cells. Peripheral blood was collected from healthy subjects (20–40 years old), PBMCs were isolated using Ficoll‐Hypaque Plus, and T cells were stimulated using 1 μg/ml of anti‐CD3/CD28 antibodies. Following 3‐day cultures, T cells were harvested, counted and GLUT‐1 expression was quantified on both unactivated and activated CD4+ and CD8+ T cells by flow cytometry. GLUT‐1 expression increased significantly, on CD4+and CD8+ T cells (20‐fold and 10‐fold, respectively; P<0.05) following activation. Our results demonstrate that CD4+ and CD8+ T cells up‐regulate the surface expression of GLUT‐1 upon activation. These results suggest that increased glucose uptake via the GLUT‐1 transporter is a common feature of all activated T cells and may be a target for therapy to enhance T cell function.

Grant Funding Source: Penn State internal funding
Dietary spray‐dried plasma influences intestinal morphology, growth performance, and organ weight of mated female mice under transport stress
Authors: Song Minho , Lee Jeongjae , Liu Yanhong , Soares-Almeida Juliana A. , Che Tung M. , Campbell Joy M. , Polo Javier , Crenshaw Joe D. , Chun Ju Lan , Seo Seongwon , Pettigrew James E.
Published: June 2021

Spray‐dried plasma (SDP) enhances gut barrier function and our previous data showed it attenuated the negative effects of inflammation caused by transport stress on pregnancy rate. We hypothesized that SDP can influence gut morphology, growth, and organ weight of mated female mice under transport stress. Mice (C57BL/6 strain) were shipped from Bar Harbor, ME to Urbana, IL on the day the vaginal plug was found [gestation day (GD) 1], arriving at the laboratory in IL on GD 3. Mice were randomly assigned to diets [0 (CON), 1, or 8% SDP]. Randomly selected mice were euthanized each day from GD 3 to 7 to collect duodenum, jejunum, ileum, and colon samples for gut morphology, and liver, spleen, kidneys, and lungs for organ weights, and to measure body growth rate. SDP increased (P < 0.05) ratio between villus height and crypt depth of all small intestinal sections (12–38%) by increasing villus height (4–22%) and decreasing crypt depth (5–11%), and increased (P < 0.05) crypt depth of colon (14–20%), compared with CON. SDP increased (P < 0.05) number of goblet cells in all intestinal sections (15–35%) compared with CON. SDP increased (P < 0.05) growth rate (21–33%) and liver weight (9–12%) compared with CON. No difference was found between 1% and 8% SDP. These data support the hypothesis that SDP influences intestinal morphology, growth, and organ weight of mated female mice under transport stress.
Differences and similarities in feeding behaviors between Guatemalan mothers and their children
Authors: Gatica Giovanna Laura María , Ramírez-Zea Manuel , Letona Paola , Caballero Benjamin
Published: June 2021

Objective

To identify feeding behaviors in mothers and their offspring.

Methods

Mothers and children (4th–6th grade) were recruited (91 pairs) in low‐income urban schools from Guatemala City. A validated 24‐hr food list, which allows assessing food frequency at each meal, was administered to each mother and child in 3 days of the same week. A Monday was included in all cases, to get data from the previous Sunday. Overweight/obesity classification for mothers and children were made according to WHO standards.

Results

The ten food items most consumed by mothers and children differed just in one food item (snacks was ranked 6th place in children, while boiled black‐beans was ranked 10th in mothers). Taking into account food items that were consumed at least once a week (31 for the mothers and 36 for the children), principal component analysis showed different food patterns between the mother and her child. Girls consumed more non‐fried vegetables, fruit, and fruit juice while boys consumed more artificial juice. Mothers with normal BMI consumed more sweet bread compared to overweight/obese mothers, who consumed more soda.

Conclusions

Daily food consumption between mothers and their children were similar and the variability was explained by particular food patterns that included similar and different foods.

