bluuebunny/crossref_metadata_2025 huggingface dataset:
No association between garlic intake and risk of colorectal cancer
Authors: Meng Shasha , Zhang Xuehong , Giovannucci Edward L , Ma Jing , Fuchs Charles , Cho Eunyoung
Published: June 2021

Background

Although experimental studies have suggested that garlic intake may reduce cancer risk, limited prospective studies have evaluated garlic intake and colorectal cancer (CRC) incidence.

Methods

We followed 76,208 women in the Nurses’ Health Study and 45,592 men in the Health Professionals Follow‐up Study up to 24 years and examined cumulative garlic intake and garlic supplement use in relation to CRC risk. Information on garlic intake and supplement use was assessed using a validated food frequency questionnaire and Cox proportional hazard regression model was used to estimate the multivariable relative risk (MV‐RR) and 95% confidence intervals (95% CIs).

Results

We found no association between garlic intake and CRC risk; the MV‐RR (95% CI) associated with garlic (1 clove or 4 shakes) intake ≥1/day compared with <1/month were 1.21 (0.94–1.57; p‐trend=0.14) for women and 1.00 (0.71–1.42; p‐trend=0.89) for men in 2,368 incident CRC cases. The MV‐RRs (95% CIs) of CRC for garlic supplement use, which was used in 6% of the participants in each study, were 0.72 (0.48–1.07) for women and 1.22 (0.83–1.78) for men.

Conclusion

Our data do not support a beneficial role of garlic intake or garlic supplement use in colorectal carcinogenesis. Intake of garlic or use of garlic supplement is not warranted to reduce colorectal cancer risk.

Funding: This study was supported by research grant CA136950, CA87969, and CA55075 from the National Institutes of Health.

Grant Funding Source: NIH
Trends in food intakes of U.S. adults: Findings from the Continuing Survey of Food Intakes by Individuals 1994–1996 and What We Eat in America, NHANES 2001–2002 and 2009–2010
Authors: Sebastian Rhonda S , Enns Cecilia Wilkinson , Goldman Joseph D , Moshfegh Alanna J
Published: June 2021

With the upsurge in diet‐related chronic diseases including obesity and diabetes in recent decades, it is of interest to determine if there are any concurrent changes in food consumption patterns. Using data from the Continuing Survey of Food Intakes by Individuals 1994–1996 (n = 9,872), What We Eat in America (WWEIA), NHANES 2001–2002 (n= 4,744), and WWEIA, NHANES 2009–2010 (n = 5,762), this study sought to identify trends in food intakes of U.S. adults age 20+ years. Regression analyses were used to estimate intake of major food groups and subgroups that were based on WWEIA Food Categories. For each food (sub)group, a t‐test of the β statistic representing survey period identified significant trends in intake (p<.001). A few trends were found in major food group intakes, such as increases in fruits and alcoholic beverages and decreases in vegetables. Among food subgroups, intakes rose for cheese, yogurt, nuts/seeds, pizza, candy, non‐citrus juice, and diet beverages and fell for whole/reduced‐fat milk, non‐fried potatoes/potato mixtures, and citrus juice. Although some of the trends that occurred suggest greater adherence to national dietary guidance, these improvements were offset by higher intakes of other foods that contribute large amounts of empty calories from added sugars, solid fats, and alcohol. Funding source: ARS/USDA.
Maternal serum 25‐hydroxyvitamin D and fetal growth in a multicenter US study
Authors: Gernand Alison D , Simhan Hyagriv N , Baca Katharyn M , Bodnar Lisa M
Published: June 2021

We examined the association between maternal serum 25‐ hydroxyvitamin D [25(OH)D] levels and measures of newborn size. Data came from a randomized controlled trial of aspirin for the prevention of preeclampsia in women at high risk for the disorder in 13 US medical centers. A total of 574 women delivered singleton, live births at term and had a serum sample drawn at ≤26 weeks gestation. Serum 25(OH)D was assayed with LC/MS‐MS. Multivariable linear and logistic regression models were used to assess the association between 25(OH)D and birth weight, length, head circumference, and risk of small‐for‐gestational‐age after adjustment for maternal prepregnancy BMI, race/ethnicity, age, gestational age at blood draw, and treatment allocation. Median birth weight was lower for women with 25(OH)D <30 nmol/L vs. ≥50 nmol/L (3200 g vs. 3395 g, p=0.004). After adjustment for confounders, 25(OH)D <30 nmol/L vs. ≥50 nmol/L was associated with a 168 g decrease in birth weight (95% CI: −301, −34). Furthermore, 25(OH)D 30–49 nmol/L vs. ≥50 nmol/L was associated with a 0.63 cm decrease in length (95% CI: −1.16, – 0.11). No association was observed for head circumference or risk of small‐for‐gestational‐age. Our results suggest that maternal vitamin D status may influence physiologic fetal growth in full term infants of high risk mothers. Supported by the National Institutes of Health grant HD056999 (PI: Dr. Bodnar).
Development of a Figural Drawing Scale for Body Shape Assessment in Chinese
Authors: Li Edmund T. S. , Cheng Tiffany P. M.
Published: June 2021

