bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Leukocyte Antigen (HLA) Genotype and Probiotics Modify the Association between Early Infant Feeding and Islet Autoimmunity in the TEDDY Study
Authors: Uusitalo Ulla , Lee Hye‐Seung , Aronsson Carin Andren , Vehik Kendra , Yang Jimin , Hummel Sandra , Silvis Katherine , Lernmark Åke , Rewers Marian , Hagopian William , She Jin‐Xiong , Simell Olli , Toppari Jorma , Ziegler Anette G. , Akolkar Beena , Krischer Jeffrey , Virtanen Suvi , Norris Jill ,   The TEDDY Study Group
Published: November 2023

Several studies have examined the duration of breastfeeding and timing of introduction of complementary foods in the development of type 1 diabetes (T1D) related islet autoimmunity (IA). The results have varied by country. This is the first attempt to study the association in a multi‐country setting.

The Environmental Determinants of Diabetes in the Young (TEDDY) study prospectively follows 8676 children with T1D‐associated human leukocyte antigen (HLA) DR‐ and DQ‐alleles in the U.S., Finland, Germany, and Sweden. Blood samples were collected every 3 months from birth to evaluate the primary outcome of IA, defined as appearance of one or more of the islet cell antibodies to insulin, glutamic acid decarboxylase or insulinoma antigen‐2, confirmed in two laboratories at two consecutive visits. The age at introduction of solid foods was categorized: < 4, 4–5.9, and ≥ 6 months. Reported probiotic exposure via infant formula or via dietary supplements during the first year of life was collected. Children followed at least 1 year (n=7481) were included in the study. We used Cox regression model adjusting for country, family T1D status, HLA‐DR‐DQ genotype, and sex of the child to study the risk of IA.

Significant interactions between the HLA‐DR‐DQ genotype, probiotic exposure before 6 months, and timing of solid foods (pinteraction=0.036) and gluten containing cereals (pinteraction=0.015) in relation to IA risk were detected. Early introduction of any solid food was associated with increased risk of IA (<4 months versus ≥6 months: HR 2.46; 95% CI 1.43, 4.22, p=0.001, and 4–5.9 months versus ≥ 6 months: HR 2.32; 95% CI 1.36, 3.94, p=0.002) if the child had DR3/4 genotype and had not received probiotics during the first 6 months of life. However, if probiotics were given before 6 months of age the risk of IA was lower among those with DR3/4 genotype: introduction of solid foods before 4 months of age: HR 0.44; 95% CI 0.20, 0.99, p=0.047 and at 4–5.9 months: HR 0.35; 95% CI 0.16, 0.77, p=0.009, respectively, when compared with introduction of solid foods at 6 months or later.

We conclude that timing of solid food introduction may be associated with IA in TEDDY in a manner linked to HLA genotype and to the exposure of probiotics during the first 6 months.

Support or Funding Information

Funded by NIH, NIDDK, NIAID, NICHD, NIEHS, JDRF and CDC.
Serum Trimethylamine N‐Oxide (TMAO), Diet, and Colorectal Cancer Risk
Authors: Guertin Kristin A , Graubard Barry I , Goedert James J , Li Xin‐Min , Hazen Stanley L , Sinha Rashmi
Published: November 2023

Objective

Choline and carnitine are two compounds that are found in human diets and can be metabolized to trimethylamine (TMA) and, ultimately, to Trimethylamine N‐Oxide (TMAO) by the gut microbiota. There is evidence that individuals with higher serum TMAO concentrations may have subclinical inflammation and greater risk of deleterious health outcomes, including heart disease. There is limited research on the potential role of TMAO in colorectal risk, and no studies that consider dietary intake of choline‐ and carnitine‐containing foods simultaneously with TMAO and colorectal cancer risk. We evaluated the association between serum TMAO and colorectal cancer risk (total, proximal, distal, and rectal colorectal cancers) in a nested case‐control study.

Methods

Within the Alpha‐Tocopherol Beta‐Carotene (ATBC) Study, a cohort of male smokers, we incidence‐density matched 644 incident colorectal cancer cases (median 14 years after baseline) and 644 controls. We selected baseline fasting serum samples and used liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) to measure TMAO and stable isotope dilution LC‐MS/MS to measure carnitine and choline. Spearman correlations describe the association between these serum biomarkers and the association between the biomarkers and self‐reported dietary intake of choline‐ and carnitine‐containing foods (e.g., meat, eggs). Logistic regression models estimate the adjusted odds ratios (OR) and 95% confidence intervals (CI) for colorectal cancer by quartile of each serum biomarker and, separately, by choline‐ and carnitine‐containing foods.

Results

Colorectal cancer risk was not associated with serum TMAO [ORhighest vs. lowest quartile=1.20; 95% CI=0.86–1.68], carnitine [ORhighest vs. lowest quartile=1.03; 95%CI=0.73–1.44], or dietary intake of choline‐ and carnitine‐containing foods. Individuals with higher serum choline had greater risk of colorectal cancer, with about a 3‐fold greater risk for the highest compared to lowest quartile in models adjusted for age, batch, and smoking [OR (95% CI)=3.38 (2.37–4.80); P‐trend<0.0001]; results were similarly robust for all anatomical subsites (P‐trend<0.0001 for proximal, distal, and rectal colon cancers).

Conclusions

This study provides strong evidence that alterations in choline metabolism, here investigated as higher serum choline concentrations, may be a marker of increased colorectal cancer risk. There was no evidence of an association between serum TMAO and risk of colorectal cancer. The strength of the associations between choline and colorectal cancer risk and the consistency of the risk estimates across all examined anatomical subsites strongly supports a role of altered choline metabolism in colorectal cancer pathogenesis.

