bluuebunny/crossref_metadata_2025 huggingface dataset:
Serum‐derived bovine immunoglobulin isolate (SBI) as a potential therapy for inflammatory bowel disorders (IBD)
Authors: Klein Gerald L , Weaver Eric , Shaw Audrey Lynn , Cohn Jeffrey
Published: June 2021

Cytokine‐mediated dysfunction of the intestinal mucosal barrier is observed during the active phase of IBD. SBI may reduce its impact by reducing excessive pro‐inflammatory cytokine expression, such as TNFα, INFγ, and interleukins (eg, IL‐1, IL‐6). For example, mdr1a knockout mice prone to intestinal mucosal barrier alterations (a genetic model for IBD) who received SBI had lower levels of IL‐6 and IL‐17 in the mucosa during the onset of colitis.

Research demonstrates that SBI survives initial digestive processes, improving gastrointestinal (GI) function and morphology in a clinically significant way. Animal studies confirm SBI improves cellularity of the lamina propria of the small intestine, resulting in improved mucosal barrier function, reduced inflammation and diarrhea, and improved dietary protein utilization through increased absorption, reduced permeability and mitotic activity, increased villus heights, with improved weight gain and bone density. SBI decreased watery bowel movements and increased nutrient absorption/retention in HIV patients, and was well tolerated. Early studies in patients with IBS‐D, malnutrition or HIV enteropathy demonstrated improved GI symptoms. In toto, the accumulated evidence suggests SBI is beneficial for inflammatory GI diseases such as Crohn's and ulcerative colitis and demonstrates strongly the need for additional research. Study funded by Entera Health.
Vitamin D (VitD) Supplementation of Mother and Infant vs. Mother Alone
Authors: Wagner Carol L , Howard Cynthia M , Hulsey Thomas C , Ebeling Myla , Lawrence Ruth A , Hollis Bruce W
Published: June 2021

Objective

To assess safety & effectiveness of higher dose maternal vitD alone compared w/maternal & infant supplementation of 400 IU/day.

Methods

From 2006–2011, fully lactating women/infants at 1‐month postpartum living in Charleston, SC and Rochester, NY were randomized to 400, 2400 or. 6400 IU vitD3/d. Infants of Control mothers received 400 IU vitD3/d while infants of 2400 IU and 6400 IU grps received placebo.

Results

476 mothers/infants were enrolled. No differences betw tx grps in mat/infant baseline 25(OH)D (p=0.8). 2400 IU arm was stopped in 2009: higher % infants with deficiency (25(OH)D<20 ng/mL) at 4 mos. Of remaining 2 grps, maternal 25(OH)D (ng/mL) changed from 34.0 to 30.1 in 400 IU grp (p=0.04) and from 37.2 to 59.5 in the 6400 IU grp (p<0.0001); between grp (p<0.0001). Infant 25(OH)D in the 400 IU grp increased from 17.4 to 45.2 (p<0.0001) and from 17.8 to 43.2 in the 6400 IU grp (p<0.0001); between infant grps (p=0.4). No differences in safety profiles of women and infants by tx.

Conclusions

Women in 6400 IU grp had improved vitD status compared to 400 IU/d grp; and their infants’ vitD status was equivalent to those infants receiving 400 IU/d. When mother is vitD sufficient, her milk is sufficient, and her BrF infant will be sufficient without further supplementation. Findings have implications re: vitD supplementation recommendations during lactation.
Anti‐angiogenic activity of alpha7‐nicotinic receptor antagonists in human small cell lung cancer
Authors: Thornhill Brent A , Brown Kathleen C , Lau Jamie K , Dom Aaron M , Witte Theodore R , Hardman W Elaine , Luo Haitao , Chen Yi C , Dasgupta Piyali
Published: June 2021

Small cell lung cancer (SCLC), a highly angiogenic tumor displays a 90% association with smoking. Nicotine promotes angiogenesis in lung cancer via alpha7‐nicotinic acetylcholine receptors (α7‐nAChRs). We conjectured that α7‐nAChR antagonists should suppress nicotine‐induced angiogenesis and may have potential applications for the therapy of SCLC. Here we show that the synthetic α7‐nAChR antagonist MG624 suppresses nicotine‐induced proliferation of human microvascular endothelial cells from the lung (HMEC‐L). MG624 displayed potent anti‐angiogenic activity in the Matrigel, rat aortic ring, and chicken chorioallantoic membrane models and in human SCLC tumors xenografted in athymic mice. The administration of MG624 did not cause any discomfort, gross toxicity or lethargy in mice. Chromatin immunprecipitation assays show that MG624 decreases the recruitment of Egr‐1 on FGF2 promoter thereby decreasing its transcription and suppressing nicotine‐induced angiogenesis. Currently, our laboratory is testing the anti‐angiogenic activity of other well characterized α7‐nAChR antagonists like MLA and COG. Our results suggest that α7‐nAChR based anti‐angiogenic agents may be useful for the therapy of human SCLC.
High prevalence of Chlamydophila Psittaci subclinical infection in Italian patients with Sjögren's syndrome and parotid gland B‐cell lymphoma.
Authors: Fabris Martina , Dolcetti Riccardo , Pasini Elisa , Quartuccio Luca , Pontarini Elena , Ponzoni Maurilio , Scott Cathryn Anne , Curcio Francesco , De Vita Salvatore
Published: June 2021

To investigate Chlamydophila psittaci (Cp) subclinical infection in Sjögren's syndrome (SS), seventy‐four consecutive Caucasian SS patients (55.4 ±13.4 yrs; 4M/70F) were studied. Among them, 18 with salivary gland MALT B‐cell NHL, 20 with myoepithelial sialoadenitis (MESA), and 36 without lymphoproliferative disorders (LPD). The presence of Cp DNA was assessed in peripheral blood of all patients and 225 healthy donors by previously reported methods (Fabris, Clin Exp Rheumatol 2011). Paired salivary gland samples were also investigated in 34 patients. Cp DNA was detected in 11/74 (14.9%) SS patients vs. 1/225, 0.4% controls (p<0.0001). Cp was detected at higher frequency in MALT NHL (6/18, 33.3%) compared to MESA (3/20, 15%) and no‐SS‐associated LPD (2/36, 5.6%), (p=0.02). A similar percentage of Cp positivity was detected in blood vs. salivary glands, with higher frequency in major than in minor salivary glands (5/18, 27.8% vs. 1/17, 5.9%; p=ns). Our results suggest that Cp infection could be involved in a subset of patients with SS developing lymphoma. When also considering that Cp infection can be treated, additional studies appear worthwhile.
In vivo murine lymph node imaging with multispectral photoacoustic technology and clinical contrast agents
Authors: Heinmiller Andrew , Hupple Clinton , Needles Andrew , Theodoropoulos Catherine , Wood Ronald W
Published: June 2021

Here we describe the use of a commercially available photoacoustic (PA) imaging system (Vevo LAZR, VisualSonics, Toronto) to detect methylene blue (MB) and indocyanine green (ICG) in the left and right iliac lymph nodes after injection into the synovial space of the left and right knee joints.

