bluuebunny/crossref_metadata_2025 huggingface dataset:
Phosphoserine Aminotransgerase I Is A predictor of Early Recurrence and Poor Prognosis of Resected Stage I Non‐small Cell Lung Cancer That Induces Metastasis via MMP1 Activation
Authors: Chan Yung‐Chieh , Liu Yu‐Peng , Su Chia‐Yi , Jan Yih‐Hua , Yang Yi‐Fang , Chang Yu‐Chan , Lai Chu‐Ching , Hsaio Michael
Published: June 2021

Stage I non‐small cell lung cancer (NSCLC) is potentially curable after completely resection. However, recurrence may occur in 30 % of the stage I cases and influence prognosis. It is important to identify biomarkers that can predict the early recurrence in Stage I lung cancer patients. By using publicly available microarray database, we have identified Phosphoserine Aminotransferase I (PSAT1) which overexpression is associated with poor prognosis (P<0.001) with hazard ratio of 2.09 (P=0.00048) in stage I and II of lung cancer patients. The immunohistochemical staining was carried out with 104 specimens of NSCLC, which showed higher PSAT1 expression level in tumor tissue than adjacent non‐tumor tissue (P<0.011). High PSAT1 immunoreactivity was relevant to shorter disease free survival (P=0.034) in early stage adenocarcinoma. Two way complementary overexpression and knockdown of PSAT1 in low invasive and high invasive lung cancer cell promotes invasion and migration, respectively. Furthermore, high‐throughput promoter screening showed significant activation of MMP1 promoter (10‐fold, P<0.01) by PSAT1 expression.

MMP1 knockdown in PSAT1 overexpressed cells inhibited invasion, suggested that MMP1 is a downstream target of PSAT1 in lung cancer cells. In summary, our results indicate that PSAT1 is a novel predictor in stage I NSCLC, which promote cell invasion through the activation of MMP1 pathway.
Pheophorbide‐a prevents vascular endothelium dysfunction and down‐regulates AGEs‐induced proinflammatory cytokines.
Authors: Nam Mi‐Hyun , Oh Ji‐Sun , Hong Chung‐oui , Son Da‐Hee , Hong Seung‐Teak , Seol Hae‐Min , Lee Kwang‐Won
Published: June 2021

During hyperglycemia, nonenzymatic glycation occurs through the reaction of aldehyde groups of reducing sugars and free amino groups of proteins. Excessive intracellular reactive oxygen species (ROS) derived from advanced glycation end products (AGEs) may play a key role of inflammation on endothelium. ROS enhance the expression of specific adhesion molecules on the surface of endothelium, and it up‐regulates the monocyte adhesiveness. Glycapaldehyde induced AGEs (Glycol‐AGEs) have been reported to express pro‐inflammatory cytokines such as tumor necrosis factor‐alpha (TNF‐α) and interleukin 1‐beta (IL‐1β), and it might be one of the important factors in the pathogenesis of initial stage of atherosclerosis. Pheophorbide a showed not only a scavenging activity of the intracellular ROS, but also monocyte adhesiveness inhibitory activity on THP‐1 and HUVEC co‐cultivation system. The mRNA levels of inflammation related genes such as MCP‐1 and IL‐6 were significantly decreased by pheophorbide a, and AGEs‐stimulated TNF‐α and IL‐1β were down‐regulated with it. In these results, we demonstrated that the pheophorbide a has a significant ROS scavenging activity, monocyte adhesive inhibitory activity, and down regulation activity of cytokines related to inflammation on endothelium. We expect that the pheophorbide a might potent candidate compound to control endothelial cell dysfunctions.
Insulin receptor isoform B is downregulated in intestinal stem cells and tumors, limits colon cancer cell growth, and promotes differentiation
Authors: Bortvedt Sarah , Mah Amanda , Van Landeghem Laurianne , Simmons Jim , Santoro M. Agostina , Lund P. Kay
Published: June 2021

Insulin receptor (IR) has been little studied in intestinal epithelium, despite evidence that obesity and hyperinsulinemia promote intestinal dysfunction and cancer. In other organs, two IR splice variants mediate metabolism (IR‐B) or growth (IR‐A). This study examined IR‐A and IR‐B in intestinal stem cells (ISC), progenitors, and differentiated lineages isolated from Sox9‐EGFP reporter mice, normal tissue (N) and tumors (T) of ApcMin/+ mice, and colorectal cancer cells (CRC). Effects of IR‐B overexpression were tested in Caco2 CRC. We hypothesized that IR‐B predominates in differentiated normal lineages or differentiated CRC, limits CRC proliferation, and promotes differentiation.

Results

IR‐A predominates in ISC and progenitors; IR‐B predominates in differentiated lineages. IR‐B is downregulated in T vs N of ApcMin/+ mice and aggressively growing CRC. In Caco2 CRC, IR‐A and IR‐B are both expressed during exponential growth, but IR‐B and an IR‐B splicing enhancer are dramatically upregulated during spontaneous differentiation. Overexpression of IR‐B in Caco2 CRC dramatically reduced proliferation, but accelerated and enhanced expression of the differentiation marker sucrase. We conclude that maintained or upregulated IR‐B expression may be critical to differentiation of intestinal epithelial cells or CRC and protective against colon cancer growth. Grants: F31AG040943, NIH DK040247–19.
Endothelial dysfunction driven by mitochondrial reactive oxygen species – proof of concept studies in CypD −/− mice
Authors: Daub Steffen , Kroeller‐Schoen Swenja , Steven Sebastian , Kossmann Sabine , Scholz Alexander , Oelze Matthias , Xia Ning , Treiber Nicolai , Scharffetter‐Kochanek Karin , Li Huige , Schulz Eberhard , Wenzel Philip , Münzel Thomas , Daiber Andreas
Published: June 2021

There is growing body of evidence for a crosstalk between different enzymatic sources of oxidative stress. With the present study we sought to determine the underlying mechanisms of this crosstalk.

