bluuebunny/crossref_metadata_2025 huggingface dataset:
Theory of Gastric Ventilation: Hypercapnia Stimulates Gastric Acid Secretion in the Rat
Authors: Held Heather E. , Landon Carol S. , Dean Jay B.
Published: June 2021

We recently described gastric ventilation, in which hypercapnia stimulates gastric acid and bicarbonate secretion in the gastric lumen, where it produces CO2 that may be vented through esophagus. This study attempts to correlate respiratory and gastric responses to CO2. Rats were implanted with gastric catheters. A pH probe sat in the stomach while animals were in a head‐body plethysmograph. Animals breathed air and air +5% CO2 while awake (n=4) and anesthetized (n=1). While awake, hypercapnia decreased gastric pH (mean ± SE 3.52 ± 0.35 in air; 3.42 ±0.39 in CO2) and tidal volume (VT, 1.12 ± 0.1 ml in air; 0.83 ± 0.1 ml in CO2) but increased frequency of breathing (f, 98.0 ± 6.2 breaths/min in air; 140.4 ± 7.5 breaths/min in CO2) and minute ventilation (VE, 71.7 ± 8.1 ml/min in air; 124.1 ± 15.1 ml/min in CO2). Isoflurane blunted the hypercapnic ventilatory response (VT 1.35 ml in air, 0.66 ml in CO2; f 41.2 breaths/min air, 36.6 breaths/min CO2; VE 42.9 ml/min in air, 24.0 ml/min in CO2) and the gastric response (pH 1.27 in air, 1.29 in CO2). This shows that systemic pH affects gastric acid secretion, which may explain the observed comorbidity of hypercapnia and gastrointestinal disease. (ONR)
Non‐invasive measure of corticosterone in food restricted rats
Authors: Amara Catherine E , Cole Deborah C , Bellem Astrid , Civitarese Robert A , Boonstra Rudy
Published: June 2021

Fecal corticosterone (CORT) metabolites (FCM) have been measured in laboratory animals and wildlife populations because they avoid handling‐induced stress response. Here we test whether FCM provide a measure of acute and chronic stress induced by an adrenocorticotropic hormone (ACTH) challenge and food restriction (FR), respectively in 4 month old female Wistar rats. Blood and fecal samples were collected before and after 7 days of FR to 88% baseline body mass (FR n = 10) or ad libitum feeding (CON n = 8) and after ACTH challenge. Enzyme immunoassays showed greater FCM after FR compared with baseline (419.1 ± 250.5 vs. 294.6 ± 145.6 ng.g−1). Serum CORT increased from baseline in response to ACTH challenge in both FR (79.7 ± 45.5 ng.ml−1vs. 610.6 ± 253.4 ng.ml−1) and CON (62.2 ± 46.2 ng.ml−1vs. 227.2 ± 78.1 ng.ml−1), with a greater response in FR. There was no difference in FCM measured at 2 h intervals over the 24 h period post ACTH injection between groups and the increase in 24h‐average FCM post ACTH only reached significance for CON (384.1 ± 29.1 vs. 521.5 ± 26.3 ng.g−1) and not FR (419.1 ± 250.5 vs. 523.8 ± 405.2 ng.g−1). Thus FCM provide a measure of the response to a chronic stress, however this measure is less sensitive than serum CORT for describing the acute response. These data support the use of FCM for measurement of long‐term adaptive response to physiologic stressors in rats and demonstrate increased adrenal sensitivity in FR. Funded by Kinesiology & Physical Education
N‐acetylcysteine improves skeletal muscle fatigue resistance in a model of peripheral arterial insufficiency
Authors: Roseguini Bruno , Souccar Caden , Han Sang
Published: June 2021

Oxidative stress plays a major role in the genesis of skeletal muscle dysfunction in patients with Peripheral Arterial Disease (PAD). We tested the hypothesis that administration of the antioxidant N‐acetylcysteine (NAC) would improve isolated skeletal muscle fatigue resistance in a mice model of PAD. Male Balb/c mice (n=15) were subjected to bilateral ligation of the femoral artery and after 2 weeks of recovery, received daily intraperitoneal injections of either NAC (150 mg/kg, n=7) or saline (n=8) for 15 days. At the end of the treatment, the extensor digitorium longus (EDL) and soleus (SOL) muscles were excised for analysis of the contractile function and fatigue resistance in vitro (32°C). Maximal specific force was not different between groups for both the EDL (NAC: 37.3±1.4 N/cm2; Saline: 36.1±1.5 N/cm2) and the soleus (NAC: 35.5±1.2 N/cm2; Saline: 34.1±1.2 N/cm2). In the soleus muscle, force after 10 min of submaximal tetanic stimulation (60 Hz, 300 ms trains, 0.3 trains/s) was higher (p<0.05) in NAC‐treated animals (45±3% of the initial value) when compared to controls (30.3 ±3%). No differences were found in fatigue development between groups in the EDL muscle (NAC:35.7±1.9%; Saline 37.5±1.1%). These results indicate that NAC treatment substantially improves fatigue resistance in the soleus muscle in a model of peripheral arterial insufficiency.

