bluuebunny/crossref_metadata_2025 huggingface dataset:
Epigenetic regulation of physiological cardiac hypertrophy by O‐GlcNAc and histone deacetylases in diabetes
Authors: Cox Emily J , Marsh Susan A
Published: June 2021

Exercise causes physiological cardiac hypertrophy, and type 2 diabetes blunts hypertrophic signaling. Diabetic hearts have elevated levels of O‐linked β‐N‐acetylglucosamine (O‐GlcNAc), which post‐translationally modifies a complex of mSin3A and histone deacetylases (HDACs) 1 and 2. mSin3A/HDAC1/2 regulates hypertrophy and is targeted to genes by REST and O‐GlcNAc transferase (OGT). We examined the role of O‐GlcNAc in these mechanisms in 8‐week‐old diabetic (db/db) and non‐diabetic C57BL/6J mice randomized to exercise or sedentary groups for 4 weeks. Db/db mice had higher heart weight to tibia length (5.4±0.1 vs 4.7±0.1 mg/mm), fasting glucose (10.1±3.2 vs 5.5±1.5 mmol/L), and body weight (39.1±2.5 vs 20.1±1.2 g). Db/db hearts had higher levels of O‐GlcNAc and OGT, but lower mSin3A, HDAC1, and HDAC2. Conversely, HDAC2 expression and HDAC1/2 activity were higher in db/db hearts, and α‐actin and BNP expression were lower. In db/db hearts, immunoprecipitation showed that mSin3A was more O‐GlcNAcylated and more associated with REST, and OGT was less associated with HDAC2. Exercise corrected these changes, showing that diabetes and exercise have interacting effects on the epigenetic control of hypertrophy.
Professional Skills Courses Increase Trainee Skills in Writing, Reviewing, Networking, and Poster Presentations
Authors: Lowy Melinda Elaine , Matyas Marsha Lakes
Published: June 2021

APS developed professional skills training courses [Writing and Reviewing for Journals (WR), Presentation Skills (PS)] for live or online use with trainees, specifically targeted at underrepresented trainees. Courses were tested as two live and one online course, with an average of 34 trainees in each. Underrepresented minorities comprised 51% of trainees and included Hispanics (32%), African‐Americans (15%), Native Americans (2%), and Native Hawaiians (2%). Graduate students (66%) and postdoctoral fellows (34%) participated in WR, whereas mainly graduate students (95%) participated in PS because of content. Both courses received high overall ratings (WR, 4.8; PS, 4.8; scale of 1–5 where 5 is highest rating) regardless of whether live or online. There were no rating differences between trainees regardless of course of study, minority status, or education level. All components of courses (pre‐workshop assignments, plenary sessions, small group work, and interactive exercises) were rated highly by all participants (3.7 – 5.0). Impacts of both courses on trainees were significant. WR trainees self‐rated their skills at an average of 37.4 before vs. 59.5 pts after the course (70 pts possible, P<0.001). PS trainees self‐rated their skills at 43.1 before and 65.7 pts after the course (P<0.001). These are effective instructional courses for trainees from diverse groups. Supported by NIGMS Grant 5T36GM073062–03.
The Lateral Thoracic Artery Revisited
Authors: du Plessis Maira , Tubbs R Shane , Loukas Marios
Published: June 2021

Although the variations in the origin of the branches of the axillary and subclavian arteries have been documented, little information regarding the highly variable lateral thoracic artery (LTA) is available. Descriptions of the LTA variations may decrease intraoperative problems during surgery of the lateral aspects of the thorax such as reconstructive plastic surgery and modified radical mastectomy. The aim was therefore to review the anatomy of the LTA and to offer an accurate account of its variability. The entire course and distribution of the LTA was examined in 420 formalin‐fixed adult human cadavers. The LTA was found in 96.7% of our specimens, showing great morphological variability and classified the variations according to origin into 6 types. The most common was where the LTA arose from the thoracoacromial trunk (Type I, 70%). In decreasing order of incidence were origins from; the axillary artery (Type II, 17.6%), the thoracodorsal artery (Type III, 5.2%), and the subscapular artery (Type IV, 4%). Additionally, multiple LTAs were present (Type V, 3.2%) and complete absence of LTA (Type VI, 3.3%) was observed. Despite variations in the origin our dissections demonstrated that the ultimate tissue distribution of the LTA remained typical (90%) and descended on the lateral border of the pectoralis minor. Our results illustrate the need for re‐evaluation of the branches of the thoracoacromial trunk with possible consideration that the LTA arises from it instead of directly from the axillary artery. We hope that the information supplied by this study will provide useful information to anatomists and surgeons alike.

Grant Funding Source: N/A
Possibility of natural healing to anterior cruciate ligament tear
Authors: Takayanagi Kiyomi , Kanemura Naohiko , Kokubun Takanori , Maejima Hiroshi , Nishikawa Yuichi , Ihara Hidetoshi
Published: June 2021

An acute tear of the ligament generally heals conservatively. Whereas, it is thought that the tear of the anterior cruciate ligament (ACL) does not heal by the conservative treatment. There are many studies about the factors that ACL does not heal conservatively.

The purpose of the research was to create a rat model of natural healing of ACL and to examine the healed ligament histologically and mechanically.

After cutting ACL, the artificial ligament was penetrated the bone tunnel of tibia tuberosity and back of the distal femur, and controlled the abnormal movement of the knee joint. We had confirmed the continuity of the healed ACL in all rats with histological evaluation. Running of the healing ligament had been normally approximated. Compared to the normal ACL, the healing ligament was thick, and scar tissue had grown into joint intercondylar fossa. Healing ACL was about half strength compared to the normal.

