bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Sucromalt consumption on mental and physical energy questionnaire responses
Authors: Berger Alvin , Bell Margie , Kanter Mitch
Published: June 2021

We evaluated whether consumption of the low glycemic carbohydrate Sucromalt (SULT) may benefit mental performance. Forty‐four subjects consumed a standardized dinner, then beverages containing 75 g dextrose or SULT at 7:30 AM. Subjects self‐reported on 19 individual and 4 composite questions related to physical or mental energy, hunger, and sleepiness (“POMS Brief Test”). Results were analyzed with repeated measures (RM‐) ANOVA, using ranks for non‐normally distributed data. RM‐ANOVA for change from baseline yielded changes (p<0.10, unadjusted for all tests), in favor of SULT for: Composite Physical Energy State; Composite Mental Fatigue State; Physical‐Fatigue; Physical‐Vigor; Physical‐Exhaustion; Mental‐Fatigue; and Mental‐Vigor. RM‐ANOVA for the Incremental Area Under the Curve (IAUC) yielded additional differences in favor of SULT for: Composite Physical Fatigue State; Composite Mental Energy; Physical‐Energy; Physical‐Pep; and Mental‐Energy. Changes were most pronounced 4–5 h post‐consumption in favor of SULT for all composite questions; and 5/6 Physical‐ and 4/6 Mental individual questions (following baseline adjustment, Wilcoxon Signed Rank test). In addition to benefits for blood sugar and weight control, SULT‐containing beverages may thus additionally provide cognitive benefits.
A preliminary investigation of nutritional status and attentional performance of 3‐month‐old infants
Authors: Aubuchon‐Endsley Nicki Lynn , Stoecker Barbara , Thomas David G. , Kennedy Tay S. , Froese Chandra , Bugh Lauren , Bezabih Afework M.
Published: June 2021

The present study investigated the statistical predictive utility of indicators of nutritional status on heart‐rate‐defined phases of attention in 3‐month‐old infants. The former included infant anthropometric variables; maternal blood cadmium, lead, and hemoglobin concentrations; and maternal serum transferrin receptor concentrations. A multiple regression analysis using indicators of nutritional status as predictors and attentional performance as the criterion was statistically significant (R = 0.69, p = 0.04). Maternal cadmium concentrations were the only statistically significant individual predictor (partial r = 0.66, p < 0.001). Additional t–tests demonstrated significant differences in attentional performance between those with the highest quartile of cadmium concentrations versus all other infants (t(36) = 3.00, p < 0.005), which was not attenuated by high iron status (t(36) = 2.40, p = 0.02). This supports the use of maternal nutritional status variables to statistically predict early infant attention abilities. This research was supported by National Research Initiative Grant 2008‐35200‐18779 from the USDA National Institute for Food and Agriculture.

Grant Funding Source: National Research Initiative Grant 2008‐35200‐18779 from the USDA National Institute for Food and Agriculture
Adherence to a Mediterranean type dietary pattern may reduce cognitive decline
Authors: Tangney Christy C , Kwasny Mary J , Li Hong , Evans Denis A , Morris Martha Clare
Published: June 2021

Background

Many foods abundant in the traditional Mediterranean diet, such as vegetables, fish and wine have been associated with slower cognitive decline. We investigated whether a diet consistent with this traditional pattern among black and white older (≥65 years of age) adults in the ongoing Chicago Health and Aging Project (CHAP) is associated with cognitive changes.

Methods

CHAP participants (n=3790) with valid baseline dietary measurement and 2 or more cognitive assessments were evaluated for adherence to a Mediterranean diet pattern based on the MedDiet score of Panagiotakos and coworkers (2007). Scores from 0 to 5 were assigned to daily intake of food items belonging to each of 11 components for a total possible score of 55. Linear mixed models were used to examine the association of MedDiet scores to change in cognitive function. Cognitive function was estimated as the sum of standardized scores on four cognitive tests assessed every three years. Mean follow‐up was 7.6 years.

Results

Mean (± standard deviation) MedDiet score for participants was 28.2 ± 4.7. Whites had higher energy‐ adjusted MedDiet scores (29.3 ± 4.8) when compared to those of blacks (27.5 ± 4.4, p <0.0001). Higher MedDiet scores were associated with slower rates of cognitive decline (β = +0.0014 per 1‐point increase, SE= 0.0004, p=0.0004) after adjustment for age, sex, race, education, cognitive activities, and energy.

Conclusion

Greater adherence to a Mediterranean diet may reduce the risk of cognitive decline with older age.

Support provided by RO1 AG021972
Involvements of Akt and phospholipase D‐mediated phosphatidic acid in the regulation of mTOR signaling during immobilization and remobilization of rat skeletal muscle
Authors: Lee Yeon‐Sook , You Jae‐Sung , Park Mi‐Na
Published: June 2021

Skeletal muscle activity is known to determine its mass by regulating protein synthesis through Akt‐mTOR‐p70s6k signaling. It has been revealed that mTOR and its downstream signaling is also activated by phosphatidic acid (PA) in mechanically stimulated skeletal muscle. This study was conducted to investigate, during altered skeletal muscle usage, how phospholipase D (PLD)‐mediated phosphatidic acid (PA) level is modulated, and that is involved in Akt signaling to p70s6k. Eleven‐weeks‐old male rats were subjected to unilateral hindlimb immobilization (Im) for 10 days and allowed to recover by remobilization (Rm) for 3 or 13 days. Non‐immobilized muscles were served as an internal control. Immobilized gastrocnemius was significantly atrophied (muscle mass loss), but during Rm its mass had been gradually recovered. Deactivated Akt signaling to p70s6k induced by Im was also recruited by Rm. Specifically changes in activation of p70s6k were more dramatic than that of Akt during Im and Rm periods. The level of PA was reduced during Im with impaired activity of PLD. However, Rm increased the activity of PLD and level of PA, and even greater than control muscle at the 3 day. These results suggest that mTOR signaling during inactive or active skeletal muscle use may be regulated by PLD‐mediated PA level as well as Akt activation. Supported by the Korea Research Foundation (KRF‐2008‐C00071).
Measuring Food Security Using a Self‐Administered Mail Survey Method among Older Georgians Requesting Older Americans Act Nutrition Program: Georgia Advanced Performance Outcomes Measures Project
Authors: Lee Jung Sun , Johnson Mary Ann , Brown Arvine , Nord Mark
Published: June 2021