Support‐ Fapesp 2011/18197–0 (B.R.), 2010/51344–3 (CS)
Vasopressin V1a receptor and pressor responses for food‐seeking behavior in fasting mice
Authors: Sumiyoshi Eri , Masuki Shizue , Nose Hiroshi
Published: June 2021

We have reported that cerebral activation at the onset of voluntary locomotion suppressed baroreflex control of heart rate (BR) and increased arterial pressure (AP) in wild‐type mice (WT) but the responses were abolished in V1a receptor (V1aR) knockouts (KO). Here, we examined if the similar responses occurred during food seeking in food‐deprived WT (n=9) and KO (n=10) under LD cycle (7:00/19:00). BR was determined from cross‐correlation function (CCF) between changes in heart rate and AP every 4 s. Cerebral activity was determined from the power density ratio of θ to δ wave band (θ/δ) on EEG every 4 s. Their behavior was recorded with a CCD camera from 19:00–21:00. For 3 consecutive days, mice were fed ad libitum (FED), food deprived (FD), and refed (REF), respectively. Food was removed on day 2 and restored on day 3 at 18:00. We found in WT that CCF between θ/δ and locomotor activity during 19:00–7:00 was stable in FED, while it increased after the onset of FD and peaked at 21:40, higher than FED from 19:00–5:00 (P<0.001), and returned to FED level after REF (P=0.2). The same response occurred in CCF between θ/δ and BR, also peaking at 21:40 after FD (P<0.05). Moreover, only food‐seeking behavior increased after FD, twofold higher than FED (P<0.01), and returned to FED level after REF (P=0.3). However, none of these occurred in KO (P>;0.1). Thus, V1aR is involved in motivated behavior through link between cerebral activity and BR.
Impaired Autonomic Regulation during Exercise in Obese Zucker Rats
Authors: Lu Silu , Xiang Lusha , Clemmer John , Mittwede Peter , Hester Robert
Published: June 2021

Obesity is associated with altered autonomic function and impaired exercise performance. We hypothesized that sympathetic nerve activation and/or vagal withdrawal is impaired during exercise in obese Zucker rats. Oxygen consumption (VO2), mean arterial pressure (MAP), and heart rate (HR) were measured before and during graded treadmill exercise in lean (LZ) and obese Zucker rats (OZ). The following day, experiments were repeated in the same rats following treatment with methoctramine hydrate (MH, 0.15 mg/kg; i.v.), a selective M2 receptor antagonist to block cardiovascular parasympathetic activation. Basal MAP and HR were not different between LZ and OZ. In control animals, exercise increased the MAP, HR and VO2, in both LZ and OZ. MAP increased with exercise to similar levels in all groups. OZ exhibited blunted increases in HR and VO2 as well an impaired VO2max. MH treatment increased basal VO2 in both LZ and OZ. With MH treatment, OZ exhibited a blunted increase in HR and no increase in MAP. MH treatment caused similar increases in the VO2 and HR in LZ and OZ during exercise, with the difference in HR between LZR and OZR still sustained. MH has no effect on VO2max in all groups. These results suggest a blunted parasympathetic regulation of HR under resting conditions in obesity. The autonomic response during exercise is impaired due to blunted sympathetic activation. (Supported by NIH HL‐89581, HL‐51971, AHA postdoctoral fellowship, AHA‐12SDG12050525)
High blood pressure but normal oxidative stress in D3 dopamine receptor deficient mice
Authors: Wang Xiaoyan , Johns John , Asico Laureano , Armando Ines , Jose Pedro
Published: June 2021

To determine if renal oxidative stress is involved in the regulation of blood pressure (BP) upon disruption of D3R, we measured the BP (telemetry) and indices of oxidative stress in D3R−/−, D3+/− and their wild type (D3+/+) mice on normal (0.8% NaCl) or high (4% NaCl) salt diet for one week. Systolic BP (SBP) was higher in D3‐/‐ and D3+/− relative to D3+/+ mice on normal salt diet. NOX1 (101±10 % of D3+/+ mice), NOX2 (101±8) and NOX4 (120±16) was similar in D3−/− and D3+/+ mice. CuZnSOD expression was increased (135±9, P<0.05, n=5), while the EcSOD and MnSOD were unchanged in D3−/− relative to D3+/+ mice. Nitrotyrosine (NT) was not altered in D3−/− (107±30). Urinary isoprostane excretion tended to be lower in D3−/− (445±106 pg/mg of creatinine) than D3+/+ (852±213), but the difference did not reach statistical significance. In D3+/− mice, NOX1 and NOX4 were unchanged but NOX2 (70±8, n=6) was slightly decreased. NT was similar in D3+/− and D3 +/+ mice. On high salt diet, SBP was increased more than 10% over baseline in D3−/− mice but not in D3+/+ mice. NOX1, NOX2, and NOX4 were not altered while NT was decreased (49±8, P<0.05, n=4) and EcSOD was increased (145±1) in D3−/− relative to D3+/+ mice. Thus, the hypertension and salt sensitivity observed in D3−/− and D3+/− mice are not associated with increased renal oxidative stress and compensatory mechanisms may be involved in maintaining normal production of ROS.
Reducing parathyroid hormone is essential for correcting cortical bone deficiencies associated with chronic kidney disease
Authors: Newman Christopher , Moe Sharon M. , Chen Neal X. , Brown Drew M. , Gattone Vincent H. , Allen Matthew R.
Published: June 2021

Chronic kidney disease (CKD) results in an increased fracture risk, partially due to elevations in parathyroid hormone (PTH). Bisphosphonates (BP's) reduce fracture risk by preventing bone loss, but the lack of data in CKD has limited their use. Using the BP zoledronic acid (ZOL) and calcium (CA) (known to lower PTH), the present study used a rat model of CKD to examine the impact of PTH levels on the skeletal abnormalities associated with kidney disease. Skeletally mature male rats with CKD (n=56) were given vehicle, ZOL, CA, or ZOL&CA for 10 weeks. Age‐matched male rats without CKD (n=14) served as controls. Outcomes were determined by serum PTH, femoral cortical porosity, and femoral mechanical testing. In untreated CKD animals, PTH levels were 12 times higher than in normal animals. Animals treated with CA and ZOL&CA had lower PTH values, whereas rats with ZOL alone did not. Compared to normal animals, untreated CKD rats had 41‐fold higher cortical porosity and lower bone strength (−31%). After correcting for differences in porosity, reductions in strength remained (−27%), indicating that CKD adversely impacts bone quality. Animals treated with CA and ZOL&CA had cortical porosity and bone strength similar to normal controls. ZOL alone did not alter porosity or strength relative to untreated CKD rats. These data indicate that normalization of PTH is necessary for controlling cortical bone abnormalities in CKD.

