bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of chromogranin A by hsa‐miR‐107 in hypertension and nephropathy (1136.19)
Authors: Vaingankar Sucheta , Mir Saiful , Zhang Kuixing , Hightower C. Makena , Miramontes‐Gonzalez Jose , O'Connor Daniel
Published: June 2021

Chromogranin A (CHGA) is a major protein in secretory vesicles of neuroendocrine cells, co‐released with catecholamines. Genetic variation in the CHGA 3’‐UTR region is associated with hypertensive nephropathy. This study documents the biological mechanism underlying the genetic association of marker and trait.

Methods and Results: An African‐American population case/control study showed that individuals carrying the CHGA 3’‐UTR variant C+87T (rs7610) haplotype are likely to develop end stage renal disease. Computational analysis showed that the variant is in a micro‐RNA (hsa‐miR‐107) recognition motif, with a superior match for the risk (T) allele.

In vitro expression studies revealed that CHGA mRNA expression is inversely dependent on miR‐107 abundance in cells and tissues. The effects of miR‐107 were seen with transfected chimeric luciferase/CHGA 3’‐UTR (C+87 versus +87T) reporters in cell lines and corroborated in studies of coupled in vitro transcription/translation of each 3’‐UTR/C+87T allele in CHGA cDNA.

In vivo, we exploited mice with a “humanized”CHGA locus (T/T homozygotes) generated by BAC transgenesis. These mice treated with miR‐107 antagomir yielded prolonged, significant falls in SBP/DBP compared to the wild‐type mice.

Conclusion: The 3’‐UTRmRNA:miRNA‐107 (cis:trans) interaction plays a role in the clinical manifestations of the CHGA 3’‐UTR associations with BP and nephropathy.

Grant Funding Source: Supported by National Institutes of Health (SMV & DTO'C)
Parallel changes in the urinary excretion of ANG II and angiotensinogen and kidney ANG II levels in slowly progressive ANG II‐dependent hypertension (1136.20)
Authors: Sigmon David , Seth Dale , Sato Akemi , Davis Porcha , Navar L , Mitchell Kenneth
Published: June 2021

Previous studies have demonstrated that the urinary excretion of ANG II and AGT, and kidney ANG II levels are markedly elevated in ANG II‐dependent malignant hypertension associated with marked renal injury. The present study was performed to determine the temporal changes in urinary ANG II and AGT excretion and kidney ANG II levels in a model of slowly progressive ANG II‐dependent hypertension in Cyp1a1‐Ren2 transgenic rats with inducible expression of the Ren2 renin gene. Male Cyp1a1‐Ren2 rats (n=6/group) were induced to develop slowly progressive ANG II‐dependent hypertension by dietary administration of indole‐3‐carbinol (I3C, 0.15% wt/wt) for 14 days. Conscious systolic blood pressures (SBP) increased from 140±8 to 176±9 mmHg (P<0.05) by day 4 of I3C induction, whereas the urinary excretion of ANG II and AGT both increased by day 3 of I3C induction (32.1±3 to 64.8±6 fmol/hr and from 123±25 to 437±75 ng/day, P<0.05 in both cases). Both urinary ANG II and AGT excretion continued to increase (P<0.01) and averaged 120±5 fmol/hr and 1384±514 ng/day, respectively, by day 13 of induction. Similarly, kidney ANG II levels were increased by day 13 of induction (1042±219 vs. 349±96 fmol/g, P<0.05). SBP, urinary ANG II and AGT excretion and kidney ANG II levels promptly decreased to control levels following termination of dietary I3C administration (to 127±2 mmHg, 60±5 fmol/hr, 83±34 ng/day, and 473±185 fmol/g, respectively, P<0.01 in all cases). The present findings demonstrate that the urinary excretion of ANG II and AGT increases progressively, and that total kidney ANG II levels are increased, following induction of ANG II‐dependent hypertension and decrease within 3 days to control levels following termination of dietary I3C administration. The data also indicate that the intratubular generation of ANG II increases progressively following induction of ANG II‐dependent hypertension and decreases promptly to control levels following termination of induction of ANG II‐dependent hypertension.