Food security is one of the key outcomes in Older Americans Act Nutrition Program (OAANP), however it is unknown how best to measure food security to evaluate this program's impact on a large scale. We measured food security in all new OAANP participants and waitlisted people in Georgia between July and early November, 2008 (n=4,731) using the self‐administered mail survey method used in the ongoing Georgia Performance Outcomes Measures project. We used a modified 6‐item US Household Food Security Survey Module (HFSSM) with a 30 day reference period and two reminder postcards. About 33% of those identified completed the survey (n=1,594, mean age 74.6±9.5, 68.6% female, 30.6% black). Most of the respondents (91%) completed all 6 food security questions while 26 did not respond to any question. Infit and outfit statistics for each of the 6 questions and overall Rasch model fit were within an acceptable range. Overall psychometric properties observed in our food security data were comparable to the national food security statistics provided by USDA. Our findings suggest that food security can be reasonably measured by a mail survey and a short form of HFSSM in older adults requesting OAANP. Such methodology can be also used to estimate the extent of food insecurity and help guide program and policy decisions to meet the nutrition assistance needs of vulnerable older adults. This study was funded in part by US AoA and USDA ERS.
Dietary fat intake in Europe
Authors: Elmadfa Ibrahim , Nowak Verena , Kornsteiner‐Krenn Margit
Published: June 2021

The quantity and quality of fat have a major impact on coronary heart diseases (CHD), which contribute to morbidity and mortality. The objective of this study was to analyze fat intake data from the European Union. Data were collected in the context of the European Nutrition and Health Report (ENHR) 2009. Total fat intake (TFI) data were available from 22 countries, saturated fatty acids (SFA) from 18, and monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) from 15 countries.

There was a great difference in the TFI ranging from 28.4 (Portugal, men) to 47.2 (Greece, women) %E. TFI of only 5 countries were in agreement with the FAO/WHO 2008 intake goals. Hence, only Portugal had a TFI < 30%E and the Czech Republic, Denmark, Finland, and Sweden < 35%E; Germany, Ireland, Italy, and Poland were at the borderline (~ 35%E). The remaining 11 countries had high TFI > 35%E. For SFA only Portugal reached the recommendation of < 10%E. The intake of MUFA ranged from 10.9 to 22.9%E. The PUFA intake varied from 4.7 to 9.2 %E. Only 7 countries showed a higher intake (> 6%E) which is targeted to prevent CHD.

In summary, the majority of the countries showed an unfavorably high total fat (> 35%E) and SFA intake, whereas the PUFA intake should be increased. There is potential to improve the quantity and quality of fat intake.

The ENHR 2009 was funded by the EC under the Public Health Program 2006, agreement nr. 2006128.
Quality of reporting dietary supplement research in scientific publications and other observations
Authors: Saldanha Leila G , Sempos Christopher T , Costello Rebecca B
Published: June 2021

From 2001 to 2008 the Office of Dietary Supplements engaged annually in the process of identifying 25 exemplary original research papers on dietary supplements and disseminating these papers through the Annual Bibliography of Significant Advances in Dietary Supplement Research. Over the 8‐yr period, more than 2000 papers from over 50 peer‐reviewed journals were evaluated by over 50 scientific reviewers to identify the 200 papers that appeared in the bibliography. The papers covered a wide array of research. An instrument to evaluate the types of journals publishing dietary supplement research, study design, quality of reporting, study outcome, and funding sources was developed for this purpose. Journals for the 200 papers were grouped into three categories: clinical/medical, nutrition, and other. Of the 200 papers, 45.5% appeared in clinical/medical journals, 25% in nutrition, and 29.5% in other journals. 25% of the studies used diseased populations, 60% high‐risk and 20.5% healthy, 24.5% could not be categorized. 62% provided the chemical composition of the test materials. Only 14.5% verified the composition of the test materials. The quality of reporting studies is critical as it enables other researchers to duplicate the findings and it affects the ranking of studies in systematic reviews. The final evaluation will be reported at EB10. Funding: Office of Dietary Supplements/NIH.
Portal Vein Ligation Accelerates Tumor Growth in Ligated, but not Contralateral Lobes
Authors: Sakai Nozomu , Clarke Callisia N , Tevar Amit D , Edwards Michael J , Lentsch Alex B
Published: June 2021

BACKGROUND

Portal vein embolization (PVE) is a widely accepted procedure to induce hypertrophy of contralateral lobes prior to extensive liver resection. However, the manner in which PVE alters liver and/or tumor growth is unknown. We investigated the mechanisms of liver growth after portal vein ligation (PVL) as compared to partial hepatectomy (PH), and evaluated these effects on the growth of liver tumors.

METHODS

Mice were subjected to PVL, PH, or sham surgery. Liver histology, activation of transcription factors, and expression of cytokines and growth factors involved in liver regeneration were evaluated. In some experiments, murine colorectal carcinoma cells were injected via the portal vein.

RESULTS

Liver regeneration of non‐ligated lobes after PVL was similar to that occurring after PH and was accompanied by similar changes in transcription factors, cytokines and growth factors. In comparison, ligated lobes had greatly increased activation of transcription factors, cytokines and growth factors, showed no proliferation and atrophied. When liver tumors were measured, the ligated lobes showed significantly greater tumor growth than non‐ligated lobes or PH.