We found that the ruptured anterior cruciate ligament was healed by joint control kinematically.
LIMBAL STEM CELL DEFICIENCY MODEL IN RABBITS FOR EXPERIMENTAL CORNEAL SURFACE REGENERATION
Authors: Oliva Joan , Bardag‐Gorce Fawzia , Hoft Richard , Wood Andrew , Pan Derek , Sota Hiroyuki , Niihara Yutaka
Published: June 2021

In this study, we present a modified technique for creation of limbal stem cell deficiency in rabbits. Twenty percent isopropanol was used to completely remove rabbit corneal epithelium, and then superficial lamellar limbectomy (developed by other investigators) was performed to excise most or all limbal epithelial stem cells. Rabbit eyes were examined at least once every month for 3 months with slit lamp, fluorescein staining, impression cytology (IC), and western blot analysis after LSCD creation. Slit lamp exams indicated that the severity of corneal vascularization and conjunctivalization did not stabilize until 2–3 months after LSCD creation. IC showed many macrophages on the corneal surface at one month, reduced but still significant macrophage infiltration at two months, and very few macrophages with many fibroblasts on the central corneal surface at 3 months. Conjunctival marker MUC5AC was present on the central cornea at 3 months. In addition, the corneal epithelium stained negative for K3 and K12 and positive for K13 after 3 months, indicating significant conjunctivalization of the corneal surface epithelium. These results indicate that our technique is effective in creating LSCD and conjunctivalization of the rabbit corneal surface. This model will be used to study regeneration of normal corneal epithelium with cultured autologous oral mucosal epithelial cell sheet grafts.
Association Of CTLA4 Gene 49A/G Polymorphism With The Incidence of Type 1 Diabetes Mellitus In the Iranian Kurdish Population
Authors: Ahmadi Slahadin , Rostamzadeh Jalal , Khosravi Darya , Shkiba Nadia , Nabavi Hassan
Published: June 2021

Cytotoxic T‐lymphocyte‐associated antigen‐4 (CTLA‐4) has an inhibitory function on T cells and is critical for the induction of peripheral tolerance. CTLA‐4 +49 G allele affects the CTLA‐4 function and has been reported to be correlated with a higher risk of various autoimmune diseases including type 1 diabetes (TIDM). The present study was conducted to investigate the association between the polymorphism of the CTLA‐4 exon 1 +49 A/G and susceptibility to TIDM in Sanandaj population, a Kurdish city in the west of Iran.

The +49 A/G polymorphism was analyzed in 64 patients with TIDM, 60 patients with type 2 diabetes (T2DM ) and 107 control subjects with no history of autoimmune disease using PCR‐RFLP method. All populations (controls, T2DM and TIDM) were in Hardy‐Weinberg equilibrium ( p; 0.892003, 0.75897 and 0.168063, respectively). There was significant CTLA‐4 +49 G allele difference between TIDM with control (p=0.002163) and T2DM subjects ( p=0.008504). No significant difference was found between T2DM and control subjects regarding the distribution of the CTLA‐4 +49 G allele.

Our findings revealed a significant association of CTLA‐4 exon 1 49 A/G polymorphism with the increase risk of T1DM in the Iranian Kurdish population.

Financial support for this research was provided by the Research Department of the Kurdistan University of Medical Sciences, Kurdistan, Iran.
Miniaturization of commercial swine
Authors: Sough Ashley Marie , Piedrahita Jorge
Published: June 2021

Swine are used throughout the biomedical community as translational research models, due to their physiological similarity to humans. However commercial swine are bred for consumption, therefore bigger is better, which is not always beneficial to the scientific community, in terms of maintenance costs. To generate pigs of reduced size, the current study utilized RNA interference to knockdown the high mobility group AT‐hook 2 (HMGA2) gene. Three short hairpin RNA sequences (shRNA; SH1, SH2, and SH3) targeting HGMA2 were inserted into the pLKO.1 vector, a vector used by the RNAi Consortium to produce their shRNA library. A scrambled shRNA was used to control for off‐target effects. Expression levels of HMGA2 mRNA in porcine fetal fibroblasts were analyzed by quantitative real‐time polymerase chain reaction and the experiment was replicated three times. Results indicate that SH1 and SH3 were able to significantly (p<0.05) reduce the level of HMGA2 mRNA by 71% and 78%, respectively. Future work will focus on whether the knockdown of HMGA2 is proportional to adult pig size, since Zhou et al. produced a pygmy mouse model by deleting the HMGA2 gene. This research is funded by grants from NIH: 1R21OD010553–01A1 and PAR‐06–553, respectively.
Nonspecific Phospholipase C Activity from Listeria Monocytogenes is Activated by Phagosome Acidification
Authors: Huang Qiongying , Roberts Mary
Published: June 2021

Broad‐range, phosphatidylcholine‐preferring phospholipase C (PC‐PLCLM) from Listeria monocytogenes is one of the critical virulence factors mediating the dissolution of vacuolar membranes so that the intracellular pathogen can escape into the cytosol and spread to neighboring cells. We have expressed the enzyme in E. coli and characterized its kinetics with a variety of substrates. The amino acid sequence of PC‐PLCLM is highly homologous to Bacillus cereus PC‐PLCBC. Both are metalloenzymes with identical residues coordinating to the Zn2+ ligands. Intriguingly, the PC‐PLCLM exhibits an acidic pH optimum (pH5~6) compared to the well‐studied PC‐PLCBC which is more efficient at basic pH values. This is significant as PC‐PLCLM activity is needed to aid in Listeria escape from acidified vacuolar pH. The optimal efficiency at acidic pH results from dramatically reduced Km values but moderate changes in kcat at lower pH. In addition, unlike PLCBC, PC‐PLCLM can cleave sphingomyelin. Mechanistic details for the acidic pH optimum and substrate specificity are probed with mutagenesis and a combination of biochemical and biophysical approaches. This study is supported by the National Institutes of Health GM60418.
E. coli peritonitis initiates D‐series Resolvin‐metabolome
Authors: Gao Fei , Dalli Jesmond , Claria Joan , Serhan Charles N.
Published: June 2021