Support or Funding Information

Intramural Research Program of the National Cancer Institute of the National Institutes of Health
Comparing the Impact of Saturated Fatty Acids from Different Dairy Sources on Plasma Lipids: Preliminary Data from a Randomized Controlled Trial
Authors: Tessier‐Grenier Maude , Brassard Didier , Rajendiran Ethendhar , She Yongbo , Ramprasath Vanu , Gigleux Iris , Levy Emile , Tremblay Angelo , Jones Peter , Couture Patrick , Lamarche Benoît
Published: November 2023

Objective

To investigate the impact of saturated fatty acids (SFA) from different dairy products, namely cheese and butter, on plasma lipid levels in men and women.

Methods

This multicenter randomized crossover trial study involved 90 men and women with abdominal obesity and relatively low HDL‐C levels from the Quebec City and Winnipeg metropolitan areas. Subjects were randomized to sequences of 5 diets of 4 weeks each, with a 4 week washout between diets. The 5 isocaloric diets were the following: 1) a diet rich in SFA from cheese (CHEESE); 2) a diet rich in SFA from butter (BUTTER); 3) a diet rich in mono‐unsaturated fatty acids (MUFA); 4) a diet rich in poly‐unsaturated fatty acids (PUFA); and 5) a low‐fat, high carbohydrate diet (CHO). The macronutrient, calcium and sodium content of each diet is shown in Table 1. All foods were provided to participants in the context of this feeding study.

Preliminary results

Preliminary results are presented for a total of 57 participants recruited in Quebec City. There was no significant change in anthropometric measurements among the five diets. BUTTER significantly raised serum LDL‐C and HDL‐C (+7.1% and +8.0% respectively, p<0.01) while CHEESE only raised serum HDL‐C (+5.3%, p<0.05) compared with CHO. Serum LDL‐C concentrations were higher with BUTTER than with CHEESE (+2.5%, p=0.02) while serum TG were higher with CHEESE than with BUTTER (+9.5%, p=0.007). Lowest serum LDL‐C concentrations were seen after the MUFA and the PUFA diets.

Conclusion

Preliminary data from this carefully controlled feeding study suggest that the cardiometabolic change induced by consumption of SFA may be influenced by its food source. SFA from cheese may have less of an effect on serum LDL‐C but may be associated with increased plasma TG levels compared with SFA from butter. This suggests that the food matrix may influence the impact of constituent nutrients on health. The impact of such findings in terms of cardiovascular risk needs to be investigated further.

Support or Funding Information

Dairy Research Cluster Initiative (Dairy Farmers of Canada, Agriculture and Agri‐Food Canada, the Canadian Dairy Network and the Canadian Dairy Commission)

Macronutrient (% energy), calcium and sodium content of experimental diets

CHEESE
BUTTER
MUFA
PUFA
CHO

Lipids (%)
32
32
32
32
25

SFA (%)
13
13
6
6
6

MUFA (%)
14
14
21
14
14

PUFA (%)
5
5
5
12
5

Carbohydrates (%)
52
52
52
52
59

Proteins (%)
16
16
16
16
16

Calcium (mg/2500 kcal)
1261
812
812
812
842

Sodium (mg/2500 kcal)
2482
2480
2479
2479
2485
Comparison of Different Tissue Clearing Methods and 3D Imaging Techniques for Visualization of GFP‐expressing Mouse Embryos and Embryonic Hearts
Authors: Kolesova Hana , Capek Martin , Janacek Jiri , Sedmera David
Published: November 2023

Aims

To find a suitable tissue clearing protocol preserving GFP fluorescence for whole mount imaging of embryonic and adult hearts of transgenic mice.

Methods

We tested various published organic solvent‐based or water‐based clearing protocols claiming to preserve GFP fluorescence in central nervous system: tetrahydrofurane dehydration and dibenzyl ether protocol (DBE), SCALE, CLARITY, and CUBIC to evaluate their use for embryonic heart tissue. Tissue transparency and preservation of GFP fluorescence was evaluated in hearts of Cx40:GFP knock in mice at stages: ED10.5 – ED18.5, and adults. We also tested several imaging techniques and evaluated their use for embryonic hearts visualization. We compared an Upright Single Photon Confocal Microscope (Olympus) with 4×, 10×, and 25× objectives after mounting the specimens into cavity slides or custom imaging chambers with an Optical Projection Tomography (OPT, custom made) and with a Single Plane Ilumination Microscopy (SPIM) (constructed according to the OpenSPIM project).

Results

Despite careful control of all critical parameters, DBE clearing protocol did not preserve GFP fluorescence; in addition, it caused considerable tissue shrinking and deformation. The SCALE clearing resulted in good tissue transparency up to ED12.5; at later stages and in the adults the useful depth of imaging was limited by tissue light scattering. The CLARITY method considerably improved tissue transparency at later stages, but also decreased GFP fluorescence intensity. The CUBIC method resulted in very well cleared specimens even at the adult stages, and GFP fluorescence was also preserved. In addition, it decolorized the blood and myocardium by removing iron from the tissues. High‐resolution images were thus obtained even from deep tissue layers with a confocal microscope with a Scale View immersion 25× objective. Simultaneously, intact specimens were imaged with OPT and SPIM that have lower resolution, but are more suitable for observing large samples.

Conclusions

From the tested methods, the CUBIC protocol turned out to be the best for whole mount GFP cardiac specimens. SCALE technique is also suitable for younger embryos. Confocal microscopy is needed for detailed view, while OPT and SPIM enable to observe larger samples. These protocols will be used for three dimensional reconstructions of normal and abnormal specimens to visualize the developing cardiac conduction system and coronary vasculature.