CONCLUSIONS

The data suggest that PVL results in acceleration of tumor growth in ligated but not contralateral lobes. Liver regeneration is similar in non‐ligated lobes and PH.
The role of mechanical forces in stem cell differentiation to vascular lineage
Authors: Nikmanesh Maria , Shi Zhong‐Dong , Tarbell John M
Published: June 2021

The overall goal of our study is to evaluate the role of wall shear stress (WSS) and cyclic strain (CS) on the development of vascular cells from mouse embryonic stem cells (MESCs) differentiated in vitro. The aim is to generate an early marker of endothelial cells (ECs), platelet endothelial cell adhesion molecule‐1 (PECAM‐1+ cells), from ESCs differentiated in vitro, subjecting these cells to WSS and CS, and analyzing the functionality of these cells by testing their permeability to solutes of varying molecular weight and expression of key components of the inter‐endothelial tight junctions. If we are successful, the results will make an important contribution to vascular tissue regeneration.

Results

1) We have been able to sort PECAM‐1+ cells from the rest of the MESC culture; 2) The endothelial‐like cells display border staining for VE‐cadherin; 3) ES‐derived endothelial cells form multidimensional tubular structures on Matrigel; and 4) ES‐derived endothelial cells uptake acetylated low density lipoprotein (ac‐LDL).

Conclusion

Our sorting for PECAM‐1+ cells has been successful and these cells express hallmark of the endothelial phenotypes. We can use these cells for evaluating the role of WSS and CS on their development into mature endothelial cells.

Supported by: NIH/NHLBI 1 RO1 HL086543‐01 and State of New York Department of Health (NYSTEM)
Maturation and growth of the bladder wall in a rodent model of organ regeneration
Authors: Burmeister David M , AbouShwareb Tamer , Soun Jennifer , Link Kerry , Tan Josh L , Olson John L , Andersson Karl‐Erik , Christ George J
Published: June 2021

We previously established a model of organ regeneration in which the rodent bladder regenerates to the original volume after subtotal cystectomy (STC; removal of ~70% of the bladder). In this study, we further examined the growth and maturation of the bladder wall over time from 1–8 weeks post‐STC. Female F344 rats underwent STC with age‐matched control rats run in parallel. MRI scans revealed a thinner bladder wall 1 week post‐STC (281.96 μM) compared to pre‐STC levels (493.53 μM) that improved greatly by 8 weeks (427.76 μM). In vivo urodynamics revealed bladder hyperactivity after afferent nerve activation via intravesical administration of 30 μM capsaicin. Vesicular acetylcholine transporter staining demonstrated the presence of motor nerves. Immunohistochemistry for proliferating cell nuclear antigen revealed proliferating cells 1, 2, and 4 weeks post‐STC. Uroplakin 3 staining showed mature urothelial cells 1 week post‐STC. Positive immunostaining was also observed for smooth muscle alpha actin, myosin heavy chain and desmin; with the greatest staining closest to the plane of excision. These studies establish that regeneration of the urinary bladder involves regeneration of both sensory and motor nerves with progressive maturation of the bladder wall. The goal of these studies is to uncover the mechanistic basis of bladder regeneration in adult mammals and use this knowledge for clinical applications.
Hyaluronic acid/collagen (HA/CN) assay for epithelial mesenchymal transformation (EMT) in cardiac valvulogenesis
Authors: Bick Alexander , DeLaughter Daniel , Khademhosseini Ali , Barnett Joey V.
Published: June 2021

An in vitro assay system was devised for the study EMT in developing cardiac tissue. Explants of isolated chick atrioventricular (AV) canal cushions were placed on hydrated HA/CN gel lattices. Similar to the CN gel assay, cells grew out of the explant from the endocardium and across the gel surface morphologically transitioning from epithelial cells to mesenchymal cells. This process mimics in vivo embryonic heart valvulogenesis. We first optimized the HA/CN gel composition, finding that a 50 mg/mL HA, 1.5 mg/mL CN produced the highest explants viability and were 1000x more mechanically robust than 3D collagen gels with similar endothelial viability. Explants on CN were viable up to 4 days, while HA/CN explants were viable for up to six weeks. We compared the HA/CN to CN gels assay in terms of specificity and gain of function experiments. Though cells began to morphologically undergo EMT on HA/CN gels, they rarely invaded. While further optimization is required before HA/CN can fully recapitulate AV cushion EMT, the enhanced stability and longevity of HA/CN provides a powerful tool to potentially derive novel cardiac cushion endothelial cell lines.
Association of ABCB1 and NR1I2 polymorphisms with response to carbamazepine in Malaysian epilepsy patients
Authors: Haerian Batoul Sadat , Mohamed EHM , Lim KS , Tan HJ , Raymond AA , Tan CT , Wong CP , Wong CW , Zain SM , Roffeei SNM , Mohamed Z
Published: June 2021

The human nuclear pregnane X receptor (PXR;NR1I2) which regulates the expression of ABCB1 gene which encodes phosphoglycoprotein, an efflux pump for carbamazepine (CBZ). The aim of this study was to investigate whether ABCB1 C3435T and NR1I2 A7635G variants significantly impact the response of epilepsy patients to CBZ monotherapy. ABCB1 C3435T and NR1I2 A7635G genotyping were performed on 168 Malaysian epilepsy subjects (76 drug‐responsive and 92 drug‐resistant) receiving CBZ monotherapy by PCR‐RFLP. The results for C3435T revealed a genotype distribution of CC, CT, TT: 39.5%, 43.4%, 17.1% for drug‐resistant patients compared to CC, CT, TT: 29.3%, 58.7%, 12.0% for drug‐responsive patients. The genotypes distribution of A7635G in the drug‐resistant patients for AA, AG, and GG were 13.0%, 39.1%, 47.8% and in drug‐responsive patients 10.5%, 57.9%, 31.6%, respectively. Furthermore, among drug‐resistants and drug‐responsive patients, the frequency of carriers of T allele and was 41.3.1% and 38.8% and for G allele 67.4% and 60.5%, respectively. The association of genotype and allele frequencies in C3435T and A7635G with response to CBZ was nonsignificant. In conclusion, the allelic and genotype analysis in Malaysian epilepsy patients suggests that the NR1I2 A7635G and ABCB1 C3435T polymorphisms are not associated with resistance to CBZ.
Regulation of estrogen sulfotransferase (SULT1E1) expression by proliferation state in MCF10A human breast epithelial cells: Potential role of microRNAs
Authors: Fu Jiaqi , Kocarek Thomas A , Dombkowsi Alan , Cukovic Daniela , Runge‐Morris Melissa
Published: June 2021