Resolvin D1 (RvD1, 7S, 8R, 17S‐trihydroxy‐4Z, 9E, 11E, 13Z, 15E, 19Z‐docosahexaenoic acid) and D2 (RvD2, 7S, 16R, 17S‐trihydroxy‐4Z, 8E, 10Z, 12E, 14E, 19Z‐docosahexaenoic acid) are novel potent anti‐inflammatory and pro‐resolving lipid mediators derived from omega‐3 docosahexaenoic acid. In this study, we report the temporal regulation of RvD1 and RvD2 metabolic products in murine inflammatory exudates during E.coli peritonitis. Both RvD1 and RvD2 are further converted to novel products by human neutrophils and macrophages, as well as in murine exudates. The structures of these novel RvD1 and RvD2 derived products were determined using both GC‐MS and LC‐MS‐MS. The new RvD2 products elucidated included 7‐oxo‐RvD2, 16‐oxo‐RvD2, 22‐hydroxy‐RvD2, and 8, 9‐dihydro‐RvD2. Direct comparisons of the bioactions of select RvD2 metabolic products indicated that 7‐oxo‐RvD2 and 8, 9‐dihydro‐RvD2 displayed reduced activiy (i.e., anti‐inflammatory actions in vivo, and reduced ability to inhibit human neutrophil chemotaxis and macrophage phagocytosis in vitro), whereas 16‐oxo‐RvD2 was essentially inactive. Together, these findings indicate that, during inflammation and its controlled resolution circuits, specific enzyme systems inactivate via metabolizing pro‐resolving signals, i.e. resolvins, to permit the coordinated return to homeostasis.
ASSOCIATION BETWEEN FOOD PURCHASE AND BREASTFEEDING STATUS OF LOW‐INCOME FAMILIES IN WOMEN, INFANTS, AND CHILDREN (WIC)
Authors: KIM HYUN , KIM JUHEE
Published: June 2021

Low‐income mothers and especially those who participate in WIC have lower breastfeeding rates compared to other groups. This may be problematic for these mothers, as breastfeeding can help reducing overall health costs and improve health outcomes for mothers and their infants. This study focuses on the role of breastfeeding status on the food purchase behaviors of WIC participants. Since breastfeeding mothers tend to be more health‐conscious, this paper will test whether breastfeeding WIC participants make healthier food choices than non‐breastfeeding WIC participants. The paper analyze the food purchase patterns of low‐income mothers enrolled in the WIC program (n = 110). The measures of food purchase include food purchase frequency, food expenditures by food group and type of food outlets. The analyses using multivariate regression models are based on food receipts obtained during a one‐month period, as well as survey from mothers having a child or children enrolled in a WIC program in Eastern Illinois in 2009. We will describe the patterns of food outlet use and food purchases to understand food shopping or eating out behaviors of WIC families, stratified by their breastfeeding practice status. Since WIC participants view the cost and difficulty of food selection as barriers to their purchase, this study can evaluate their food purchase behavior based on their demographic characteristics.

(Funding: National Research Foundation of Korea Grant (NRF‐2011–330‐B00190)

Grant Funding Source: National Research Foundation of Korea Grant (NRF‐2011–330‐B00190)
Increasing access to fresh produce by partnering urban farms with corner stores: a pilot study in a low‐income urban setting
Authors: Gudzune Kimberly Anne , Welsh Claire , Lane Elisa , Chissell Zach , Anderson Elizabeth , Gittelsohn Joel
Published: June 2021

Residents in low‐income areas often rely on corner stores for food, but these storeowners seldom stock fresh produce. Many urban farms are looking to provide better access to produce for low‐income consumers. Our objective was to partner two urban farms with two corner stores in Baltimore, MD to distribute fresh produce. In summer 2012, we conducted a pre‐post evaluation using semi‐structured interviews coupled with distribution and sales data. Partnership #1 paired a 6‐acre urban farm with a store that served commuters. Produce was delivered all 5 weeks with the mean total quantity of produce delivered being 38 (SD 16) items. On average each week, 6 (SD 2) different types of produce were brought into the store, of which 2 (SD 2) types sold out. Partnership #2 paired a ¼‐acre urban farm with a store that served local residents, and was promoted by the neighborhood association. Produce was delivered all 5 weeks with the mean total quantity of produce delivered being 77 (SD 12) items. On average each week, 10 (SD 1) different types of produce were brought into the store, of which 6 (SD 2) types sold out. Partnering urban farms and corner stores for produce distribution is feasible. For future programs to be successful, strong community support appears vital to sustain produce sales.

Grant Funding Source: NICHD U54HD070725
Examination of Chondroitin Sulfate Molecular Weights on In Vitro Anti‐inflammatory Activity
Authors: Surapaneni Lahari , Haley‐Zitlin Vivian , Bodine Ashby , Jiang Xiuping , Brooks James
Published: June 2021

Objective

The purpose of this study was to investigate if anti‐inflammatory activity of chondroitin sulfate (CS) is affected by molecular weight.

Methods

Murine macrophage cells were pre incubated with 8 different CS samples (mol. wt. 7 kDa to 35 kDa) for an hour followed by addition of bacterial lipopolysaccharide (LPS). Following further incubation for 4 hours, the anti‐inflammatory activity of CS samples was assessed by measurement of the ability of CS samples to inhibit expression of a panel of inflammatory cytokines; tumor necrosis factor‐alpha (TNF‐α), Interleukin‐1 beta (IL‐1β) and Interleukin‐6 (IL‐6). These inflammatory markers were quantitatively measured using ELISA. Inhibition of nitric oxide (NO) production was determined using Greiss reagent.

Results

At concentrations 5μg/ml and 15 μg/ml, CS samples significantly inhibited expression of LPS induced TNF‐α and IL‐1β. Expression of IL‐6 was inhibited by selected CS samples at 15 μg/ml concentration but not at 5μg/ml. No CS samples inhibited the expression of NO. Statistical analysis revealed no correlation between molecular weight and the inflammatory markers examined.