In an earlier study, we adopted the Stunkard nine‐figure scale to examine body shape dissatisfaction in Hong Kong adults. Due to ethnic specificity, a set of Chinese‐based body shape silhouette would be more helpful in understanding the weight‐related concepts of this population. The present study aimed to develop a set of self‐rating figural scale for Chinese adults and compared the discrepancies between the actual BMI (measured) and the BMI represented in the chosen figure (perceived) of the Stunkard scale and the new scale. The new figure dimensions were based on published anthropometric database from China and Taiwan. Figure #5 represents the mean BMI (21.9 kg/m2) and figures #1 and 9 anchor respectively the 3rd and 97th percentile of the population BMI. In a preliminary study, male (n=53) with BMI ranging between 15.9–28.0, indicated strongly that the new scale better represents the body proportion and shape of Chinese male when presented with both scales in a random and blinded fashion (χ2 =18.132, P < 0.001). They were asked to select the figure that best match their current body shape. The BMI difference (actual minus perceived) was −2.47 with the Stunkard scale and 0.083 with the new scale (t=12.806, P < 0.001). Hence when provided with the Stunkard scale, subjects tended to pick a figure that is larger than actual. Further study is warranted to validate the newly developed figural drawing scale.
Effect of Sugars‐Containing Beverages on Satiety and Short‐Term Food Intake in Normal Weight and Overweight/Obese Girls
Authors: Bellissimo Nick , Bennett Lorianne , Poirier Kelly , Hurton Evelyn , Luhovyy Bohdan , Nunes Fernando , Constantino Monika , Gladanac Branka
Published: June 2021

The effect of commercial beverages on satiety and short‐term food intake (FI) has received little investigation in girls. The purpose of this study was to determine the effect of commonly consumed sugars‐containing beverages on FI regulation in normal weight (NW) (n=12; 15–85th BMI percentile) and overweight (OW)/obese (OB) girls (n=11; >;85th BMI percentile). On 4 separate mornings and in random order, girls (n=23) received 350 ml of either a fruit drink, carbonated cola, 1% chocolate milk or a water control 2 h after a standardized breakfast of milk, cereal, and orange juice. FI (mean kcal ± SEM) from an ad libitum pizza meal was measured 60 min later. Only 1% chocolate milk decreased FI (746 ± 64) compared with the water control (935 ± 64; P<0.001) in NW girls, but none of the beverages decreased FI in OW/OB girls. In the pooled sample (n=23), FI was decreased by carbonated cola and 1% chocolate milk compared with the water control, but the fruit drink failed to result in a statistically significant decrease in FI. Caloric compensation scores for the fruit drink, cola, and 1% chocolate milk were 76%, 80%, and 86% in NW, and 45%, 74%, and 59% in OW/OB girls, respectively. Prospective food consumption (P<0.05) and desire‐to‐eat (P<0.05) scores, when corrected for the energy content of the beverages, were lowest after 1% chocolate milk compared to both cola and fruit drink, and cola, respectively. In conclusion, the effect of sugars‐containing beverages on short‐term FI in girls was dependent on the interaction between macronutrient composition and body weight status.
A NON‐INVASIVE TOOL FOR PREDICTING HUMAN ZINC DEFICIENCY
Authors: Harland Barbara , Edmonds Janet Harland
Published: June 2021

PHYTATE (also known as Phytic Acid or Inositol hexaphosphate (IP6)), is a form of phosphorus found in grains, beans, nuts, and a variety of other plant‐based foods. It is frequently consumed in over‐abundance throughout the world where the vegetarian/vegan diet is predominant. Phytate binds very tightly with minerals, especially zinc, causing the inhibition of absorption/re‐absorption of the di‐valent cations. Zinc is essential for numerous functions in the body, such as providing structural integrity to an enzyme as a cofactor, and as an essential component of zinc‐finger proteins. Zinc deficiency is common in regions where phytate consumption is high. A phytate:zinc molar ratio defines homeostasis of zinc from foodstuffs. When the phytate:zinc molar ratio is greater than 10, a zinc deficiency is defined. Phytate HPLC methods have been used to determine the amount of phytate in foods. Available zinc values contribute to the calculation of the pertinent phytate:zinc molar ratio for each food. This study presents analytical phytate:zinc values for several common foodstuffs.

Funding for this research is from Howard University Nutritional Sciences Department and College of Medicine, Washington, DC.
Processed purple‐fleshed potato prevents and protects against high‐fat diet elevated oxidative stress and inflammation markers in vivo in a pig model
Authors: Radhakrishnan Sridhar , Kim Sung Woo , Reddivari Lavanya , Vanamala Jairam
Published: June 2021

Little information exists if purple‐fleshed potatoes (PP), rich in anti‐oxidant and anti‐inflammatory anthocyanins, even after processing, retain their health benefiting properties in vivo. We hypothesized that processed PP will suppress high‐fat diet (HFD) elevated oxidative stress/inflammation biomarkers in a pig model. Thus, we performed two studies; a prevention study where 64 pigs, 3wk post‐weaning, consumed one of the eight diets; low‐fat (LFD), HFD and HFD supplemented with white‐fleshed potato (WP) or PP (raw/baked/chips; 10% w/w) for 13wk; and a protection study where pigs (12 wk on HFD) consumed HFD containing 10/20% PP or WP chips for additional 5wk (n=8). Both PP and WP diets (raw/baked/chips) suppressed urinary oxidative stress markers, 8‐isoprostane (8IP) and DNA adduct 8‐OHDG; and serum levels of TNF‐α (ELISA) compared to HFD control (P=0.038). Potato diets elevated colonic mucosal glutathione ratio (GSH:GSSG) compared to HFD control, however, greatest enhancement (P=0.046) was seen in raw PP consuming pigs. In the protection study, biomarkers of oxidative stress and inflammation, including colonic mucosal expression of COX‐2 and IFN‐γ (qRTPCR), and systemic levels of 8IP and TNF‐α were suppressed (P=0.049) by PP diets compared to control. These findings indicate that PP, even after processing, prevents and protects against HFD elevated colonic‐systemic oxidative stress/inflammation.