Results

NADPH oxidase (Nox) activation and triggered oxidative burst was assessed by chemiluminescence or fluorescence based assays and translocation of cytosolic Nox subunits in response to mitochondrial reactive oxygen species (mtROS) formation in human leukocytes. mtROS induced Nox activation was prevented by inhibitors of the mitochondrial permeability transition pore (mPTP), protein kinase C or Nox activity. mtROS induced Nox‐dependent oxidative burst was absent in white blood cells with p47phox deficiency (regulatory cytosolic Nox2 subunit) or with cyclophilin D deficiency (regulatory mPTP subunit), whereas this crosstalk was amplified in white blood cells with mitochondrial manganese superoxide dismutase deficiency (MnSOD+/−).

Conclusions

Our data show that mtROS trigger the activation of phagocytic and cardiovascular NADPH oxidases, which may have fundamental consequences on immune cell activation and progression of angiotensin II‐mediated hypertension.

Research support

The present work was supported by generous financial support by the Johannes Gutenberg University and Medical Center Mainz (MAIFOR and Forschungsfonds grants to A.D.), the German Research Foundation (DFG WE 4361/3–1 to P.W. and DFG KFO142 SCHA 411/15–2 to K.S.‐K.) and the Stiftung Mainzer Herz to all authors.
Improvement of periodontal healing after periodontal surgeries supported by nutritional supplement drinks
Authors: Lee Jaeri , Cho Hyun‐Ji , Park Jung‐Chul , Kim Chang‐Sung , Park Yoo‐Kyoung
Published: June 2021

The aims of the study were to evaluate the effects of the nutritional supplement drinks on the periodontal health and tooth mobility over time after periodontal surgery. Twenty three patients after flap surgery were randomly assigned to two groups. The intervention group consumed 3 cans of nutritional supplement drinks per day for 8 wks, while the control group did not receive any. Clinical periodontal measurements such as plaque index (PI), gingival index (GI), tooth mobility and oral health impact profile 14 (OHIP‐ 14) were measured at baseline, 1, 4, and 8 wks. Also, anthropometric measurements, the simplified nutritional appetite questionnaire (SNAQ) and dietary intake assessment were performed at baseline and 8 wks. The PI values were significantly improved in both groups. One week after surgery, the GI values were reduced significantly only in the intervention group (P<0.05) and tooth mobility were increased significantly in both groups (P<0.05), but less in the intervention group. In both groups, OHIP‐ 14 scores, anthropometric measurements and the mean SNAQ score did not differ significantly after intervention. At 8 wks, the intake of protein, vitamin A, B1, niacin was increased in the intervention group, and the intake of vitamin A, C, niacin was higher in the intervention group than those in the control group. This result demonstrated that nutritional supplementation improves early periodontal healing after surgery.
A food liking survey to assess n‐3 PUFA status in healthy postmenopausal women
Authors: Hutchins‐Wiese Heather L , Duffy Valerie B , Watkins Bruce A , Li Yong , Kenny Anne M
Published: June 2021

Diets rich in long chain (LC) omega (n)‐3 polyunsaturated fatty acids (PUFA) are beneficial in the prevention of age‐related conditions (i.e. heart disease). Rapid dietary screening for LC n‐3 PUFA (eicosapenataenoic and docosahexaenoic acids, the primary fatty acids in fish) status is lacking. To address this need, we tested the use of fish questions from a food liking survey compared to a food frequency questionnaire (FFQ) to predict serum LC n‐3 PUFA levels in 48 health postmenopausal women (average age 77y). LC n‐3 PUFA were determined by gas chromatography of fatty acid methyl esters and observed higher levels in supplement users vs. non‐users (4.94±1.2 vs. 7.27±2.38, p<.001); 1 in 3 reported supplemental fish oil use. Women reported an average of 1–2 fish servings/week for the FFQ that took nearly 1 hr to conduct. The liking survey took ¼ hr to complete; 1/3 reported a strong liking. While fish liking correlated with intake frequency, only liking correlated with LC n‐3 PUFA levels (rho=.57, p<.001). In linear regression, fish liking and supplement use were significant contributors to explain 46% of the variance of LC n‐3 PUFA; FFQ fish intake did not. These pilot data suggest use of simple fish liking and supplement questions as a proxy for serum LC n‐3 PUFA status in older women. Validation in other populations is needed to aid clinicians to rapidly screen for low LC n‐3 PUFA levels and associated health conditions. Funding sources: JoAnne Smith Memorial Fund and USDA Hatch Funds.
Towards effective food security interventions for HIV+ individuals: Impact of home delivered meals
Authors: Jin Sarah , Tavarez Isy , Boshnack Nick , Cole Chris , Fang Xindi , Shao Stephanie , Girguis Mariam , Gibson Britton , Humphries Debbie
Published: June 2021

Nutrition interventions are increasingly recognized as pivotal to vulnerable HIV positive (+) patients.

Objective

To determine whether HIV+ individuals receiving home delivered meals have different rates of viral suppression when compared to HIV+ individuals who are not receiving home delivered meals.

Methods

Two chart reviews were conducted, comparing rate of viral suppression among HIV+ participants in Caring Cuisine (CC), a home delivered meals program, with HIV+ case management clients who did not receive home delivered meals. The first was a cross‐sectional study of current clients (n=164). The second was a five year retrospective matched case‐control study of clients with multiple measures of viral load at different time points with respect to initiation of CC participation (n=45 CC participants; n=90 for non‐CC participants).

Results

Cross‐sectionally, CC clients had higher rates of viral load suppression (58.5% vs. 38.5%, p=<0.01) despite being older (p <0 .01) and with lower annual incomes (p<0.05). After six months of receiving home delivered meals, CC participants were more likely to have viral load suppression (OR=2.11; p=0.038) than matched non‐participants.