The MCF10A cell line represents a model of the normal human breast epithelial cell. Estrogen sulfotransferase (SULT1E1), a major estrogen‐inactivating enzyme, is up‐regulated in confluent relative to preconfluent MCF10A cells. The mechanism responsible for SULT1E1 up‐regulation in confluent MCF10A cells was investigated. Transient or stable tranfection of reporter constructs containing up to 7 kb of the SULT1E1 5′‐flanking region failed to recapitulate confluency‐induced SULT1E1 up‐regulation. The half‐life of SULT1E1 mRNA in preconfluent MCF10A cells was estimated to be ~4h based on analysis of SULT1E1 mRNA levels in MCF10A cells that were replated after reaching confluency. Microarray analysis revealed that 85 microRNAs were significantly up‐regulated and 24 microRNAs were down‐regulated in confluent relative to preconfluent MCF10A cells. Computational analysis of the SULT1E1 3′‐untranslated region identified 4 putative candidate binding sites for confluency‐regulated microRNAs. The confluency‐related decrease in two of these microRNAs (miR‐221 and miR‐100*) corresponded with the confluency‐induced increase in SULT1E1 mRNA observed in MCF10A cells. These results support a role for the post‐transcriptional up‐regulation of SULT1E1 mRNA expression in confluent relative to preconfluent MCF10A cells. Supported by NIH grant (ARRA) ES016373 (MRM) and EHS Center grant ES06636.
Role of Toll‐like receptor signaling in hepatocytes in regulating the gene expression of drug metabolizing enzymes
Authors: Ghose Romi , Guo Tao
Published: June 2021

In inflammation, drug metabolism and clearance is altered due to suppression of drug metabolizing enzymes (DMEs) and their regulatory nuclear receptors (NRs: PXR, CAR and RXRα). Inflammatory responses are mediated by cytokines produced by activation of Toll‐like receptors (TLRs) on liver macrophages or Kupffer cells (KCs). Cytokines act on hepatocytes to suppress gene expression. However, hepatocytes also express TLRs, and we hypothesize that TLRs on hepatocytes are directly involved in suppression of hepatic DMEs. Primary mouse hepatocytes from TLR4+/+ (C3HeB/FeJ) and TLR4‐mutant (C3H/HeJ) mice were treated with LPS (1 μg/ml) or saline. RNA and protein levels were analyzed by real‐time PCR and immunoblotting, respectively. The purity of the hepatocytes was assessed by CD68 staining for KC contamination. RNA levels of the major DMEs, Cyp3a11 and Ugt1a1 were reduced ~60% by LPS in TLR4+/+, but not TLR4‐mutant hepatocytes. The cell‐signaling components, JNK and NF‐κB, known regulators of NRs and DMEs were activated in TLR4+/+ and not TLR4‐mutant cells. Nuclear protein levels of RXRα were rapidly reduced after LPS treatment of TLR4+/+, but not TLR4‐mutant hepatocytes. Treatment of hepatocytes with the KC inhibitor, Gadolinium chloride did not attenuate the effect of LPS. Thus, both cytokine‐dependent and independent pathways are involved in the regulation of hepatic gene expression in response to LPS signals.
Nanoparticles attenuate P‐glycoprotein function in A549 cells
Authors: Salomon Johanna J , Ehrhardt Carsten
Published: June 2021

P‐glycoprotein (P‐gp) is a well characterised membrane transporter relevant to drug disposition and multi‐drug resistance in cancer. Here we studied the effect of nanoparticles (NP) on uptake and release of the P‐gp substrate, rhodamine 123 (Rh123), in human alveolar epithelial cells (A549).

For uptake experiments cell culture medium was removed and replaced with Rh123 (10 μM) in Krebs‐Ringer Buffer (KRB) also containing the different NP formulations (see below). In release studies cells were loaded with Rh123 for 30 min and subsequent release of the substrate from the cells into fresh KRB with NP was measured. NP studied were either carboxylated (CNP, 100 nm, −50 mV), amino‐modified (ANP, 50 nm, 60 mV) or latex‐based (LNP, 100 nm, −50 mV). In addition, selected pharmacological inhibitors of P‐gp function were investigated.

Rh123 uptake was increased in the presence of CNP and LNP in a time‐dependent manner. No effect of ANP on Rh123 uptake was observed. The release of Rh123, however, was significantly (p<0.05) attenuated in the presence of CNP and ANP, but not LNP.

Our findings suggest that nanomaterials interact with membrane transporter function in a complex fashion and hence, might influence disposition of xenobiotics as well as endogenous substrates.

This work was supported by a Strategic Research Cluster grant (07/SRC/B1154) from Science Foundation Ireland.
Acetaminophen‐Induced Alterations in Hepatic Mitochondrial Manganese Superoxide Dismutase (MnSOD; SOD2) Activity in Mice
Authors: Agarwal Rakhee , MacMillan‐Crow Lee Ann , Rafferty Tonya , Saba Hamida , Hinson Jack A
Published: June 2021