Grant Funding Source: This work was supported by National Research Initiative Grant 2009–55200‐05197 from the USDA‐NIFA.
In vitro immunomodulatory effect of Ipomoea batatas L. tuber bioactives
Authors: Chen Chiao‐Ming , Li Sin‐Chung , Liao Yen‐Yin , Liu Jen‐Fang , Shyur Lie‐Fen , Chen Peng‐Ru , Hsieh Wan‐Chen , Lin Pin‐Sing , Chen C‐Y. Oliver
Published: June 2021

Bioactives in Ipomoea batatas L. (sweet potatoes) have shown to exhibit radical‐scavenging activity, antimutagenicity, potential chemopreventive properties, and antidiabetic effects. However, bioactions of constituents in the Simon No. 1 (S1) and Tainong 66 (T66) sweet potatoes have not been fully elucidated. Thus, the objective of this study was to investigate the immunomodulatory effect of bioactives extracted from the S1 and T66 tubers harvested from the Chiayi Agricultural Experiment Station, Agricultural Research Institute, Taiwan. Bioactives in the tubers were first extracted with ethanol. After solvent removal with lyophilization, the resulting powder was further fractionated using liquid‐liquid partition to yield ethyl acetate (EAF) and water (WF) fraction. Mice splenocytes were harvested and treated with EAF or WF at concentrations ranging 0–200 μg/mL (w/v) for 24–48 h. EAF of the S1 and T66 increased cell proliferation in a dose‐dependent manner, but not WF. EAF of the T66 and S1 at 200 μg/mL enhanced cytotoxic activity of splenocyte NK cell by 24 and 41%, respectively, compared to control group at an effector cells/target cells ratio of 100. In conclusion, our results showed that bioactives in sweet potatoes exhibit immunomodulatory activities in a cultivar dependent manner. Further studies are warranted to examine underlying mechanism(s) for the modulations and characterize bioactives.
High Glucose Induced Monocyte‐to‐Macrophage Differentiation: Role of AMPk
Authors: VASAMSETTI SATHISH BABU , Rao Padmapriya , Kotamraju Srigiridhar
Published: June 2021

Diabetes‐induced vascular dysfunctions are on the upsurge and earlier studies have correlated increased inflammatory events as a result of hyperglycemia, plays a major role in the development of diabetes‐associated vascular dysfunctions including atherosclerosis. Monocyte‐to‐Macrophage differentiation has been implicated as a key event during the progression of atherosclerosis. This process induces an inflammatory environment by generating proinflammatory cytokines. In this study, we tested the hypothesis that high glucose levels which mimic diabetic conditions can augment monocyte to macrophage differentiation and also promote the release of inflammatory mediators. It was found that glucose dose‐dependently increased PMA‐induced THP‐1 monocyte differentiation which is coincided by the production of proinflammatory cytokines like IL‐1â, MCP‐1 and MMP‐9. Interestingly, High glucose–induced monocyte differentiation as well as proinflammatory cytokine production was attenuated by AMPk activators like metformin. In conclusion, our results show that high glucose levels cause increased monocyte activation via altered regulation of AMPk signaling cascade and thereby may promote a proatherogenic setting.
Relative neutralization of unfractionated heparin and various Enoxaparins by platelet factor 4 in platelet rich plasma. Therapeutic implications
Authors: Syed Daniel , Iqbal Omer , Hoppensteadt Debra , Jeske Walter , Fareed Jawed
Published: June 2021

Endogenous platelet factor 4 released from blood platelets is capable of neutralizing both heparin and enoxaparin (ENOX). The purpose of this study is to investigate the differences in the neutralization profile of heparin and ENOX preparations by PF4. Citrated whole blood was centrifuged to obtain platelet rich (PRP) and platelet poor (PPP) plasma. Both were supplemented with three batches of heparin, generic enoxaparin (GENOX) and branded enoxaparin in a concentration of 0–10 ug/ml. Clot based, APTT, Heptest, Thrombin time and amidolytic anti‐Xa and IIa were measured. The anticoagulant responses in the aPTT, Heptest and TT were found to be markedly reduced in the PRP samples, >;70% neutralization was observed in these assays. Similarly in the anti‐Xa and IIa assays, PRP samples showed a reduction in the inhibitory effects, which were >;50% in both assays. The GENOX and ENOX exhibited a lesser neutralization effect in the APTT and Heptest assays in comparison to the heparin. The neutralization profile ranged 20–30% in the clot based assays. In the amidolytic assays, the anti‐IIa activity was more efficiently neutralized. The mean neutralization was >;70% for the anti‐IIa. In the anti‐Xa assay, the GENOX's showed less neutralization (ranging from 20–30%). These results suggest that endogenous platelet factor 4 is capable of differentially neutralizing both the heparin and ENOX in an assay dependent fashion.
VP16‐HCF interaction is required for herpes simplex virus exit from latency in neurons in vivo
Authors: Sawtell Nancy M , Thompson Richard L
Published: June 2021

A mechanistic understanding of viral and host functions regulating herpes simplex virus (HSV) latency and reactivation remains a major goal. We have shown that the interaction of VP16 and Oct‐1 is required very early during entry into the lytic cycle from the latent state and have proposed a VP16‐dependent model of reactivation in vivo. HCF‐1 has important roles in promoting HSV replication and a model has been proposed in which stress induces translocation of HCF‐1 into the neuronal nucleus and reactivation is regulated by direct recruitment of HCF to viral IE genes. To investigate whether HCF‐1 can precipitate viral gene expression in latently infected neurons in the absence of binding to VP16, a mutation in VP16 at Y364A, shown previously to disrupt VP16‐ HCF interaction (but not interaction with Oct‐1) was engineered into HSV‐1. Three independent isolates and a genomically restored virus were examined in vitro and in the mouse ocular model. Our findings demonstrate that the interaction of VP16 with HCF‐1 is required for efficient infection at low moi, efficient viral replication in vivo, and exit from latency/entry into the lytic cycle in trigeminal ganglion neurons in mice following hyperthermic stress. In the absence of VP16 binding, nuclear translocation of HCF is not sufficient to initiate the viral lytic cycle, demonstrating the key role of VP16 in regulating in vivo reactivation.
Role of CDX2 protein expression in the diagnosis of Barrett's esophagus.
Authors: Abdelbaqui Maisoun , Coppola Domenico
Published: June 2021

Background

CDX2 is a nuclear homeobox transcription factor not expressed in normal esophageal and gastric epithelium but has been reported to highlight the intestinal metaplasia of the esophagus. In this study we investigate the utility of CDX2 in detecting Barrett's esophagus as compared to the conventional Alcian blue pH 2.5 special stain (AB).