The PA imaging system generated light from a tunable laser (680 – 970 nm) which was delivered through fiber optic bundles integrated into a linear array transducer (LZ‐250, fc = 21 MHz), driven by a linear stepper motor for 3D imaging. Ultrasound image‐guided injections of 25ul injections of MB (10 mg/ml) and ICG (5 mg/ml) were performed by inserting a 30 gauge needle inserted into the synovial space of the left and right knee, respectively. Two‐dimensional regions of interest surrounding the left and right iliac lymph nodes and the abdominal aorta provided dye and blood absorption spectra.

Comparison of pre‐injection and post‐injection multiplexed images clearly demonstrated ICG and MB in the left and right iliac lymph nodes, respectively. The absorbance spectra of the dyes demonstrated characteristic results with clear differences apparent for MB (peak at 680nm), ICG (peak at 800nm) and typical blood spectra.

Photoacoustic imaging thus can be used to identify sentinel lymph nodes. With dyes targeted to biomarkers for cancer cells, the method could also be used to identify metastases there and elsewhere. Photoacoustic imaging should also be useful in the study of lymphatic function in other disease processes such as lymphedema, arthritis, and parasitic infections.

All research was conducted and funded by VisualSonics, Inc.
Maternal pre‐gravid body mass index and adiposity influence umbilical cord gene expression at term in AGA infants
Authors: Thakali Keshari , Saben Jessica , Andres Aline , Shankar Kartik
Published: June 2021

While maternal obesity is associated with unfavorable maternal and fetal outcomes, the influence of maternal obesity on fetal gene expression is less clear. Umbilical cords (UC) from 12 lean (pre‐gravid BMI < 25) and 10 overweight/obese (OB, pre‐gravid BMI ≥25) women without gestational diabetes were collected at term and utilized for gene expression analysis using Human Primeview microarrays (Affymetrix). Although offspring birth weight and infant adiposity (at 2‐wk) did not differ between groups, expression of 232 transcripts was affected in UC from OB compared to lean mothers. GSEA analysis revealed an up‐regulation of genes related to metabolism, stimulus and defense response and inhibitory to insulin signaling in the OB group. We confirmed that Egr‐1, periostin, FosB, and NEDD9 mRNA expression was induced in UCs from OB moms, while endothelin receptor B, Klf10, Peg3 and Egln3 expression was decreased. mRNA expression of Egr‐1, FosB, MEST and SOCS1 was positively associated (p<0.05) with mother's 1st trimester body fat mass (%). Metabolic and hormonal assessments were also assayed in mother's serum at 30 weeks of gestation and in UC plasma. Our data indicate that maternal obesity per se causes changes in UC gene expression favoring inflammation and insulin resistance, possibly predisposing infants of overweight moms to become overweight later on in life. Funding USDA ARS‐CRIS 6251–51000‐007–04S.
TRANSCRIPTION FACTOR EGR‐1 IN MOLECULAR MECHANISMS OF EXPERIMENTAL GASTRIC ULCERS
Authors: Beregoviy Sergii , Kukharskyy Vitaliy , Bernyk Ksenia , Beregova Tetyana , Tolstanova Ganna
Published: June 2021

Egr‐1 is a key gene & transcription factor in experimental duodenal ulceration (Khomenko T et al. 2006). Role of Egr‐1 in gastric ulcer (GU) pathogenesis is unclear. We tested the hypotheses that Egr‐1 might play a role in the molecular mechanisms of experimental GU. GU was induced in male rats (150–190 g) by: immobilization (IMO) stress for 6, 12 & 24 hr; IMO stress combined with water‐immersion (IMO‐WI) for 20 min, 1 & 3 hr; ethanol (1 ml 96%, per os) for 20 min & 1 hr; aspirin (10 mg/100g, per os) for 20 min, 1 & 3 hr. Rats were euthanized immediately after ulcerogenic compound exposure. IMO‐WI stress induced rapid early up‐regulation of Egr‐1 by 1.6‐ & 1.5‐folds (p<0.05) at 20 min & 1 h and down‐regulation ‐ at 3 hr. IMO stress increased Egr‐1 levels at 6 hr (2.1‐folds, p<0.05) with gradual down‐regulation by 24 hr. Ethanol & aspirin didn't change Egr‐1 levels. IMO‐WI stress but no aspirin & ethanol increased phosphorylation of Erk1/2 by 2.2; 2.6; 2.5‐folds at 20 min, 1 & 3 hr (p<0.01), respectively & didn't change ‐ p38. Erk2 was the major isoform expressed & phosphorylated.