Grant Funding Source: 5P30GM103337
Disruption of the WNK/SPAK pathway attenuates angiotensin II, but enhances aldosterone‐induced hypertension (1136.3)
Authors: Cervantes‐Perez Luz , Castañeda‐Bueno María , Vázquez Norma , Arroyo Isidora , Bobadilla Norma , Alessi Dario , Gamba Gerardo
Published: June 2021

It is known that angiotensin II (AngII) hypertensive effect is dependent on the intrarenal actions of this peptide hormone. On the other hand, we have proposed that AngII effect on the renal NaCl cotransporter, NCC, requires integrity of the WNK/SPAK pathway. We thus tested the effect of AngII or aldosterone administration on blood pressure in SPAK knockin mice in which the T243A mutation prevents the WNK activation of SPAK and thus, NCC activation.

AngII or aldosterone was infused at 1440μg/kg/d or 700μg/kg/d, respectively, in SPAK wild type and SPAKT243A knockin mice for 14 days by osmotic minipumps. Aldosterone treated mice were exposed to NaCl drinking water (1%) during the hormone administration. Arterial blood pressure was assessed by radiotelemetry. Electrolyte excretion after diuretic challenge and NCC protein expression and phosphorylation was assessed on day 14. Our results show that hypertensive effect of AngII was blunted by at least half in SPAK‐knockin mice, together with absence of AngII induced NCC phosphorylation. In contrast, the hypertensive effect of aldosterone was enhanced. In wild type mice blood pressure was not significantly modified during aldosterone administration, while in SPAK‐knockin mice hypertension was developed. Aldosterone treatment was associated with increased response to amiloride, but not to thiazide diuretics, and no increase in NCC phosphorylation was observed. Thus, our work suggest that AngII induced hypertension requires, at least in part, NCC activation via WNK/SPAK signaling pathway, while aldosterone induced hypertension depends on ENaC activation that is WNK/SPAK independent. The SPAK‐KI mice emerges as a useful model to distinguish between the AngII vs aldosterone effects on distal nephron.
Nephron‐specific disruption of Cullin3 causes salt‐sensitive hypotension (1136.7)
Authors: McCormick James , Terker Andrew , McCully Belinda , Meermeier Nicholas , Yarbrough Bethzaida , Park Hae , Yang Chao‐ling , Singer Jeffrey , Ellison David
Published: June 2021

Familial Hyperkalemic Hypertension (FHHt) is caused by mutations in genes that regulate activity of the renal sodium chloride cotransporter (NCC), including the kinases WNK1 and WNK4, and Cullin3 (Cul3) and KLHL3, members of a ring ubiquitin ligase complex. To examine the physiological roles of Cul3 and how mutations in it cause FHHt, we generated Cul3‐/‐ mice. An inducible system that disrupts Cul3 specifically along the entire nephron was used since complete Cul3 disruption is embryonic lethal. Western blots showed nearly complete absence of Cul3 in Cul3‐/‐ kidneys. Cul3‐/‐ mice displayed volume contraction, hypochloremia and alkalosis, and elevated plasma aldosterone. Blood pressure, measured by radiotelemetry, was similar in both groups on normal NaCl diet, but during NaCl restriction, Cul3‐/‐ mice displayed a progressive reduction in blood pressure. After 9 days, the 24h mean blood pressure pressure was 18 mmHg lower in Cul3‐/‐ mice. Expression of total and phospho‐NCC were slightly elevated in Cul3‐/‐ mice on normal diet, but dietary NaCl restriction did not cause their levels to increase to the same extent as it did in wild types. WNK4 expression was dramatically increased in Cul3‐/‐ mice on normal and low NaCl diets, while WNK1 and WNK3 were slightly elevated. In conclusion, these data suggest that loss of Cul3 activity along the nephron does not mimic FHHt, but leads to a mild salt‐wasting phenotype.

Grant Funding Source: KO1 DK076617
Impacts of UT‐A2 inhibition on urine composition: a mathematical model (1137.8)
Authors: Moss Rob , Layton Anita
Published: June 2021