Conclusion

CS samples exhibited selective inhibition of inflammatory cytokines but no correlation was noted between the molecular weights and anti‐inflammatory activity of these samples.

Grant Funding Source: Pharmavite, Inc.
Sex Specific Differences in the Predictive Value of Cholesterol Homeostasis Markers and 10‐Year CVD Event Rate in Framingham Offspring Study
Authors: Matthan Nirupa Rachel , Zhu Lei , Pencina Michael , D'Agostino Ralph B. , Schaefer Ernst J. , Lichtenstein Alice H.
Published: June 2021

Data are inconsistent on factors influencing plasma cholesterol homeostasis marker levels and their value in predicting subsequent CVD events.

We assessed the relationship between markers of cholesterol absorption (campesterol, sitosterol, cholestanol) and synthesis (squalene, desmosterol, lathosterol), and 10‐year CVD incidence in FOS participants without CVD at baseline and not taking lipid lowering medications (N=2616). The primary endpoint was “hard” coronary heart disease (HCHD: coronary death and myocardial infarction [MI]), and secondary endpoint was full CVD (HCHD + stroke, coronary insufficiency, angina pectoris, PAD and CHF). In cross‐sectional analysis, significant differences by sex, age, BMI, BP and smoking status were observed. In both women and men, lower cholesterol absorption was associated with higher triglyceride and lower HDL‐C levels, while lower cholesterol synthesis was associated with higher LDL‐C levels (p<0.05). In women only, lower cholesterol synthesis and absorption were associated with higher non‐HDL‐C levels. Using Cox proportional hazards model correcting for standard CVD risk factors, squalene levels were associated with lower HCHD in women (HR=0.70 [0.5–0.9]). In contrast, squalene (HR=1.40 [1.1–1.8]) levels were associated with higher HCHD in men (p‐value <0.0001 for interaction). The cholesterol absorption markers were not predictive of HCHD or full CVD in either sex.

These data suggest significant sex differences in the 10‐year prognostic value of cholesterol synthesis markers and mortality as well as MI incidence.

NIH‐HL‐074388, NO1‐HC‐25195 and USDA #: 58–1950–4–401
Sodium, potassium, sodium to potassium ratio and hypertension in China, 1991–2009
Authors: Du Shufa , Batis Carolina , Wang Huijun , Zhang Bing , Popkin Barry M
Published: June 2021

Little is known about patterns and trends and patterns of sodium, potassium, sodium to potassium (Na/K) ratio and their health effects in China. Our study examines these trends and their health impact using China Health and Nutrition Survey data (1991–2009) for 16,869 adults aged 20–60 years. Detailed dietary data was collected by three consecutive 24–hour recalls with weighing of all foods and condiments. Blood pressure was measured with standard mercury sphygmomanometers. Flexible parametric models for survival‐time data were used to estimate the hazard ratios. Recent intakes of sodium, potassium, and Na/K ratio had strong dose‐response associations with incident hypertension: hazard ratios (95%CI) were 1.27 (1.05, 1.52), 1.47 (1.20, 1.80), 2.03 (1.63, 2.52), 2.16 (1.72, 2.71) for sodium, 0.83 (0.71, 0.97), 0.94 (0.80, 1.10), 0.74 (0.63, 0.87), 0.66 (0.56, 0.78) for potassium, and 1.12 (0.93,1.35), 1.27 (1.06,1.53), 1.64 (1.37,1.97), 2.14 (1.79,2.55) for Na/K ratio for the second to fifth quintiles compared to the lowest group respectively. These findings suggest that adults in China have low potassium intake and high Na/K ratio and both increase the risk of hypertension. Replacing sodium with potassium in salt to control and prevent hypertension in China should be considered among other public health and clinical prevention options.
Trend in calories consumed in the US diet over the last decade: Analysis of What We Eat In America (WWEIA)/National Health and Nutrition Examination Survey (NHANES) 2001–2010
Authors: Fulgoni Victor L , Schorin Marilyn , O'Neil Carol E , Nicklas Theresa A
Published: June 2021

WWEIA data and the food groupings within were used to assess energy intake and sources of energy in the diets of those 2+ years of age (N=42,151) over the last decade. Regression analyses were conducted to determine a p‐for trend across the five NHANES data releases (2001–2010). Covariate adjusted energy intake was 2132 ± 9 kcal/d and major contributors of energy (mean ± SE, kcal/d) in the diet across the decade were Grains (749 ± 6); Meat, Poultry, Fish and Mixtures (390 ± 4); Sugars, Sweets and Beverages (337 ± 4), and Milk and Milk Products (246 ± 2). Energy intake decreased across the five NHANES releases and there was a significant (p<0.0001) p‐for trend of −25 kcal/d per 2 yr cycle release (−44 in those 2–19 yrs and −17 in those 20+ yrs). The largest decrease was from Sugars, Sweets and Beverages (−15 kcal/d per release, p<0.0001). Smaller decreases (kcal/d per release) in Fats, Oils and Salad Dressings (−3; p<0.0001) and Vegetables (−3; p<0.05) were also seen. One of the largest decreases (p<0.0001) in calories was in Soft Drinks (Carbonated) of −13 kcal/d per release (−15 in those 2–19 yrs and −12 in those 20+ yrs). There were smaller decreases (p<0.01) in Cakes, Cookies, Pies, Pastries (−4 kcal/d per release), Milks and Milk Drinks (−3 kcal/d per release), Sugars and Sweets (−2 kcal/d per release), Cereals, Not Cooked (−2 kcal/d per release) and Salad Dressings (−2 kcal/d per release). Support: USDA and American Beverage Association.
Flaxseed Oil Supplementation Increases Long‐chain Omega‐3 and Omega‐6 Fatty Acids in Plasma
Authors: Cheatham Carol L , Sheppard Kelly W , Lupu Daniel S , Niculescu Mihai D
Published: June 2021