Conclusion

Egr‐1 plays a role in the early stages of stress‐induced GU development vs. aspirin or ethanol; Erk1/2 might regulate Egr‐1 activity during stress‐induced GU.
RGS 2 and RGS 4 Differentially Modulate G Protein Coupled Receptor Signaling in the Mouse Aorta
Authors: Mabe Nathaniel W. , Seeley Sarah , Fehrman Lauren , Garber Paige , Neubig Richard R. , Rorabaugh Boyd R.
Published: June 2021

Regulator of G protein signaling (RGS) proteins terminate G protein signaling by enhancing the rate of GTP hydrolysis. We compared the ability of RGS 2 and RGS 4 to modulate the actions of several G protein coupled receptor agonists in isolated mouse aortic rings. Genetic deletion of either RGS 2 or RGS 4 significantly enhanced the efficacy of phenylephrine‐induced contraction but had no effect on the potency of phenylephrine. The serotonin‐induced contractile response was unaffected by deletion of RGS 2, but the efficacy of serotonin was significantly attenuated by deletion of RGS 4. This effect of RGS 4 deletion was reversed by L‐nitroargenine methyl ester or by denuding the aorta. Deletion of RGS 2 or RGS 4 had no effect on the efficacy or potency of carbachol‐induced aortic relaxation. Angiotensin II and clonidine produced no aortic contraction irrespective of the presence of RGS 2 or RGS 4. These data demonstrate that RGS 2 and RGS 4 differentially modulate aortic responses induced by phenylephrine, serotonin, and carbachol. Furthermore, RGS 4 modulates serotonin‐induced contraction through a mechanism that is dependent upon the presence of an intact endothelium and functional nitric oxide synthase. (This work was supported by GM39561).
Activation of G protein‐coupled estrogen receptor 1 (GPER) induced coronary vasodilation by activation of MLCP via cAMP/PKA pathway
Authors: Yu Xuan , Li Fen , White Richard E. , Stallone John N. , Heaps Cristine L. , Han Guichun
Published: June 2021

Activation of GPER exerts a protective effect in hypertension and ischemia‐reperfusion models, and relaxes arteries in vitro. However, our understanding of the mechanisms of GPER‐mediated vascular regulation is far from completed. We have investigated whether GPER‐mediated relaxation of porcine coronary arteries involves cAMP/PKA signaling. In these vessels, the selective GPER agonist, G‐1, increased cAMP production in a concentration‐dependent manner. Myograph studies demonstrated that inhibiting the activity of adenylyl cyclase (AC) with100 μM SQ‐22538 inhibited 0.3–3 μM G‐1‐induced relaxation of coronary arteries preconstricted with 1 μM PGF2α. Activating AC with forskolin also induced a concentration‐dependent relaxation. Furthermore, G‐1‐induced coronary vasodilation was attenuated by inhibiting PKA with either 5μM Rp‐8‐CPT‐cAMPS or 5μM PKI. To determine downstream signals of the cAMP/PKA cascade, we measured myosin light chain phosphatase (MLCP) activity in these artery rings and cultured smooth muscle cells with Western blot by detecting phosphorylation of myosin‐targeting subunit protein‐1 (pMYPT‐1), the MLCP regulatory subunit. Treatment with G‐1 decreased pMYPT‐1induced by PGF2α. Both Rp‐8‐CPT‐cAMPS and PKI attenuated the decrease of pMYPT‐1 by G‐1. These findings demonstrate that cAMP/PKA signaling is involved in GPER‐mediated coronary vasodilation by activating MLCP.
High levels of serotonin modulate oxidative balance in brain's area related with anxiety behavior?
Authors: da Silva Aline Isabel , Freitas Cristiane , Nascimento Luciana , Galindo Ligia C.M. , Manhaes‐de‐Castro Raul , Lagranha Claudia , de Souza Sandra Lopes
Published: June 2021

There is abundant evidence that manipulating of serotonin system can affect innumerous behavior. Anxiety behavior's test had shown contradicted reports, some studies had shown that higher levels of serotonin increase anxiety and other shows no significant effect. In addiction, studies are showing that some behaviors alteration are linked to the reactive oxygen specie production which may induce oxidative stress. With this our aim was evaluate the effect manipulation of serotonin system during the lactation time on anxiety behavior and oxidative balance in hippocampus. The treatment was carryout during the lactation time using 10mg/Kg B.W. (i.p injection) in female Wistar rats. In vivo was evaluated anxiety behavior using elevated plus‐maze (EPM) test and in vitro we evaluate the levels of oxidative stress (Malondialdeyde‐MDA), antioxidant enzyme (superoxide dismutase‐SOD, catalase activity‐CAT and glutathione S‐transferase‐GST)on hippocampus. Our data suggest that modulation of serotonin levels by fluoxetine during developmental period decrease levels of oxidative stress accompanied by a reduction in anxiety levels. The present study receives financial support from FACEPE and CNPq.
Effect of cilostazol on patients with intermittent claudication: role of endothelial progenitor cells
Authors: Tejerina Teresa , Ramajo Marta , Navarro‐Dorado Jorge , Medina Ursula , Rydings Manuel H , Perera Miriam , Puras Enrique , Serrano‐Hernando Francisco J , Redondo Santiago
Published: June 2021

Cilostazol is an emerging vasodilatory drug which has been approved for the symptomatic treatment of intermittent claudication (CI). On the other hand, a growing body of evidence highlights the importance of endothelial progenitor cells in vascular repair. We aimed to study the effect of cilostazol on endothelial progenitor cells (EPCs) levels and clinical markers of a cohort of 24 patients with CI at baseline and after 6 months of treatment with the drug. EPCs were measured as CD34+/KDR+ and CD34+/CD144+ mononuclear cells by flow cytometry. We found that patients treated with cilostazol had an increase of CD34+/KDR+ from 0.053±0.009% to 0.128±0.029%, p=0.0161, whereas CD34+/CD144+ rose from 0.178±0.032% to 0.228±0.060, p=0.47. Moreover, a subgroup of patients who discontinued cilostazol had a trend towards lower EPC number. Cilostazol treatment was associated to a better clinical profile (maximum walking distance and ankle‐brachial index). However, in the whole cohort EPCs numbers did not show a significant correlation compared to these two clinical outcomes. In conclusion, cilostazol was associated to a better clinical profile and an increase of CD34+/KDR+ EPCs in patients with CI.