Conclusion

Home delivered meals may be an effective ancillary treatment for vulnerable HIV+ patients. Further research is needed to determine efficacy and effectiveness of this approach. Research Support: Yale School of Medicine
Dietary Patterns are Associated with Obesity in Young Women
Authors: Lindgren Jessica A. , Hartman Terryl J. , Mitchell Diane C. , Coffman Donna L. , Van Horn Linda , Dorgan Joanne F.
Published: June 2021

Dietary pattern (DP) analyses have been successfully used to study the effects of total diet on health outcomes. This cross‐sectional study used latent class analysis to derive DPs in a cohort of 250 young adult women (25–29y) originally enrolled between 8–10y into the Dietary Intervention Study in Children. Using repeated 24‐ hour dietary recall data, 15 food group variables were used to derive five DPs designated as “Western”, “Healthy”, “Low Energy”, “Alcohol”, and “Non‐drinking” patterns. Preliminary multivariable analyses suggest that body fatness (p= 0.002), waist circumference (p=0.004), and dietary energy density from food only (ED; p<0.001) were significantly different across DPs. Women in the “Western” pattern, which was characterized by high intakes of red meat, refined grains, and starchy vegetables, had the highest mean body fatness (40.3 ± 9.6%), waist circumference (91.1 ± 11.1 cm), and ED (2.15 ± 0.36 kcals/g). Women in the “Healthy” DP, which was characterized by high intakes of fruit, whole grains, and dark green and yellow vegetables, had a significantly lower mean ED (1.46 ± 0.39 kcals/g) than each other DP (all p<0.05). These results suggest that DPs may be associated with obesity.

Grant Funding Source: R01CA104670
Nutrition education integrates culture via storytelling in cooking workshops for community nutritionists
Authors: Winham Donna M , Quiroga Seline Szkupinski , Underiner Tamara , Woodson Stephani , Todd Megan
Published: June 2021

For nutrition education to be effective in changing behavior, the message must be culturally meaningful, and the recipient must have sufficient self‐efficacy and desire to alter practices. We conducted a 6‐session cooking workshop led by a chef/performance artist with 14 nutrition students and lay health workers in January 2012. The feasibility study purpose was 1) to assess the perceived effectiveness of storytelling and creative performance in heightening awareness about culture and food; and 2) to evaluate changes in self‐efficacy and knowledge among the participants. Validated questionnaires to measure self‐efficacy, health beliefs, healthy lifestyle behaviors and dietary intakes were administered before and after the workshop series. Quantitative results indicated a high degree of self‐efficacy at start and end. Changes in the perceptions of healthy food costs, availability of healthy foods, and the time to buy healthy foods showed positive trends, but were not significant. Statistically significant pre‐post differences were found in: perceptions of ability to exercise and to engage others through storytelling; frequency of eating meals with family; and healthy eating beliefs. Qualitative interviews conducted one month later on experiences and changes since the workshop supported improvements in perceptions and behaviors. Creative activities may enhance nutrition education skills.
Storage of Leftover Unpasteurized and Pasteurized Human Milk
Authors: Meng Ting , Fogleman April , Cohen Ronald , Sakamoto Pauline , Allen Jonathan
Published: June 2021

The current human milk storage guidelines recommend that leftover HM be discarded immediately or soon after an infant is finished feeding. There is limited research to support this recommendation. The objective was to observe the difference in bacterial growth in leftover unpasteurized human milk (HM) and leftover pasteurized milk (PHM). Five mother‐baby dyads participated in the study. Each baby was fed 1 – 2 ounces of milk from a bottle prepared with the mother's pasteurized milk and the mother's unpasteurized milk. Leftover milk was collected in sterile containers and analyzed for bacterial growth at specific time points during storage. Acceptable levels of bacteria were chosen as 10^5 colony‐forming units per milliliter (CFU/ml). HM was stored from 6 to 9 hours and PHM from 9 to 24 hours at 24°C before reaching 10^5 CFU/ml. HM was stored from 4 to 6 days and PHM was stored at least 6 days at 4°C before reaching 10^5 CFU/ml. More research is needed with a larger number of participants and focusing on the growth of specific strains of pathogenic bacteria in leftover human milk so that consumer recommendations can be made. The data show that bacteria originating from the mother are greater than bacteria originating from contamination by an infant.
Individual amino acids and insulin sensitivity: relationships determined by race and gender
Authors: Ingram Katherine Heimburger , Thalacker‐Mercer Anna , Garvey W. Timothy
Published: June 2021

The purpose of this study was to examine relationships between fasting serum amino acid levels and insulin sensitivity in adults of African (AA) and European (EA) descent. In 63 EA (41F, 22M) and 60 AA (32F, 28M) adults, insulin sensitivity was assessed by hyperinsulinemic clamp (glucose disposal rate, GDR) and serum levels of 16 amino acids by tandem mass spectrometry. Univariate ANOVA revealed reduced alanine and serine, and higher methionine, in EA, compared to AA. Correlations controlled for age, sex, and BMI revealed negative relationships between GDR and leucine/isoleucine, valine, and glutamate/glutamine in only EA. In contrast, serine, methionine, proline, histadine, ornithine, and arginine were positively associated with GDR in AA. Glycine was positively associated with GDR in both EA and AA. Further stratified by gender, strong relationships between GDR and leucine/isoleucine, valine, and citruline were seen in EA women, but not EA men. Likewise, serine was positively associated with GDR in AA women. Proline and citruline were associated in AA men. Stepwise multiple regression indicated that the best predictor of GDR is leucine/isoleucine in EA and glycine in AA. Our results reveal racial and gender differences in circulating amino acids, indicating likely differences in protein metabolism between these groups. Supported by NIH (DK‐038765, DK‐083562, PO1 HL‐55782), Veterans Affairs Merit Review.
Validity of a novel digital approach for dietary recall in athletes
Authors: Baker Lindsay , Heaton Lisa E. , Nuccio Ryan P. , Stein Kimberly W.
Published: June 2021