Grant Funding Source: NIH
Chromosome Fragile Sites Signature and Genome Instability in Human Epithelial cells
Authors: Hosseini Seyed Ali , Saldivar Joshua Charles , Horton Susan , Sun Jin , Heerema Nyla A , Huebner Kay
Published: June 2021

Common chromosome fragile sites (CFSs) are highly sensitive to genotoxic stress and perturbation of replication, resulting in deletions within genes located at these sites. CFSs at 3p14.2, 16q23, 4q, 7q, and 6q are the most frequently expressed in lymphoblasts but CFSs have not yet been mapped in epithelial cells. We are defining CFSs in epithelial cells derived from breast tissue to determine if they represent the sites most frequently altered in cancer cells of epithelial organs from which most human cancers derive. We observed that in MCF10A noncancerous breast epithelial cells, chromosome regions 16q23, 2q, 12q, 1p most frequently exhibited chromosome breaks typical of CFS after aphidicolin treatment. This suggests that, as observed for cells of fibroblast origin, CFSs in epithelia are epigenetically determined. In parallel, we have focused on the role of Fhit protein, encoded at the CFS at 3p14.2, on chromosome stability and association with nucleotide pyrimidine pool imbalance. We investigated chromosome instability in epithelial cells deficient for Fhit expression vs Fhit‐expressing cells and observed increased aneuploidy, sister chromatid exchanges, chromosome breaks and gaps in Fhit‐deficient cells. Moreover, Fhit deficiency leads to thymidine kinase 1 down regulation, resulting in pyrimidine pool disequilibrium and accounting, in part, for genome instability.
Transcription Factor Sp1 Promotes Chromatin Remodeling at DNA Double‐Strand Breaks
Authors: Beishline Kate , Koduri Sravanthi , Clifford Jane
Published: June 2021

DNA damage is intimately associated with cancer and aging. The DNA damage response (DDR) is complex, with pathways that sense lesions, signal cell‐cycle arrest, and repair damaged DNA to prevent mutations. Transcription factor Specificity Protein 1 (Sp1) is phosphorylated in response to DNA damage and recruited to sites of Double Strand Breaks (DSBs). We have determined Sp1 is required for DSB repair. The N‐terminal 182 amino acids of Sp1 are sufficient for localization to DSBs and while it retains no transcriptional activity, this domain is sufficient to rescue the repair defect in Sp1‐depleted cells, suggesting a direct role in repair. We have shown a unique interaction between Sp1 and the MRN complex member, Nbs1, which likely promotes its recruitment. Sp1 mediates chromatin remodeling at promoters and our data suggest that Sp1 plays a similar role at DSBs. We have shown that the Sp1 is required for recruitment of the acetyl‐transferase p300 to break sites, which acetylates histone H3 on Lysine 18. This modification, and others promote nucleosome displacement at break sites to allow for repair factor recruitment. Sp1 expression is modulated in multiple cancers and investigating the role of Sp1 in DNA repair is essential for a mechanistic understanding of damage signaling. This study will provide insights into genomic instability, as well as novel therapeutic targets.
ENU‐Induced Mutagenesis in Grass Carp (Ctenopharyngodon idellus) by Treating Mature Sperm
Authors: Zou Shu‐Ming , Jiang Xia‐Yun
Published: June 2021

N‐ethyl‐N‐nitrosourea (ENU) mutagenesis is a useful approach for genetic improvement of plants, as well as for inducing functional mutants in animal models including mice and zebrafish. In this present study, mature sperm of grass carp (Ctenopharyngodon idellus) were treated with a range of ENU concentrations for 45 min, and then wild‐type eggs were fertilized and resulting embryos scored for survival and dominant effects. The results indicated that the proportion of embryos with morphological abnormalities at segmentation stage or dead fry at hatching stage increased with increasing ENU dose up to 10 mM. Choosing a dose that was mutagenic, but provided adequate numbers of viable fry, an F1 population was generated from 1 mM ENU‐treated sperm for screening purposes. The ENU‐treated F1 population demonstrated large variations in growth during the first year. Analysis of DNA from 15 F1 ENU‐treated individuals for mutations in partial coding regions of igf‐2a, igf‐2b, mstn‐1, mstn‐2, fst‐1and fst‐2 loci revealed that most ENU‐treated point mutations were GC to AT or AT to GC substitution, which led to nonsense, nonsynonymous and synonymous mutations. The average mutation rate at the examined loci was 0.41%. These results indicate that ENU treatment of mature sperm can efficiently induce point mutations in grass carp, which is a potentially useful approach for genetic improvement of these fish.
Amplification of human DNA for probing DEB‐induced damage via a quantitative polymerase chain reaction
Authors: Chuang Edward , Millard Julie T.
Published: June 2021

Diepoxybutane (DEB) is a bifunctional alkylating agent linked to the high incidence of certain cancers in industrial workers exposed to butadiene. DEB forms interstrand cross‐links in duplex DNA, inhibiting replication and transcription. Since PCR mimics DNA replication, we can use it to assess cross‐linking by measuring the amount of DNA available for amplification. Our ultimate goal is to determine whether DNA structure affects DEB reactivity in vivo by monitoring damage through a quantitative polymerase chain reaction (PCR) at regions that differ in their degree of chromatin condensation. To date, we have optimized the conditions for traditional PCR within human DNA at nuclear loci of about 10 kb in length. We have also obtained CT values, using real‐time PCR with SYBR green dye. When our real‐time PCR conditions are optimized, we can use the difference between amplification of control, unmodified DNA versus DNA from DEB‐treated cells to determine the lesion frequency at the loci of interest. These findings are important because they will further characterize the mode of action of these compounds and thus help determine the carcinogenic risk for workers in the butadiene industry. This project was supported by grants from the National Center for Research Resources (5P20RR016463–12) and the National Institute of General Medical Sciences (8 P20 GM103423–12) from the NIH.
Detection of pathological tau conformers in cerebrospinal fluid
Authors: March Zachary McDowell , Gupta Sharad , Colby David W.
Published: June 2021