FISS (Health Research Fund, PI080920) and Red Tematica de Investigacion Cardiovascular RECAVA (RD06/0014/1007), Spanish Ministry of Health and Ferrer Group.
Chemopreventive Role of Metformin during DMBA–induced Breast Carcinogenesis in MCF10A Cells through CYP1A1‐AhR Signaling Pathway
Authors: Maayah Zaid Hussain , AlHaider Abdulqader A. , Korashy Hesham M.
Published: June 2021

Recent studies have established that metformin (MET), an oral hypoglycemic drug, possesses antioxidant activity and effective against different types of cancer. Although several postulated mechanisms were reported, the role of the cytochrome P450 1A1 (CYP1A1)‐aryl hydrocarbon receptor (AhR) signaling pathway in the protective effect of MET against breast cancer has not been reported before. Therefore, we examined the capacity of MET to inhibit CYP1A1 gene expression and DNA adducts formation in human breast cells MCF10A by dimethylbenz[a]anthracene (DMBA), a well‐known breast chemical carcinogen. Our results showed that DMBA‐induced DNA adduct was completely prevented by MET as evidenced by decrease in DNA repair genes and 8‐hydroxy‐2′‐deoxyguanosine level. Importantly, MET inhibited the induction of CYP1A1 mRNA, protein and activity levels by DMBA, suggesting a direct role of CYP1A1. Mechanistically, this was strongly correlated with MET ability to inhibit AhR‐dependent luciferase activity, but there was no effect on AhR activation, as evidenced by binding assay, suggesting that MET is not an AhR ligand. In conclusion, the present work provides the first evidence that MET inhibits the DMBA‐mediated carcinogenicity by inhibiting the expression of CYP1A1 through an AhR ligand‐independent mechanism. This work was supported by the Deanship of Scientific Research grant no. RGP‐VPP‐141.
Pro‐apoptotic agents induce ATP release from leukemia/lymphoma tumor cells via pannexin‐1 dependent and pannexin‐1‐independent mechanism
Authors: Boyd‐Tressler Andrea Michelle , Dubyak George R
Published: June 2021

Extracellular ATP released from dying tumor cells treated with chemotherapeutic drugs plays a significant role in modulating antitumor immune responses. However, the mechanisms by which ATP is released from such tumor cells remain poorly defined. Physiological apoptosis induced by Fas signaling can elicit ATP release via pannexin 1 (panx1) channels gated by caspase 3/7 (casp3/7)‐mediated proteolytic cleavage of the panx1 C‐terminus. We used human Jurkat leukemia and murine EG7 lymphoma cells to test whether the pro‐apoptotic drugs doxorubicin (Dx) and staurosporine (STS) elicit the same casp3/7>; panx1>; ATP release pathway. Cells were treated with Dx or STS for 1–18 h in the presence or absence of the pan‐caspase inhibitor zVAD and then assayed for accumulation of extracellular adenine nucleotides (ANex). Apoptotic progression was monitored by accumulation of active casp3/7 and cleavage of PARP. Apoptotic progression in response to STS or Dx were characterized by similar kinetics and efficacies in both tumor cell models. STS and Dx‐induced ATP release from Jurkat cells was markedly suppressed by zVAD. In contrast, STS and Dx‐induced ATP release from EG7 cells was completely insensitive to zVAD despite very high expression of panx1 protein in this tumor model These findings indicate that the ability of pro‐apoptotic drugs to activate the casp3/7>; panx1>; ATP release pathway in hematopoietic cancer cells is tumor‐model specific and is not well‐correlated with panx1 protein expression. (Support: NIH‐R01‐GM3637).
Apoptotic mechanism of novel anticancer agents is mediated by MAPKs in breast cancer cells
Authors: McLean Lancelot Stuart , Brantley Eileen
Published: June 2021

Aminoflavone and 5F203 are two potential candidate anticancer agents. While their mechanism of action is not fully elucidated, aspects of their anticancer activity differ from currently available agents. Even though the aryl hydrocarbon receptor (AhR) signaling pathway is the primary known mechanism of action of these agents, other signaling pathways may also be involved. Our objective was to investigate the apoptotic mechanism of AF and 5F 203 in MDA‐MB‐468 and T47D breast cancer cell lines. One group of cells were treated time dependently with vehicle or AF or 5F203. A second group of cells were pretreated with MAPK inhibitors, followed by AF or 5F203. Cells were then analyzed for MAPK phosphorylation, apoptosis and ROS formation. Our results showed that AF and 5F203 induced phosphorylation of ERK1/2, JNK1/2 and p38 in MDA‐MB‐468 and T47D breast cancer cell lines. We also showed that ERK, JNK and p38 inhibitors attenuated the apoptotic effect of AF and 5F203. Additionally, AF and 5F203‐induced ROS production was decreased by the MAPK inhibitors. These results suggest that the MAPK signaling pathway is involved in the mechanism of action of AF and 5F203 in MDA‐MB‐468 and T47D cells.

These studies were supported in part by the School of Dentistry and Medicine at Loma Linda University
Oncolytic adenoviral therapy enhanced by targeting cyclin E overexpression and inducing autophagy
Authors: Cheng Pei‐hsin , Rao Xiao‐Mei , McMasters Kelly M. , Zhou Heshan Sam
Published: June 2021

Oncolytic virotherapy is a new potential approach in cancer therapy. Here we aim to enhance the efficacy of this drug platform with two approaches. First, we used the cyclin E promoter to control the E1a gene of a tumor‐specific oncolytic adenovirus (Ad). So far, most tumor‐specific promoters are relatively weak and only Ad dl1520 with its native E1a promoter has been approved for the cancer treatment. We have shown that the activity of cyclin E promoter is high in cancer cells and further enhanced by Ad infection. Thus, we constructed a novel oncolytic Ad‐cycE which specifically targets cancer cells and self‐enhance during the viral replication process. Ad‐cycE is competent to induce oncolysis in multiple cancer cells comparable to the wild‐type Ad and Ad dl1520. Our animal studies also illustrated the promising efficacy of Ad‐cycE on repressing lung tumors growth. Secondly, we applied autophagy to improve the oncolytic Ad‐cycE replication. We have demonstrated that Ad‐induced autophagy helps Ad replication and is related to cell death. Here we further revealed that an autophagy inducer rapamycin increased adenoviral E1A expression of Ad‐cycE and increases virus production in the human lung cancer cells. Our studies provide a new insight in vector development and demonstrate the novel roles of autophagy in adenovirus replication that may benefit future combination virotherapy. Supported by NIH Grant R01 CA129975.
Effects of Fluoxetine and Aspirin on Bleeding Time and Liver When Used Alone or in Combination in Rats
Authors: Shrestha Sachita , Sookvanichsilp Nongluck
Published: June 2021

Objectives

To investigate the effect of fluoxetine and aspirin, alone or in combination, on bleeding time and hepatotoxicity.