Support or Funding Information

Supported by 13‐12412S from the Czech Science Foundation, Ministry of Education PRVOUK P35/LF1/5, institutional support RVO:67985823, and Charles University UNCE.
Assessment of an Educational Intervention on Blood Pressure Knowledge and Risk Factor Practices in Collegiate Football Athletes
Authors: Armstrong Sarah , Jackson Loreto , Sharp Julia , Cason Katherine , Haley‐Zitlin Vivian
Published: November 2023

Hypertension is a serious health problem affecting approximately one‐third of the adult population in the US. While athletes work to maintain high levels of fitness, they too, are affected by this disease. American–style football is the most popular sport in which adolescents within the US participate. This translates into many individuals pursuing collegiate football activities. Few studies have examined blood pressure and football relationships. However, an increase in blood pressure was observed within the first season of playing football, with a diagnosis of hypertension in >10% of individuals assessed. Interestingly, some training‐related behaviors associated with athletes’ regimens to enhance performance such as intense resistance training, a relative lack of aerobic exercise, and weight gain have been suggested to play a role in professional football players’ diagnoses of hypertension and left ventricular hypertrophy, as well as premature CVD‐related mortality. Lifestyle choices have been identified that increase the risk for hypertension. Education of college‐age athletes regarding these risk factors has the potential to result in better blood pressure control. Sixty‐six college‐aged football athletes were queried on blood pressure knowledge and diagnostic criteria along with selected dietary habits and training activities via a pre‐test. An educational intervention in the form of a Fact Sheet was delivered via paper copy (most preferred method) and e‐mail per the preferences expressed by the athletes. A post‐test was delivered to assess changes in knowledge of hypertension and change in hypertension‐related dietary practices. Pretest results indicated variability in the knowledge of the collegiate football athletes regarding hypertension and a need for educational intervention. The pretest and post‐test results will provide evidence of the effectiveness of the intervention.

Support or Funding Information

NA
Characterizing Six Commercially‐Available Teas for Polyphenol Content and Antioxidative Capacity in Relation to Steep Time
Authors: McAlpine Michael D. , Ward Wendy E.
Published: November 2023

Globally, tea is the most commonly consumed beverage after water and is associated with many health benefits. These health benefits are often attributed to the unique profile and abundance of polyphenols in tea. In vivo, polyphenols have been shown to exhibit antioxidant activity and are thus capable of inhibiting free radical production. The purpose of this study was to characterize six popular and commercially available teas in terms of total polyphenolic content (TPC) and relative antioxidant capacity after varying the steep times. Dragonwell (DW), Japanese Sencha (JS), English Breakfast (EB), Golden Monkey (GM), Green Rooibos (GR), and Red Rooibos (RR) loose leaf teas were individually steeped in water for 5 or 10 minutes at the manufacturer's recommended temperature (79°C for DW, JS; 96°C for EB, GM, GR, RR) to extract polyphenols. TPC was determined using Folin‐Ciocalteau's reagent with Gallic acid as a standard; and antioxidative capacity was determined by the ability of a normalized amount of polyphenols (1μg/ml) from each tea to scavenge the free radical 2, 2‐diphenyl‐1‐picrylhydrazyl (DPPH). TPC was observed to be significantly greater (p<0.05) after 10 minutes of steeping than it was after 5 minutes for each of the teas. However, 66–73% of polyphenols measured at 10 minutes were extracted in the first 5 minutes (% of total TPC extracted: DW=66.08±6.36, JS=73.37±3.08, EB=71.74±4.74, GM=65.40±2.69, GR=68.31±3.17, RR=71.14±5.95) regardless of tea type. After 5 minutes of steeping, significant differences (p<0.05) in TPC were observed among different teas (JS~EB>DW>GM~GR~RR). Different teas with their respective polyphenol profiles also varied (p<0.05) in their ability to inhibit the free radical DPPH (% inhibition: DW>JS~EB~GM>GR>RR). In summary, TPC varies greatly among types of tea, but the majority of polyphenols are extracted after 5 minutes of steeping regardless of tea type. The wide range in the ability of the different teas to scavenge free radicals, even after normalizing for TPC, indicates an important role for the specific polyphenol profile and potential different health benefits among teas.

Support or Funding Information

NSERC
“Genetic Haplotypes and Family History of Alzheimer's Disease Modify Homocysteine and Influence Neurovascular Pathology”
Authors: Webb Joe L , Willette Auriel A
Published: November 2023

Objective

To delineate novel relationships between Alzheimer disease (AD) factors, such as Translocase of the Outer Mitochondrial Membrane ‐ kD 40 (TOMM40) genotype, AD family history (FH), the vascular risk factor homocysteine, and vascular neuropathology.

Methods

We performed linear mixed model regression on longitudinal imaging and biomarker data from the Alzheimer's Disease Neuroimaging Initiative (ADNI) cohort. Among 210 healthy adults with 1 or 2 Apolipoprotein ɛ4 (APOE4) alleles (mean age 73.8), we stratified by TOMM40 genotypes S/L (n=84), L/L (n=49), L/VL (n=68). Main effects and interactions between TOMM40 and FH were conducted on neurovascular outcomes, including white matter hyperintensities (WMH), infarctions, as well as total intracranial volume. Vascular risk factors were similarly tested, including homocysteine and total cholesterol. Models were adjusted for age, gender, body mass index and hyperglycemia status.