In overdose the analgesic/antipyretic acetaminophen (APAP) is hepatotoxic. Toxicity is mediated by hepatic metabolism to a reactive metabolite (NAPQI). Following low doses NAPQI is detoxified by glutathione (GSH); however, in overdose GSH is depleted, NAPQI covalently binds to proteins, and oxygen/nitrogen stress occurs. Toxicity is believed to occur by mitochondrial dysfunction. MnSOD inactivation occurs during many oxidant stress mediated diseases; thus, we examined MnSOD in APAP toxicity. MnSOD is a critical mitochondrial antioxidant enzyme in preventing oxygen/nitrogen stress within the mitochondria. Mice were treated with 300 mg/kg APAP. GSH was significantly reduced by 65% at 0.5 h and remained reduced from 1–4 h returning to control levels by 6 h. Serum ALT (hepatotoxicity) did not significantly increase until 4 h and was 2290 IU/L at 6 h. MnSOD activity was significantly reduced by 25% at 0.5 h and was 50% at 1 h and 2 h. In a dose‐response study performed at 1 h, GSH was significantly depleted by 62% and 80% with nonhepatotoxic doses of 50 and 100 mg/kg APAP, respectively. No further GSH depletion occurred with 200 and 300 mg/kg APAP. MnSOD activity was reduced by 28, 32 and 57% with 50, 100 and 300 mg/kg APAP respectively. Thus, MnSOD activity is dramatically decreased following toxic and nontoxic doses of APAP, and MnSOD inactivation precedes induction of hepatic toxicity. Supported by NIH grant 1 R01 DK079008.
Early life exposure of rats to citalopram, a selective reuptake inhibitor (SSRI) antidepressant, results in structural abnormalities in the adrenal gland
Authors: Tucci Michelle A. , Benghuzzi Ham , Swilley Sharonda , McDowell Roderick , Lin Rick , Simpson Kim , Paul Ian
Published: June 2021

Thirty percent of infants exposed in utero to SSRIs display behavioral and physiological abnormalities in the first few weeks after birth including increased incidence of pulmonary hypertension. Rats exposed from postnatal day (PN) 1–21 to SSRIs display persistent neurobehavioral abnormalities which include severe deficits in social interactions and expression of serotonergic neurons. The adrenal glands function as a reserve in response to stress. Chronically elevated cortisol may induce clinical depression by interfering with central serotonergic neurotransmission. SSRIs were developed to restore serotonin homeostasis. However, exposure to SSRIs can increase glucocorticoids in females and decreases in LH, FSH, and testosterone (T) in males. Therefore, adrenal gland morphology in male and female rats exposed to an SSRI as neonates was evaluated. Long Evans rats (30 male, 30 female) were treated from PN 1–21 with 0.1 mL s.c. of either saline or citalopram (5, 10 or 20mg/Kg). Animals were sacrificed at PN‐180. No changes in glomerulosa zone areas were seen. However, decreases in fasiculata areas were evident in female rats, and the reticularis area was increased in the male rats. In conclusion, data collected from this study indicate than early life exposure to citalopram results in morphological abnormalities in adrenal structure which may result in disturbances in corticosteroid levels in female rats and alterations in LH, FSH, and T in male rats. Supported by RR 017710 to IAP, KLS, and MH 084194 to RCSL.
The effects of Ro 64‐6198 and diazepam on antinociception and remifentanil self‐administration in rhesus monkeys
Authors: Podlesnik Christopher A. , Ko Mei‐Chuan , Winger Gail , Woods James H.
Published: June 2021

The aim of this study was to compare the behavioral effects of a synthetic nonpeptidic NOP (nociceptin/orphanin FQ Peptide) receptor agonist, Ro 64‐6198, with a benzodiazepine, diazepam, in rhesus monkeys. Ro 64‐6198 (0.001–0.01 mg/kg, i.v.) produced antinociception against an acute noxious stimulus (50 degrees C water), but diazepam (0.32–3.2 mg/kg, i.v.) did not produce such effects at any dose tested. Higher doses of Ro 64‐6198 (0.32 mg/kg, i.v.) and diazepam (1.0–5.6 mg/kg, i.v.) decreased lever pressing maintained by intravenous self‐administration of the mu‐opioid agonist, remifentanil. Although neither drug increased nonreinforced responding with complete consistency, such effects were more frequent with diazepam. These effects of Ro 64‐6198 on lever pressing were blocked by an NOP‐receptor antagonist, J‐113397, but not by the benzodiazepine antagonist, flumazenil. Conversely, the effects of diazepam on lever pressing were blocked by flumazenil but not J‐113397. These findings suggest the effects of Ro 64‐6198 on operant lever pressing are mediated by NOP receptors and that higher doses are required compared to those producing antinociception. Moreover, Ro 64‐6198 was less likely to disinhibit nonreinforced operant responding than diazepam, suggesting diazepam might produce greater anxiolytic‐like effects than Ro 64‐6198 in rhesus monkeys.
Morphine regulates dopaminergic system via miR‐133b and Pitx3 in zebrafish embryos
Authors: Sanchez‐Simon Fatima Macho , Zeng Yan , Loh Horace H. , Lae Ping‐Yee , Rodriguez Raquel E.
Published: June 2021

Current studies were initiated to address whether morphine could affect neural development via its regulation of miRs. By microarray analysis, we show that morphine decreases miR‐133b expression. miR‐133b targets Pitx3, a transcription factor that activates genes involved in dopaminergic differentiation, such as TH and DRD1. Exposure of 24 hpf zebrafish embryos to morphine resulted in a decrease in miR‐133b and an increase in Pitx3, TH and DRD1 mRNA levels. Injection of the 3′UTR region of Pitx3 did not alter miRNA‐133b expression, and increased the expression of Pitx3, TH and DRD1. When ZfMOR (zebrafish mu opioid receptor) was silenced in embryos via morpholino injection, miR‐133b level increased and the expression of Pitx3 decreased. Parallel morphine regulation of miR‐133b level can be observed in primary neuron cultures from rat hippocampus. Embryo treatment with MAPK inhibitors U0126 or PD98059 increased miR‐133b level and decreased Pitx3 level. These studies suggest that the dopaminergic system development could be controlled by morphine‐mediated activation of ZfMOR. As the dopaminergic system is essential in the development of addiction, our findings contribute to the better understanding of the pathway that leads to addictive disorders.