The urea transporter UT‐A2 is expressed in the inner‐stripe segments of the thin descending limb of Henle in superficial nephrons (SDL), and in the initial inner medullary (IM) segments of the descending limb in juxtamedullary nephrons. It was previously assumed that some of the urea ascending from the IM in the ascending vasa recta was “recycled” in the SDL, which run in close proximity to the vascular bundles in rats and mice. However our model simulations suggest that when urea is actively secreted into the pars recta, a hypothetical scenario has yet to be demonstrated experimentally, there is net urea reabsorption along the SDL. We use a dynamic model of the urine concentrating mechanism to investigate the hypothesis that the role of the SDL UT‐A2 lies in the transient buildup of urea and osmolality gradient of the IM, rather than in the maintenance of the steady‐state gradients. This motivation for this hypothesis is that the transit time for urea reabsorbed in the SDL to return to the IM via the descending vasa recta is significantly shorter than for urea to traverse the thick ascending limb and subsequently be reabsorbed in the terminal collecting duct. Also, during the generation of the IM urea gradient, active urea secretion into the pars recta of the long loops may yield a transepithelial gradient favorable for urea delivery by the long thin descending limbs into the interstitium, thereby adding to the urea delivered to the IM interstitium through the terminal collecting duct. This research was supported in part by NIH grant DK‐89066.

Grant Funding Source: NIH DK089066
DNA methylation in genes related with cartilage, obesity, and thrombosis in LCPD patients (1138.7)
Authors: Lee Gyuhwi , Park Kwang‐Won , Jo Inho , Song Hae‐Ryong , Park Yoon Jung
Published: June 2021

LCPD (Legg‐Calve‐Perthes disease) is a kind of osteonecrosis of the hip found only in children. It has been associated with few genetic mutations and rather with obesity and thrombosis. This study was designed to investigate whether epigenetic alteration might be related with its etiology. We analyzed DNA methylation in genes commonly involving cartilage, obesity, and thrombosis in blood from patients and healthy donors (HD) using the quantitative MassArray analysis. Total 6 amplicons covering the promoters of Col2A1, FII, FV, and FVIII genes were measured for DNA methylation. All genes, except FVIII, showed no difference between LCPD and HD. Also, the difference in FVIII was significant, but was only about 1%, which might not result in any changes physiologically.

Therefore, the results suggested that the genes might be disturbed in the pathogenesis of LCPD, at least not by epigenetic alterations, although this study was, to our knowledge, the first trial to investigate the involvement of epigenetic alterations in LCPD.

Grant Funding Source: supported by Korean NRF (22A20130012143) & Korean HealthTech R&D(HI11C0388)
Oral cavity morphometric evaluation in 7,12 dimethylantracene administration (1139.11)
Authors: Cabeças José Ricardo , Costa Eduardo , Alves Gonçalo , Jesus Patrícia , Cabrita António
Published: June 2021

Many injuries which occur in human body is mainly caused by the action of xenobiotics present in the environment and coming into contact with the body.舲 The xenobiotics they induce a good morphological identifiable damage, also cause functional alterations of varying severity. The action of xenobiotics on the body depends on many factors among which we highlight: the chemical nature of the molecule, its concentration in the environment, the repeated contact, the gate of entry and the physiological activity of the organism. DMBA is a toxic present in the contaminated environment, in some foods and also in tobacco smoke 7. The aim of this work is to evaluate the influence of DMBA in morphological changes of the periodontal ligament.舲 In this study was used 20 male Wistar rats with eight weeks old at the beginning of the study. The rats were distributed into two groups of 10 animals each. Group I, the control group, was maintained for 8 weeks without any inducing manipulation changes. Group II was subjected to the administration of 7,12‐dimethyl‐benzanthracene (DMBA) three times a week, for 4 weeks at a dose of 20 mg / kg in each takes.舲 At the end of the administration's, the animals were sacrificed and their necropsy was made.

In group I, the value of R1 was 0.07 ± 0.07 and R2 was 0.79 ± 0.25, not checking the presence of DMBA in adipocytes. The distribution of R2 values in this group was as follows: 15.38% small, 69.23% medium,10.99% large and 3.30% very large.

In group II, the value of R1 was 0.07 ± 0.04 and R2 was 0.84 ± 0.35. The distribution of R2 values in this group was as follows: 15.07% small, 63.01% medium, 8.22% large and 12.33% very large. The animals in this group was detected the presence of DMBA in adipose tissue at all.