Methods

The antiplatelet effect of fluoxetine (10mg/kg, orally) alone or combination with aspirin (25 mg/kg, orally) after 4‐week treatment in male Wistar rats (200–250g) was determined using established rat's tail bleeding model. Hepatotoxic biomarkers and liver histological patterns were evaluated. The experimental protocol was approved by the Institutional Animal Care and Use Committee.

Results

Bleeding time tended to be prolonged by aspirin but not significantly compared to control. Fluoxetine did not change the bleeding time. Treatment with concomitant aspirin and fluoxetine exhibited a trend to shorten the bleeding time. Serum ALT and ALP but not total bilirubin the aspirin group seemed to be higher than in other groups although significant differences could not be detected. Concomitant aspirin and fluoxetine could prevent the increase in these two enzyme levels. Adverse alterations in liver morphology were observed in all treated groups compared to control.

Conclusion

Fluoxetine exhibits a trend to shorten the prolonged bleeding time and prevents the increase in serum ALT and ALP in aspirin treated rats suggesting possible drug interaction upon the concomitant administration. This study was supported by Thailand International Development Co‐operation Agency
The balance between neural and hemodynamic factors is abolished in hypertensive men
Authors: Hart Emma , Burchell Amy , Ratcliffe Laura , Baumbach Andreas , Nightingale Angus , Paton Julian FR
Published: June 2021

Bristol, Bristol, United Kingdom In normotensive men there is a balance among sympathetic nerve activity (SNA), peripheral resistance (TPR) and cardiac output (Q). Thus, the net effect that high resting levels of SNA has on blood pressure (BP) is minimal. SNA is elevated in resistant hypertensive men, despite treatment with three or more medications. We hypothesised that there would be an imbalance between SNA, TPR and Q in resistant hypertensive men. In 10 (age; 53 ± 4) resistant hypertensive men, we measured muscle SNA (peroneal microneurography), BP (cuff and Finapres) and heart rate (HR, ECG) during 10 minutes of supine rest. Stroke volume was calculated using Modelflow analysis on the Finapres BP waveform. Q was calculated as stroke volume × HR. MSNA was positively associated to TPR (r = 0.59, P<0.05), but not Q (r = 0.13). Thus mean BP was directly related to MSNA (r = 0.56, P<0.05). In summary, despite treatment with more than 3 anti‐hypertensive medications, high MSNA was related to high TPR, which was not balanced by lower Q (as it is in normotensive men). Consequently, sympathetic activity may continue to play a role in resistant hypertension despite treatments targeting the sympathetic nervous system. British Heart Foundation funded.
Maternal choline supplementation programs offspring choline metabolism in a mouse model of Down syndrome
Authors: Yan Jian , Ginsberg Stephen D , Powers Brian , Alldred Melissa J , Saltzman Arthur , Strupp Barbara J , Caudill Marie A
Published: June 2021

Maternal choline supplementation (MCS) induces lifelong cognitive benefits in the Ts65Dn mouse, a model of Down syndrome and Alzheimer's disease. To gain insight into the mechanism(s) underlying these beneficial effects, we conducted a study to test the hypothesis that MCS alters brain choline metabolism in adult Ts65Dn offspring. Ts65Dn dams were randomly assigned to a control diet or one supplemented with additional choline (1.1 g/kg or 5.0 g/kg choline chloride) throughout pregnancy and lactation. After weaning, the offspring consumed standard rodent chow. At ~6–7 mo of age, deuterium labeled methyl‐d9‐choline (1.6 mM) was administered as a tracer for 8 wks in the drinking water of the female offspring (Ts65Dn and disomic littermates) and brain tissues were harvested. Choline‐supplemented Ts65Dn mice had higher (P < 0.05) enrichment of all examined choline metabolites (e.g., d9‐acetylcholine) and a higher (P < 0.05) enrichment ratio of d3‐choline/d3‐glycerophosphocholine (GPC), d3‐GPC/d3‐phosphatidylcholine (PtdCho), and d3‐choline/d3‐PtdCho in the basal forebrain, hippocampus, and neocortex compared to unsupplemented Ts65Dn mice. No effect of MCS was detected in normal disomic littermates. These data suggest that MCS produces lifelong changes in choline metabolism in female Ts65Dn offspring with evidence of enhanced uptake and use of choline (and its methyl groups) in several brain regions.

Funded by HD057564, AG14449, and the Alzheimer's Association
Acute Renal Medullary Interstitial Infusion of either saline or 2% albumin indistinctly rises RIHP and RMBF in the rat
Authors: Sánchez‐Briones Maria Eugenia , López‐Rodríguez Juan Francisco , Calvo‐Turrubiartes Miriam Zarahí , Rodríguez‐Martínez Manuel
Published: June 2021

The effects of an acute renal medullary interstitial infusion (RMII) of 100 μl of either 0.9% saline (Sal) or 2% albumin in saline (Alb) on the time course (30 min) of renal interstitial hydrostatic pressure (RIHP, subcapsular), renal medullary blood flow (RMBF, L‐D), urinary nitrogen oxides excretion (UNOxV, Griess) and urinary flow rate (V) were assessed in the left denervated kidney of anesthetized male Wistar rats under control of renal perfusion pressure at 100 mmHg. After a 10 min control period (CP) and a 15s RMII period, three 10 min post‐infusion periods (P‐IP) of urinary collections were generated. The results are mean ± SEM; ^ p<0.05 vs. CP.

Group
CP
RMII
P‐IP

10min
20 min
30 min

RIHP (mmmHg)
Sal
3.0
3.8
3.9
5.3
6.8

n=17
± 0.4
± 0.5
±0.5^
± 0.7^
± 0.8^

Alb
3.0
3.2
3.5
4.0
4.8

n=17
± 0.3
± 0.3^
± 0.3^
± 0.5^
± 0.7^

RMBF (%Δ)
Sal
0.0
2.0
10.7
12.5
8.2

n=17

± 3.4
± 3.3^
± 4.0
± 4.3

Alb
0.0
2.2
9.1
6.6
8.7

n=17

± 5.0
± 4.5^
± 4.7
± 5.2

There were no differences between groups in UNOxV nor in V through time.