Grant Funding Source: NHLBI HHSN 268200900028C
Age of introduction of ten sentinel complementary foods as reported for 6 to 12 month old, low‐income infants in metropolitan, Quetzaltenango (Western Highlands), compared to Guatemala City and Santo Domingo Xenacoj (Central Highlands)
Authors: Vossenaar Marieke , Campos Raquel , Hernandez Liza , Solomons Noel W
Published: June 2021

Background

The WHO recommends exclusive breastfeeding (EBF) during the first 6 mo of life. Evidence complied in earlier studies of CeSSIAM documented that mixed feeding often occurs earlier.

Objective

To compare the pattern of age of introduction of commonly consumed foods in the western highlands with what has been reported from the central highlands of Guatemala.

Methods

Age of introduction of 10 sentinel foods was queried by means of structured face‐to‐face interviews in 6 to 11 mo old infants living in 2 urban areas (Quetzaltenango, n=128 and Guatemala City, n=64) and 1 rural area (Santo Domingo Xenacoj, n=64).

Results

Items most frequently reported as introduced before 6 mo in Quetzaltenango were vegetables (28%), followed by fruits (27%), Incaparina® (23%), potatoes (20%), and sweet rolls (17%). These rates of premature introduction are higher than reported in a poor neighborhood in Guatemala City with a respective: fruits (18%); vegetables (17%); potatoes (14%); Incaparina® (13%); and rice (11%). The hierarchy in a Mayan village was: potatoes (15%); vegetables (13%); Incaparina® (13%); rice (12%); and oatmeal (11%).

Conclusion

Different patterns of early introduction of food and beverage items contribute to the precocious occurrence of mixed feeding in the first semester of infancy in distinct regions of Guatemala.

Funded by the Nestlé Foundation, Switzerland
Relations between iron status and cognitive measures in Indian adolescents
Authors: Scott Samuel Pew , Wenger Michael J. , Murray-Kolb Laura E. , Udipi Shobha A. , Ghugre Padmini S. , Boy Erick , Haas Jere D.
Published: June 2021

Iron deficiency (ID) is the most common nutrient deficiency, affecting an estimated 25% of the global population. However, the functional effects of ID in children between the ages of 12–16 years, a period of life during which behavior and cognition continue to develop, are largely understudied. In the current 7‐month double‐blind intervention, pearl millet biofortified with iron was tested for its efficacy to improve iron status in rural Maharashtrian children (n=247; 12–16 y; female=73, male=174). Here, we present analysis of baseline data. A random subsample (n=150) underwent testing of cognitive performance using 5 measures to evaluate simple reaction time, response inhibition, low‐ and high‐level attentional control, semantic memory, and recognition memory. Blood measures of iron status included hemoglobin, ferritin and transferrin receptor. Inflammation was assessed via C‐reactive protein and alpha‐1‐acid glycoprotein. Relations between iron status and performance on cognitive tasks were examined.
Weight status in HIV‐positive women after termination of a drug and lipid‐based nutrient supplement intervention is predicted by food availability and health
Authors: Jordan-Bell Elizabeth , Adair Linda , Flax Valerie , Chasela Charles , Kayira Dumbani , Daza Eric , Tembo Martin , Chitsulo Phindile , Jamieson Denise , van der Horst Charles , Bentley Margaret
Published: June 2021

This study examined weight status in HIV‐positive Malawian mothers after they completed an antiretroviral (ARV) and nutritional supplement intervention. Women in the Breastfeeding, Antiretrovirals, and Nutrition study (n=2369) received either lipid‐based nutrient supplements (LNS) to prevent maternal depletion or no LNS and were further randomized to receive ARVs to prevent HIV transmission or no ARVs from delivery through 24 wks of exclusive breastfeeding and 4 wks of rapid weaning. All women received a 2 kg/wk family maize supplement and enhanced standard of care for 48 wks. Women on ARVs entered rapid weaning with a lower mean BMI than the other groups and had negligible weight change (<0.3 kg) in the subsequent 24 wks, while women in the LNS, no ARVs group lost weight (−1.0 kg) post‐supplementation. Termination of ARVs and LNS had no serious consequences for maternal weight status: <4% of women lost >10% weight from wks 24 to 48. A longitudinal random effects model controlling for 24 wk BMI showed that vomiting (p<0.01), diarrhea (p<0.01), oral thrush (p<0.05), hungry season (p<0.01), and famine year of 2005 (p=0.02) were negatively associated with weight, while CD4 count (p<0.01) was positively associated with weight. Thus, the key predictors of weight status in this population were food availability and health status.