DMBA in the studied conditions does not significantly interfere with the periodontal ligament in morphological terms.
Postprandial blood glucose level reducing effect of low molecular weight chitosan oligosaccharide, GO2KA1, in healthy individuals (1139.12)
Authors: Jung‐Yun Lee , Hwang‐Yong Choi , Se‐Woon Lee , Yu‐Ri Kang , Sung‐Hoon Jo , Emmanouil Apostolidis , Young‐In Kwon
Published: June 2021

We investigated the effects of chitosan oligosaccharide (GO2KA1) on the level of postprandial blood glucose in adult subjects with normal blood glucose level. Postprandial blood glucose levels were tested for 2 hours after sucrose administration with or without 500 mg of GO2KA1. GO2KA1 significantly lowered the mean, maximum and minimum level of postprandial blood glucose at 30 and 90 min after meal. Additionally, GO2KA1 treatment reduced the AUC and Cmax while Tmax was significantly increased. Our findings suggest that GO2KA1 reduces postprandial blood glucose rise due to slow absorption of glucose in the small intestine, via carbohydrate hydrolyzing enzymes inhibition.
Expression of cytochrome P450 genes in human cord blood hematopoietic stem and progenitor cells (1139.7)
Authors: Xu Shuoqi , Ding Xinxin , Jiang Yongping , Wang Yanan , Ren Zhihua
Published: June 2021

The human cytochrome P450 (CYP) superfamily comprises 57 genes, which encode enzymes important in the biotransformation of numerous endogenous and exogenous compounds. The aim of this study is to characterize the expression of all CYP genes in CD34(+) human cord blood hematopoietic stem and early progenitor cells (CBHSPCs), as a first step toward assessment of their biological functions in these cells. CD34(+) CBHSPCs were purified from umbilical cord blood via antibody affinity chromatography, using magnetically‐assisted chemical separation. Purity of CD34(+) cells was assessed using fluorescence‐activated cell sorting. RNA was isolated from purified CD34(+) cells for RNA‐PCR analysis, using primers specific for each of the 57 CYPs. Of the 57 CYPs, only 14 were detected by RNA‐PCR in CD34(+) CBHSPCs, including CYP2E1, CYP2U1, CYP2R1, CYP2S1, CYP2J2, CYP4F2, CYP20A1, and CYP27A1. CYP identification was confirmed by sequencing analysis of the detected, mRNA‐derived, PCR products. Preliminary quantitative analysis of CYP transcripts using real‐time RNA revealed preferential expression of multiple CYPs, such as CYP2R1, in CD34(+) CBHSPCs, compared to total lymphocytes or CD34(‐) cells from the cord blood. Additional studies on detection of CYP protein expression are underway.

Grant Funding Source: Supported by State Scientific Key Projects for New Drug Research and Development(2011ZX09102‐010‐04)
Identification of orphan receptor GPR88, GPR99, GPR107 and GPR124 in Wistar Kyoto and spontaneous hypertensive rats (1140.13)
Authors: Calderon‐Zamora Loranda , Leon‐Sicairos Nidia , Canizalez‐Roman Adrian , Hong Enrique , Villafaña Santiago
Published: June 2021

Hypertension is a chronic disease characterized by a continuous increase of blood pressure, this is one of the most important cardiovascular diseases although there are many treatments, hypertension has been associated with 9.4 million deaths per years, therefore is necessary to study new mechanisms that could contribute to explain the hypertension. Recently, a new class of receptor has been reported that are called orphan receptors because the ligand is not known, these receptors have been associated with some pathologies. For that reason, the aim of this work is to study the distribution of orphan receptors GPR88, GPR99, GPR107 and GPR124 in kidney, heart and brain in wistar kyoto (WKY) and spontaneously hypertensive rats (SHR). We used WKY and SHR male rats with 10‐12 weeks old and we measured weight, blood pressure, cardiac frequency and mRNA from kidney, heart and brain by RT‐PCR. The results obtained showed that SHR presents higher values of systolic blood pressure compared than WKY, and that the levels of orphan receptors GPR88, GPR99, GPR107 and GPR124 are expressed in both strains. In conclusion, this study suggests that GPR88, GPR99, GPR107 and GPR124 are expressed in both strain and could be associated in the hypertension process.