Conclusions

Acute RMII of either 0.9% saline or 2% albumin indistinctly rises RIHP and RMBF in the anesthetized rat, without affecting UNOxV nor urinary flow rate at all. Supported by IPL's financial resources.
Vacuolar proton pump a4 subunit is critical for inner ear development and renal function
Authors: Picard Nicolas , Hennings Jean‐Christopher , Huebner Antje , Stauber Tobias , Maier Hannes , Brown Dennis , Jentsch Thomas , Vargas‐Poussou Rosa , Eladari Dominique , Huebner Christian
Published: June 2021

Renal distal tubular acidosis (dRTA) has been associated with human ATP6V0A4 (a4 subunit of the proton pump) gene mutations. This disease is characterised by acidic blood pH, low blood bicarbonate and potassium concentrations, and often, hypercalciuria leading to nephrocalcinosis. Some patients with ATP6V0A4 mutations also develop late onset senso‐neurinal deafness.

We generated a mouse model deficient for the a4 subunit. Survival rate of KO mice was dramatically reduced and did not exceed 7–8 weeks of age. Mouse deficient for the Atp6v0a4 gene (a4−/−) recapitulated most of the renal impairment seen in patients with dRTA since they exhibited life‐threatening metabolic acidosis consecutive to a marked reduction in renal acidification capacity. This was associated with complete hearing loss associated with a strong enlargment of the endolymphatic sac of the inner ear. In the kidney, a4 subunit has been localised in the proximal tubule and in the intercalated cells of the collecting duct which are two renal segments involved in acid‐base balance. Histo‐morphological analysis of a4−/− mouse kidneys revealed a profound lysosomal remodelling in the proximal tubule associated with an abnormal trafficking of the proton pump in both proximal tubule and intercalated cells. Their low molecular weight proteinuria was explained by a general proximal tubule dysfunction. We conclude that A4 subunit is a mandatory component for the assembly of the vacuolar proton pump ATPase complex and is critical for inner development, distal acidification and proximal tubule absorptive capacity.
Testosterone increases muscle sympathetic nerve activity in young healthy males
Authors: Miner Jonathon Cory , Miner Jennifer Ann , Kaplan Paul F. , Minson Christopher Todd
Published: June 2021

A positive correlation between levels of testosterone and muscle sympathetic nerve activity (MSNA) has been found in women with polycystic ovarian syndrome, and is associated with cardiovascular risk factors. However, testosterone has been shown to be cardio‐protective in male rats. To date, we are unaware of any studies that have tested the direct impact of testosterone administration on MSNA in young, healthy males. The purpose of the current study was to examine the acute effects of short‐term testosterone administration on MSNA on this demographic.

Methods

MSNA (via microneurography), heart rate, and beat‐by‐beat blood pressure were recorded following a venous blood draw for baseline testosterone levels in four young, healthy male subjects. After assessing resting measures, each subject self‐administered 120 mg/day of transdermal testosterone for seven days and were then retested. We analyzed MSNA frequency (bursts min−1) and incidence (bursts 100 heartbeats−1).

Results

Testosterone level increased in all subjects (676±47 ng/dl to 774±16 ng/dl). Increased levels of total testosterone tended to increase MSNA frequency, (11±3 to 15±1, p=0.10) and incidence (22±4 to 27±3, p=0.18).

Conclusion

Short‐term, transdermal testosterone administration tends to increase MSNA frequency and incidence in young, healthy males. Supported by NIH Grant HL081671.
Impaired hypoxic vasodilation in healthy older adults: role for altered sympatho‐adrenal control of vascular tone
Authors: Richards Jennifer Clarke , Crecelius Anne R. , Kirby Brett S. , Garcia Leora J. , Luckasen Gary J. , Larson Dennis G. , Dinenno Frank A.
Published: June 2021

We tested the hypothesis that peripheral vasodilation to graded systemic hypoxia (90, 85, 80% SaO2) is impaired in older healthy adults due to attenuated β‐adrenergic mediated vasodilation and elevated α‐adrenergic vasoconstriction. Forearm blood flow was measured (Doppler ultrasound) and vascular conductance (FVC) was calculated in 12 young and 8 older adults to determine the dilatory response to hypoxia in control, local (brachial artery catheter) β‐blockade (propranolol), and combined α+β blockade (phentolamine+propranolol) conditions. Compared to young, older individuals exhibited impaired vasodilation to hypoxia (peak Δ+35±8% vs. +9±6 %ΔFVC P<0.05). Following β‐blockade, older adults actively constricted (−14±5%ΔFVC) whereas the young response was not significantly impacted. α+β blockade increased the dilatory response to hypoxia in both groups, however the dilatory response remained significantly greater in young compared with the old (+58±11% vs.+17±11%ΔFVC P<0.05). Our findings indicate that hypoxic vasodilation is significantly impaired in older adults and this can be attributed to lower β‐receptor mediated dilation, elevated α‐adrenergic constriction and lower local vasodilatory signaling. These findings may have implications for understanding regional blood flow control in aging patient populations that exhibit systemic and/or local hypoxia.

Supported by HL102720
Role of NOS in early‐stage BBB disruption following transient focal cerebral ischemia
Authors: Jiang Zheng , Li Chun , Arrick Denise M. , Yang Shu , Sun Hong
Published: June 2021

Background

Early‐stage BBB disruption following ischemic stroke is an antecedent event to infarction and hemorrhagic transformation. We determined the role of NOS in early‐stage BBB disruption using a new model of transient focal cerebral ischemia.

Methods

A cranial window was prepared over the left parietal and temporal cortex of C57BL/6J mice. To induce ischemia, the MCA was ligated at M2 segment just proximal to its bifurcation for 2 hours. Reperfusion was induced by releasing the ligation. L‐NAME, L‐NPA, 7‐NI and aminoguanidine alone or with papaverine were topically perfused to the cranial window during the reperfusion. The cerebral vasculature was imaged at 5, 10, 30, 60, 90, 120, 150 and 180 minutes of reperfusion and diameter change in all branches of the MCA, anastomosis and terminal branches of the collateral arteries were calculated. BBB disruption was assessed by measuring extravasation of Evans Blue (EB) and sodium fluorescein (Na‐F) at 3 hours of reperfusion.