Research funded by the Spanish MEC (SAF2007‐61581) and the NIH, USA (DA007339).
UTP induced inotropic effects in the human atrium
Authors: Neumann Joachim , Simm Andreas , Silber Rolf Edgar , Gergs Ulrich
Published: June 2021

In humans, UTP levels are increased during myocardial infarction. UTP can act via G protein‐coupled P2Y receptors, which are further divided into P2Y2, 4, 6, 11, 12, 13, 14 receptors. The inotropic effects of UTP in the human heart have hitherto not been studied. Therefore, we looked for contractile effects in isolated electrically driven (1 Hz) atrial preparations from patients undergoing cardiac bypass surgery. In these preparations UTP (cumulatively applied as 10, 30, 100 μM) concentration‐dependently exerted a positive inotropic effect. At 100 μM UTP (the highest concentration investigated) force of contraction was increased by 21 ± 6% of pre‐drug value (n = 10, p < 0.05). The effect of UTP (100 μM), non‐cumulatively applied, was more pronounced (by 53%, n = 7, p < 0.05) and was stable up to 20 min. The effect could be washed out and could be elicited again, arguing against tachyphylaxis. The inotropic effect occurred without changes in time parameters of contraction. The positive inotropic effect of UTP was not attenuated by pre‐incubation with inhibitors of adenylyl cyclase activity (SQ22536, 10 μM) or phospholipase C (U73122, 10 μM). In summary, we describe inotropic effects of UTP on force of contraction in the human atrium, which occur independently of cAMP and IP3. The exact receptor subtypes need to be elucidated.
Molecular modeling study about consequences of human β1 adrenoceptor Ser49Gly and Gly389Arg polymorphisms
Authors: Correa‐Basurto Jose , Ciprés Fabiola , Trujillo‐Ferrara Jose G , Soriano‐Úrsua Marvin Antonio
Published: June 2021

Two human β1 adrenoceptor (Hβ1AR) polymorphisms have been widely studied (Ser49Gly and Gly389Arg). In this study, Hβ1AR 3‐D models with/without these polymorphisms were built and refined. For studying ligand binding process, docking simulations was carried out using a set of known ligands. The binding sites and affinity values on Hβ1AR structures were analyzed. Additionally, glycosylation and phosphorylation were in silico‐evaluated. Results showed differences in ligand Hβ1AR affinity and recognition mode. Besides, conformational disposition of extracellular and intracellular loops was different among structures. The former could be related to differences in glycosylation and phosphorylation processes.
Activation of Calcitonin Receptor‐like Receptor by Membrane‐anchored Ligands
Authors: Hsu Sheauyu , Chang Chia Lin , Park Jae‐Il
Published: June 2021

G protein‐coupled receptors (GPCRs) are the most important pharmaceutical targets, and more than 40% of drugs in use today modulate GPCR signaling. A major hurdle in the development of therapies targeting GPCRs is the drug candidate's nonselective actions in multiple tissues. The ability to control GPCR signaling spatially would provide a venue for developing therapies that require targeted GPCR signaling. Here, we show that the fusion of a GPCR co‐receptor RAMP1 with the calcitonin gene related peptide (CGRP), or calcitonin, transforms the RAMP1 from a co‐receptor to bonafide membrane‐anchored ligands (CGRP‐RAMP1 and CAL‐RAMP1). The CAL‐RAMP1 selectively activates the calcitonin receptor (CR), whereas, the CGRP‐RAMP1 activates both the calcitonin receptor‐like receptor (CLR) and CR. Unlike a free peptide, the anchored CGRP‐RAMP1 and CAL‐RAMP1 ligands confine their activities to individual cells. In addition, our study suggests that the interaction of a free CGRP peptide with RAMP1 could lead to the generation of a unique interface that would directly activate CLR in a stochastic manner, and the activating interface may resemble the anchored ligands characterized here. Taken together, our finding thus provides a novel class of GPCR ligands that function exclusively in an autocrine manner, and the proof‐of‐concept demonstration for future development of targeted therapies aimed at select class B GPCRs.
UDP‐glucose acting at P2Y14 receptors is a mediator of mast cell degranulation
Authors: Gao Zhan‐Guo , Ding Yi , Jacobson Kenneth A.
Published: June 2021

UDP‐glucose (UDPG), a glycosyl donor in the biosynthesis of carbohydrates, is an endogenous agonist of the G protein‐coupled P2Y14 receptor. RBL‐2H3 mast cells endogenously express a P2Y14 receptor at which UDPG mediates degranulation as indicated by β‐hexosaminidase (HEX) release. Both UDPG and a more potent, selective 2‐thio‐modified UDPG analog, MRS2690 (2‐thio‐UDPG), caused a substantial calcium transient in RBL‐2H3 cells, which was blocked by pertussis toxin, indicating the presence of the Gi‐coupled P2Y14 receptor, supported also by quantitative detection of abundant mRNA. Expression of the closely related P2Y6 receptor was over 100 times lower than the P2Y14 receptor, and the P2Y6 agonist 3‐phenacyl‐UDP was inactive in RBL‐2H3 cells. P2Y14 receptor agonists also induced [35S]GTPγS binding to RBL‐2H3 cell membranes, and phosphorylation of ERK1/2, P38 and JNK. UDPG and MRS2690 concentration‐dependently enhanced HEX release with EC50 values of 1150 ± 320 and 103 ± 18 nM, respectively. The enhancement was completely blocked by pertussis toxin and significantly diminished by P2Y14 receptor‐specific siRNA. Thus, mast cells express an endogenous P2Y14 receptor, which mediates Gi‐dependent degranulation and is therefore a potential novel therapeutic target for allergic conditions.
Sodium channel activator‐induced neuronal development is NMDA receptor dependent and involves a Ca2+/calmodulin kinase (CaMK)‐CREB‐BDNF signaling pathway
Authors: George Joju , Murray Thomas F
Published: June 2021