Support: CDC (U48‐DP000059‐01); Gates Foundation (OPP53107); CPC (NICHD 5 R24 HD050924)
Bioavailable carotenoid and cyanogen content of biofortified cassava after food preparation
Authors: La Frano Michael R , LaPorte Delphine , Burri Betty J
Published: June 2021

Vitamin A deficiency is common where cassava is a dietary staple, because the diet contains few pro‐vitamin A carotenoids (pVAC) or preformed retinol. Biofortifying cassava so that it is rich in pVAC is an attractive alternative to food fortification or providing vitamin A supplements for preventing vitamin A deficiency, especially in rural areas that are difficult to reach by these interventions. We are investigating the effect of feeding single doses of carotenoid biofortified cassava, with or without oil, on the carotenoid and vitamin A concentrations of healthy adult women fed a low fat, low vitamin A, and otherwise low carotenoid diet. The biofortified variety of cassava contained 12 μg/g total carotenoid and 8 μg/g beta‐carotene, compared to 0.67 μg/g typical non‐biofortified cassava. All cassava, including biofortified varieties, contains cyanogenic glycosides that form cyanide when the plant is cut or processed. The biofortified cassava variety we obtained originally contained 282 ug/g (fresh weight) cyanogen, a harmful amount. Therefore it had to be detoxified before it could be used for human feeding trials, by methods that did not also destroy carotenoids. We explored the effects of repeated 2 hour soakings in distilled water on carotenoid and cyanogen concentrations on peeled, chopped, cassava. The cyanogen content decreased approximately 3X with each soaking, attaining the low level of 1.1 after the 4th soaking, Soaking, washing, and boiling for 30 min caused small (7 – 19%) decreases in carotenoid concentrations, Thus, soaking and boiling can effectively remove cyanogens while retaining carotenoids in cassava. This research was supported by HarvestPlus.
Palmitic acid induces but docosahexaenoic acid (DHA) inhibits inflammasome‐mediated secretion of IL‐1β in monocytes by modulating the activation of Toll‐like receptor 2 (TLR2)
Authors: Snodgrass Ryan G , Huang Shurong , Choi Il-Whan , Rutledge John C , Hwang Daniel
Published: June 2021

Activation of proinflammatory pathways in monocytes leads to enhanced inflammation in various tissues which in turn can promote the development of many chronic diseases. Unlike other cytokines, IL‐1β production is regulated at both transcriptional and posttranslational levels. TLR activation induces expression of pro‐ IL‐1β that needs to be processed by activated inflammasome to release bioactive IL‐1β. High saturated fat consumption is associated with increased postprandial inflammation. However the mechanism remains unclear. Thus, we determined whether 1) a major dietary saturated fatty acid palmitic acid (C16), as well as lipolysis products of postprandial triglyceride‐rich lipoproteins (TGRL) containing palmitic acid as the major fatty acid, induce IL‐1β expression and stimulate inflammasome‐mediated IL‐1β secretion in monocytes, 2) whether this process is TLR‐mediated and, 3) whether DHA inhibits this process. Results show that C16 and TGRL lipolysis products induce but DHA inhibits the expression and secretion of IL‐1β by reciprocally modulating TLR2 activation as assessed by recruitment of TLR2 and adaptor molecules into lipid rafts (faithful readout for direct activation of TLR2). Together, these results suggest that inflammasome‐mediated inflammatory responses in blood monocytes can be dynamically modulated by types of dietary fat we consume.