Grant Funding Source: SIP‐20130041/COFAA
Modulation of the cannabinoid‐1 receptor by angiotensin II in astrocytes isolated from spontaneously hypertensive rats (1140.8)
Authors: Haspula Dhanush , Clark Michelle
Published: June 2021

Objective: To determine whether a mediator of neuroprotection, the Cannabinoid Type 1 receptor (CB1R), is altered by Angiotensin (Ang) II in astrocytes. Background: Along with an elevated Ang II level, neuroinflammation in the brainstem (BS) has been shown to shift the balance towards an augmented sympathetic drive, the latter being associated with hypertension. Astrocytes play a central role in the bidirectional modulation of inflammation Methods: BS astrocytes isolated from Spontaneously Hypertensive Rats (SHR) were used as a hypertension model to determine whether Ang II induces CB1R protein and mRNA expression. The results were compared to normotensive Wistar rats. The cells were treated with 100nM Ang II in a time‐dependent manner, and the effect of Ang II on CB1R protein and mRNA levels were measured using Western blotting and qt‐PCR techniques, respectively. Results: In Wistar cells, Ang II increased the CB1R protein and mRNA levels in a time‐dependent manner. In SHR astrocytes, Ang II down‐regulated or had no effect on CB1R protein and mRNA levels. Compared with Wistar astrocytes, the CB1R protein and mRNA levels in SHR astrocytes were significantly different. Conclusion: Our findings suggest that the CB1R is differentially regulated in SHR versus normotensive astrocytes. This interaction may serve as a potential target to restore an elevated sympathetic drive in hypertension.

Grant Funding Source: NSU PFRDG 2013‐14: Grant # 335309
CYP2C9*8 frequency distribution in Puerto Ricans: implications for warfarin dosing (1141.10)
Authors: Claudio‐Campos Karla , Orengo‐Mercado Carmelo , Berrios Lorena , Pagan Robert , Renta Jessicca , Cadilla Carmen , Duconge Jorge
Published: June 2021

Warfarin is an oral anticoagulant that requires individual monitoring since serious adverse events are common. CYP2C9 encodes for the enzyme mainly responsible of S‐warfarin’s metabolism. Polymorphisms in CYP2C9 have been previously found to be strongly associated with observed warfarin dose variability in different populations, but not in Caribbean Hispanics. Caribbean Hispanics originated as a result of a complex admixture among Caucasians, Africans and Amerindians ancestors‐a characteristic that should be considered for warfarin management. The rare loss‐of‐function CYP2C9*8 allelic variant is reportedly more prevalent among individuals with African heritage. Since Puerto Ricans has a significant contribution of African ancestry in their genetic backgrounds, this cross‐sectional study was aimed to determine the frequency of CYP2C9*8 in a cohort of 150 Puerto Rican patients undergoing warfarin therapy. DNA specimens were extracted and genotyped for the CYP2C9*8 using a PCR‐based Taqman genotyping assay. We found 3 heterozygous for the CYP2C9*8 variant in our study cohort, corresponding to a minor allele frequency of 1% (95%CI: 0.0026‐0.031). The observed frequency met Hardy‐Weinberg equilibrium. Allele frequency in our cohort was found to be significantly lower than that from a previous report in African‐Americans (0.01 versus 0.047, respectively, p=0.045 by two‐tailed z‐test), with a carrier frequency of 1 in 50 (Puerto Ricans) versus 1 in 11 (African‐Americans). Due to the CYP2C9*8 prevalence found among Puerto Ricans, we concluded that this variant should be included in any pharmacogenetic‐guided algorithm for warfarin dose predictions in this population. Approved by University of Puerto Rico, Medical Sciences Campus Institutional Review Board protocol A4070109.

Grant Funding Source: Supported by NIH grants# 5SC2HL110393 (NHLBI) and grant# G12RR‐03051 (RCMI‐ NCRR).
A potential role for human UDP‐glucuronosyltransferase 1A4 promoter SNPs in the pharmacogenomics of Tamoxifen and its derivatives (1141.13)
Authors: Dates Centdrika , Greer Aleksandra , Bratton Stacie , Koroth Edavana Vineetha , Kadlubar Susan , Radominska‐Pandya Anna
Published: June 2021

Tamoxifen (Tam) is a selective estrogen receptor modulator used to inhibit breast tumor growth. Tam can be directly N‐glucuronidated via the tertiary amine group or O‐glucuronidated after cytochromes P450 mediated hydroxylation. In this study, the glucuronidation of Tam and its hydroxylated and/or chlorinated derivatives [4OHTamoxifen (4OHTam), Toremifene (Tor), and 4OHTor] has been studied using recombinant human UGTs from the 1A family and human hepatic microsomes (HLM). Recombinant UGT1A4 was found to catalyze the formation of N‐ glucuronides of Tam and its derivatives and to be the most active UGT isoform toward these compounds. Therefore, it was hypothesized that single nucleotide polymorphisms (SNPs) in the promoter of UGT1A4 have the potential to significantly decrease the glucuronidation rates of Tam metabolites in the human liver. In vitro assays using 64 genotyped HLM were used to determine the association between UGT1A4 promoter and coding region polymorphisms and the glucuronidation rates of Tam, 4OHTam, Tor, and 4OHTor. Significant decreases in enzymatic activity were observed in microsomes for individuals heterozygous for ‐163G<A and ‐217T<G. These alterations in glucuronidation could lead to prolonged circulating half‐lives and potentially modify the effectiveness of these drugs in the treatment of breast cancer.