Misfolded tau is a pathological hallmark of over 20 phenotypically distinct neurodegenerative disorders, including Alzheimer's disease (AD). Researchers are beginning to speculate that misfolded tau may spread throughout the brain from a single locus like a prion; whether or not tau aggregates can cross the cell membrane remains unclear. To investigate the presence of misfolded tau in cerebrospinal fluid (CSF) of patients with AD, we developed an amyloid seeding assay for tau. Our findings indicate that misfolded tau molecules migrate in the CSF through the extracellular space, supporting the possibility that misfolded tau may spread through the brain like a prion.
Structural and functional importance of third‐circuit hydrogen bonds within hemolysin A
Authors: Apolinario Michael Angelo , Weaver Todd Michael
Published: June 2021

In this study, hemolysin A was used as a model to investigate the β‐helix structure‐function (SF) paradigm. The research implements a truncated form of hemolysin A termed HpmA265. HpmA265 facilitates uni‐directional, concentration‐dependent and biphasic template‐plate assisted hemolytic activation of full length HpmA. Prior x‐ray crystallographic results reports that the HpmA265 structure consists of a three‐sided, right‐hand parallel β‐helix, adjoining two and four‐stranded antiparallel sheets and four inner core solvent molecules. The specific objective of research examined a complex bonding network surrounding the third circuit of HpmA265 and explored its contributions to thermal stability and hemolytic activity. A series of phenylalanine and alanine site‐directed mutants (SDMs) were used to investigate the importance of an inner core hydrogen‐bonding network between Gln125, Tyr134, and an embedded solvent. Additionally, a cross‐strand carboxyamide ladder between circuits 2 and 4 was targeted by both alanine and lysine replacements. Circular dichroism spectroscopy and template‐assisted hemolytic assays were conducted to ascertain thermodynamic and functional differences between these HpmA265 SDMs. Collectively, the alanine SDMs reported a 6 – 12 °C lowering of the melting point (Tm) with minimal change in hemolytic activity. However, the phenylalanine SDMS led to a lower Tm and a decrease in hemolytic activity. The difference in alanine and phenylalanine hydrophobicity appears to play a crucial role in the β‐helix SF paradigm. The research supported by NSF‐RUI 1050435 (to TW).
Electron transfer without metals. How amino acid radicals (mis)behave
Authors: Koppenol Willem H. , Mahmoudi Leila , Kissner Reinhard , Nauser Thomas
Published: June 2021

Tyrosine participates in electron transfer reactions in biology. The reaction mechanism of ribonucleotide reductase involves electron transfers between Tyr, Trp, and Cys. The Tyr radical may also be formed as the result of reaction with HO•. Tyrosine is the ultimate electron donor in a pecking order consisting of Met, Trp, and Tyr ≈ Cys. As the range of published electrode potentials for Trp at pH 7 is +0.56 – +1.05 V, and +0.68 – +0.94 V for Tyr, we set out to determine a consistent set of electrode potentials. By cyclic voltammetry, we measured E°′(TyrO•,H+/TyrOH) = +0.98±0.02 V and E°′(Trp•,H+/TrpH) = +1.00±0.01 V, thus ΔE°′ = 0.02±0.02 V (K = 2) for electron transfer between Trp•and Tyr; by pulse radiolysis, we found K = 9, corresponding to ΔE°′ = 0.05 V. We recommend that the presently accepted values of E°′ for Trp (+1.05 V) and Tyr (+0.94 V), be replaced with +1.01 V and +0.97 V, respectively. For nitrotyrosine, we found +1.11±0.02 V by cyclic voltammetry. Pulse radiolysis experiments described in the literature suggest that the value of E°′(Cys•,H+/CysH) is similar to that of the Tyr couple; Schöneich and coworkers, and we have shown, however, that the S‐centered radical is in rapid equilibrium with C‐centered radicals in Cys and neighbouring amino acids (10.1021/tx3000494 and 10.1021/tx300045f).

Supported by the Swiss National Science Foundation and the ETH.
Anti‐cancer effect of TRAIL in combination with caffeic acid phenethyl ester in human hepatocellular carcinoma SK‐Hep1 cells
Authors: Kim An Keun , Kim Eun Young
Published: June 2021

Tumor necrosis factor‐related apoptosis inducing ligand (TRAIL) has been considered as a promising candidate for cancer therapy since it selectively induces apoptosis in various malignant cells but not in normal cells. However, some cancer cells are resistant to TRAIL. Here, we investigated the sensitizing effect of caffeic acid phenethyl ester (CAPE), a phenolic ingredient of honeybee propolis, on TRAIL‐resistant SK‐Hep1 cells. Combined treatment of TRAIL with CAPE synergistically inhibited cell proliferation, and induced apoptosis, whereas either agent alone induced limited cytotoxicity and apoptosis in SK‐Hep1 cells. The synergistic effect of CAPE and TRAIL combination on SK‐Hep1 apoptosis was examined by annexin V staining and poly (ADP‐ribose) polymerase (PARP) cleavage. Moreover, combination of CAPE and TRAIL resulted in significant activation of effector caspases. Furthermore, z‐VAD‐fmk, a pan‐caspase inhibitor, blocked the enhanced apoptosis induced by the combination of CAPE and TRAIL, suggesting the involvement of caspase‐dependent pathway. CAPE also increased the expression of death receptor 5 (DR5). The enhanced cell death and caspase activiation by co‐treatment with CAPE and TRAIL were markedly reduced by human recombinant DR5/Fc chimera protein. Taken together, our results indicate that CAPE enhances TRAIL‐induced apoptosis by caspase activation through induction of DR5.