Grant Funding Source : USDA‐ARS WHNRC Program Funds and NIHDK 064007
Polymorphisms in regulator of protease B (RopB) alter disease phenotype and strain virulence of serotype M3 Group A Streptococcus
Authors: Olsen Randall J , Laucirica Daniel R , Watkins Melanie E , Feske Marsha L , Garcia-Bustillos Jesus R , Vu Chau , Cantu Concepcion , Shelburne Samuel A , Fittipaldi Nahuel , Kumaraswami Muthiah , Shea Patrick , Flores Anthony R , Beres Stephen B , Lovgren Maguerite , Tyrrell Gregory J , Efstratiou Androulla , Low Donald E , Van Beneden Chris A , Musser James M
Published: June 2021

Whole‐genome sequencing of serotype M3 group A Streptococcus (GAS) strains recovered from oropharyngeal and invasive infections in Ontario recently showed that the gene encoding regulator of protease B (RopB) is highly polymorphic in this population. To test the hypothesis that ropB is under diversifying selective pressure among all serotype M3 GAS, we sequenced this gene in 1,178 strains collected from different infection types, geographic regions and time periods. The results confirmed our hypothesis by identifying 84 distinct ropB alleles and discovered a significant association between ropB mutations, decreased secreted protease activity of its major regulatory target SpeB, and oropharyngeal infection. Additionally, compared to the parental wild‐type strain, isoallelic strains with ropB polymorphisms were significantly less virulent in mice. These studies provide a model strategy for applying whole‐genome sequencing followed by deep single‐gene sequencing to generate new insight to the rapid evolution and virulence regulation of human pathogens.
Whole Genome Sequence Analysis of a B. cereus Strain Causing a Fatal Anthrax‐Like Pulmonary Infection
Authors: Long S. Wesley , Wright Angela M , Beres Stephen B , Consamus Erin N , Flores Anthony R , Barrios Roberto , Richter G. Stefan , Oh So-Young , Garufi Gabriella , Maier Hannah , Drews Ashley L , Stockbauer Kathryn E , Cernoch Patricia , Schneewind Olaf , Olsen Randall J , Musser James M
Published: June 2021

A 39‐year‐old male presented with a rapidly progressive hemorrhagic anthrax‐like pneumonia. Within 3 days, the overwhelming systemic infection proved fatal. Microbial culture of antemortem and postmortem specimens grew pure cultures of Gram‐positive rods that were provisionally identified as Bacillus species. An autopsy was performed, and we characterized the genome of a strain isolated from the patient's blood within days using next‐generation sequencing technology. In addition, the genomes of sixteen strains recovered from different anatomic sites and time points were fully sequenced to assess intra‐host genetic variation. Whole genome analysis demonstrated that the fatal infection was caused by a previously unknown strain of B. cereus that was closely related to, but genetically distinct from, B anthracis. The exceptional virulence of this B. cereus strain was due in part to carriage of a pXO1‐like plasmid that encodes the tripartite anthrax toxin. The sequence data revealed the molecular determinants leading to the exceptional virulence of this organism, helped guide the institutional infection control response, and provided new insight to host‐pathogen interactions. This case exemplifies the value of autopsy and whole genome sequencing technologies to laboratory medicine.
Iron and copper in the pathogenesis of Friedreich's ataxia
Authors: Mazurkiewicz Joseph Edward , Koeppen Arnulf H , Yu Devin , Ramirez Liane , Qian Jiang , Parsons Patrick J , Yang Karl , Chen Zewu , Feustel Paul J
Published: June 2021

Friedreich's ataxia (FRDA) causes progressive atrophy of the large neurons of the dentate nucleus (DN). Possible contributing factors to FRDA pathogenesis are the high iron (Fe) concentration in the DN and failure to clear the metal from the damaged tissue. Fe, copper (Cu), and zinc (Zn) content in the DN was imaged and quantified by X‐ray fluorescence of PEG‐embedded tissue from 10 FRDA patients and 13 controls. In normal DN, Fe occurred predominantly in central white matter and hilus whereas Cu and Zn were present at peak levels in the gray matter. The location of maximum Fe signal did not change in FRDA DN, but the Cu region broadened overlapping extensively with the Fe‐rich region. Maximal metal concentrations did not change. The most abundant immunochemical reaction (ICR) for ferritin was in oligodendroglia of the central white matter of the DN. Ferritin‐ reactive oligodendroglia in the DN hilus of FRDA were much smaller than normal and more numerous. ICR for Cu, Zn‐superoxide dismutase (SOD) and Cu++‐transporting ATPase α‐peptide (ATP7A, Menkes disease protein) was present in microglia and vessel walls, respectively, but FRDA did not cause a distinct change in ICR from the normal state. The shift of Cu into the Fe‐rich white matter may cause greater oxidative injury. Supported by: NIH‐NS069454‐02; Friedreichˈs Ataxia Research Alliance; National Ataxia Foundation; and Neurochemical Research, Inc
Spatial variation in susceptibility to chronic wasting disease in white‐tailed deer
Authors: Manjerovic Mary Beth , Green Michelle L. , Kelly Amy C. , Mateus-Pinilla Nohra E. , Shelton Paul , Novakofski Jan
Published: June 2021