Materials

Patients who underwent upper endoscopy to rule out Barrett's at the Florida Digestive Health Specialists and at the Moffitt Cancer Center within the past 12 months were selected. H&E biopsies slides and AB stains were reviewed. Cases were categorized as follows: 108 cases with histologically evident goblet cells (BE), 48 cases without histologic evidence of goblet cells (NBE), 43 cases containing esophageal glands (EG), and columnar blue cells (CB). These cases were stained for CDX2 following the manufacturer recommendations.

Results

We reviewed a total of 199 esophago‐gastric biopsies from 186 patients. All 108 BE (100%) were positive for AB and 102 (94.4%) were positive for CDX2. 89.6% of 48 NBE were negative for AB and CDX2. 100% of 43 cases showing EG and CB were AB falsely positive. These cases were CDX2 negative. The positive predictive value in detecting intestinal metaplasia for CDX2 was 95.6% and for AB it was 71.5%.

Conclusion

Adjunct of CDX2 immunostain to AB helps in the evaluation of esophago‐gastric biopsies performed to rule out BE.
Chronic Toll‐like receptor 9 activation mediates heightened vascular contractility via attenuated NOS activity in isolated aortic segments
Authors: McCarthy Cameron , Goulopoulou Styliani , Wenceslau Camilla , Matsumoto Takayuki , Webb R. Clinton
Published: June 2021

Toll‐like receptor 9 (TLR 9) is a pattern recognition receptor of the innate immune system. Activation of TLR 9 in immune cells leads to the release of pro‐inflammatory cytokines. TLR 9 is also expressed in the vasculature; however, its role in vascular function remains to be clarified. We hypothesized that chronic administration (three i.p. injections within five days) of TLR 9 agonist [synthetic oligonucleotide (ODN 2395); 0.1 μg/i.p.], would augment aortic contractility in female, 13–15 weeks old, Sprague Dawley rats. Concentration response curves were performed ex vivo using the myograph to norepinephrine (NE; 10−9–3×10−5 M), in the presence of either nitric oxide synthase (NOS) inhibitor L‐NNA (10−4 M; 30 min), or cyclooxygenase inhibitor indomethacin (10−5 M; 30 min). ODN 2395 amplified the contractile response to NE [Emax (% max KCl), ODN 2395: 108±4 vs. Veh: 94±5]. This difference was normalized in the presence of L‐NNA [Emax (% max KCl), ODN 2395: 141±6 vs. Veh: 134±10], but not indomethacin [Emax (% max KCl), ODN 2395: 105±8 vs. Veh: 89±8]. These data illustrate that NOS activity was decreased in the ODN 2395 treated animals, as NOS inhibition normalized the potentiated contractile response to ODN 2395. Therefore, attenuation of NOS activity by TLR 9 activation could subsequently contribute to the development and progression of vascular diseases such as hypertension.

Research Support: Society for Women's Health Research (SWHR)
Overexpression of the large‐conductance, Ca2+‐activated K+ (BK) channel shortens action potential duration in HL‐1 cardiomyocytes
Authors: Rhee Sung W , Song Li , Steephan Mathew , Rusch Nancy J , Stimers Joseph R
Published: June 2021

Hereditary long‐QT syndrome (LQTS) is caused by mutations in ion channel genes resulting in a prolonged action potential duration (APD) in cardiomyocytes. Complications include syncope and sudden cardiac death. Large‐conductance, Ca2+‐activated K+ (BK) channels mediate repolarization of many cell types, but they are not expressed in the plasma membrane of cardiomyocytes. We hypothesized that introduction of BK channels may shorten APD in cardiomyocytes as a potential therapy for LQTS. HL‐1 atrial myocytes were transiently transfected with BK plasmids bicistronically expressing hBK and mCherry or control plasmids expressing only mCherry. Expression of the hBK transgene was confirmed by Western blot, immunocytochemistry and by detecting iberiotoxin (Ibtx)‐sensitive BK current in patch‐clamped cells. Western blots showed that the expression of other ion channels including KV4.3, KV7.1, NaV1.5, and CaV1.2 did not significantly change after BK transfection, whereas KV11.1 was down‐regulated by 20% (n=5). APD at 50% repolarization was 3.7±0.5 ms in BK‐transfected cells compared to 6.3±0.6 ms in control‐transfected cells. Ibtx (100 nmol/L) restored the shortened APD in hBK‐transfected cells to 7.9±1.5 ms (n=5–14). Thus, BK channels can exert a powerful repolarizing influence in HL‐1 myocytes, and may be a useful therapy to reverse LQTS. AHA SDG 0830060N (SWR) and NIH R01HL093526 (NJR)
A single exposure to morphine induces long‐lasting sensitization: age‐related differences in mice
Authors: Koek Wouter
Published: June 2021

Given evidence for age‐related differences in effects of drugs of abuse, surprisingly few studies have explored effects of opioids in adolescents (versus adults). This study compared locomotor sensitization in adolescent and adult mice after a single exposure to morphine.