Our animal work indicates postnatal maternal alpha‐linolenic acid (LNA, 18:3 n‐3) supplementation alters FADS2 methylation (Niculescu et al., 2012) and increases neurogenesis in the pups’ dentate gyrus (Niculescu et al., 2011) contingent on gestational LNA levels. Based on these models, we posit that humans who convert more LNA into longer‐chain fatty acids may be better able to utilize the precursor fatty acids in flaxseed oil, which is less expensive and more commercially viable than sources of preformed docosahexaenoic acid (DHA, 22:6 n‐3). We asked if flaxseed oil supplementation would up‐regulate LCPUFA production. 16‐month‐olds were randomized to control (1288 mg corn oil) or treatment (1200 mg flaxseed oil). Plasma was sampled at baseline and after 4m supplementation. There were no differences in dietary intake of fatty acids. Changes in fatty acid levels in plasma were assessed using ANOVAs. We found those whose baseline plasma LNA level was undetectable, suggesting efficient utilization of LNA, had higher docosapentaenoic acid (DPA, 22:5 n‐3) at baseline and higher arachidonic acid (ARA, 20:4 n‐6) and lower n‐6 to n‐3 fatty acid ratios after supplementation (p < .05). Supplementation differentially altered ARA levels (Figure 1) with main effects of intervention (p<0.01) and baseline LNA (p<0.01). These results suggest postnatal supplementation can up‐regulate LCPUFA pathways in humans.
Pregnancy‐specific up‐regulation of zinc (Zn) absorption does not occur in indigenous Mayan population
Authors: Hambidge K Michael , Mazariegos Manolo , Westcott Jamie E , Miller Leland V , Close Kelly , Goco Norman , Kemp Jennifer F , Lei Sian , Krebs Nancy F
Published: June 2021

Absorption of Zn has been reported to be up‐regulated in late pregnancy (Donangelo, Nutrients, 2012) which has implications for estimating the effect of pregnancy on Zn dietary requirements. The objective of this study was to undertake a longitudinal study of Zn absorption pre‐pregnancy, first and third trimester in 17 indigenous Guatemalan women who were participating in a low phytate maize study with maize as the principal dietary component. Fractional absorption of Zn (FAZ) was determined using extrinsic labeling with a dual isotope tracer ratio method; duplicate diets were analyzed to determine total daily Zn intake (TDZ); total absorbed Zn (TAZ, mg/d) calculated from FAZ × TDZ. As there was only one difference between low‐phytate and control groups for one variable at one time, data were subsequently combined and significance across time points determined. Phase differences (mean (SD)) are summarized in the table:

For both FAZ and TAZ there was a significant difference across the 3 time points, attributable to the Δ between preconception and early pregnancy phase for both FAZ and TAZ. A Spearman correlation for TAZ in the 1st trimester phase showed that this was attributable to a lower TDZ. In conclusion, the results of this study did not support the concept of up‐regulation of Zn absorption specific to late pregnancy. Supported by Eunice Kennedy Shriver NICHD/NIH #HD04065.
Acute phase proteins are more elevated in acute respiratory infections than in diarrhea.
Authors: Contreras‐Manzano Alejandra , Villalpando Salvador , Lamadrid Héctor , Shamah‐Levy Teresa
Published: June 2021

Introduction

Differences in the responses in the secretion of lymphocytes and limphokynes by diarrhea and acute respiratory infections (ARI) predict differences in the secretion of c reactive protein (CRP) and alpha glycoprotein (AGP.)

Objective

To compare the magnitude of the acute inflammation as by CRP and AGP in a group of school age children suffering episodes of diarrhea or ARI (duration and symptoms)

Material y methods

Morbidity data from the prior 14 d and final blood samples to measure CRP and AGP were obtained. The values of CRP, AGP and ferritin were log transformed. The evaluation of inflammation for diarrhea and ARI was made by linear regression models with random effects.

Results

One episode of ARI produce greater CRP response than one of diarrhea (0.637, p=0.01) and two or more symptoms of ARI produced a greater CRP and AGP response than diarrhea (0.357 and 0.810, p<0.01, respectively). An increase of ferritin was seen in both infections when CRP and AGP were elevated (Ratios infection/no infection: 1.93 [1.68–2.22] and 1.84 [1.56–2.18], respectively, (p<0.001).

Conclusion

Our data demonstrate that ARI has a greater acute response as by CRP and AGP changes and by number of symptoms than diarrhea.
Dietary nucleotides and yeast extract affect the composition and metabolic activities of the swine gut microbiota in a new in vitro fermentation model
Authors: Doo Eun‐Hee , Chassard Christophe , Lacroix Christophe
Published: June 2021

Nucleotides (NT) are important compounds to many biological processes but data on the exact role of NT on the gut microbiota remain unclear. In this work we investigated the effects of dietary NT in a medium formulated to simulate the pig ileal chyme on gut microbiota composition and activity using a novel continuous in vitro intestinal fermentation models mimicking the proximal colon. Pig fecal microbiota was immobilized in gellan‐xanthan beads and used as a continuous inoculum in a novel intestinal model platform, operated with multiple parallel reactors to compare treatments on the exact same microbiota composition. Daily effluent samples were collected and analyzed to investigate the composition (qPCR, pyrosequencing) and metabolic activities (HPLC) of the microbiota.

We measured significant shifts of microbial diversity, microbiota composition, and activity of effluent samples induced by dietary NT and YE treatments. In particular Lactobacilli (+1.4 Log10) and propionate concentration (+18.7 %) significantly increased in 7.5 g/l high NTYE compare to the control medium. Our data suggest beneficial effects of dietary NT in YE on pig gut microbial balance, associated with enhanced propionate production which could reinforce the barrier effect of the gut microbiota and increased populations of beneficial microbes.