Chronic wasting disease (CWD) is a prion disease that leads to neurodegeneration and death in cervids. CWD management in Illinois has been based on selectively culling deer herds in CWD infected and high risk areas with the target goal of decreasing infectious foci. Previous research found associations between susceptibility to CWD and polymorphisms in the prion protein gene (PRNP). The objective of this study was to use nucleic acid sequences of the prion gene, in particular codon 96, to examine genetic profiles of deer throughout northern Illinois. Using muscle tissue collected during the CWD management program, we paired positive and negative samples stratified by age and sex and amplified the coding region for the mature prion protein (using previously published primers). Sequences were aligned in Sequencher v.5.0 and compared to a PRNP consensus sequence derived using the GenBank database. All statistical analyses were performed with SAS v.9.1. We compared the frequency of coding polymorphisms and found a correlation between disease status and total number of polymorphisms per animal. Thus, how genetic heterogeneity relates to CWD susceptibility and resistance can affect the natural response of individual animals to CWD exposure. This information can help guide management of deer populations with the additional goal of increasing herd immunity as we decrease CWD foci. Research funded by IDNR and UIUC‐OVCR.
Loss of Cardiac Muscle Ring Finger‐1 Augments Right Ventricular Hypertrophy Following Chronic Hypoxia‐Induced Pulmonary Hypertension
Authors: Paffett Michael , Lucas Selita , Anderson Tamara , Nysus Monique , Norenberg Jeffery , Willis Monte , Campen Matthew
Published: June 2021

Exposure to chronic hypoxia (CH) induces elevated pulmonary artery pressure/resistance, leading to an eventual maladaptive right ventricular hypertrophy (RVH). Muscle RING‐finger (MuRF)1 is a muscle specific protein has been shown to play a role in left ventricular hypertrophy and altered cardiac bioenergetics in pressure overloaded hearts, however little is known about the contribution of MuRF1 impacting RVH in the setting of CH. Therefore, we hypothesized that MuRF1 deletion would enhance RVH compared to their wild‐type (WT) littermates following CH‐induced pulmonary hypertension. Thus, we assessed right ventricular systolic pressure (RVSP), right ventricle to left ventricle plus septal weight ratio (RV/LV+S) and hematocrit (Hct) following a 3 week isobaric CH exposure. CH induced pulmonary hypertension, measured by increases RVSP, RV/LV+S and Hct, WT mice compared to normoxic WT mice. Normoxic WT and MuRF1‐null mice exhibited no significant differences in RVSP, RV/LV+S or Hct. CH‐induced increases in RVSP were also similar between WT and MuRF1‐null mice; however, RV/LV+S and Hct were significantly elevated in CH‐exposed MuRF1‐null mice compared to WT. In conclusion, these results suggest an atrophic role for MuRF1 regulating the magnitude of right ventricular hypertrophy and polycythemia following CH‐induction of pulmonary hypertension.
Anti‐proliferative effects of carvacrol on human prostate cancer cell line, LNCaP
Authors: Patel Bhushankumar , Shah Vichiksha R. , Bavadekar Supriya A.
Published: June 2021