Results

L‐NAME abolished reperfusion‐induced vasodilation in all branches of the MCA, anastomosis and terminal branches of the collateral arteries, whereas L‐NPA, 7‐NI and aminoguanidine only inhibited the vasodilation in large size branches of the MCA and anastomosis. On the other hand, L‐NAME, L‐NPA, 7‐NI and aminoguanidine all reduced extravasation of EB and Na‐F. In addition, papaverine abolished L‐NAME‐induced reduction in extravasation of Na‐F.

Conclusions

All three types of NOS may contribute to early‐stage BBB disruption directly and indirectly via NOS‐mediated hyperemia following transient focal cerebral ischemia.
Role of Fibrinogen in Traumatic Brain Injury
Authors: Muradashvili Nino , Tyagi Reeta , Benton Richard L. , Lominadze David
Published: June 2021

Elevated level of fibrinogen (Fg) ‐ hyperfibrinogenemia (HFg) exacerbates circulatory complications during inflammatory diseases such as stroke, hypertension, traumatic brain injury (TBI). We already showed that HFg alters vascular reactivity and compromises endothelial cell layer integrity causing a leakage of plasma substances into the interstitium. We tested the hypothesis that TBI increases Fg deposition and cerebrovascular permeability in injury penumbra through activation of matrix metalloproteinase‐ 9 (MMP‐9).

Permeability of pial venules and Fg deposition in injury penumbra formed after a mild cortical contusion injury (CCI) were studied in wild type (WT, C57BL/6J) and MMP‐9 gene knockout (MMP9−/−) mice. Fg deposition and formation of caveolae, examined by the extent of co‐localization of caveolin‐1 and plasmalemma vesicle‐associate protein‐1 were assessed in brain cryo‐sections. In each group, BSA leakage, caveolae formation, and Fg deposition were increased after CCI compared to those in respective sham‐operated mice. However, they were greater in WT than in MMP9−/− mice.

These data suggest that blood level of Fg is elevated after TBI and its increased deposition intensifies cerebrovascular protein leakage mainly through caveolar transcytosis via activation of MMP‐9. The results of our study point to a new therapeutic targets that can be used to assist the post‐TBI healing process.
Lactational DEHP Exposure‐Impaired Motor Coordination and Motor Skill Learning in Adolescent Rats
Authors: Yueh Shu‐Chieh , Yang Rei‐Cheng , Wang Dean‐Chuan
Published: June 2021

Di‐(2‐ethylhexyl) phthalate (DEHP) migrates from the polyvinyl chloride (PVC) devices to the environment and is considered as an environmental contaminant. The purpose of this study is whether neonatal DEHP exposure affects functions of the cerebellum. Sprague‐Dawley female rats were orally fed with DEHP (10 mg/kg/day) or corn oil everyday during lactational period. After weaning (postnatal day 22), the male pups were divided into 2 groups: control and DEHP. At 8 weeks old rats were trained on the rotarod to evaluate the motor coordination and then trained on a runway to evaluate the cerebellar‐dependent motor skill learning. After a 7‐day motor skill learning paradigm, rats were sacrificed and their cerebella were isolated for detection of Purkinje cells‐specific marker calbindin D28K, an important calcium buffer in the neurons, by Western blotting. The results show that motor coordination and motor skill learning were both impaired in the DEHP group compared to the Control group. The expression of calbindin D28K was increased significantly in the DEHP group, indicating a disturbed calcium homeostasis in the cerebellum of DEHP‐exposed rats. These data demonstrate that neonatal DEHP‐exposure may induce cerebellar dysfunction in adolescent rats.
Control of feeding behavior by CRH 2 receptors in the paraventricular nucleus of the hypothalamus of adrenalectomized rats
Authors: Rito‐Domingo Melisa , Alonso Verónica Elsa López , Santos Karla Araceli Reyes , Segundo Gabriela Ambrocio , García Karina Cruz , Díaz Juan Manuel Mancilla
Published: June 2021

The Corticotropin releasing hormone (CRH), play an important role in the normal maintenance of food intake, body weight and energy metabolism. CRH is a powerful inhibitor of feeding. Urocortin 2 (Ucn 2) a CRH like peptide mediate this effect through CRH 2 receptors expressed in the paraventricular nucleus of the hypothalamus (PVN), but it is not yet know the behavior mechanism and effect on glucose and triglycerides, that acts in the presence of agonist Ucn 2 and antagonist Antisauvagine 30 (Asv 30). The aim of the present study was determinate the role of CRH 2 receptors in food intake feeding behavior and in concentration of glucose and triglycerides of adrenalectomized Wistar rats. 34 male rats of Wistar strain of 250–300 g were accustomed to a diet of separate sources of protein, carbohydrate and fat, then were implanted a cannula on right PVN and a second surgery to remove the adrenal glands bilaterally (ADX) or sham surgery (sham). After the post‐surgery recovery period of 5 days, the rats were assigned to a group and a treatment, sham (vh+vh), ADX (vh+vh), ADX Agonist (vh+Ucn 2), ADX antagonist (Asv 30+vh) and ADX pretreatment (Asv 30+Ucn 2). A record of continuous duration of 60 min was made for the analysis of the behavioral satiety sequence (BSS). The results showed that administration of Ucn 2 reduced carbohydrates intake and promotes the transition of feeding and resting in the BSS, The pretreatment with Asv‐30 delayed the transition of feeding and resting. The glucose and triglycerides concentration decrease with the administration of Ucn‐2 and the antagonist reverse this effect. In conclusion the results of this study suggest that CRH 2 receptors of the PVN are important for the regulation of feeding, glucose and triglycerides in adrenalectomized rats.