This study was supported by a grant from the Worldwide Research Alltech, Inc. in USA.

Grant Funding Source: Nutrition (ASN): ASN Graduate Student Research Award Competition, supported by the Nutritional Sciences Council
Pathophysiologic and metabolomic analyses in a model for closed abdomen septic swine treated with negative pressure therapy (NPT)
Authors: Norbury Kenneth Carl
Published: June 2021

This study evaluated the impact of NPT in a porcine model of sepsis using pathophysiologic and metabolomic analyses. NPT consisted of polyurethane foam dressing with a visceral protective layer that extended into the paracolic gutters. NPT at −125 mmHg evacuated ascites into a canister. Injury was induced in 10 female swine via intestinal ischemia/reperfusion and induction of sepsis with a fecal suspension into the peritoneal cavity. After 10 hours, NPT was applied to 5 animals, and the abdomen closed, initiating treatment for up to 38 hours. Five control animals received no NPT. Animals were monitored throughout the study and terminal histopathology was performed. Biochemical and metabolomic analyses were performed on blood samples taken at intervals throughout the study. Inflammatory responses were measured by proteomic analysis of peritoneal and plasma samples. Findings are summarized below.

Organ System
Parameter
Control
NPT

I/R
Lactate
↑
↓

Hemodynamic
CVP
↑
↓

SvO2
↓
↑

Pulmonary
PaO2:FiO2 & Cstat
↓
↑

Acute alveolar congestion; interstitial edema and congestion
↓
↑

Renal
BUN & Creatinine
↑
↓

Inflammatory
TNF‐α
↑
↓

Metabolomic
myo‐inositol (a storage reservoir for proinflammatory arachidonic acid)
↑
↓

Long chain fatty acid biosynthesis
↓
↑

In this swine model of ischemic/reperfusion injury and sepsis, NPT had a beneficial effect on reversing the effects of injury compared to a control group that did not receive NPT.

Study funded by KCI.
A novel in vitro three‐dimensional bioprinted liver tissue system for drug development
Authors: Robbins Justin Baker , Gorgen Vivian , Min Peter , Shepherd Benjamin R. , Presnell Sharon C.
Published: June 2021

Despite efforts to improve the ability to identify the toxicity of therapeutic compounds, the attrition rate for both experimental and approved drugs remains very high. Cardio‐ and hepatotoxicity remain primary reasons for late stage failures and post‐market withdrawals. Therefore more robust human, in vitro models of these organ systems are needed. We have developed a bioprinted three‐dimensional (3D) liver system that captures several key features of in vivo tissue, in a multi‐well format suitable for drug screening. Using NovoGenTM bio‐printing technology we have fabricated 3D liver constructs containing architecturally‐ and physiologically‐relevant features for two hepatic cell lines and primary hepatocytes, within standard multi‐well culture plates. Bioprinted, 3D hepatic neotissues were further enhanced in complexity with the addition of endothelial and hepatic stellate cells. Biochemical studies demonstrate that several critical liver functions are present including cytochrome P450 activity. Tight junction protein expression was observed throughout the 3D tissue. Analysis of cell death and proliferation following in vitro maturation revealed the constructs were viable. These results demonstrate a flexible bioprinting method to rapidly fabricate multi‐cellular 3D liver constructs in a multi‐well format enabling both drug screening and interrogation of liver biology.
Evaluation of the VPT‐60 to measure peripheral nerve sensitivity
Authors: Gibson Janelle , Blackburn Mollie , Breen Paul , Latman N. S.
Published: June 2021

The loss of peripheral nerve sensitivity can result in serious morbidity (including limb amputation) and mortality. Monitoring peripheral nerve sensitivity is important in avoiding these consequences.

The purpose of this study was to evaluate the capability of an experimental VPT‐60 to accurately determine peripheral nerve sensitivity. Peripheral nerve sensitivity measurements from the VPT‐60 were compared to measurements of the two clinical reference standards, Semmes‐Weinstein monofilaments (SWM) and the Bio‐Thesiometer (BT).

Peripheral nerve sensitivity and temperature of the index finger tip of 41 subjects, with ages ranging from 20–87 years, and mean SWM size range of 2.4–4.2 were measured. Finger tip temperature ranged from 71.4–88.2°F, and the force exerted by the finger on the vibrating probe ranged from <1.0 to 82 grams. Each subject's nerve sensitivity was measured 3 times with each of the 3 instruments.

The correlation coefficient (r) of VPT‐60 and SWM was 0.75 and between the VPT‐60 and the BT was 0.86. The r between SWM and BT was only 0.51. As the nerve sensitivity decreased, the sensitivity of the VPT‐60 to detect changes increased. Reliability of the VPT‐60, SWM, and BT was 0.88, 0.78, and 0.74 respectively. In conclusion, this study indicates that the VPT‐60 provides results that are as capable as, if not more capable than, the gold standards in determining peripheral nerve sensitivity.
Evaluation of Ion Torrent's AmpliSeq Cancer Hotspot Panelv2 on FFPE tissues
Authors: Tunkey Christopher D. , Amos Christopher I. , Wells Wendy A. , Tsongalis Gregory J.
Published: June 2021

Objective

We routinely perform clinical molecular genetic testing using PCR‐based methods to detect BRAF, KRAS and EGFR variants in human cancers. Clinically actionable findings are reported to oncologists who can then personalize management strategies for any given patient. In this study we evaluated precision and accuracy of the Ion Torrent AmpliSeq Cancer Hotspot Panelv2 (46 genes).

Methods

Six known mutation positive FFPE tissues were selected (2 BRAF, 2 KRAS, 2 EGFR mutants). Normal kidney and colon FFPE tissues were used as controls. 10ng of starting material from each sample was used to prepare 8 barcoded libraries. The libraries were multiplexed and run through on 3 separate OneTouch reactions and then processed on 3 separate 316 chips over a period of two weeks.