Carvacrol, a monoterpene phenol, is known to be present in the volatile oils of Thymus vulgaris, Carum copticum and Origanum vulgare, and has been shown to have anti‐proliferative effects on various human cancer cell lines, such as breast cancer, hepatoma, leukemia, non‐small cell lung cancer, and cervical cancer. However, its effect on prostate cancer cells has not yet been determined. The objective of this research was to investigate the anti‐proliferative effects of carvacrol on an androgen‐sensitive, metastatic human prostate cancer cell line, LNCaP. In this study, cells were treated with increasing concentrations of carvacrol, ranging from 10−7M to 3×10−3M, and viability was determined after 48 and 96 hr using the XTT cell proliferation assay. Carvacrol inhibited cell viability in a dose‐dependent manner, and the resultant IC50 values were found to be 135 μM and 11 μM, respectively. Data from phosphatidylserine externalization assays suggest that carvacrol causes induction of apoptosis in these cells. This is the first study that has identified the anti‐proliferative effects of carvacrol in prostate cancer cells, and results suggest that the compound may be valuable as a potential therapeutic agent for prostate cancer. This study is supported by the Division of Pharmaceutical Sciences at Long Island University, Brooklyn, NY.
Adenosine A2B receptor blockade slows growth of bladder and breast tumors
Authors: Cekic Caglar , Sag Duygu , Linden Joel
Published: June 2021

The accumulation of high levels of adenosine in tumors activates adenosine A2A and A2B receptors on immune cells to suppress tumor rejection. We evaluated the effects of intratumor injection of a non‐selective adenosine receptor (AR) antagonist, aminophylline (AMO) and, for the first time, a selective A2BAR antagonist, ATL801. AMO and ATL801 slowed the growth of MB49 bladder and 4T1 breast tumors in syngeneic mice, and reduced by 85% metastasis of breast cancer cells from mammary fat to lung. Based on receptor deletion experiments, the effect of AMO injection was unexpectedly attributed to A2BAR and not to A2AAR blockade. AMO and ATL801 significantly increased tumor levels of IFNγ, the interferon‐inducible CXCR3 ligand, CXCL10, and tumor‐infiltrating CXCR3+ T cells. AMO decreased endothelial cell precursors within tumors. Tumor growth inhibition by AMO or ATL801 was eliminated in CXCR3−/− mice and in RAG1−/− mice that lack mature T cells. Bone marrow chimera experiments demonstrated that CXCR3 and A2BAR expression on bone marrow cells are required for the anti‐tumor effects of AMO. In RAG1−/− mice A2BAR blockade enhanced CD86 expression on CD11b‐ DCs. The data suggest that blockade of A2BARs enhances DC activation and CXCR3‐dependent anti‐tumor responses.
Metformin Attenuates Doxorubicin‐induced Cardiotoxicity In Rats
Authors: AlHaider Abdlqader A. , Korashy Hesham M.
Published: June 2021

The clinical use of doxorubicin (DOX), a potent chemotherapeutic agent, is limited by its cardiotoxic side effects that are attributed to free radical generation. Recent studies have established that metformin (MET), an oral antidiabetic drug, possesses an antioxidant activity. However, whether it can protect against DOX‐induced cardiotoxicity has not been reported before. The goal of the present study was to assess the extent to which MET can prevent cardiotoxicity induced by DOX. Results of our study in a rat model of DOX‐induced cardiotoxicity show that DOX treatment caused significant increases in serum levels of lactate dehydrogenase (LDH) and creatine phosphokinase isoenzyme MB (CK‐MB), indicators of cardiac injury. In addition, DOX therapy decreased cardiac glutathione, an endogenous antioxidant; CoA‐SH, an activator in mitochondrial energy providing systems. These biochemical changes were associated with myocardial histopathological and ultrastructural deteriorations. Co‐treatment with MET eliminated all DOX‐induced increases of LDH and CK‐MB and prevented GSH, CoA‐SH and ATP depletions. MET therapy also prevented all the myocardial histopathological and ultrastructural deteriorations. These findings demonstrate, that MET successfully prevents DOX‐induced cardiotoxicity.
Akt kinase inhibition facilitates p53 protein‐dependent mitochondrial damage responses after DNA damage
Authors: Franke Thomas , Shostak Gregory , Grabham Peter
Published: June 2021