Many aspects of activity‐dependent neuronal development such as dendritic arborization, spinogenesis and synaptogenesis are mediated through NMDA receptor [NMDAR]‐dependent Ca2+ signaling mechanisms. Brevetoxin‐2 (PbTx‐2), an activator of voltage‐gated sodium channels, mimics activity‐dependent control of neuronal development by elevating intracellular sodium levels and augmenting NMDAR function. PbTx‐2 enhanced cerebrocortical neuron spinogenesis and synapse density. Morphological analysis showed that PbTx‐2 enhanced spinogenesis by stimulating dendritic filopodia formation. Pharmacological evaluation showed that the PbTx‐2 induced neuronal plasticity was mediated by signaling mechanisms downstream of CaMKK. PbTx‐2 exposure produced phosphorylation and activation of CaMKI (Thr177) and CaMKII (Thr286). PbTx‐2 also induced ErK 1/2, but not Akt, phosphorylation. CREB is known to be an activity‐regulated transcription factor, and PbTx‐2 exposure increased CREB phosphorylation at Ser‐133. Assessment of brain derived neurotrophic factor (BDNF) transcription with RT‐PCR demonstrated that BDNF mRNA expression level increased in a time and PbTx‐2 concentration dependent manner. These findings suggest that the sodium channel activator influence on neuronal developmental involves NMDAR‐mediated CaMK signaling with downstream activation of CREB‐dependent transcription of BDNF.
Lipopolysaccharide (LPS) hyperpolarizes the guinea‐pig tracheal epithelium (Epi) by increasing the activities of the epithelial sodium channel (ENaC) and the Na,K‐pump
Authors: Dodrill Michael W. , Fedan Jeffrey S.
Published: June 2021

The administration of LPS (4 mg/kg, i.p.), which is involved in several airway inflammatory diseases, causes hyperpolarization of tracheal Epi at 18 h. This study set out to understand the mechanism(s) by which LPS causes hyperpolarization. In the isolated, perfused trachea preparation (IPT), LPS had no effect on epithelial ENaC or Na,K‐ATPase transcription or ENaC abundance, but doubled Na,K‐ATPase protein levels. In the presence of basolateral amphotericin B (AB, 7.5 μM) to short‐circuit the Na,K‐pump, apical amiloride (10 μM) inhibited transepithelial potential (Vt) to a greater extent in the LPS group compared to control. In the presence of apical AB to short‐circuit ENaC, the resulting Vt, due to the Na,K‐pump, was greater in the LPS group relative to control. In the Ussing chamber, readmission of K to the Epi Na‐loaded by K omission led to a greater hyperpolarization after LPS compared to control. Apical trypsin (100 U/ml) added to activate ENaC, increased Vt to a similar degree in control and LPS‐treated groups. Thus, LPS hyperpolarizes the Epi by increasing the activities of both ENaC and the Na,K‐pump. This may reflect alteration in their regulation. Funded by NIOSH
Src kinase‐dependent gating properties of single Ca2+ channels are altered by tyrosine nitration in colitis
Authors: Ross Gracious R , Kang Minho , Akbarali Hamid I
Published: June 2021

Reduced L‐type Ca2+ ‐channel (LTCC) function contributes to colonic dysmotility during inflammation. We have previously shown that nitration of tyrosine residues within the carboxy terminus of LTCC (Y1837 and Y2134) impairs c‐Src kinase regulation during colitis. We examined the gating properties of single LTCC, using cell‐attached patch clamp mode, in mouse colon smooth muscle cells (control vs colitis‐TNBS) and CHO cells transfected with hCaV1.2b (wild type) or mutated hCaV1.2b (Y1837F:Y2134F). Inflammation significantly reduced LTCC ensemble average current (−0.33 vs −0.22 pA, P=0.05), without affecting the single‐channel properties: mean amplitude (−2.4pA), slope conductance (23 vs 27pS) or the open‐time constants (τ1: 1.6 vs 1.2; τ2: 6.1 vs 3.9 ms). However, the availability (slow gating) of LTCC was significantly reduced by inflammation (0.19 vs 0.06; P=0.02) without affecting the open probability of modal gating. Similar effects were seen with inhibition of c‐Src kinase by PP2 or by tyrosine nitration induced by SIN‐1+ONOO− combination in smooth muscle cells. Mutation of tyrosine residues (Y1837/2134F) in hCaV1.2b, which prevents c‐Src kinase regulation, resulted in diminished channel availability. This study demonstrates that src kinase‐ phosphorylation enhances the availability of LTCC, that is impaired by tyrosine nitration during colonic inflammation. Supported by NIH DK046367.
ARF1 GTPase modulates α2B‐adrenergic receptor‐mediated ERK1/2 activation
Authors: Dong Chunmin , Wu Guangyu
Published: June 2021

ADP‐ribosylation factors (ARFs) belong to the superfamily of Ras‐related small GTPases and modulate multiple intracellular trafficking processes. Here we report that ARF1 is required for ERK1/2 activation by α2B‐adrenergic receptor (α2B‐AR). We demonstrated that α2B‐AR associated with ARF1 in an agonist‐dependent fashion as measured by co‐immunoprecipitation. Interestingly, α2B‐AR activation upon agonist stimulation also markedly activated the small GTPase ARF1. GST‐fusion protein pull down assays combined with progressive deletion and site‐directed mutagenesis studies further showed that double Trp residues (diW) located in the third intracellular loop mediated α2B‐AR interaction with ARF1. Mutation of the diW motif dramatically disrupted the agonist‐dependent association of α2B‐AR with ARF1 and attenuated α2B‐AR‐mediated ERK1/2 activation, without altering the cell surface transport and internalization of α2B‐AR. These data demonstrate that the diW motif‐directed agonist‐dependent α2B‐AR interaction with the small GTPase ARF1 modulates ERK1/2 activation by the receptor (R01GM076167).
Characterization of the α1D‐Adrenergic Receptor Signalosome
Authors: Lyssand John Sverre , DeFino Mia C , Tang Xiao‐bo , Wacker Jennifer L , Gardner Richard G , Adams Marvin E , Hague Chris
Published: June 2021