Results

FH * TOMM40 interactions showed that L/L and L/VL genotypes respectively exercised “beneficial” effects for FH negative (FH−) and “detrimental” effects for FH positive (FH+) participants. In FH+, for example, risk alleles were associated with higher serum homocysteine (P=0.024), total cholesterol (P=0.005), infarction occurrence (P=0.045), and greater WMH burden (P = 0.032). Total intracranial volume was also smaller (P<0.001). Homocysteine levels mediated FH * TOMM40 associations with neurovascular measures.

Conclusions

These results suggest that FH modulates TOMM40's effects on vascular markers and neurovascular pathology of AD in APOE4 participants. These interactions between FH and TOMM40 influence on neuropathology may be mediated via homocysteine suggesting that TOMM40 genotype could serve as a novel predictor for AD and that homocysteine may be one potential mechanism.

Support or Funding Information

This study was supported in part by the National Institutes of Health award number K99AG047282, an Iowa State University Stewart Research Award and an Iowa State University University Honor's Program Research Award. Role of Funding: The funding sources had no role in the design or conduct of the study; collection, management, analysis, and interpretation of the data; preparation, review, or approval of the abstract.
THE ROLE OF INSULIN RESISTANCE AND SIRT‐1 IN CARDIAC REMODELING INDUCED BY CIGARETTE SMOKE
Authors: Azevedo Paula Schmidt , Pires Vanessa , Santos Priscila , Gonçalves Andrea , Okoshi Marina , Fernandes Ana Angelica , Roscani Meliza , Polegato Bertha , Paiva Sergio , Zornoff Leonardo , Minicucci Marcos
Published: November 2023

The aim of this study is to investigate if insulin resistance and SIRT‐1 participate of cardiac remodeling induced by TS.

Methods

20 animals were allocated into 2 groups: group exposed to cigarette smoke (CS) and group not exposed to cigarette smoke (C). After 2 months the animals were subjected to: a) echocardiographic study; b) energy metabolism enzymes evaluation; c) Western blot for Akt, AMPK, SIRT‐1 and GLUT‐4. Cotinine and insulin was measured by ELISA kit. The HOMA‐index was calculated. The CS group showed larger left atrium size and lower ejection fraction than those of group. Serum cotinine was positive in CS exposed group (P<0,001). HOMA‐index showed insulin resistance in CS group (P<0,05). As regards the analysis of energy metabolism enzymes, phosphofructokinase (PFK); P = 0.003), lactate dehydrogenases (LDH) (P = 0.017) were greater in the TS group and citrate synthase P <0.001), complex I ( P <0.001), complex II (P <0.001), ATP synthase (P <0.001) were lower in the TS group. Akt/Aktp was similar between the groups. AMPK was reduced in CS group (p<0,001), SIRT‐1 increased in CS group (P<0,001) and GLUT‐4 (P=0,47) was similar.

Conclusion

Cigarette smoke induced insulin resistance, that may have reduced AMPK and probably compromised glucose metabolism and energy transformation. SIRT‐1 increases in oxidative stress set and may promote hypertrophy and apoptosis. Therefore, SIRT‐1 is also a possible metabolic pathway involved in CS induced remodeling.

Support or Funding Information

Supported by FAPESP
Recommendations of the 2015–2020 Dietary Guidelines for Americans
Authors: Casavale Kellie O. , Stoody Eve Essery , Rihane Colette , Olson Richard
Published: November 2023

Objective

The US Departments of Health and Human Services and Agriculture released the 8th edition (2015–2020) of the Dietary Guidelines for Americans (DGA) in January 2016.

Methods

The DGA is required under the National Nutrition Monitoring and Related Research Act of 1990 and informs the development of federal food, nutrition, and health policies and programs.

Results

The 2015–2020 DGA focuses on shifts in eating patterns to align current dietary intake with recommendations and highlight multiple strategies across all segments of society to promote healthy eating and physical activity behaviors. In abbreviation, the 5 overarching Guidelines are to follow a healthy eating pattern across the life span; focus on variety, nutrient density, and amount; limit calories from added sugars and saturated fats and reduce sodium intake; shift to healthier food and beverage choices; and support healthy eating patterns for all. Key Recommendations provide further guidance on how individuals can follow the five Guidelines and state that a healthy eating pattern accounts for all foods and beverages within an appropriate calorie level and includes a variety of vegetables from all of the subgroups (dark green, red and orange, legumes/beans and peas, starchy, and other); fruits (especially whole fruits); grains (at least half of which are whole grains); fat‐free or low‐fat dairy (including milk, yogurt, cheese, and fortified soy beverages); a variety of protein foods (including seafood, lean meats and poultry, eggs, legumes/beans and peas, and nuts, seeds, and soy products); and oils. A healthy eating pattern also limits saturated fats and trans fats, added sugars, and sodium. Key Recommendations that are quantitative are provided for several components of the diet of particular public health concern that should be limited: consume less than 10% of calories per day from added sugars; consume less than 10% of calories per day from saturated fats; consume less than 2,300mg/d of sodium; and if alcohol is consumed, it should be consumed in moderation(up to 1 drink per day for women and up to 2 drinks per day for men and only by adults of legal drinking age). The DGA also recommends to meet the Physical Activity Guidelines for Americans.

Conclusion

Everyone has a role in supporting healthy eating patterns. Concerted efforts among health care professionals, communities, businesses and industries, organizations, governments, and other segments of society are important to support individuals and families in making dietary and physical activity choices that align with the DGA. Health care professionals can help individuals identify how they can modify and improve their dietary patterns. Educational materials for both professionals and consumers will be available in 2016.