Results

There was 100% concordance between previous PCR results and the corresponding variants identified in each of the mutant patient samples using the Ion Torrent panel. The BRAF, EGFR and KRAS mutations were consistently identified in each of the triplicate NGS reactions.

Conclusions

Samples chosen based on a diversity of important clinical results were selected as a test set to evaluate the performance of the AmpliSeq Cancer Panel Hotspotv2. Every sample included in an Ion Torrent run with a known mutation status has been confirmed when run on the PGM. We also demonstrated the ability of this platform to produce consistent variant calls between runs and offer better coverage of clinically important gene regions.
Stabilization of gene expression in whole blood: high quality RNA with room temperature handling of human blood samples.
Authors: Liberal Vasco , Stassinopoulos Angela , Whitney Scott , Wilkinson Steven , Huang Winnie , Muller Rolf , Muller‐Cohn Judy
Published: June 2021

Gene expression profiles from blood samples are being increasingly used to diagnose and monitor disease progression and assess patient responses to treatment. This requires reliable preservation of RNA in samples during collection, transport and storage. The highly dynamic transcription profiles can change rapidly after blood collection, potentially affecting interpretation of gene expression, and ultimately dictate inadequate treatment. Biomatrica has developed a collection device for whole blood, which stabilizes the RNA in blood cells and allows for ambient temperature sample handling and storage. We have also developed a robust RNA purification method that coupled with the RNAgard® Blood Tube provides a complete solution for blood collection, storage and RNA purification. We present data validating higher yields and greater quality of RNA while maintaining a gene expression profile similar to the time of collection. Comparative testing shows that blood in Biomatrica's RNAgard Blood Tube retains high quality RNA for much longer storage periods, and exhibits significantly less transcriptome variability than a leading competitor. Biomatrica's RNA preservation and purification solution can serve as the front end for research and molecular diagnostics and is more effective in delivering accurate gene expression patterns and prove of extreme value for better patient treatment.
9‐phenanthrol inhibits TMEM16A channels
Authors: Burris Sarah , Neeb Zachary P. , Jaggar Jonathan
Published: June 2021

Intravascular pressure stimulates several smooth muscle cell TRP channels, including TRPM4, leading to membrane depolarization and vasoconstriction (the myogenic response). Previous studies using 9‐phenanthrol (9‐phen), a TRPM4 channel inhibitor, suggest that TRPM4 is essential for pressure‐induced depolarization and constriction (Gonzales et al, AJP Cell 2010). We have recently shown that transmembrane protein 16A (TMEM16A), a Ca2+‐activated chloride channel, contributes to pressure‐induced depolarization and constriction (Bulley et al, Circ Res 2012). To investigate these apparently dissimilar results, we tested the hypothesis that 9‐phen is also a TMEM16A channel blocker. Patch‐clamp electrophysiology indicated that 9‐phen inhibited whole‐cell currents generated by recombinant TMEM16A channels expressed in HEK293 cells. Analysis of tail‐currents indicated that 9‐phen reduced TMEM16A currents in a concentration‐ and voltage‐dependent manner. IC50s for 9‐phen were 15 and 10 μM at −100 and +100 mV, respectively. Cell‐attached patch recordings indicated that 9‐phen (10 μM) reduced single TMEM16A channel open probability, amplitude, and mean open time, and increased mean closed time. Taken together, our data suggest that 9‐phen inhibits TMEM16A channels, in part by acting as a pore‐blocker. NHLBI/NIH
Inhibition of chemotherapy‐induced neuropathic pain with S1P receptor modulators
Authors: Salvemini Daniela , Janes Kali , Doyle Timothy , Bryant Leesa , Stockstill Katherine , Chen Zhoumou , Cuzzocrea Salvatore , Kamocki Krzysztof , Snider Ashley , Obeid Lina , Petrache Irina
Published: June 2021

Chemotherapy‐induced peripheral neuropathy (CIPN), accompanied by chronic neuropathic pain, is the major dose‐limiting toxicity of numerous antitumor agents. We now show that paclitaxel‐induced neuropathic pain is associated with increased sphingosine 1‐phosphate (S1P) formation in spinal cord acting through S1P1 receptor‐mediated NF‐κB and MAPKs activation and proinflammatory cytokine release. Inhibition of spinal cord S1P‐to‐S1PR1 signaling with the S1PR1 antagonist, W146, or FTY720, a FDA‐approved functional S1PR1 antagonist for treating multiple sclerosis, attenuated neuroinflammation and neuropathic pain. The effects of FTY720 occurred at non‐immunosuppressive doses without interfering with antitumor actions and extend to oxaliplatin and bortezomib‐induced neuropathic pain. Our findings identify the key role of the S1P‐to‐ S1PR1 pathway in CIPN and provide a first mechanistic insight into the S1P‐mediated biochemical pathways; establishing a pharmacological basis for S1PR modulators as chemotherapeutic adjuncts to block CIPN. Supported by Saint Louis University President Research Funds, Saint Louis University Cancer Center, Lymphoma and Leukemia Society Translational Research Grant.
Isoform‐selective glucuronidation of triclosan
Authors: Jackson Erin N , Schneider Jennifer , Faux Laura Rowland , James Margaret O
Published: June 2021