Activated PI3K/AKT signaling is frequently found in human cancer and contributes to chemotherapeutic resistance. By examining the biochemical consequences of treating human cancer cells with chemotherapeutic drugs, we found that PI3K/AKT inhibition sensitized cancer cells to DNA damage‐inducing drugs to facilitate caspase activation and apoptosis induction. The synergistic effects of combining chemotherapeutics with pathway inhibition were specific to PI3K/AKT inhibition and not observed after ERK and mTOR inhibition. The effects required the mitochondrial pathway of apoptosis induction. Caspase activation was best predicted by the extent of mitochondrial damage leading to cytochrome c release and required ongoing protein translation. A similar synergy resulting from combination treatment was not seen in p53‐deficient cancer cells unless p53 expression was restored by using an inducible p53 mutant protein. Our findings suggest that non‐genomic consequences of p53 induction on mitochondrial integrity play a facilitating role in determining the outcome of PI3K/Akt inhibition on chemotherapeutic treatment, and support a critical role for mitochondrial damage during pro‐apoptotic drug responses.
The role of GPR30 in breast cancer cell migration
Authors: Doran Heather , Ross Ruth A
Published: June 2021

Some cells, although lacking the canonical estrogen receptors (ERs) still display rapid non‐genomic signalling in response to estrogen. Recently, a G protein coupled receptor; GPR30 (or GPER) and an alternative splice variant of the traditional ERα (ERα36) have been proposed as alternative estrogen receptors that may mediate these non‐genomic effects. This study investigated the role of GPR30 and ERα36 in breast cancer cell migration. Using a combination of cell migration assays, actin polymerisation measurements and immuno‐fluorescence we have demonstrated that SKBr3 cells (a ER receptor negative, breast cancer cell line) migrate towards the GPR30/ERα36 agonist G‐1; an effect that is attenuated by G‐15, a GPR30/ERα36 antagonist and pERK inhibitors, but not by ROCK inhibitors. We have also demonstrated that G‐1 activates a rapid signalling pathway involving changes in the actin cytoskeleton. The findings may progress the development of new therapeutics targeting ER negative breast cancer tumors.

This research was sponsored by Selcia Limited (Ongar, Essex) and the University of Aberdeen
A corticotropin‐releasing factor (CRF) receptor antagonist prevents bladder pathology associated with either social stress or partial bladder outlet obstruction (pBOO)
Authors: Wood Susan K , McFadden Kile V , Zderic Stephen A , Bhatnagar Seema , Valentino Rita J
Published: June 2021

Barrington's nucleus (BN) regulates micturition through projections to preganglionic parasympathetic neurons. The stress‐related neuropeptide, CRF has an inhibitory influence in this pathway. We previously reported that social stress increases CRF mRNA in BN and causes bladder dysfunction resembling pBOO. The present study examined the ability of a CRF1 receptor antagonist to alter social stress‐ or pBOO‐induced bladder dysfunction. Male rats were exposed to 7 days (30 min/day) of social stress using the resident‐intruder model or had pBOO surgery. Rats were treated with vehicle or a CRF1 antagonist (NBI‐ 30775, 20 mg/kg/day, sc). Urodynamic function was recorded in unanesthetized rats using cystometry 72 h after the 7th stress exposure or the 16th day after pBOO. Social stress and pBOO increased bladder capacity, micturition volume, and intermicturition interval compared with controls or shams and these effects were prevented by NBI‐30775. One distinction between stress‐induced urodynamic changes and pBOO is that increased micturition pressure is only present in pBOO due to increased resistance at the bladder neck. Importantly, NBI‐30775 did not block this increase in pressure. These studies implicate a role for CRF in stress‐ and pBOO‐induced urodynamic changes and suggest that CRF1 antagonists may be useful therapeutic agents for the treatment of urinary disorders. DK052620
Developing zebrafish models of depression?: Effects of reserpine on zebrafish behavior and physiology
Authors: Kyzar Evan , Roth Andrew , Green Jeremy , Gaikwad Siddharth , Monnig Louis , Kalueff Allan
Published: June 2021