Support or Funding Information

None
Effect of Baking on the β‐carotene Content of Orange Flesh Sweetpotato (Ipomoea batatas) Puree Bread and OFSP Flour Bread
Authors: Muzhingi Tawanda , Mbogo Daniel , Low Jan , Magnaghi Antonio , Heck Simon , Gule Simon
Published: November 2023

Orange Fleshed Sweetpotato (OFSP) is a biofortified crop rich in provitamin A carotenoids, and is being promoted a sustainable food‐based strategy to address vitamin A deficiency in sub Saharan Africa. This study was aimed at evaluating the provitamin A carotenoid retention in bread made using OFSP flour to substitute for 10% of wheat flour, and bread made using OFSP puree to substitute for 45% of wheat flour. Samples of OFSP flour, OFSP puree, bread and ingredients for baking were obtained from local bakers in Kenya. The OFSP flour, OFSP puree, dough and breads were extracted for carotenoid by KOH saponification and hexane extraction. The extracted carotenoids were analyzed in an HPLC system with C30 Carotenoid S column and eluted in methanol, water and methyl tert butyl ether gradient profiles. The major carotenoids of OFSP were all‐trans β‐carotene, and its isomers (9‐cis β‐carotene and 13‐cis β‐carotene). Bread made with OFSP puree had the highest all‐trans β‐carotene content of 3.14 mg/100g dry weigh (dw) compared to 0.08mg/100g dw of OFSP flour bread. Adjusting for moisture content the OFSP puree bread will provide about 507μg all‐trans β‐carotene/slice (39g) of bread. Therefore, two slices of OFSP puree bread will provide 42.25 Retinol Activity Equivalence (RAE), which meets almost 30% of the RDA for vitamin A in children aged 1–3 years old.

Support or Funding Information

DFID (CIP SUSTAIN) and BMGF (CIP SASHA 2)
Control of NF‐κB activity, inflammation and stemness properties in cancer cells by Copper Metabolism (Murr1) Domain‐Containing 1
Authors: Yeh Da‐Wei , Lai Chao‐Yang , Liu Yi‐Ling , Lu Chih‐Hao , Chuang Tsung‐Hsien
Published: November 2023

It has been shown that persistant activation of NF‐κB in cancer cells contributes to inflammation, expansion of tumor‐initiating cancer stem‐like cells (CSCs), and tumor progression. In contrast, NF‐κB activation in CSCs can elevate expression and release of pro‐inflammatory mediators into the tumor microenvironment and further increase stemness and inflammatory conditions in cancer cells. Nevertheless, the underlying molecular controls are still ill‐defined. In this study, we used bioinformatic analysis to show the upregulation of NF‐kB‐target pro‐inflammatory gene and downregulation of Copper Metabolism (Murr1) Domain‐containing 1 (COMMD1) during the enrichment of stemness of SAS, an oral cancer cell line in tumor sphere culture. The 3′‐UTR of COMMD1 mRNA consists of miR‐205 target site. Parallel studies with SAS and non–small‐cell lung cancer cells, H460 and D121 indicated that miR‐205 reduces COMMD1 expression and the expression of miR‐205 was upregulated upon NF‐kB activation. COMMD1 effectively restrained inflammatory stimuli‐induced NF‐κB activation, cytokine production, as well as leukocytes migration. The lentiviral shRNA‐mediated knockdown of COMMD1 increased the expression of stemness genes, sphere‐forming capacity, and potential for anchorage‐independent growth in cancer cells. Moreover, study with cancer animal model showed that COMMD1 knockdown enhances tumorigenesis and tumor growth, as well as expanded population of CD11b+ tumor‐associated leukocytes and CD117+ stemness‐enriched cancer cells. These results suggest that the miR‐205‐COMMD1‐NF‐κB axis forming a positive feedback loop for amplifying inflammatory and stemness‐associated properties of cancer cells that further promote pro‐inflammatory tumor microenvironment.
Thiostrepton inhibits Toll‐like receptors 7, 8 and 9 mediated inflammatory responses
Authors: Chuang Tsung‐Hsien , Lai Chao‐Yang , Yeh Da‐Wei , Lu Chih‐Hao , Liu Yi‐Ling
Published: November 2023

Toll‐like receptors (TLRs) play a key role in sensing pathogen‐associated molecular patterns for initiating immune responses against microbial infections. Ten TLRs are identified in human cells. Of them, TLRs 7, 8 and 9 (TLR7‐9) comprise a subfamily. Activation of these three TLRs has been linked to the pathogenesis of autoimmune diseases including psoriasis. Thus antagonists of these TLRs are being investigated for their therapeutic applications on these diseases. In an attempt to develop novel TLR7‐9 inhibitors, we searched the Gene Expression Omnibus database for gene expression profiles in cells treated with bortezomib which is known as an inhibitor for TLR7‐9. These profiles were then used as an input to screen the Connectivity Map database for chemical compounds with similar functions as bortezomib. The result suggest that the antibiotic thiostrepton could has similar function as bortezomib in inhibition of TLR7–9. Further studies showed that thiostrepton effectively inhibits TLR7–9 activation in cell‐based assays and dendritic cells. In contrast to bortezomib, thiostrepton is less cytotoxic to dendritic cells, and its inhibitory activity is more specific to TLR7–9. Thiostrepton inhibits TLR9 localization in endosomes. In different animal models, thiostrepton attenuated LL37‐ and imiquimod‐induced psoriasis‐like inflammation. These results indicated that thiostrepton is a novel inhibitor for TLR7–9.
Routine, Automated RNA in situ Hybridization Profiling of Target Candidates and Safety Markers in Preclinical Animal Model Tissue Panels using RNAscope® LS Assay on Leica Biosystems’ BOND RX
Authors: Fang Li , He Ming‐Xiao , Kim Daniel , Franks Tania , Roy Marc , Bunker Chris , Luo Yuling , Ma Xiao‐Jun , Park Emily
Published: November 2023

Determining tissue expression profiles of therapeutic target candidates is essential for safety assessment and the potential for adverse events. Immunohistochemistry is often not possible due to lack of reagents with the required specificity and sensitivity coupled with extensive time in attempts to validate IHC. RNAscope® in situ hybridization (ISH) technology is a universal assay able to detect and characterize tissue distribution of any target mRNA. RNAscope ISH has molecular sensitivity, high specificity and robust performance in FFPE tissue. Fully automated RNAscope ISH is performed on Ventana Discovery and Leica Biosystem Bond RX instrumentation; detection of marker RNAs is achieved in 8 hours and visualized via bright field microscopy. Marker RNAs can be quantified with cellular resolution using digital image analysis software.