Adolescent (postnatal day 29) and adult male C57BL/6J mice were treated with saline or morphine (1–100 mg/kg, i.p.) (session 1), and morphine (10 mg/kg)‐induced locomotion was assessed 3–36 days later (session 2). Morphine during session 1 dose‐dependently enhanced the locomotor‐stimulating effects of 10 mg/kg morphine during session 2. The minimum significant dose was 3.2 mg/kg in adults and 32 mg/kg in adolescents, and the maximal enhancement was 200% in adults and 150% in adolescents. Irrespective of age, the enhancement increased with the length of the intersession interval, attained a maximum at 1 week, and remained unchanged at 2 and 5 weeks. In adults, but not in adolescents, the enhancement was largest when both sessions were conducted in the same environment.

These results show that a single exposure to morphine induces dose‐, delay‐, and context dependent sensitization in adults. In adolescents, morphine induces sensitization less potently, less efficaciously, and less dependent on the context than in adults, but its sensitizing effects are as long‐lasting as in adults, and last into adulthood.

Supported by DA23261
Sphingosine 1‐phosphate receptor 1 mediates morphine‐induced antinociceptive tolerance and hyperalgesia
Authors: Doyle Timothy M. , Bryant Leesa , Cuzzocrea Salvatore , Salvemini Daniela
Published: June 2021

The clinical utility of morphine is hampered by the development of antinociceptive tolerance and hyperalgesia. We have reported these conditions arise in part from ceramide‐sphingosine‐1‐phosphate (S1P)‐dependant proinflammatory processes in tolerant rats, but the signaling pathways are not clear. We now show in rats chronically treated with morphine that the S1P signaling is dependent on the activation of the S1P receptor 1 (S1PR1). Systemic FTY720, a functional antagonist of S1PR1 that inhibits S1P signaling through S1PR1 internalization and degradation, attenuated NF‐κB and MAPK activation, TNFα and IL1β release in spinal cord, and completely prevented the development of tolerance and hyperalgesia in a dose‐dependent manner. The role of S1PR1 signaling was further supported by the attenuation of tolerance and hyperalgesia with intrathecal administration of selective S1PR1 antagonist, W146, but not its inactive enantiomer, W140, and by the induction of hyperalgesia in normal rats by intrathecal SEW2870, a selective S1PR1 agonist. Collectively, our findings identify a role of the S1P‐to‐S1PR1 pathway in morphine tolerance and hyperalgesia and, since FTY720 is FDA‐approved for the treatment of multiple sclerosis, may provide a fast‐track to clinical application of S1PR modulators as adjuncts to opiates. Supported by NIH/NIDA [R01 DA024074] and APS Future Leaders in Pain Research Grant.
Bone marrow inflammation precedes hexahydro‐1‐nitroso‐3, 5‐ dinitro‐1, 3, 5‐triazine (MNX) induced delayed myelosuppression in rats.
Authors: Jaligama Sridhar , Kale Vijay M. , Wilbanks Mitchell S. , Perkins Edward J. , Meyer Sharon A.
Published: June 2021

MNX, an environmental nitroreduced product of munitions RDX contaminates military sites. Our previous studies identified bone marrow (BM) as target of acute oral exposure to MNX in rats in the form of delayed loss of BM Granulocyte Macrophage Colony Forming Cells (GM‐CFCs) and concomitant decrease in blood granulocytes (NOAELs 24, 47 mg/kg resp) at 14 days after exposure. To address whether delayed effects of MNX are due to persistence of early hematological effects or are late‐onset due to required expression period, female Sprague‐Dawley rats were orally gavaged with MNX from 0 to 94 mg/kg and different toxicological endpoints were evaluated over a time course at 2, 7, 10, 12, and 14d. A significant increase in blood granulocytes and circulating levels of RANTES, a leukocyte chemokine, indicate that an acute inflammatory response occurs at 2d after exposure to MNX. Also, persistent BM macrophage infiltration in MNX (94 mg/kg) treated rat iliums (24h, 2d and 10d) and activation of NFkB signaling pathway in BM cells at 10d were observed. Further, increase in adherent BM mesenchymal stromal cell colonies constituting macrophages, endothelial cells and fibroblasts was observed in MNX treated rats. Collectively, these data suggest that myelosuppression is delayed until 10d presumably due to development of inhibitory effects of preceding BM inflammation on myelopoiesis. (Support: DoD/CDMRP, US Army Corps of Engineers)
Extracellular adenosine modulates neutrophil host‐pathogen interactions via activation of the A3‐adenosine receptor
Authors: Corriden Ross , Briddon Steve , Hill Stephen J , Nizet Victor
Published: June 2021

Extracellular nucleotides/nucleosides are important mediators of immune cell function that activate a diverse array of membrane receptors. Of these, the A3‐adenosine receptor (A3AR) stands out as a key regulator of neutrophil behavior. Here, we use a fluorescent A3AR ligand in conjunction with flow cytometry and confocal microscopy to show that endogenous A3ARs aggregate in highly polarized plaque‐like microdomains that modulate human neutrophil chemotaxis. In addition to regulating chemotaxis, A3ARs promote the formation of membrane projections that enable neutrophils to tether and “reel in” bacteria to the cell membrane. Exposure of neutrophils to adenosine, particularly in the presence of the A2a‐selective inhibitor ZM 241385, enhances their ability to kill Staphylococcus aureus and phagocytose Staphylococcus aureus bioparticles. Our results reveal new properties of human neutrophils and identify the A3AR as a potential target for modulating the innate immune function of these cells.
Careers beyond the bench ‐ an overview
Authors: Zuk Dorit
Published: June 2021

Individuals trained in biomedical research follow many career paths, extending well beyond the traditional path of conducting research in academia. A recent report from a working group of the Advisory Committee to the NIH Director tasked with examining the future of the biomedical research workforce in the US (http://acd.od.nih.gov/bwf.htm) showed that fewer than 50% of US‐trained individuals who hold a PhD in biomedical research pursue a research or teaching career in academia. Many conduct research in industry or government settings, while others take positions in science‐related non‐research occupations. Careers paths in science policy and scientific editing will be discussed as examples of such occupations.
Determination of the human SULT1C3 mRNA sequence in LS180 cells
Authors: Duniec‐Dmuchowski Zofia , Runge‐Morris Melissa , Kocarek Thomas A.
Published: June 2021