We measured the agreement of our novel, digital dietary analysis tool for athletes (DATA) with the 24‐h dietary recall interview using the USDA 5‐step multiple‐pass method (INTERVIEW). Fifty‐six athletes (14–20 yr; 42 male, 16 female) completed DATA and INTERVIEW in a randomized counter‐balanced order (back‐to‐back on the same day). Intraclass correlations (ICC) and paired comparisons (expressed as % difference between methods) with 95% confidence intervals were used to determine statistical agreement between DATA and INTERVIEW. The ICC's between DATA and INTERVIEW were significant for energy (0.86, 0.76–0.92), carbohydrate (0.89, 0.81–0.94), total fat (0.75, 0.58–0.86), protein (0.89, 0.81–0.93), water (0.85, 0.75–0.91), sodium (0.85, 0.75–0.91), iron (0.83, 0.72–0.90), and calcium (0.91, 0.85–0.95). In addition, the differences between methods were not significant for energy (−1.1%, −9.1 to 7.7%), carbohydrate (0.2%, −7.1 to 8.0%), total fat (−6.1%, −17.8 to 7.4%), protein (−2.7%, −11.3 to 6.7%), water (−1.2%, −9.1% to 7.3%), sodium (−3.2%, −13.3 to 8.1%), iron (−8.4%, −18.3 to 2.7), and calcium (−0.4%, −10.0 to 10.3%). The DATA digital program is a valid 24‐h dietary analysis tool and can be used by sports health professionals looking for a quick, user‐friendly, athlete‐specific alternative to conventional 24‐h dietary recall methods. This study was funded by the Gatorade Sports Science Institute.
Daidzein Metabolizing Phenotypes and Nutritional Metabolomics Profiling After a Soy Intervention: A Pilot Study
Authors: Reverri Elizabeth J. , Slupsky Carolyn M. , Steinberg Francene M.
Published: June 2021

Soy is a well‐characterized food containing the highest concentration of isoflavones. One isoflavone, daidzein, is metabolized by gut microbiota to ODMA and equol in some individuals. Variable clinical effects have been described following soy consumption and equol production status is postulated to be a contributing factor. Nutritional metabolomics is an emerging field that comprehensively characterizes biological samples. This pilot trial aims to use metabolomic approaches to assess biomarkers of soy intake (polyphenol metabolites) and biomarkers of effect (endogenous metabolites). We hypothesize that daidzein metabolizing phenotypes will be associated with significant changes in the endogenous metabolite profile. Subjects in this randomized, controlled, crossover study received either soy food or control food daily for 4 weeks, separated by a 2 week washout. Equol status was analyzed in urine by LC‐MS/MS and nutritional metabolomics, in urine and serum by NMR. Subjects include 17 adults (12F, 5M) at cardiometabolic risk with mean age 56 and BMI 31. Preliminary results confirm compliance to the intervention and reveal 47% equol producers and 71% ODMA producers. The metabolomic results will identify metabolic responses associated with daidzein metabolizing phenotypes in adults with cardiometabolic risk factors and contribute to the understanding of variable responses to soy consumption.
The effect of beta‐carotene on migration and invasion in SK‐N‐BE( 2)C neuroblastoma cells
Authors: Kim Yoo‐Sun , Lee Hyun‐Ah , Kim Yuri
Published: June 2021

Neuroblastoma (NB) is the extracranial tumor of children. The invasive or metastatic NBs have poor clinical outcome, therefore, to understand the mechanism(s) that modulate cancer cell invasion is important to develop more effective chemotherapeutic agents. While Beta‐carotene (BC), the most active vitamin A precursor, has been previously shown to exert various antioxidant and anticancer effects, the anti‐metastatic effects of BC on NB cells remains poorly understood. The aim of this study is to investigate the anti‐metastatic effects of BC on highly malignant SK‐N‐BE( 2)C NB cells. We found that BC attenuated the migratory and invasive capabilities of the cells. The protein expression and enzymatic activity of matrix metalloproteinase‐2 (MMP2) was also suppressed by BC under normoxia or hypoxia determined by western blotting and gelatin zymography assay respectively. BC down‐regulated the expression of hypoxia‐inducible factor 1α (HIF‐1α), a well‐known tumor metastasis regulator and further down‐regulated its down‐stream genes, vascular endothelial growth factor (VEGF) and glucose transporter‐1 (GLUT‐1). In conclusion, the present study provided the first evidence that BC may be utilized as an effective chemotherapeutic agent by regulating invasion potential, thereby controlling the metastasis of neuroblastomas.

[This research was financially supported by the National Research Foundation of Korea (Project Number is 2012–0008169)]

Grant Funding Source: National Research Foundation of Korea
Genome‐wide association study on coffee intake in Korea population
Authors: Lee Jae Kyung , Sung Joohon , Kim Kyunga , Lee Jung Eun
Published: June 2021

The objective of this genome‐wide association study (GWAS) is to identify genetic variants for coffee intake in Korean adults. The Korea Association Resource (KARE) study included 8,842 participants aged 40–69 years who resided in Ansung or Ansan. Coffee intake was assessed in 2001 by using a validated food frequency questionnaire. A total of 352,228 SNP markers were directly genotyped with Affymetrix Genome‐Wide Human SNP array 5.0 Genotypes. GWAS was performed by using PLINK v1.07 (http://pngu.mgh.harvard.edu/~purcell/plink/) with adjustment for area, age, and sex. We used coffee intake transformed by Box‐Cox procedure as quantitative traits in the multivariate linear regression model or classified into non‐drinker and ≥2 cups/d of coffee intake in the multivariate logistic regression model. A total of 8,697 participants were included in the analyses. rs11066280 and rs2074356 located with threshold of P<1.0×10−9 in chromosome 12 open reading frame 51 (C12orf51) gene reached genome‐wide significance in both linear and logistic models. When we examined analyses in Ansung or Ansan population separately, we found that significance of those two SNPs with threshold of P<1.0×10−6 still remained. Those SNPs were associated with alcohol consumption, waist to hip ratio, and blood pressure in Korean adults. We found that rs11066280 and rs2074356 in C12orf51 were associated with coffee intake in Korean adults.
Effects of γ‐glutamyl hydrolase modulation on CpG promoter DNA methylation and gene expression in human colon cancer cells
Authors: Kim Sung‐Eun , Hinoue Toshinori , Kim Michael , Sohn Kyoung‐Jin , Cho Robert , Weisenberger Daniel , Laird Peter W. , Kim Young‐In
Published: June 2021