Reserpine is well‐known for its ability to deplete brain monoamines and cause depression‐like symptoms in humans and animals. Zebrafish (Danio rerio) are rapidly emerging as a promising, high‐throughput model for translational neurobehavioral research. Here, we have examined the acute (20 min) and long‐term (7 day) effects of reserpine exposure on zebrafish behavior and physiology. Several behavioral paradigms (the novel tank, light–dark box, social preference and shoaling tests) were used to characterize the effects of these treatments. Reserpine did not evoke overt behavioral effects acutely, but affected activity 7 days later, resulting in robust hypolocomotion without affecting anxiety‐related top/bottom preference. Fish treated chronically with reserpine for 2 h/day for 7 days displayed marked hypolocomotion, including an overall reduction in distance traveled. The global reduction in activity without altering anxiety strikingly resembles motor retardation observed in clinically depressed patients. Additionally, elevations in cortisol and the gene c‐fos were found in fish treated with reserpine. The long‐term depression‐like effects of reserpine in zebrafish parallel its effect in both rodent and clinical studies, emphasizing the utility of zebrafish models to study a wide spectrum of drug‐evoked affective states. This study was supported by Tulane University intramural funds.
Contribution of β2 subunit containing nicotinic acetylcholine receptors to anxiety‐like behaviors in male mice using a novel model of anxiety
Authors: De Jesus Angelica , Anderson Shawn M , Brunzell Darlene H.
Published: June 2021

To test the hypothesis that inhibition of beta 2 subunit containing nicotinic acetylcholine receptors (β 2*nAChRs) promotes anxiolysis, we compared β2*nAChR null mutant (β2KO) and heterozygous (Het) mice to wild type (WT) littermate controls in the light‐dark assay and a novel bottle inversion assay (BIA) where water access was restricted. Behavioral measures included bottle‐interaction, rearing, digging, and social pursuit of a cage mate. In support of our hypothesis, there was a non‐significant trend for beta2KOs and Hets to show shorter latencies than WT mice to leave the dark chamber. BIA revealed a significant reduction in social pursuits of β2KOs and Hets in comparison to their WT counterparts (p's < 0.05). There was a similar reduction of this behavior following injection of 1 mg/kg midazolam in WT (p < 0.05) mice, validating social pursuit as a measure of anxiety‐like behavior in this task. These findings suggest that inhibition of β2*nAChRs may promote anxiolysis and identify β2*nAChRs as a potential therapeutic target for anxiety disorder. This research was supported by a Jeffress Memorial Trust research grant J‐951 and R01 DA031289, P50 DA005274, R25 GM090084, and T32 DA00702734.β
A fluorescence microplate screen assay for the detection of neurite outgrowth and neurotoxicity using antibodies against βIII‐tubulin
Authors: Popova Dina , Jacobsson Stig O.P.
Published: June 2021

The majority of environmental and commercial chemicals have not been evaluated for their potential to cause neurotoxicity. Retinoic acid‐induced neurons from mouse embryonal carcinoma cells is a rapid and sensitive model to assess chemical‐induced neurotoxicity, and we have investigated if neuron specific anti‐βIII‐tubulin antibodies are useful in a microplate assay of neurite outgrowth. By incubating the P19‐derived neurons with the primary anti‐βIII‐tubulin antibody and a secondary Alexa Fluor 488‐conjugated followed by measuring the fluorescence in a microplate reader, a time‐dependent increase in anti‐βIII tubulin immunofluorescence was observed ‐ the relative fluorescence units increased by 4.3‐fold from 2 to 10 days in culture. The results were superimposable to those obtained by semi‐automatic tracing of neurites in fluorescence microscopy images of βIII‐tubulin‐labeled neurons. The sensitivity of the neurite outgrowth assay using a microplate reader to detect neurotoxicity produced by methyl mercury chloride (0.05–1 μM) and okadaic acid (1–50 nM), was significantly higher than a cell viability assay measuring intracellular fluorescence of calcein‐AM. In conclusion, the fluorescence microplate assay for the detection of neurite outgrowth by measuring changes in βIII‐tubulin immunoreactivity is a rapid and sensitive method to assess chemical‐ or toxin‐induced neurotoxicity.