The human SULT1C subfamily consists of SULT1C2, SULT1C4, and one putative gene, SULT1C3, which has been identified by computational analysis of the human genome but has not been reported as expressed in any human tissue. An unusual feature of the predicted SULT1C3 gene is that it contains a duplication of the last two exons, with the upstream copies designated as exons 7a and 8a and the downstream copies designated as exons 7b and 8b. The NCBI reference sequence for SULT1C3 mRNA (NM_001008743) begins at the predicted translation start site, ends with exons 7b and 8b, and does not include exon 7a or 8a. We used primers corresponding to sequences in exons 6 and 7b to survey SULT1C3 expression in human tissues and amplified a largerthan‐expected RT‐PCR fragment from small and large intestinal mRNA as well as from LS180 human colorectal adenocarcinoma cells. The cloned fragment contained exons 7a and 8a in addition to the expected portion of exon 7b. We then used 5′‐and 3′‐RACE to determine the entire SULT1C3 mRNA sequence in LS180 cells. These analyses indicated that the SULT1C3 gene contains a non‐coding exon ~7.1 kb upstream of the first coding exon and that the major SULT1C3 transcript contains exons 7a and 8a rather than 7b and 8b. We conclude that SULT1C3 is expressed in human intestinal cells but that its 3′‐exons are not those indicated for the reference sequence. This work was supported by NIH grants HL050710 and ES005823.
Effect of microsphere encapsulation on peptide clearance from the respiratory tract
Authors: McKenna Nicole Renae
Published: June 2021

Microspheres harbor great potential as an effective means of targeted delivery to specific tissues for use in cancer treatment and vaccinations. Both encapsulation volume and size were examined in this research. Microspheres were created by the encapsulation of EP67‐FITC in Poly Lactic‐co‐Glycolic Acid (PLGA). Different sizes (360–370 nm and 8.8 ìm) and volumes (15 ìl and 50 ìl of the same 50ìg dose) of microspheres were investigated to determine their effect on localization and clearance in the respiratory tract of female BALB‐c mice. Results indicate that the PLGA encapsulated EP67‐FITC was more rapidly cleared than unencapsulated EP67‐FITC. Also, the 50 ìl volume of unencapsulated EP67‐FITC localized to the stomach of the mice while encapsulated EP67‐FITC did not. This research shows a clear advantage to PLGA encapsulation and additionally to smaller particles (360–370 nm). This publication was made possible by grants from the National Center for Research Resources (5P20RR016469) and the National Institute for General Medical Science (NIGMS) (8P20GM103427), a component of the National Institutes of Health (NIH) and its contents are the sole responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH.
Comparison of Gel and Injection Delivered Carprofen for Post‐Operative Pain Management in Mice
Authors: Giuvelis Denise , Palmer Jay , Bergquist Ivy , Harding Lisa , Brazeau Daniel , Bilsky Edward
Published: June 2021

Post‐operative pain management in laboratory animals can be a challenge from both a delivery and cost perspective. The purpose of this study was to determine if carprofen could be effectively delivered to mice using dietary gel technology. Efficacy was assessed by measurement of a reduction in pain behaviors following ovariectomy in mice. Control or carprofen‐formulated gels were made available to animals ad libitum 24 hrs prior to surgery and throughout the recovery period. Subcutaneous carprofen injections were given 30 min prior to surgery and at 24, 48 and 72 hrs post‐surgery. Groups receiving carprofen had quicker recoveries of bodyweight compared to vehicle injections. Behavioral pain assessment included open‐field measurement of locomotor activity and tactile hypersensitivity (abdominal von Frey). The carprofen gel group had similar reductions in pain behaviors compared to animals treated with conventional injections of the analgesic. Pharmacokinetic measurements of carprofen plasma levels indicated that the gel and injection delivery methods yielded equivalent plasma levels of the analgesic at all time points. These studies suggest that the gel technology represents a convenient and cost effective alternative to traditional post‐operative pain management in mice. This work was supported by a contract from ClearH2O to the University of New England.
Neuronal nitric oxide synthase (nNOS) inhibition and regional sympathetic nerve discharge: implications for peripheral vascular control
Authors: Copp Steven W. , Hirai Daniel M. , Sims Gabrielle E. , Musch Timothy I. , Kenney Michael J. , Poole David C.
Published: June 2021

Objective

Selective nNOS inhibition with systemically‐administered S‐methyl‐L‐thiocitrulline (SMTC) in conscious rats elevates mean arterial pressure (MAP) and reduces skeletal muscle and renal blood flow (Copp et al. J Physiol 588: 1321–1331, 2010). We tested the hypothesis that this SMTC‐induced hindlimb skeletal muscle and renal vasoconstriction occurs consequent to increases in lumbar and renal sympathetic nerve discharge (SND) in preference to local vasomotor modulation.

Methods

MAP (femoral artery catheter) and lumbar and renal SND (direct nerve recordings) were measured in 8 anesthetized sino‐aortic denervated rats for 20 min each following, in random order, saline and SMTC (0.56 mg/kg) infusions.

Results

SMTC increased MAP (peak ΔMAP: 52±9 mmHg, p<0.01) compared to saline which persisted for 10 minutes and then returned to baseline values. Lumbar and renal SND were not different between saline and SMTC conditions (p>;0.05) and the differences between saline and SMTC were not different from zero (peak Δlumbar SND: 2±7 %; peak Δrenal SND: 11±9 %, p>;0.05).