γ‐Glutamyl hydrolase (GGH) plays an important role in folate homeostasis by catalyzing hydrolysis of polyglutamylated folate into monoglutamates, thereby facilitating export of folate from cell. We investigated whether GGH modulation would affect CpG promoter methylation and gene expression profiles in human HCT116 colon cancer cells. Illumina HT‐12 and Infinium Methylation assay were performed in the GGH‐modulated HCT116 cells, and Ingenuity Pathway Analysis was used for functional analysis. The mRNA expression of selected genes, expression of which was epigenetically regulated was confirmed by qRT‐PCR. We identified 153 differentially expressed genes (most involved pathways: cellular movement and cell death) associated with GGH overexpression, and 321 differentially expressed genes (most involved pathways: cell death and cell cycle) associated with GGH inhibition. 905 and 1869 genes showed altered CpG promoter methylation associated with GGH overexpression and inhibition, respectively. An integrated analysis revealed 5 and 16 genes, expression of which was regulated by CpG promoter methylation changes associated with GGH overexpression and inhibition, respectively. Thus, GGH modulation can affect differential gene expression and CpG promoter DNA methylation involved in important biological pathways and some of the observed altered gene expression appear to be epigenetically regulated.

Grant Funding Source: CIHR Fund 14126
Lower natural killer cell cytotoxicity in smokers may be mediated by epithelial cells
Authors: Muller Loretta , Brighton Luisa E. , Jaspers Ilona
Published: June 2021

Cigarette smoking is a risk factor for viral infections, yet the mechanisms underlying this susceptibility remain unknown. Natural killer (NK) cells play an important role in fighting influenza and signals from epithelial cells (EC) likely regulate NK cell activity. We have previously shown that markers of cytotoxicity are reduced in nasal NK cells from smokers after influenza infection. This study investigated if human EC from smokers and nonsmokers differentially affect NK cell activity in the context of influenza infection.

EC were infected with influenza and subsequently co‐cultured with NK cells. NK cells were also incubated with apical washes and basolateral supernatants of EC to identify the role of soluble mediators.

At baseline, co‐culturing NK cells either directly with smoker EC or stimulating with basolateral supernatants from these cells reduced markers of cytotoxicity. Similarly, after influenza infection markers of cytotoxicity as well as levels of IFN‐γ, IL‐4 and granzyme B were decreased in NK cells incubated with basolateral supernatants of smoker EC.

Co‐culturing NK cells with EC from smokers reduced markers of cytotoxicity in NK cells through a potential mechanism involving EC‐derived soluble mediators. Thus, EC‐mediated modification of NK cell activity may contribute to the higher susceptibility to influenza infections seen in smokers.

The work was supported by NIH, EPA and FAMRI.
Knockdown of CCR1 suppresses expression of Intercellular Cell Adhesion Molecule‐1 induced by hypoxia/reoxygenation in endothelial cells and reduces infarct sizes after myocardinal ischemia/reperfusion
Authors: Kim Jiyoung , Park Keun Hyung , Lee Tae Hoon
Published: June 2021

It has been reported that CCL15, a CCR1 ligand, induces proatherogenic factors and has been implicated in atherosclerosis. The present study showed that exposure of human endothelial cells (ECs) to hypoxia and reoxygenation (H/R) resulted in a rapid increase in expression of ICAM‐1 and CCR1 ligands. Our results revealed that knockdown of CCR1 attenuated expression of ICAM‐1 in ECs after H/R. Stimulation of ECs with CCL15 increased expression of ICAM‐1 compared with those in non‐stimulated cells predominantly via CCR1. Our data showed that CCL15 increased cell adhesion of monocytoid cells to endothelial cells. Pretreatment of these cells with CCR1 siRNA prevented the CCL15‐induced expression of ICAM‐1 and monocyte adhesion to ECs. We further investigated expression of CCR1 ligands and ICAM‐1 in vivo after myocardinal ischemia and reperfusion (MIR). Enhanced expression of ICAM‐1 and CCR1 ligands has been demonstrated in rat hearts in vivo after MI‐R. Direct injection of CCR1 siRNA onto the ventricular wall reduced expression of ICAM‐1 in rat hearts activated by MI‐R. Staining of hearts revealed that silencing of CCR1 markedly reduced the infarct sizes after MI‐R. Our results indicate that CCR1 and its ligands play important roles in inflammatory responses after MI/R via upregulating expression of cell adhesion molecules.

This work was supported by a grant from National Research Foundation of Korea.
Thymoquinone enhances coronary flow and reduces infarct size in Langendorff‐perfused rat hearts through attenuation of inflammatory responce and apoptosis
Authors: Ismail Raed S , El‐Sayed El‐Sayed M , Mansour Ahmed M , El‐Mahdy Mohamed A , Zweier Jay L
Published: June 2021