We present RNAscope methods for routine detection of very low copy RNAs, typical of most targets in normal tissues, as well as applications for detection of cell‐type, proliferation and apoptosis markers in animal tissues used in safety assessment and toxicity studies. We demonstrate robust performance in 25 tissues from rat, dog and cynomolgus monkey using probes to low‐, medium‐ and high‐expression housekeeping gene RNAs (POLR2A, PPIB and UBC). Specific signals and well‐maintained morphology was achieved efficiently for all 25 tissues. Based on optimized epitope retrieval conditions, tissues were grouped into two multi‐tissue microarrays for high‐throughput marker analysis. We examined and present tissue‐specific expression patterns of endothelial marker CD31, macrophage marker CD68, proliferation markers Ki‐67 and Cyclin E1, and apoptosis molecules, Puma, Fas/CD95 and DR5.

The presentation demonstrates RNAscope as a method that can be replicated in any lab and suitable for efficient and reliable detection of any target candidate or safety/toxicity biomarker mRNA from any species for interrogation in any tissue.
Oxidative stress and diabetic wound healing
Authors: Tie Lu , Shi Yun‐Di , Li Xue‐Jun
Published: November 2023

Objectives

Refractory wounds in diabetic patients constitute a serious complication that often leads to amputation with limited treatment regimens. Oxidative stress has been proposed as an important pathogenic factor in diabetic wound complications. The present study was designed to determine the protective effect of the GTP cyclohydrolase I (GTPCH), the rate‐limiting enzyme of BH4 synthesis; soy isoflavone genistein and Ganoderma lucidum polysaccharide (Gl‐PS) on diabetic wound healing.

Methods

Streptozotocin‐induced diabetic mice with full‐thickness excisional wounds were used.

Results

The rate of wound closure in diabetic mice was delayed compared to normal wild‐type (WT) mice, which was rescued in GTPCH transgenic (Tg‐GCH) diabetic mice. In WT diabetic mice, cutaneous eNOS activity and nitrite levels were decreased compared to WT mice, paralleled by increased levels of superoxide anion (O2.−) level. In Tg‐GCH diabetic mice, nitrite levels were potentiated and O2.− level was suppressed. Genistein treatment dose‐dependently rescued the delay of wound closure in diabetic mice and increased the mean perfusion rate. Genistein significantly normalized the elevated O2− production and reversed the attenuated nitrite level by inhibition of forkhead box O transcription factor 1 (FoxO1) levels. Gl‐PS treatment could also rescue the delay of wound closure in diabetic mice. Gl‐PS normalized the increased mitochondrial O2.− production and nitrotyrosine formation by inhibition of MnSOD nitration and the redox enzyme p66Shc expression in diabetic wound tissues.

Conclusion

GTPCH, genistein and Gl‐PS rescued the delayed wound healing and improved wound angiogenesis in diabetic mice, at least in part, by suppression of oxidative stress.

Support or Funding Information

This work was supported by the National Natural Science Foundation of China No. 81373405 and 30901803, Beijing Higher Education Young Elite Teacher Project (No. YETP0053), Leading Academic Discipline Project of Beijing Education Bureau (No. BMU20110254) and the Fund of Janssen Research Council China (JRCC2011).
Inhibition of STAT3 phosphorylation by sulforaphane reduces the lipopolysaccharide‐induced adhesion molecule expression in vascular endothelial cell
Authors: Ahn Hee Y , Cho Young S
Published: November 2023

Intercellular adhesion molecule 1 (ICAM‐1) and vascular cell adhesion molecule 1 (VCAM‐1) play key roles in the initiation of vascular inflammation. In this study, we explored whether sulforaphane, a dietary phytochemical, can inhibit the expression of ICAM‐1 and VCAM‐1 in human umbilical vein endothelial cells (HUVEC) stimulated with lipopolysaccharide (LPS), and the mechanisms involved. Sulforaphane prevented the LPS‐mediated increase in ICAM‐1 and VCAM‐1 expression, (P<0.01) in HUVEC. Sulforaphane also prevented the LPS‐mediated increase in the phosphorylation of signal transducer and activator of transcription 3 (STAT3) (P<0.01). Stattic, a STAT3 inhibitor, reduced the LPS‐induced expression of ICAM‐1 and VCAM‐1, and STAT3 phosphorylation (P<0.01). STAT3 small interfering RNA treatment reduced the LPS‐induced expression of ICAM‐1, VCAM‐1 and STAT3 (P<0.01). Sulforaphane reduced LPS‐mediated THP‐1 monocyte adhesion to HUVEC (P<0.01).

These data provide insight into the mechanism through which sulforaphane partly reduces the expression of ICAM‐1 and VCAM‐1 on the vascular endothelial cell by inhibiting STAT3 phosphorylation.