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2010–0010538).
Inhibition of Usp14 Stimulates the Proteolytic Degradation and Clearance of Misfolded Proteins Associated with Neurodegenerative Diseases
Authors: Villella Adriana T , Malhotra Jyoti , Bhalla Akhil , Nokes Eva , Hurtado‐Lorenzo Andres , Calamani Barbara , Soper James , Gao Benbo , Giuliano Kenneth , Hafiz Mohmmad , Foley Megan , Tait Brad , Haeberlein Markus , Saha Anjanabha , Reinhart Peter , King Randall , Finley Daniel , Lo Donald , Garza Dan
Published: June 2021

Proteasomes play a key role in the degradation of polyubiquitinated proteins. The proteasome‐associated de‐ubiquitinating enzyme (DUB) Usp14 slows the degradation of some ubiquitin‐protein conjugates by removing ubiquitin chains prior to substrate degradation. We have carried out proof‐of‐concept (PoC) experiments demonstrating that siRNA‐mediated knockdown of Usp14 leads to reductions in α‐synuclein levels in HEK293 cells, induced pluripotent human stem cell‐derived neurons (iPS neurons) and rat brain slices. Compound‐mediated inhibition of Usp14 using selective inhibitors was also shown to reduce α‐synuclein protein levels in both iPS neurons and rat brain slices. Further, these results have been extended to other disease‐related client proteins. These results suggest Usp14 inhibition as a new therapeutic approach for the treatment of protein aggregation diseases through proteasome‐mediated clearance of disease‐associated proteins. The use of small molecules to manipulate proteasomal activity is one of several strategies being used at Proteostasis Therapeutics to treat a number of neurodegenerative and orphan diseases through modulation of the proteostasis network responsible for the folding, trafficking, and degradation of misfolded and aggregated proteins.
Effect of exposure to volatile organic compounds (VOCs) on inflammatory response in gas station attendant.
Authors: Corazzini Roseli , Gonçalves Giovana , Bonetti Tatiana , Garcia Diego Martinez , Fonseca Fernando Luiz Affonso , Carvalho Cristina
Published: June 2021

Introduction

Volatile Organic Compounds (VOCs) exposition causes multiple chemical sensitivity reactions in human. The exposures of human to VOC environments are associated with an enhanced inflammatory response. The occupational exposures of VOCs and inflammatory responses in gas station attendant not have been reported.

Aim

Evaluate the inflammatory markers plasma concentrations in gas station attendants.

Materials and Methods

20 gas station attendants who are occupational exposed to high VOC levels (VOC group), and 16 controls individuals (CT group) who are not occupational exposed to VOC. Serum inflammatory markers interleukin (IL)‐8, IL‐1β, IL‐6, IL‐10, IL‐12 and tumor necrosis factor‐alpha (TNF‐α), were measured by flow cytometry using the Human Inflammation Kit BD Cytometric Bead Array (CBA).

Results

Individuals were 20 to 70 years of age, and those on VOC group had a mean of 12 years working at gas station. Multivariate linear regression model showed that VOCs exposure influenced significantly IL‐12 plasma concentration in gas station attendants (Coef. 1170.44; p=0.007) adjusted to age and smoking. To confirm that result, the IL‐12 concentration was significantly higher on VOC group (1609±250) than CT (720±247; p=0.018).

Conclusion

Our findings show that occupational exposed to VOC from gas stations are associated with increased levels of inflammation marker related to cytotoxic response, which may be related to reactions to chemicals in human.
AssuriTEA® Wellbeing Improves Antioxidant Status in Healthy Male Subjects
Authors: Capodice Jillian , Herrlinger Kelli , Chirouzes Diana , Katz Aaron , Ceddia Michael
Published: June 2021

Green and black teas have known anti‐oxidant and anti‐inflammatory properties. The aim of this randomized, doubleblind, placebo‐controlled clinical trial was to evaluate the effect of a water‐extracted green and black tea blend (AssuriTEA® Wellbeing [ATW]) on serum antioxidant status, vital signs, and tolerability in healthy males. Men, ages 25–70, were randomized to receive placebo, 250 mg, 500 mg, or 1000 mg ATW daily for 28 days. At baseline (BL) and day 28, serum antioxidant capacity was evaluated through ferric iron reducing antioxidant power (FRAP) and cellular antioxidant protection (CAP‐e). Forty‐three subjects were evaluable (>;80% compliant). Subjects (56.8 ± 2.8 years [mean ± SEM]) demonstrated improved antioxidant status as measured by FRAP at all doses vs. placebo (p < 0.05). CAP‐e was improved (p = 0.032) at 1000 mg ATW dose vs. placebo. At day 28, fasting serum glucose levels were significantly higher in the placebo group vs. 250 mg ATW (p = 0.023) with a trend observed at 1000 mg ATW (p=0.054). There was a significant improvement in systolic blood pressure (BP) in 250 mg ATW at day 28 vs. BL (p = 0.048). In the 1000 mg ATW, there was a significant decrease in systolic BP and a trend toward decrease in diastolic BP at day 28 vs. BL (p = 0.017, p = 0.067). There were no abnormal changes in hematology or hepatic function. ATW was well tolerated and resulted in improved serum antioxidant status, blood glucose and systolic BP.
Comparison of daily flavonoid intake and major food sources in 1990 with 2006 in Nurses’ Health Study and Health Professional Follow‐Up Study
Authors: Yang Meng , Kim Hyun Ja , Chun Ock K. , Sampson Laura , Cassidy Aedin , Willett Walter C. , Cho Eunyoung
Published: June 2021

Emerging studies have reported flavonoid intake in relation to risk of chronic disease, but few studies have investigated the change of flavonoid intake in U.S. adults over time. The present study aimed to compare the daily intakes of total flavonoid, its subclasses, and major food sources in 1990 with that in 2006 in prospective studies. A total of 78,884 women from the Nurses’ Health Study and 37,882 men from the Health Professionals Follow‐Up Study completed food frequency questionnaires in 1990 and 2006. Daily intakes of total flavonoid and its subclasses were calculated by using an updated USDA database. Averaged total flavonoid intake in 2006 was higher than that in 1990 in women (357 mg/d vs 353 mg/d, P=0.025) and men (389 mg/d vs 353 mg/d, P<0.0001). Similar increase was found for dietary flavonols, flavones, anthocyanins and polymers, while intakes of flavanones and flavan‐3‐ols decreased. Major food sources of total flavonoids and each subclass remained comparable across 16 years but contributions of each food item decreased while food diversity increased in 2006. Total flavonoid intake in 1990 was higher in older age and in groups with lower body mass index and alcohol intake, and was higher in non‐smokers than their counterparts (P<0.05). In conclusion, intakes of total flavonoid and some subclasses increased over time and major food sources remained similar.
Type III Transforming Growth Factor beta receptor regulates vascularization and osteoblast differentiation during palatogenesis
Authors: Hill Cynthia Renae , Jacobs Britni , Barnett Joey , Goudy Steven
Published: June 2021