The use of natural compounds to protect against a variety of cardiovascular diseases has been established. Thymoquinone (TQ), a constituent of the volatile oil derived from Nigella sativa seeds, possesses antioxidant, anti‐inflammatory, and vasodilating properties. TQ has demonstrated promising effects against ischemia‐reperfusion (IR) injury in different organs. However, the effect of TQ on myocardial IR injury (m‐IRI) has not been delineated. We show the efficacy and provide a mechanism of TQ effect on m‐IRI. Langendorff‐perfused rat hearts were subjected to 30 min of global ischemia followed by 120 min of reperfusion. TQ was infused for 20 min either pre‐or post‐ischemia. Isolated hearts of TQ‐treated rats were also subjected to the same IR protocols. TQ significantly enhanced coronary flow and markedly reduced the infarct size. IR‐induced increase of TNF‐α, myocardial NF‐kB protein expression, and apoptosis of cardiac muscle cells were attenuated following TQ treatment. Moreover, TQ modulated the expression of myocardial Bcl2, and Bax proteins and also markedly reduced oxidative stress markers and LDH release. This study indicates that TQ induced inhibition of the pro‐inflammatory cytokines and oxidative stress modulated myocardial NF‐kB, Bcl2, and Bax expression, protecting the heart against IR injury. We provide mechanistic insight for understanding the molecular basis and efficacy of TQ on m‐IRI. (NIH # HL63744 to JLZ; Egyptian government)
Macrophage (Mϕ) Depletion Reduced Vascular Oxidative Stress, Restored α2 Adrenergic Autoreceptor (α2AR) Function and Attenuated Blood Pressure Development in Deoxycorticosterone Acetate (DOCA)‐salt Hypertensive Rats
Authors: Thang Loc Vinh , Garver Hannah , Galligan James J
Published: June 2021

There is an increase in pro‐inflammatory Mϕ in mesenteric artery (MA) adventitia, and the α2AR regulating norepinephrine (NE) release from perivascular sympathetic nerves is impaired in DOCA‐salt model. The cause of this impairment and the contribution of Mϕ to the pathophysiology of salt‐sensitive hypertension are unknown. We tested the hypothesis that Mϕ release O2−, which disrupts α2AR function causing increased NE release and further increases in blood pressure in DOCA‐salt rats. Liposomal clodronate encapsulate (LEC) was used to deplete adventitial Mϕ in rat MA, (# Mϕ/frame, DOCA PBS: DOCA LEC, 39.96: 4.16, p<0.05). Mϕ depletion reduced vascular O2− (measured using dihydroethidium) (Δσ DOCA PBS‐DOCA LEC: 23.85, p<0.05). α2AR function was also restored in Mϕ depleted animals. To establish this we used focal nerve stimulation and amperometry with microelectrodes to measure NE oxidation currents at the adventitial surface of MA in the presence α2AR agonist, UK 14304, and antagonist, Idazoxan. (Normalized 1μM UK 14304 and Idazoxan NE current % control respectively, DOCA LEC: DOCA PBS, 0.326: 0.611, & 2.74: 2.18, P<0.05). Lastly, using radio telemetry, we found that Mϕ depletion attenuated DOCA‐salt blood pressure development. (MAP on day 25–28,DOCA LEC: DOCA PBS, 154.8mmHg: 197.4mmHg, p<0.05). These data support the hypothesis that Mϕ increased blood pressure in DOCA‐salt rats by releasing O2−, which disrupted α2AR function and enhanced sympathetic nerve activity.
Negative regulation of A1R‐mediated Akt signaling by neurabin
Authors: Chen Yunjia , Wang Qin
Published: June 2021

We have recently discovered a novel neural‐specific regulatory mechanism for fine‐tuning A1R responsiveness to adenosine. A neural tissue‐specific protein, neurabin, scaffolds the formation of a complex between the A1R and regulator of G Protein signaling 4 (RGS4) to attenuate A1R signaling (Chen Y. et al., 2012, J. Neurosci. 32(8):2683–2695). Disruption of this neural‐specific A1R/neurabin/RGS4 complex greatly enhances A1R signaling and promotes A1R‐mediated anti‐seizure and neuroprotective effects in mice. In the current study, we aim to understand how neurabin regulates A1R signaling pathways that are potentially involved in adenosine‐elicited neuroprotection. We chose to focus on the PI3K‐Akt pathway, which plays an important role in survival pathways in many cell systems and can be activated by A1R agonists in neurons. We found that in both heterologous cells and primary neurons, A1R‐mediated Akt activation was enhanced and prolonged in cells without neurabin expression. On the other hand, overexpression of neurabin reduced A1R‐mediated Akt activation. Meanwhile, A1R agonist‐dependent changes in ERK activation was not altered by the presence or absence of neurabin. These data demonstrate specific regulation of A1R‐mediated Akt signaling by neurabin, and provide a potential mechanism for enhanced neuroprotective effect by adenosine in the absence of neurabin.
Serotonin receptors modulate the response to cocaine and methamphetamine in adult Drosophila
Authors: Nichols Charles D , Bruce Indya , Becnel Jaime
Published: June 2021

The fruit fly, Drosophila melanogaster is a powerful genetic model system that exhibits many sophisticated behaviors, each mediated by neurotransmitters and systems conserved with higher order animals like humans. As in humans, the effects of abused drugs like cocaine and ethanol in the fly depend on the neurotransmitter dopamine. We have recently developed an open‐field testing assay for CNS stimulants to use in conjunction with traditional photobeam activity monitoring in order to more comprehensively test for the involvement of serotonin receptors in the behavioral responses to cocaine and methamphetamine. To test, flies were fed drug (cocaine, methamphetamine, serotonin receptor ligands) and tested. Overt activity was measured by photobeam breaks in arrays. Open field tests were performed 5 cm diameter chambers, where flies were video recorded for ten minutes and total distance and time in zone scored by Noldus Ethovision XT software. Cocaine and methamphetamine produced dose responsive changes in activity in both assays. Serotonin receptor selective drugs modified the responses to both cocaine and methamphetamine in both assays. The combination of open‐field testing with photobeam activity monitoring provides more quantifiable measures to examine activity than traditional assays alone, and highlights a role for serotonin in these behaviors.
Modification of overt behavioral effects of cocaine by nicotinic agonists
Authors: Desai Rajeev I , Bergman Jack
Published: June 2021