Previously, we reported the identification of syntrophins as novel accessory proteins for α1D‐adrenergic receptors (ARs). Syntrophins are members of the dystrophin complex, and act as scaffolds for anchoring membrane proteins to cytoskeletal proteins. To characterize the functional importance of this interaction, we examined how syntrophins contribute to α1D‐AR function in vitro and in vivo. In vitro, syntrophins increase α1D‐AR binding site density and functional coupling in HEK293 cells. In vivo, α‐ and β2‐syntrophin double knock‐out mice have significantly diminished α1D‐AR functional responses in comparison to wild type mice. Taken together, these findings suggest that syntrophins play an essential role for α1D‐AR function in vivo, presumably by anchoring α1D‐ARs in the plasma membrane as a functional signalosome, and by recruiting necessary components into the signalosome. Next, we determined what proteins syntrophins recruit to the α1D‐AR signalosome, by performing tandem affinity purification (TAP)/mass spectrometry on α1D‐AR, α/β1/β2‐syntrophin, and α‐dystrobrevin‐1/2. Our data revealed both previously identified and novel proteins recruited to the α1D‐AR/syntrophin signalosome. Current experiments are examining how these recruited proteins contribute to α1D‐AR function in vivo. Research was supported in part by PHS NRSA T32 GM07270 from NIGMS.
The Study of the Anti‐ulcer and Radical Scavenging Effects of Extracts from the Stem of Coscinium Fenestratum
Authors: Okechukwu Patrick Nwabueze , Ching Connie Boon Ai , Young Kevin Lai Kiat , Chyn Wong Hui , Akowuah Gabriel Akyirem
Published: June 2021

In this study we evaluated dichloromethane (DCM) extract obtained from C. fenestratum, for its ability to protect the gastric mucosa against injuries caused by necrotizing agents (0.3M HCl/60% EtOH), hypothermic restraint stress, and non‐steroidal anti‐inflammatory drugs (NSAID, indomethacin). Total phenoic content and free radical scavenging effects of the bioactive compounds were also determined. In the HCl/EtOH‐induced gastric‐ ulcer model, DCM extract demonstrated significant inhibition of ulcerative lesion index by 98% in relation to the positive control value 72.93 % (p< 0.05), 81% as compared to the positive control group 78.82% (p<0.05) in the NSAID/indomethacin –induced lesion model and 81.97% compared to positive control 73.77% (p< 0.05) in hypothermic‐restraint stress induced gastric acid ulcer model. The radical scavenging activity of the extract was 21.71% which was closely related to the total phenolic content of 256.67mg GAE/g dry weight. HPLC‐UV‐MS profiling of the bioactive compound in the DCM extract confirmed the presence of polyphenolic compounds. From this result we conclude that this extract possess a significant protective effect against gastric mucosa injuries caused by 0.3M HCl/60% EtOH, hypothermic restraint stress, and NSAID/indomethacin in the experimental rats. These effects may be due to the presence of polyphenolic compounds that provided radical scavenging activity.
A new role for Dysferlin in endothelial cell adhesion and neo‐vascularization
Authors: Yu Carol , Sharma Arpeeta , Trane Andy , Leung Cleo , Bernatchez Pascal
Published: June 2021

Dysferlin is a member of the Ferlin family of proteins known to regulate plasma membrane integrity and are linked to muscular dystrophy. However, the peripheral roles of Dysferlin in non‐muscle cells remain virtually unknown. By performing proteomic analysis of caveolae/lipid rafts, we reported that endothelial cells (EC) express Myoferlin and that it mediates plasma membrane translocation of vascular endothelial growth factor receptor‐2 (VEGFR‐2). Herein, we show by Northern and Western blot as well as by immunohistochemistry that EC and intact blood vessels of rodent and human origin all express Dysferlin but that its presence is not required for VEGFR‐2 signaling. Instead, loss of Dysferlin in EC results in deficient adhesion, detachment and growth arrest in sub‐confluent cells whereas confluent EC monolayers are remarkably resistant to cell detachment. In Dysferlin‐null mice, delivery of VEGF, an EC‐specific angiogenic agonist, results in blunted neo‐angiogenesis compared to WT mice. Mechanistically, loss of Dysferlin causes poly‐ubiquitination and proteasomal degradation of platelet endothelial cellular adhesion molecule‐1 (PECAM‐1/CD31). Our discovery that Dysferlin is a regulator of EC adhesion and neo‐vascularization broadens the functional scope of this family of proteins and suggest that specialization exist between the different Ferlins and their respective membrane‐based cargo.
Vasoactive effects of nanomolar curcumin in arterioles is mediated by adrenergic receptors.
Authors: Dewar Anthony M , Clark Richard A , Singer Adam J , Frame Mary D
Published: June 2021

Curcumin is a highly potent antioxidant associated with ameliorating oxidative stress and inflammation. Here we report for the first time that sub‐nanomolar curcumin has potent vasomotor activity. Arteriolar response to micropipette applied curcumin was tested in the mucosal region of the cheek pouch tissue of anesthetized (70 mg/kg pentobarbital) hamsters (N=30) using intravital microscopy. Diameter was recorded for the feed of a terminal arteriolar network (baseline 8±2um) and the arcade arteriole (22±11um) that supplied it. Curcumin (10‐12 ‐ 10‐4M, 60 s) induced a biphasic response over time and with increasing doses, inconsistent with vasomotion. An initial dilation, predominant at low doses, was followed by constriction, predominant at high doses. The fitted logEC50 and peak responses were similar for the arcade and feed. Dilation, logEC50 − 9.5±0.3, peak dilation +40±8%. Constriction, logEC50 − 8.5±0.4, peak constriction − 15±5%. Simultaneous atropine (muscarinic antagonist) or PD142893 (endothelin antagonist) had no effect. Propranolol (beta‐adrenergic antagonist) removed dilation, enhancing constriction to curcumin. Phentolamine (alpha‐adrenergic antagonist) removed constriction, enhancing dilation to curcumin. Thus curcumin modulates arteriolar diameter via the adrenergic receptors in a dose sensitive manner. (AHA 0655908T, Armed Forces Institute of Regenerative Medicine).