Cleft palate occurs in up to 1:500 live births associated with mutations in multiple genes. Palatogenesis involves a complex choreography of palate shelf elongation, elevation, and fusion. Transforming Growth Factor β (TGFβ) and Bone Morphogenic Protein 2 (BMP2) are known regulators of palatal development, but the contributions of the specific receptors remain elusive. The Type III TGFβ Receptor (TGFβR3) binds all three TGFβ ligands, inhibin, and BMP2. To address the role of TGFβR3 during palatal development we examined Tgfbr3−/−mice. These mice do not survive to e15.5 when palate fusion occurs, but at e14.5, palatal elongation and elevation had failed. Tgfbr3−/−embryos display unorganized and reduced vascularization of the palate at e13.5 and e14.5 using PECAM and Smooth Muscle Actin immunostaining which was accompanied by aberrant expression of arterial, venous, and lymphatic markers using qPCR. Analysis of osteoblast differentiation markers also identified significant reductions in alkaline phosphatase, osteocalcin, and collagen1. In addition, in vitro bone mineralization assays revealed reduced capacity of Tgfbr3−/−mouse embryonic palatal mesenchymal cells to undergo osteoblast differentiation and to form bone. These data demonstrate that TGFβR3 contributes to palatal development by regulating vascularization and osteoblast differentiation.
Glycemic and insulinemic response to four different sweeteners in healthy individuals: A double blind, randomized controlled trial.
Authors: Jenkins Alexandra , Shelke Kantha , Vuksan Vladimir , Wolever Thomas M , Campbell Janice , Ezatagha Adish
Published: June 2021

Background

The non‐nutritive sweetener, erythritol, does not change postprandial glucose levels, whether this holds true for forms which are modulated to be more intensely sweet is not known.

Objectives

Comparison of the postprandial glucose and insulin responses of two erythritol products differing in sweetness intensity.

Methods

15 healthy volunteers (6 M, 9F; 38±12 yr; 26.9±4.4kg/m2) consumed 4 test meals consisting of 24g sucrose, 24g erythritol (Swerve® Granular), 2.4g erythritol (Swerve® 10x) and 3g sucralose (Splenda®) mixed with 200ml of water. All meals were matched for sweetness level. Capillary blood samples were taken fasting and at 15, 30, 45, 60, 90 and 120 min after the start of the meal.

Results

Postprandial glucose and insulin levels were significantly lower after the non‐nutritive sweeteners compared to sucrose at 15, 30 and 45 min. Glucose levels were also lower after both erythritol products compared to sucralose at 15 min. At 90 and 120min, glucose levels were significantly higher after the non‐nutritive sweeteners compared to sucrose (p<0.001). Incremental glucose and insulin areas were significantly lower after the three non‐nutritive sweeteners compared to sucrose (p<0.0001).

Conclusions

This study confirms that irrespective of sweetness level, consumption of erythritol does not raise postprandial blood glucose or insulin levels significantly.
Reduced incidence and duration of acute respiratory infections (ARI) in children fed a follow‐up formula containing docosahexaenoic acid (DHA), a prebiotic blend of polydextrose (PDX) and galacto‐oligosaccharides (GOS), and yeast beta glucan
Authors: Scalabrin Deolinda , Min Xingming , Li Fei , Zhuang Wendy , Strong Paul , Liu Bryan
Published: June 2021

Intakes of DHA, prebiotics, yeast‐β‐glucan, and other micronutrients have been associated with reduced incidence of respiratory infections.

To determine if a follow‐up formula containing a combination of DHA, a prebiotic blend of PDX and GOS, and yeast β‐glucan reduced the incidence of ARI in children.

In this double‐blind, randomized, controlled trial, healthy children (3–4yrs) were fed 3 servings per day of a follow‐up formula (FF; n=156) containing DHA (25mg/serving), PDX and GOS (1.2g/serving), and yeast β‐glucan (12.8mg/serving), or a control powdered cow's milk (C; n=154) for 28 weeks. The incidence and duration of ARI were obtained from medical records. Incidence of ARI was analyzed with the CMH test. Duration of ARI was analyzed with ANOVA.

The FF group had fewer episodes and shorter average duration of ARI compared to the control group (Table).

When compared to cow's milk, daily intake of a follow‐up formula containing DHA, a prebiotic blend of PDX and GOS, and yeast β‐glucan over a 28 week period resulted in fewer episodes and shorter duration of acute respiratory infections.
Elevated leptin concentration is associated with low glucose, high triglycerides, high cholesterol and low HDL concentrations in Mexican women living in rural areas
Authors: Elian Shadia , Garcia Olga P , Ronquillo Dolores , del Carmen Caamaño María , Rosado Jorge L
Published: June 2021