Conclusion

In contrast to our hypothesis, nNOS inhibition with SMTC induced marked MAP elevations without altering lumbar or renal SND. Therefore, reductions in hindlimb skeletal muscle and renal blood flow observed previously reflect significant peripheral nNOS‐derived NO vascular effects. Funding: NIH AG‐041948
Algebraic Solutions for Homeostatic Mechanisms in a Minimal Closed‐Loop Cardiovascular System Model
Authors: Waters Shannon E. , Jalili Nima A. , Abuid Nicholas J. , Mohiuddin Mohammad W. , Quick Christopher M.
Published: June 2021

To characterize the interaction of the heart with the pulmonary and systemic circulations, engineers developed a minimal closed‐loop cardiovascular model, predicting cardiac output and pressures from 18 critical parameters. Despite the need for numerical solutions, it has potential to be used by clinical investigators to study the mechanical basis of altered hemodynamic status and by physiologists to study homeostatic mechanisms. To capture the complex interplay between hemodynamics and structural adaptation, we modified the standard minimal model to make structural parameters adapt to local hemodynamics. First, we linearized all pressure‐volume and pressure‐flow relationships. Then we made the simplifying assumption that end‐systolic pressure approximates mean arterial pressure. Finally, we assumed that each parameter adapts as a linear function of regional pressure. Yielding cardiac output and mean pressures, our model reproduces basic adaptive behaviors such as cardiomegaly in response to decreases in contractility and diastolic dysfunction in response to decreases in sensitivity of the adaptive process. More importantly, by judicious use of simplifying assumptions, all solutions can be expressed algebraically, which makes this fundamental model accessible to physiologists, clinical investigators, and students.
Increased Expression of Prorenin Receptor (PRR) in the NTS of Spontaneously Hypertensive Rats (SHR) May Be A Compensatory Mechanism of Hypertension
Authors: Shan Zhiying , Zubcevic Jasenka , Yuan Wei , Jun Joo Yun , Qi Yanfei
Published: June 2021

PRR expression is significantly increased in the nucleus of the solitary tract (NTS) of SHR compared to WKY controls. This observation led us to hypothesize that increased NTS PRR expression is linked to hypertension. We test this hypothesis by chronic knockdown or overexpression PRR in the NTS using AAV vector mediated gene transfer. Knockdown of PRR by AAV2‐PRR‐shRNA in SHR resulted in a 22 mmHg increase in mean arterial pressure (MAP) (shRNA: 173±5; Control: 151±6 mm Hg) along with shifting the set point of arterial pressure to a higher level. However, no changes were observed on MAP in WKY rats. Consistent with pressor effects of PRR knockdown, overexpression of PRR in SHR by AAV2‐human‐PRR induced a 19 mmHg decrease in MAP (PRR: 148±7; Control: 167±7 mmHg) compared to control animals. Finally, we measured the effect of increased PRR ligand, prorenin on blood pressure. Bilateral acute NTS injection of human renin (2 pmol/side) produced significant greater decreases in MAP and heart rate (HR) in SHR (ΔMAP: −32±5 mmHg, ΔHR; −40±10 bpm) compared to WKY rats (ΔMAP: −8±4 mmHg; ΔHR: −12±7 bpm). These effects in SHR were attenuated (70%) by prorenin handle region peptide, but were not affected by AT1 or AT2 receptor blockers.

This study suggests that increased NTS PRR expression may be a compensatory mechanism of hypertension and that it mediates an anti‐hypertensive effect in an Ang II independent mechanism. ( AHA 11SDG7420029)
Mutation of an anti‐aging gene klotho causes hypertension via upregulation of plasma aldosterone levels
Authors: Zhou Xiaoli , Lei Han , Sun Zhongjie
Published: June 2021

Klotho is a recently discovered anti‐aging gene. The purpose of this experiment is to assess if klotho deficiency affects blood pressure. Interestingly, klotho (+/−) mice demonstrated a significant and persistent increase in blood pressure starting form 3–4 months of age, indicating that klotho deficiency causes hypertension. Plasma level of aldosterone was elevated in KL(+/−) mice. Two groups of KL(+/−) mice and 2 groups of wild‐type mice were used. When BP was elevated in KL(+/−) mice, 1 group of KL (+/−) mice and 1 group of WT mice were treated with aldosterone receptor blocker, eplerenone (6 mg/kg/day), while the remaining groups received DMSO and serve as controls. Eplerenone decreased hypertension to the control level. Eplerenone also abolished KL deficiency‐induced kidney damage (glomerulus collapse, fibrosis). Klotho deficiency causes renal inflammation as evidenced by significant increases in inflammatory cytokines (IL‐6, TNFα and MCP‐1) and T cell and macrophage infiltration in kidneys of KL (+/−) mice. Eplerenone significantly attenuated KL deficiency‐induced inflammation. Notably, NCC and Sgk1 levels were increased in kidneys in KL(+/−) mice, which could be abolished by eplerenone. It is concluded that KL deficiency caused hypertension via increasing plasma aldosterone levels which led to inflammation and upregulation of NCC signaling in kidneys.
Proteomic analysis of outer and juxtamedullary cortex of non‐clipped kidneys in 2K1C hypertensive rats
Authors: Finne Kenneth , Skogstrand Trude , Tenstad Olav , Berven Frode , Hultström Michael , Vethe Heidrun , Vikse Bjørn Egil
Published: June 2021

Hypertensive damage in the non‐clipped kidney of two‐kidney, one‐clip (2K1C) rats is most pronounced in the juxtamedullary cortex (JMC), while the outer cortex (OC) is protected. In this study, 2K1C was induced with a 0.2mm silver clip in 6‐week‐old male Wistar rats that were sacrificed after 16‐weeks and compared to sham operated controls (19‐weeks). Differentially abundant proteins were identified in a hypothesis free manner using trypsin digested OC and JMC that was tagged with iTRAQ‐4plex, and analyzed on an Orbitrap Velos Pro liquid‐chromatography tandem mass spectrometer.