The antibacterial substance triclosan, 5‐chloro‐2‐(2,4‐dichlorophenoxy)‐phenol, is used in many consumer products resulting in widespread human exposure. Triclosan forms glucuronide and sulfate conjugates which are excreted in urine, however it is not known which UDP‐glucuronosyltransferase (UGT) isoform(s) catalyze triclosan glucuronidation. This study examined rates of glucuronidation of several concentrations of triclosan catalyzed by expressed human UGTs and individual human liver microsomes (HLMs) with 1 mM 14C‐UDP‐glucuronic acid as co‐substrate. The UGT isozymes studied are found in the human liver and intestine, the likely sites of triclosan glucuronidation. The results indicated UGT1A1 exhibited the highest glucuronidation efficiency (Vmax/Km). UGTs 1A3, 1A6,1A7, 1A8 and 1A9 also readily catalyzed triclosan glucuronidation, but UGTs 1A4, 1A10, 2B4, 2B7 and 2B15 were less active. Studies of triclosan glucuronidation in 23 individual HLMs were conducted at 0.01 mM and 1mM triclosan concentrations. More than 15‐fold variability in specific activity was found in the HLM samples at both concentrations of triclosan. These results suggest that clearance of triclosan by glucuronidation may be expected to vary considerably between individuals. Supported in part by R21 ES 020545 and an award from the University of Florida
Endoxifen and other metabolites of tamoxifen inhibit human hydroxysteroid sulfotransferase hSULT2A1
Authors: Squirewell Edwin , Qin Xiaoyan , Duffel Michael W.
Published: June 2021

Tamoxifen is a breast cancer therapeutic that has been shown to cause endometrial cancer in some women following treatment. hSULT2A1 is an enzyme that catalyzes the formation of an α‐sulfooxy derivative of tamoxifen that is reactive towards DNA. Moreover, hSULT2A1 functions in the metabolism of steroid hormones such as dehydroepiandrosterone (DHEA) and pregnenenolone (PREG). We hypothesize that major metabolites of tamoxifen regulate the catalytic activity of hSULT2A1, either through direct inhibition or through serving as alternate substrates for the enzyme. Our studies show that endoxifen is a potent inhibitor of hSULT2A1‐catalyzed sulfation of PREG and DHEA with Ki values of 3.5 μM and 2.8 μM, respectively. 4‐Hydroxytamoxifen and N‐desmethyltamoxifen had Ki values of 12.7 μM and 9.8 μM, respectively, for sulfation of PREG, whereas Ki values of 19.4 μM and 17.2 μM were observed for these metabolites when determining the sulfation of DHEA. A Ki of 9.1 μM was observed for tamoxifen‐N‐oxide when DHEA was used as a substrate, and this increased to a Ki value of 16.9 μM when PREG was utilized as a substrate. Subsequent assays indicated that N‐desmethyltamoxifen was a substrate for the enzyme. These results may be useful in interpreting ongoing clinical trials of endoxifen and in improving the design of related molecules. [Supported by NIH grant CA038683]
Positive feedback influence of KCa3.1 channels on stimulated endothelial Ca2+ dynamics
Authors: Taylor Mark Stephen , Köhler Ralf , Michael Francis , Qian Xun
Published: June 2021

Dynamic Ca2+ transients underlie physiologic endothelial signaling and vasoregulation. Intermediate conductance Ca2+‐activated K+ channels (KCa3.1) are primary targets of endothelial Ca2+ signals in the arterial vasculature and their ablation results in hypertension. Localized Ca2+ events target KCa3.1 channels along the myoendothelial interface to elicit hyperpolarization and vasodilation. Here, we employ genetically altered KCa3.1 knockout mice (IK1−/−), confocal imaging, and custom analysis to assess possible feedback influence of these channels on the Ca2+ dynamics themselves. Basal endothelial Ca2+ dynamics were not different between IK1−/− and wild‐type (WT) mice (p < 0.05). However, acetylcholine (Ach) stimulated Ca2+ dynamics to a much greater degree in WT than in IK1−/− endothelium (sites 376 vs. 156; p < 0.05). Removal of extracellular Ca2+ reduced Ach‐induced Ca2+ dynamics in WT to the level of IK1−/−. These findings indicate that whereas KCa3.1 channels do not directly influence basal Ca2+ activity in mouse mesenteric artery endothelium, they exert substantial positive feedback under stimulated conditions, recruiting additional new Ca2+ sites and events along the intima. Overall, our findings suggest that KCa3.1 channels amplify endothelial Ca2+ dynamics, further enhancing arterial hyperpolarization and vasodilation. Supported by NIH R01 HL085887 and S10RR027535.
Pressor and renal regional hemodynamic effects of urotensin II in neonatal pigs
Authors: Soni Hitesh , Adebiyi Adebowale
Published: June 2021

Renal expression of urotensin II (UII) and its receptor (UTR) are dependent on kidney maturation and anatomical regions. However, renal regional hemodynamic effects of UII in neonates, in whom the kidneys are functionally immature, are unclear. Here, we investigated regional hemodynamic responses to acute intrarenal administration of UII in newborn pigs. Western immunoblotting and immunofluorescence indicated that UTR is expressed in newborn pig renal microvessels and microvascular smooth muscle cells. Intrarenal bolus injections of human UII (hUII; 1–100 ng·kg−1) resulted in a dose‐dependent decrease in total renal blood flow (RBF) and an increase in mean arterial pressure (MAP) and renal vascular resistance (RVR) in newborn pigs. Cortical blood flow (CBF) measured by laser Doppler flowmetry was ~2‐fold higher than medullary blood flow (MBF) in newborn pigs. hUII dose‐dependently reduced CBF, but increased MBF in the pigs. hUII‐induced MAP elevation and hemodynamic changes were inhibited by urantide, a UTR inhibitor, but not losartan, a type 1 angiotensin II receptor inhibitor. U73122, a phospholipase C (PLC) inhibitor and 2‐aminoethoxydiphenyl borate, an inositol 1,4,5 trisphosphate (IP3) receptor blocker attenuated hUII‐induced MAP and RVR elevations, RBF and CBF reductions, but not MBF increase. These findings indicate that intrarenal administration of hUII elevates blood pressure and induces regional‐selective renal hemodynamic changes in newborn pigs. Our data also suggest that PLC/IP3 signaling pathway contributes to hUII‐induced alterations in MAP, RBF, RVR, and CBF, but not MBF in newborn pigs.