The objective of this cross‐sectional study was to evaluate the relationship between leptin concentration, fasting glucose and lipid profile in women living in rural Mexico. Weight, height, waist circumference and body composition were measured in 580 women (36.9 ± 7.6 y). A fasting blood sample was taken for the analysis of leptin, glucose and lipid profile. Prevalence of obesity and overweight was 44% and 36%, respectively. 82% of the population had high risk waist circumference (>;80 cm) and 96% had elevated body fat content (>;30%). Leptin concentration was positively associated with BMI, waist circumference and % body fat (p<0.05). Unadjusted regressions showed an inverse association between leptin and HDL cholesterol. In regressions analysis adjusted for body fat percent, BMI and age, an inverse relationship with glucose was found. Women with high levels of triglycerides (>;150 mg/dL), high cholesterol (>;200 mg/dL) and low concentrations of HDL (<50 mg/dL) had significantly higher leptin concentrations (p<0.05) compared with women with adequate lipid profile. In conclusion, higher leptin concentration is associated with low glucose and dyslipidemia and higher risk of heart disease in this population. Supported by CONACYT.
Dietary isoflavone intake modulates evoked responses to cardiometabolic stimuli in healthy volunteers
Authors: Ferguson Jane Francisca , Tuteja‐Stevens Sony , Shah Rhia Y , Patel Parth N , Rader Daniel J , Reilly Muredach P
Published: June 2021

Isoflavones, found in high concentrations in soy‐derived foods, function as phytoestrogens, have antioxidant and anti‐inflammatory activity, inhibit protein‐tyrosine kinase activity, and are thought to be atheroprotective. We hypothesized that habitual isoflavone intake modulates response to evoked endotoxemia and oral fat load.

In the Genetics of Evoked Responses to Niacin and Endotoxemia (GENE) Study, we administered low dose endotoxin (LPS 1 ng/kg) to induce inflammation in young, healthy volunteers (N=215, 50% female) and administered a separate oral fat tolerance test (OFTT). Data were analyzed using linear regression models adjusted for race, gender, age and BMI.

Dietary isoflavone intake was a significant predictor of plasma tumor necrosis factor alpha (TNFα) and interleukin 6 (IL‐6) post LPS. High (>;1.5mg/day, N=64) vs low (<1.5mg/day, N=151) isoflavone consumers had higher peak TNFα (64 vs 45 pg/ml, P=0.015) and peak IL‐6 (210 vs 160 pg/ml, P=0.06). Further, the high isoflavone group had significantly higher β‐hydroxybutyrate levels post‐OFTT (442 vs 332 μmol/L, 2 hours post fat load, P=0.02).

Habitual isoflavone consumption may modulate disease development through effects on inflammatory and metabolic responses in humans. For investigating nutritional effects on disease development, measuring evoked phenotypes may be more informative than describing resting characteristics.

The GENE Study was supported by NIH Grant UL1TR000003 as well as a NIH‐NHLBI SCCOR Project grant (P50‐HL‐083799) to MPR. JFF is supported by an AHA postdoctoral fellowship grant (12POST11840017).
Cox2 reporter gene expression and prostacylin mass spectrometry confirm vascular COX‐1 dominance for prostacyclin production
Authors: Kirkby Nicholas S , Zaiss Anne K , Urquhart Paula , Jiao Jing , Austin Philip J , Warner Timothy D , Nicolaou Anna , Herschman Harvey R , Mitchell Jane A
Published: June 2021

Prostacyclin is an endothelium‐derived anti‐thrombotic hormone, whose synthesis requires cyclo‐oxygenase (COX) activity. Two COX isoforms exist. It is widely held that COX‐2 drives prostacyclin production and that this explains the cardiovascular toxicity of COX‐2 inhibitors, but this supposition relies on indirect evidence from urinary markers. We previously demonstrated that COX‐1 is the dominant isoform for vascular prostacyclin release (Kirkby et al PNAS 2012), however, this work was criticized (Ricciotti et al PNAS 2012) because it relied on immunoassay and immunohistochemistry methods. To address this, we measured prostanoid levels by mass spectrometry in mouse plasma and found levels similar to immunoassay values, which were reduced (>;80%) by Cox1, but not by Cox2 deletion. Using a Cox2 luciferase reporter mouse we also confirmed that there is little gene expression from the aorta, other large arteries or veins. In contrast, thymus, brain and gut expressed much higher levels (Figure 1). These data are consistent with the conclusion that COX‐1 drives vascular prostacyclin release, allays previous criticisms, and puts the weak vascular COX‐2 expression in the context of other tissues. They also suggest target tissues to consider in developing much needed new ideas of how COX‐2 protects the cardiovascular system.
CB1‐Like Behavioral Effects of the Novel FAAH inhibitor AM3506
Authors: Leonard Michael Z , Kangas Brian D , Nikas Spyros P , Shukla Vidyanand G , Subramanian Kumara V , Thakur Ganesh A , Makriyannis Alexandros , Bergman Jack
Published: June 2021

The enzyme that degrades the endocannabinoid anandamide (fatty acid amide hydrolase; FAAH) has been forwarded as a therapeutic target for metabolic and other disorders. The recently developed ligand, AM3506, inhibits FAAH selectively and with high potency (IC50: 2.8nM) and, as well, shows moderate CB1 receptor affinity (Ki: 192nM). The present work was conducted to study behavioral consequences of these actions by assessing the CB1‐like discriminative‐stimulus effects of AM3506 and, separately, those of anandamide (AEA) after treatment with AM3506 in squirrel monkeys (n=4) trained to discriminate 0.01 mg/kg of the THC‐like CB1 agonist AM4054. Results show that: a) a high dose of AM3506 (1.0 mg/kg) fully substituted for AM4054, whereas AEA failed to substitute over a wide dose range (1.0–32.0 mg/kg), presumably due to its rapid degradation; b) a 100‐fold lower dose of AM 3506 (0.01 mg/kg) sufficed to reveal CB1‐like effects of 5.6–10 mg/kg AEA; c) rimonabant served to antagonize the CB1‐like effects of AEA and AM3506 in a qualitatively dissimilar manner; d) based on Schild analysis, AM3506 (pA2=5.6) has CB1 receptor‐mediated actions that differ quantitatively from those of AM4054 (pA2=6.5). Together, these data indicate that CB1 receptor activation by AM3506 occurs at much higher doses than those that inhibit FAAH and may differ qualitatively from activation by conventional CB1 agonists. (Supported by NIH DA31020; DA19205)
Antiangiogenic activity of the HSP90 inhibitor ganetespib in pancreatic cancer models
Authors: Ganji Purnachandra Nagaraju , Diaz Roberto , El‐Rayes Bassel
Published: June 2021