2714 proteins were quantified (FDR < 0.01). 269 proteins increased, and 127 decreased in abundance in 2K1C compared to control (fold change >; 1.5). The number of differential abundant proteins was higher in JMC (338) than in OC (231). 51 of the differential abundant proteins overlapped with 565 differentially expressed genes from microarray gene expression data obtained previously in the same model.

In conclusion, hypertensive kidney damage in the non‐clipped kidney of 2K1C is associated with large changes in the proteome, which are more pronounced in the JMC. Important proteins identified include Osteopontin, S100A4 and Decorin, which are known to play a role in fibrosis and tissue remodeling.

This work was funded by Western Norway Regional Health Authority and the University of Bergen.
Salt‐dependent inhibition of the epithelial sodium channel (ENaC) by Bradykinin
Authors: Mamenko Mykola , Zaika Oleg L. , Doris Peter A. , Pochynyuk Oleh
Published: June 2021

Kalikrein‐kinin system in the kidney promotes natriuresis to decrease blood pressure. Deletion of Bradykinin (BK) receptors results in salt‐sensitive hypertension indicating possible augmentation of Na+ transport and ENaC activity in the distal nephron. In this study, we tested a role of BK in adaptation of ENaC to changes in systemic salt status. Using patch clamping in split‐opened distal nephrons, we found that BK reversibly inhibits ENaC. BK had no effect on ENaC in mice lacking Bradykinin receptors (B1,B2−/−). Under normal Na+ intake (0.32%), ENaC Po was modestly elevated in B1,B2−/− compared to WT mice. High Na+ intake (2%) greatly augmented this difference. DOCA treatment similarly increased ENaC activity in both strains suggesting aldosterone‐independent nature of BK regulation of ENaC. Angiotensin converting enzyme (ACE) represents the major pathway of BK degradation. Systemic inhibition of ACE with captopril decreases ENaC activity in WT but not in B1,B2−/− mice. We further determined that captopril greatly potentiated BK signaling in the distal nephron resulting in a stronger ENaC inhibition. We concluded that BK cascade has its own specific role in blunting ENaC activity particularly under conditions of elevated Na+ intake. Augmentation of BK signaling in the distal nephron inhibits ENaC‐mediated Na+‐reabsorption contributing to the natriuretic and antihypertensive effects of ACE inhibition.
Inhibition of renal cyst progression by montelukast: an in vitro study in MDCK cells
Authors: Pathomthongtaweechai Nutthapoom , Chatsudthipong Varanuj , Muanprasat Chatchai
Published: June 2021

Renal cyst enlargement especially in polycystic kidney disease (PKD) is driven by P2Y receptor‐mediated and cAMP‐mediated chloride secretion into cyst lumen. This study aimed to determine the effect of an anti‐asthmatic drug montelukast, cysteinyl leukotriene (CysLT) 1 receptor antagonist, on renal cyst progression and to investigate underlying mechanisms of such an effect. In MDCK cyst model, montelukast (50 μM) inhibited forskolin‐induced cyst growth and cyst formation by 80%. This effect was not reversed by leukotriene D4 co‐incubation. In addition, zileuton, an inhibitor of leukotriene synthesis, had no effect on cyst growth. As analyzed by short‐circuit current measurements, acute treatment with montelukast (50 μM) inhibited ATP (P2Y agonist)‐induced chloride secretion by 50%, with no effect on cAMP‐induced chloride secretion. Similarly, prolonged treatment with montelukast (50 μM, 24 h) significantly inhibited ATP‐induced chloride secretion. Furthermore, montelukast had no effect on cell viability. Our study indicates that montelukast inhibits MDCK cyst progression by attenuating purinergic signaling in these cells. Therefore, montelukast represents a promising candidate drug for PKD therapy.

This work was supported by the Faculty of Science and Medical Scholars Program, Mahidol University.
Lovastatin attenuates cyclosporine inhibition of potassium channels in cortical collecting duct cells
Authors: Ma He‐Ping
Published: June 2021

Statins may benefit patients receiving cyclosporine A (CsA) treatment after organ transplantation. However, there is little in vitro data to support the use of statins to reduce the harmful effects of CsA such as hyperkalemia. Our cell‐attached patch‐clamp data showed that CsA inhibited an inwardly rectifying potassium channel in the apical membrane of mouse cortical collecting duct principal cells (mpkCCDc14 line) and that lovastatin attenuated the inhibition. Phosphatidylinositol‐3,4,5‐trisphosphate (PIP3) is a lipid which is known to stimulate inward rectifier potassium channels. Under normal conditions, PIP3 is synthesized in the basolateral membrane and can pass tight junctions to diffuse to the apical membrane. It appears that this diffusion is blocked by CsA because confocal microscopy showed that CsA reduced PIP3 in the apical membrane, but increased PIP3 in the basolateral membrane. Lovastatin reversed the effects of CsA on PIP3 levels. We also found that CsA increased, but lovastatin decreased, the transepithelial resistance by altering the levels of zonula occludens‐1 (ZO‐1) and cholesterol in tight junctions. Using scanning ion conductance microscopy, we showed that the apical membrane of mpkCCDc14 cells contains cellular protrusions in the area near tight junctions. CsA significantly increased the height of the protrusions. In contrast, lovastatin eliminated the protrusions and even caused a modest depression between the cells. These data suggest that lovastatin can abolish CsA inhibition of potassium channels in the apical membrane of CCD cells by altering the lateral diffusion of PIP3 which possibly controlled by both ZO‐1 and cholesterol levels in tight junctions. The present study provides one molecular mechanism for the previous suggestion that statins benefit CsA treatment of patients with organ transplantation.