Research support: DGAPA IN306711
Stress and immune function of social drinking college students before and after spring break
Authors: Sharma Shobhit , Howard Krista , Graham Reiko , Ceballos Natalie
Published: June 2021

Numerous studies have focused on the social aspects of collegiate holiday drinking; however, physiological measures are frequently overlooked. This project examined stress and immune function among collegiate social drinkers before and after spring break (SB). Saliva and questionnaires were collected from 29 students (9 males) at baseline and post‐SB. Participants were categorized as cortisol non‐responders (CNR; n=14) or cortisol increasers (CI; n=13) depending on their change in cortisol over SB (< 0.03 μg/dL change = CNR; ≥ 0.03 μg/dL change = CI). CNRs and CIs were matched on gender, age, baseline alcohol use and family history of alcoholism. Repeated measures ANOVAs examined differences over time and between groups on quantity/frequency of alcohol use (QFI), IgA and DHEAS. Time x group interactions were noted for QFI (F(1,22)=4.96; p=.04) and IgA (F(1,19)=5.27; p=.03). CNRs doubled their alcohol use over SB (p=.02) and CIs decreased in IgA over SB (p<.05). Changes in QFI and IgA were positively correlated among CIs (r=.63; p=.04) but were unrelated among CNRs. Compared to CNRs, CIs exhibited higher DHEAS levels overall. Blunted cortisol responses have been previously associated with risk for addictive disorders. The current results highlight the importance of physiological variables to the study of collegiate holiday drinking.
Longitudinal assessment of systolic and diastolic function using conventional and speckle tracking echocardiography following acute ischemia and prolonged reperfusion in the rat
Authors: Bao Weike , Wright Fe , Ao Zhaohui , Mirabile Rosanna C. , Frazier Kendall S. , Coatney Robert W. , Schnackenberg Christine G. , Jucker Beat M.
Published: June 2021

Speckle tracking echo strain imaging has not been used longitudinally to assess both systolic and diastolic function following myocardial ischemia (I)/reperfusion (R) injury. We tested the hypothesis that strain imaging can precisely detect systolic and diastolic dysfunction earlier than conventional echocardiography in rats following a 30 min I and 4 weeks R. Conventional and speckle tracking echocardiography were performed prior to I (baseline), at 48 hr and at 2 and 4 weeks after R. I/R injury reduced ejection fraction by 20%, 18% and 22%, at 48 hr, 2 and 4 weeks, respectively (all p<0.05 vs. sham, n=6–10). Global systolic circumferential strain decreased by 29%, 26% and 29%, respectively (all P<0.05); segmental systolic circumferential strain was reduced by 80%, 46% and 41% (all P<0.001). Conventional Doppler echocardiography showed no difference in IVRT and E/E OE (p=NS vs. sham) at any time points following R. In contrast, global and segmental diastolic circumferential strain rate were significantly reduced at all time points following R. At 4 weeks, I/R injury also significantly reduced cardiac reserve, markedly increased cardiac fibrosis and apoptosis, up‐regulated NCX1 and down‐regulated SERCA‐2a protein expression. Our results demonstrate that speckle tracking echocardiography provides a more sensitive technique for early detection of both systolic and diastolic dysfunction in the failing heart.
CD40‐CD40 ligand (CD40L) signaling modulates cardiac hypertrophy in angiotensin‐II induced hypertensive heart disease
Authors: Derks Wouter , Lutgens Esther , van Bilsen Marc , Heymans Stephane
Published: June 2021

The co‐stimulatory dyad CD40‐CD40L is involved in pro‐inflammatory activation of a wide range of immune and non‐immune cells. Here we investigated whether CD40‐CD40L interactions attenuate cardiac hypertrophy in hypertensive heart disease. Therefore CD40 knockout and wild type C57Bl6‐J mice were treated with angiotensin II (AngII) (2.5 mg/kg/day) for 4 weeks. AngII‐induced hypertension led to a significant increase in cardiac mass in Wt mice (+31%, P<0.01), but not in CD40 knockout mice (+10%, N.S.). Comparable changes were observed in diastolic wall thickness (echocardiography) and cardiomyocyte cross sectional area. To determine which CD40‐TRAF interactions were involved, the effect of mutations in the intracellular TRAF binding domains of CD40 was investigated. Relative to mice with intact downstream CD40 signaling, disruption of TRAF2/3/5, TRAF6 or both binding sites was associated with reduced hypertrophy (cardiac mass −24% for CD40‐TΔ2, −28% for CD40‐ TΔ6 and −26% in CD40‐TΔ2/6; p<0.05). Reduced hypertrophy was not associated with a decline in immune cell infiltration (CD45+ cells) or collagen content. Although the exact mechanism remains to be elucidated the present findings indicate that CD40‐ CD40L interactions are involved in the development of cardiac hypertrophy.
Angiotensin II‐induced myocardial hypertrophy and fibrosis are characterized by enhanced MMP‐9 and suppressed RECK expression
Authors: Siddesha Jalahalli M. , Valente Anthony J. , Sakamuri Siva S.V.P. , Chandrasekar Bysani
Published: June 2021

Chronically elevated angiotensin (Ang)‐II contributes to myocardial hypertrophy, fibrosis and adverse remodeling, conditions characterized by enhanced MMP expression. RECK (Reversion‐inducing‐cysteine‐rich protein with kazal motifs) is a novel membrane‐anchored MMP inhibitor. Here we investigated whether Ang‐II differentially regulates RECK and MMPs in vivo in mouse heart and the underlying molecular mechanisms in vitro in adult cardiac fibroblasts (CF). Continuous infusion of Ang‐II for 2 weeks induced myocardial hypertrophy, fibrosis, and enhanced MMP‐2/9 expression. In contrast, Ang‐II suppressed myocardial RECK expression. These responses were inhibited by losartan, an AT1 antagonist. Using pharmacological inhibitors and adenoviral overexpression of mutant proteins, we show that treatment of CF with Ang‐II induced AT1/Nox4 dependent ROS generation, ERK1/2 activation, and ERK/NF‐κB/AP‐1‐dependent MMP9 induction. Notably, Ang‐II suppressed RECK expression through both ERK/Sp1 activation and AP‐1/miR‐21 induction. Ang‐II induced CF migration, an effect reversed by RECK overexpression. These results demonstrate that Ang‐II induces MMPs, but suppresses RECK both in vivo and in vitro, and RECK overexpression blunts Ang‐II induced CF migration in vitro. Strategies that upregulate RECK expression may attenuate fibrosis and adverse remodeling in hypertensive heart disease.