Heat shock protein 90 (HSP90) has a central role in the regulation of transcriptional factors that modulate angiogenesis and growth. Ganetespib (G) is a functional inhibitor of HSP90 being studied in a broad range of clinical trials. We tested the hypothesis that functional inhibition of HSP90 by G can inhibit angiogenesis and growth in vitro and in vivo models of pancreatic cancer. PANC‐1, and HPAC cell lines were treated with vehicle or G (50nM) for 24 h and lysates were analyzed by Western blot. Egg CAM and matrigel plug assays were performed to quantify the effects of G on angiogenesis. Efficacy of G (100mg/kg) was assessed in mice bearing HPAC and ASPC‐1 xenograft. Western blot analyses demonstrated a significant reduction in intracellular HIF‐1á and VEGF protein levels in PANC‐1 and HPAC cells treated with G. Results from ELISA assays showed that G reduced VEGF secretion in the culture medium from both pancreatic lines. G treatment reduced angiogenesis compared to vehicle in all three models. Animals with human pancreatic tumor xenografts treated with G had significant tumor growth delay and inhibition of angiogenesis. The preclinical data demonstrates that G can inhibit pancreatic cancer growth and angiogenesis, suggesting that targeting HSP90 is a rational new approach to pancreatic cancer therapy to be explored in clinical trials.
Pharmacological stabilization of hypoxia inducible factors (HIF) enhances endothelial immunity against invasive gram positive bacterial pathogens
Authors: Beasley Federico Christopher , Keller Nadia , Nizet Victor
Published: June 2021

Bloodborne bacteria that target and invade the endothelium can cause life threatening syndromes such as meningitis and endocarditis. Even timely administration of antibiotic therapy may be ineffective if the infectious agent is a “superbug”, such as methicillin‐resistant Staphylococcus aureus (MRSA). Hypoxia‐inducible transcription factors (HIF) regulate energy homeostasis and cellular adaptation to low oxygen stress. The HIF‐1 isoform is also activated in response to infectious stimuli and augments the innate immune functions of white blood cells. Hypoxia mimicry through administration of the HIF‐stabilizing drug AKB‐4924 was previously shown to enhance the antibacterial activity of phagocytes and skin cells against MRSA, uncovering a new strategy in the treatment of superbugs. We show here that AKB‐4924 is also an effective activator of isoforms HIF‐1 and HIF‐2 in cultured endothelial cells. Pretreatment of cells rendered them more resistant to invasion by pathogenic Staphylococcus and Streptococcus, while concurrent treatment of sick cells limited the intracellular replication of these bacteria. Confirming a role of HIF in maintenance of the endothelial barrier against bloodborne disease, mice with endothelium‐targeted HIF deletions accumulated greater bacterial loads in organs including brains and hearts during bacteremic infection assays.
Adenosine A1 Receptor Biased Agonism: A Novel Approach for Cardioprotective Therapeutics
Authors: May Lauren Therese , Baltos Jo‐Anne , White Paul J , Valant Celine , Scammells Peter J , Sexton Patrick M , Christopoulos Arthur
Published: June 2021

Adenosine receptor (AR) activation represents the most powerful cardioprotective mechanism. Unfortunately, the translation of A1AR agonists into the clinic has been severely hindered due to on‐target adverse effects, including bradycardia, atrioventricular block and hypotension. GPCR biased agonism represents an exciting approach to develop adenosine therapeutics that selectively enhance cardioprotective signaling in the absence of unwanted effects. This study used a chemical biology approach and novel analytical methods to identify and profile physiologically relevant A1AR biased agonists. The reference A1AR agonists, NECA and R‐PIA, did not display significant bias for any of the signaling pathways assessed; whereas VCP746 was significantly biased away from calcium mobilization. VCP746 mediated potent cardioprotection with minimal effects on heart rate or blood pressure; enabling, for the first time, separation of desired from adverse effects. Collectively, these studies demonstrate that “fingerprinting” of biased agonism within a model system can be predictive of novel and physiologically relevant in vivo pharmacology. This research was supported by the NHMRC.
Epoxomicin Treatment before Ischemia Confers Cardioprotection through Up‐regulated Hsp22 Expression
Authors: Li Dan , Yoon Seong , Park Misun , Rashed Eman , Dai Huacheng , Park Jiyeon , Sui Xiangzhen , Vatner Dorothy , Babu Gopal
Published: June 2021

Background

Cell death is the cause of infarct formation in ischemic myocardium. In this study, we demonstrate that Epoxomicin (Epox), a proteosome inhibitor formerly used as an anti‐cancer drug, confers effective cardioprotection.

Methods and results

The C57 mice (3.5 months, n=7) were injected i.p. with Epox (0.5 mg/kg/day) or vehicle for 2 weeks, and then subjected to 30 min ischemia followed by 24h reperfusion. Epox reduced the infarct size by 50% (p<0.05), despite of similar area‐at‐risk. To investigate the mechanism involved, in vitro study was performed on neonatal rat cardiac myocytes (NRCM). NRCM were treated with Epox (0.01μM) or veh for 24 hours, and then subjected to 36h hypoxia. Epox decreased hypoxia‐induced apoptosis by 50% (n=6; p<0.01). The protection was associated with up‐regulated expression of Hsp22 and its downstream effector iNOS. Deletion of Hsp22 in vitro and in vivo both abolished the iNOS expression and cardioprotection. Transcription activation analysis showed that HSF1 was highly activated in Epox treated NRCM. The Upregulation of Hsp22 expression was abolished upon the addition of KRIBB11, a specific inhibitor to HSF1.

Conclusion

Epox upregulates Hsp22 expression through HSF1 thereby provides cardioprotection, in vivo equivalent to preconditioning, against ischemia/reperfusion injury. Our data point to Epox as a promising therapeutic agent against ischemic heart disease.