Nicotinic agonists and muscarinic antagonists previously have been shown to, respectively, mimic and accentuate the discriminative‐stimulus (Sd) effects of monoaminergic (MA) stimulants (e.g., cocaine; COC) that also increase overt scanning‐checking (S‐C) behavior in squirrel monkeys. The present studies were undertaken to examine the ability of cholinergic (ACh) ligands—nicotinic full [nicotine (NIC), (+)‐epibatidine (EPI)], partial [varenicline (VAR), (−)‐cytisine (CYT), DMAB‐anabaseine (DMAB)] agonists, and the muscarinic antagonist atropine (ATR)—to mimic or modify COC‐induced increases in S‐C behavior. Results show that COC (0.032–3.2 mg/kg), but no ACh drugs, produced dose‐related increase in S‐C behavior. Further, NIC, EPI—and, to a lesser extent, VAR and CYT—dose‐dependently attenuated, whereas atropine dose‐dependently enhanced, COC's effects on S‐C; DMAB had no effect. The similar effects of ATR in Sd and S‐C studies are consistent with established pharmacological MA‐ACh muscarinic interactions, whereas the qualitatively inconsistent effects of nicotinic drugs suggest that stimulant‐like Sd effects of nicotinic drugs do not involve prominent MA activity (supported by NIH/NIDA DA026548).
The Impact of Age on the Vasodilatory Capacity of Human Skeletal Muscle Feed Arteries
Authors: Park Songyoung , Ives Stephen J , Andtbacka Robert H.I , Gifford Jayson R , Richardson Russell S
Published: June 2021

The effect of aging on the vasodilatory capacity of human skeletal muscle feed arteries (FAs), recognized to be important regulators of muscle blood flow, has not been elucidated.

Purpose

To determine the impact of age on FA vasodilatory capacity. We hypothesized that vasodilation will be attenuated in FAs from old compared to young individuals and that this difference will be endothelium dependent.

Methods

FAs (754.5±53.3 μm) were obtained from young (33±2 yr; n=10) and old (62±2 yr; n=7) human subjects and studied using pressure myography. Vasodilation (%) was assessed in response to three stimuli 1) flow induced shear stress (Δ29 mmHg), 2) acetylcholine (ACh, 10−3M), and 3) sodium nitropusside (SNP, 10−4M), with the latter two discerning endothelium dependent and independent vasodilation.

Results

Both flow (old: 32±7 vs young: 68±11 %) and ACh (old: 73±4 vs young: 92±3 %) ‐induced vasodilation of the FAs were significantly blunted (p < 0.05) in the old compared to the young. In contrast, there was no impact of age on the vasodilation induced by SNP (old: 102±4 vs young: 112±11 %).

Conclusion

This study has documented that the endothelium‐mediated vasodilatory capacity of FAs, but not smooth muscle function, is impacted by age. Given the potential role of FAs in the regulation of blood flow, these findings may help to explain the attenuated perfusion of skeletal muscle associated with aging.
Recording of intracranial pressure in conscious rats via telemetry
Authors: Guild Sarah‐Jane , McCormick Daniel , Lim Matthew , Pauly Bob , Kirton Robert , Budgett David , Kondo Masahiro , Stehlin Ellyce , Barrett Carolyn J , Malpas Simon C
Published: June 2021

Measurement of intracranial pressure (ICP) is a critical diagnostic measure in treating head trauma, intracerebral hemorrhage and hydrocephalus. The pressure values are generally between 5–15 mmHg with >;20 mmHg considered abnormal. There is a scarcity of animal models of chronic ICP measurement in part associated with the difficulty of measuring ICP chronically with good accuracy. Previous telemetry devices utilizing fluid filled catheters have proved problematic due to movement artefacts. We implanted TRM54P telemetry devices which incorporate a solid state 2 Fr (0.6 mm) sensor in rats. The pressure sensor catheter was tunnelled subcutaneously to the skull and inserted thru the dura to lie against the brain tissue. We also performed drift assessments on the sensors over time before and after implantation and using bench measurements.
20‐HETE INDUCES REMODELING OF RENAL RESISTANCE ARTERIES INDEPENDENT OF BLOOD PRESSURE ELEVATION IN ANDROGEN‐INDUCED HYPERTENSION
Authors: Ding Yan , Wu Cheng‐Chia , Garcia Victor , Dimitrova Irina , Weidenhammer Adam , Zhang Frank , Falck John , Mei Shaojun , Capdevila Jorge , Schwartzman Michal Laniado
Published: June 2021

20‐Hydroxyeicosatetraenoic acid (20‐HETE) is a cytochrome P450‐derived arachidonate metabolite that has been shown to increase smooth muscle contractions and proliferation, stimulate endothelial dysfunction and promote hypertension. We examined if 20‐HETE contributes to vascular remodeling in androgen‐induced hypertension. In Sprague‐Dawley (SD) rats treated with 5α‐dihydrotestosterone (DHT), the increase in blood pressure (BP) is prevented by 20‐HETE inhibitor, HET0016, or 20‐HETE antagonist, 20–6,15‐HEDE. Likewise, inhibition of 20‐HETE's biosynthesis or activity abrogated DHT‐induced remodeling in renal interlobar arteries. In cyp4a14−/− mice, which display androgen‐driven and 20‐HETE‐dependent hypertension, treatment with 20–6,15‐HEDE abolished remodeling of renal interlobar arteries decreasing media thickness from 23.67±0.88 to 14.56±0.59μm and M/L from 0.29±0.03 to 0.17±0.01. To assess whether 20‐HETE induces vascular remodeling independent of BP increase, SD rats were administered reserpine (100μg/kg/day) and DHT for 21 days. Reserpine prevented BP elevation but did not affect vascular remodeling; under these conditions, treatment with 20–6,15‐HEDE attenuated vascular remodeling. This study demonstrates that 20‐HETE is a key regulator of vascular remodeling in androgen‐induced hypertension; its effect is independent of BP elevation.
Untangling Tumor Vasculature
Authors: Gruionu Gabriel , Bazou Despina , Song Johnatan
Published: June 2021