Support or Funding Information

This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(2012010913).
Characterization and Functional Effects of Multi‐site Interactions between calmodulin and the angiotensin II receptor type 1A
Authors: Clements Robert , Ehlers Kevin , VerMeer Mark , Tran Kim
Published: November 2023

The angiotensin II receptor type 1A (AT1AR) is responsible for many effects of angiotensin II (AngII). Calmodulin (CaM) is essential for many cell functions due to its interactions with many proteins. CaM has previously been demonstrated to interact with AT1AR at a.a. 305 – 327 in the fourth submembrane domain (SMD4) (Thomas WG et al., FEBS Lett. 1999;455:367–371) and a.a. 214 – 231 in the third submembrane domain (Zhang R et al., PLOS One 2013;8:e65266). The functional effects of CaM interaction with AT1AR, however, remains incompletely understood. Additionally, the interaction domains are not completely identified and characterized. We have observed that AT1R coimmunoprecipitates with CaM in vascular smooth muscle under resting conditions, association that is enhanced by stimulation with AngII or thapsigargin. Using novel FRET biosensors with EYFP‐ECFP pair flanking each of AT1AR's submembrane domains, we have identified a novel CaM‐binding domain in the submembrane domain 2 and found that full interaction between CaM and SMD3 requires a significantly longer sequence than previously reported. All three domains interact with CaM in a Ca2+‐dependent fashion, with drastically different affinities. Simultaneous determinations of biosensor responses and free Ca2+ concentrations showed that these interactions take place at significantly different Ca2+ sensitivities in the physiological range of intracellular Ca2+. Notably, SMD3 interacts with CaM with a Ca2+ sensitivity that falls in the range of resting intracellular Ca2+ concentration. Cells heterologously expressing full‐length AT1AR containing mutations that reduced CaM interactions at individual CaM binding domains on SMD2, SMD3 or SMD4, exhibit ~ 1.8‐, 2‐ or 3‐fold reductions in AngII‐stimulated ERK1/2 phosphorylation, respectively. These data demonstrate that CaM is involved in AngII signaling via direct interactions with multiple domains in AT1R and allow prediction of interactions between CaM and individual SMDs under distinct physiological scenarios.

Support or Funding Information

Iowa Osteopathic and Educational Research Funds

National Institutes of Health Grant HL‐112184
Statin Therapy Reduces Differentiation of Ventricular Fibroblast to Myofibroblasts in Human Failing Heart
Authors: ROSS GRACIOUS R , KRAFT KELSEY , EMELYANOVA LARISA , RIZVI FARHAN , HOLMUHAMEDOV EKHSON L , WERNER PAUL , TAJIK JAMIL , JAHANGIR ARSHAD
Published: November 2023

Background

Cardiac fibrosis (CF) is a major factor underlying the progression of heart failure (HF). Apart from the established lipid lowering effect, HMG‐CoA (3‐hydroxy‐3‐methylglutaryl‐CoenzymeA) reductase inhibition by statins reverses CF in various animal models of HF but its effect in human failing heart ventricular fibroblasts (hVF), responsible for fibrosis, is not known. Hence, we tested the differentiation phenotype of hVF isolated from HF patients under statin therapy (HF+Statin) for at least 1 year or not (HF‐No_Statin) and normal, as fibroblast to myofibroblast trans‐differentiation precedes fibrosis.

Methods

Primary hVF cultures obtained from HF patients undergoing left‐ventricle assist device implantation (n=4 each) and non‐diseased hVF from trauma victims (n=3) were used at passage 3 or 4. Confocal microscopy, immunohistochemistry, western blotting, Edu (5‐ethynyl‐2′‐deoxyuridine)‐incorporation assay data were obtained and analyzed by unpaired t test or one‐Way Analysis of Variance.

Results

Immunoblot (Fig. A) data showed significantly (P<0.05) higher expression of α‐smooth muscle actin (α‐SMA+) and immunohistochemistry (Fig. B) showed significantly high population of myofibroblasts (α‐SMA+) in the HF‐No_Statin group vs the control or HF+Statin groups, suggesting statin‐sensitive increased hVF differentiation in failing heart. In vitro treatment of simvastatin (0.3μM for 72 hrs) on HF‐No_Statin group hVFs significantly (P<0.05) reduced the high expression of α‐SMA, indicating a reversal effect of statins on myofibroblast to fibroblast. Edu‐incorporation data showed that the percentage of proliferating hVFs was not significantly different in failing heart without statins(82±3%) compared to control(82±4%) and HF+ Statin (69±5%), indicating that the DNA synthesis and proliferative capacity of hVFs remain unaltered in failing heart, irrespective of statin therapy, compared to normal.

Conclusion

The failing heart ventricle has a large population of differentiated myofibroblasts. Statins prevent/reverse hVF differentiation decreasing the ratio of myofibroblast to fibroblasts, which is anti‐fibrotic. The significance of statins’ anti‐fibrotic effect for HF therapeutics may be in reducing the CF progression by inhibiting/reversing hVF differentiation to myofibroblasts, thereby improving the cardiovascular outcome in HF patients

Support or Funding Information

Aurora Health Care ‐ Cardiovascular Surgery Research Award (CVSRA 2015) to Gracious R Ross and NIH R01‐HL101240 to Arshad Jahangir.
Impact of SPR to minimize CNS penetration on peripheral tissue distribution and bioavailability in mice
Authors: Zhou Xin , Cramer Jeff , Jones Tim , Hudson Loyd , Sawada Geri , Desai Prashant , Raub Tom
Published: November 2023

SPR (structure‐property relationship) to reduce CNSpenetration for LY1 was implemented to mitigate the toxicity induced by hittingthe brain target. In a 24‐hr infusion study in mice, increase in hydrogen bondacceptor/donor strength and subsequent Pgp liability (LY2) led to 5‐times lowerKpuu in the brain. Introduction of carboxylic acid (LY3) and increased polarsurface area (PSA) resulted in 25‐times lower Kpuu. Significant increase in PSA for LY4, also a Pgp substrate, did not lead to further reduction in Kpuucompared to LY2 in steady‐state. Peripheral tissue distribution did not differ for LY1 and LY2 in high and low perfusion tissues; whereas the acidic LY3, displayed lower Kpuu in various tissues, including peripheral target tissues. The pharmacokinetics study revealed that the lower permeability of LY3 and LY4 was detrimental to bioavailability after administration of compounds by oralgavage at 10 mg/kg. The results demonstrated that SPR for increased Pg pliability without significant impact on pKa and PSA would minimize brain penetration, and maintain adequate bioavailability and peripheral tissue distribution for efficacy.

Compound
MW
clogP
pKa
PSA
HBD/HBA strength
Kpuu, brain

LY1
285
1.6
2.2 (most basic)
85
5
0.5

LY2
293
2.0
2.3 (most basic)
85
5.8
0.1

LY3
304
2.2
4.0 (most acidic)2.2 (most basic)
102
8.5
0.02

LY4
339
1.7
10.0 (most acidic)2.3 (most basic)
125
9.3
0.2
Resveratrol‐Enriched Rice Protects Human Skin against UVB‐Induced Photoaging
Authors: Lee Taek Hwan , Subedi Lalita , Wahedi Hussain Mustatab , Park Yong Un , Kim Sun Yeou
Published: November 2023

Ultraviolent radiation can cause skin photoaging, skin wrinkle and pigmentations. Synthetic cosmaceuticals as well as medicines have number of side effects as well as drawbacks so natural therapies are the best way of protecting human skin and looking young and smart. Genetically engineered Resveratrol‐enriched rice (RR) has been reported to have excellent anti‐obesity and depigmenting effects in human skin however its effect in UVB induced skin aging was not studied yet. In this study, we evaluated the effects of normal rice (NR), resveratrol‐enriched rice (RR), and resveratrol (R) on ultraviolet B (UVB)‐irradiated human dermal fibroblasts (HDFs) and reconstructed skin tissue to determine the therapeutic potential of these agents in skin wrinkling and skin photoaging. We measured the cellular levels of procollagen type I (PIP‐1) and matrix metalloproteinase (MMP)‐1 using ELISA in NHDF cells after UVB irradiation. UVB‐irradiated NHDF cells that were treated with NR, RR, and R exhibited higher PIP‐1 levels and lower MMP‐1 levels than untreated cells. Among the treatments, RR was found to have the maximum potential to prevent skin damage in both, for MMP‐1 inhibition as well as PIP induction. Taken together, our findings demonstrated that the anti‐wrinkle properties of RR might be superior to those of R or NR alone. Additionally, RR possesses anti‐skin photoaging effects, via downregulating the MMP‐1 levels and upregulating the pro‐collagen production in UVB‐irradiated reconstructed skin as well. In conclusion, RR may provide a non‐toxic and convenient means of preventing UVB‐induced skin wrinkles and related diseases.

Support or Funding Information

This work was supported by a grant from the Next‐Generation BioGreen 21 Program (No. PJ01118803), Rural Development Administration, Republic of Korea
Overexpression of Tbx20 in Adult Cardiomyocyte Promotes Myocyte Proliferation and Improves Cardiac Function post Myocardial Infarction
Authors: Xiang Fuli , Guo Minzhe , Yutzey Katherine
Published: November 2023

Background

Adult mammalian cardiomyocytes (CM) have the potential to proliferate, but this is not sufficient to compensate for the massive loss of functional CMs after myocardial infarction (MI). During embryonic heart development, the transcription factor Tbx20 is required for CM proliferation, and Tbx20 overexpression (Tbx20OE) promotes fetal characteristics in adult CMs when initiated before birth in mice. We hypothesize that Tbx20OE, when induced in adult CMs after injury, improves cardiac function and repair via dedifferentiation and cell cycle re‐entry of CMs.

Methods and Results

Tbx20OE was induced in inducible CM‐specific Tbx20 overexpressing adult mice via tamoxifen. Increased CM proliferation and fetal characteristics were found in Tbx20OE hearts compared to controls without affecting cardiac function and morphology 4 weeks after Tbx20 overexpression at baseline. However, Tbx20OE in adult CM after MI significantly improved cardiac function and infarct size, and decreased myocardial hypertrophy 4 weeks post‐MI. Improved cardiac repair as indicated by increased capillary density and CM proliferation was also observed in border zone of Tbx20OE hearts compared to controls. Expression of proliferation activators (cyclin D1, E1, and IGF1) and fetal contractile proteins (ssTNI, βMHC) mRNA was increased, while negative cell‐cycle regulators (p21, Meis1) were decreased, in Tbx20OE hearts compared to controls under both baseline and MI conditions. Interestingly, multiple pro‐proliferation pathways including Akt, YAP and BMP signaling were activated in Tbx20OE adult heart at baseline. And we found p21, Meis1 and Btg2, act as direct downstream targets of Tbx20 in CM proliferation.

Conclusion(s

In conclusion, Tbx20OE in adult CM promotes cell proliferation via regulating multiple pro‐proliferation pathways and directly impair cell‐cycle repressor gene p21, Meis1 and Btg2. Overexpressing of Tbx20 in adult CM improves cardiac function and repair in mice when induced post‐MI.

Support or Funding Information

American Heart Association postdoc fellowship 15POST22060001