Tumor vasculature is highly dynamic, dense, and non‐homogeneous. These features make it difficult to identify key determinants of tumor angiogenesis and remodeling at the single vessel level. To overcome this problem, we isolated growing tumors from the host tissue, by placing a perforated PDMS disk between the tumor and the dorsal skin in transparent windows in mice. Using this model we are able to observe and quantify various processes and stages of network formation. Between days 9–14 microvascular sprouts emerged from the perforations/holes and merged to form loops in and out of holes. Later, sprouts led by tip cells separated from the perfused, looping vessels. Surprisingly, many of the sprouts migrated away from the tumor, instead connecting to sprouts from other holes or from the edge of the PDMS disk. At day 19, smooth muscle cells were associated with the larger vessel connecting two holes. Other changes were observed after treatment with a VEGF inhibitor. In the first two days, feeding vessels dilated, but within 1–2 weeks, large vessels constricted and smaller branches are pruned. This was associated with dramatic changes in flow direction, increased perfusion and flow interruption in some vessels. The experimental model offers a novel approach for identifying key steps in tumor vascularization so that more effective and specific targeting strategies can be developed. Supported by NIH R01CA149285.
High salt diet suppresses the angiogenic competency of bone marrow mononuclear cells
Authors: Karcher Jamie R. , Greene Andrew S.
Published: June 2021

Bone marrow mononuclear cells (BM‐MC) are easy to obtain in fairly large numbers and have potential to induce revascularization in ischemic tissues. Unfortunately many disease states, such as hypertension and heart failure, suppress BM‐MC function. In this study we tested the hypothesis that high salt diet (4.0% NaCl, HSD) in donor rats would suppress the BM‐MC angiogenic response in angiogenic deficient rats following 7 days of hind limb electrical stimulation (ES). Following one week on HSD, male Sprague Dawley (SD) rats undergoing ES were injected with either vehicle or BM‐MCs from SD donor rats on HSD or normal salt (0.4% NaCl, NSD). Administration of BM‐MCs from NSD donors, but not HSD donors, restored the angiogenic response in HSD recipients. AngII treatment of HSD donor rats restored angiogenic capacity and Losartan inhibited BM‐MC angiogenic competency in NSD donor rats. The angiogenic capacity of HSD BM‐MCs was recovered by maintaining the donor rats on a NSD following the initial 7 days on a HSD. Furthermore, HSD BM‐MCs had increased levels of apoptosis and the number of endothelial progenitor (CD34+/VEGFR2+) cells, which are believed to participate in revascularization, were reduced as compared to NSD BM‐MCs. This study suggests that HSD impairs the angiogenic competency of BM‐MCs due to suppression of the RAS causing increased apoptosis and decreased CD34+/VEGFR2+ cells in the bone marrow.
Role of adrenergic neurotransmission in the Median Preoptic Nucleus in experimental hypertension
Authors: Silva Elaine Fernanda , Silveira Ludmila Lima , Pedrino Gustavo Rodrigues
Published: June 2021

The present study sought to determine the involvement of the adrenergic neurotransmission in the Median Preoptic Nucleus (MnPO) in arterial blood pressure (MAP) control in normotensive and spontaneously hypertensive rats (SH). Male Wistar normotensive (NT) and SH rats (250–320g) were anesthetized with urethane (1.2 g/kg, i.v.) and instrumented for recordings of MAP, aortic blood flow (ABF) and renal (RBF). The rats were fixed in a stereotaxic apparatus and made nanoinjections (100 nl) of saline (0. 15 M) and Fentolamina (6.3 mM; α‐adrenergic antagonist). Aortic vascular conductance (AVC) and renal (RVC) was calculated as the ratio ABF or RBF/MAP and expressed as percentage of baseline. The results demonstrated that adrenergic neurotransmission blocking in MnPO resulted in a decrease in MAP (−7.7 ± 1.1 mmHg, n=7), ABF (−7.3 ± 2.0% above baseline, n=5) and RBF (−7.2 ± 2.1% above baseline, n = 7) of NT rats. In SH also observed decrease in MAP (−14.3 ± 6 mmHg, n = 6) and increase in AVC (8.3 ± 3.0% above baseline, n=5) and RVC (9.6 ± 1.9% above baseline, n=6), indicating vasodilation. These results suggest the participation of MnPO in tonic regulation of blood pressure in NT and SH rats.
Macrophage Migration Inhibitory Factor (MIF) Acts in the Paraventricular Nucleus of the Hypothalamus (PVN) to Decrease the Corticosterone Response to Stress
Authors: Erdos Benedek , Clifton Rebekah , McCowan Michael L , Sumners Colin , Scheuer Deborah A
Published: June 2021

MIF acts within PVN neurons to attenuate the excitatory actions of the neuropeptide angiotensin II, but is absent in PVN neurons of Spontaneously Hypertensive rats (SHR). Since angiotensin II in the PVN activates the hypothalamic pituitary adrenal axis resulting in corticosterone (Cort) secretion from the adrenal, we tested the hypothesis that restoration of MIF within SHR PVN neurons will attenuate the Cort response to stress. AAV2‐based viral vectors were microinjected into the PVN to express either GFP (n=6) or MIF (n=7) in adult male 11 week old SHR. Three weeks later a femoral arterial catheter was implanted under isoflurane anesthesia. At least 3 days later, rats were subjected to 60 min of restraint stress. Prior to stress, baseline plasma Cort was not significantly different between GFP‐ and MIF‐treated rats (6±1 and 4±1 μg/dl). However, during stress, plasma Cort was significantly (P<0.05) lower in MIF‐ compared with GFP‐treated rats at 15 min (36±2 vs 43±2 μg/dl) and 45 min (48±3 vs 58±2 μg/dl), and tended to be lower (P=0.057) at 60 min (48±2 vs 59±5 μg/dl). Thus, the integrated average increase in Cort during stress was significantly lower in MIF‐ compared with GFP‐treated rats (39±2 vs 44±1 μg/dl/min). We conclude that MIF acts in the PVN to attenuate the Cort response to restraint stress. Supported by HL093186 (MPI Sumners and Scheuer)