Grant Funding Source: Supported by DoD‐W81XWH1110795 and UAMS TRI (CTSA Grant Award #UL1TR000039) ABCRP funds to AR‐P
Gestational age‐dependent maternal‐fetal glyburide disposition in pregnant mice (1141.5)
Authors: Shuster Diana , Risler Linda , Liang Jason , Rice Kenneth , Shen Danny , Hebert Mary , Thummel Kenneth , Mao Qingcheng
Published: June 2021

Glyburide (Gly) is routinely used to treat hyperglycemia during pregnancy. Our objective was to determine if maternal‐fetal disposition of Gly and metabolites is dependent on gestational age in pregnant mice because similar studies cannot be done in humans. Following retro‐orbital injection of Gly, maternal plasma and fetuses were collected at 0.5‐240 min after dosing. Pharmacokinetic parameters were determined using a pseudo‐profile based bootstrap approach and compartmental/noncompartmental analyses. Maternal Gly clearance increased 2‐fold on gestation day (gd) 15 and 19 versus gd 0. Gly depletion analysis using mouse liver microsomes confirmed that gestational age‐dependent changes in Gly clearance is caused by increases in hepatic metabolism. Maternal metabolite/Gly area under the curve (AUC) ratios were low throughout gestation (~0.004), indicating an increase in both metabolite formation and clearance. Fetal/maternal Gly AUC ratios doubled from gd 15 to 19 (0.02 to 0.04), which is likely due to decreased expression of placental efflux transporter Bcrp1 on gd 19 versus 15. Our data suggest that clinical dosages of Gly may need to be doubled to treat hyperglycemia in pregnant women compared to nonpregnant women.

Grant Funding Source: Supported by the NICHD [U10HD047892], NCATS [TL1 RR025016], and NIH [T32 GM 007750]
Effects of dichlorodiphenyltrichloroethane (DDT) on myogenesis in C2C12 cells (1142.14)
Authors: Kim Jonggun , Kim Yoo , Rutherford Amanda , Yoon Kyong , Clark John , Park Yeonhwa
Published: June 2021

An organochlorine insecticide, dichlorodiphenyltrichloroethane (DDT), has been widely used on agricultural products during the twentieth century and is still present in the environment and foods. Based on emerging body of evidences indicating that exposure to persistent organic pollutants, including organochlorine insecticides, is linked to obesity and type 2 diabetes, we investigated the role of DDT on myogenesis, which may have significant implications regarding glucose uptake. C2C12 myoblasts were treated with various concentrations of DDT (0.1‐10 μM) for 8 days, during differentiation into myotubes. Myotube formation and gene and protein expressions for myogenic factors were determined. DDT reduced myotube formation dose‐dependently. DDT treatment decreased mRNA expression of MyoD, an early myogenic factor, while no effects of DDT were observed in other myogenic factors. Phosphorylation of protein kinase B (pAKT) was significantly reduced by DDT treatment. These results suggest that DDT inhibits myogenesis by affecting the early differentiation factor of myotube formation targeting upstream regulator of AKT. This may attribute to decreased muscle mass and lead to the development of type 2 diabetes.
Comparison of the prevalence of hypertriglyceridemic‐waist phenotype in university students from two regions of Mexico (1142.2)
Authors: Villalobos‐Molina Rafael , Rodríguez‐Tadeo Alejandra , Wall‐Medrano Abraham , Ramos‐Jiménez Arnulfo , Urquidez‐Romero René , Hernández Rosa Patricia
Published: June 2021

The prevalence of HW is unknown among Mexican young, although cardiovascular disease (CVD) is an outcome of it. We evaluated the prevalence for HW (hypertriglyceridemic‐waist) phenotype and related factors, in university students from Juarez City and Mexico City. 12,966 students (<25 y) from “Healthy University” Project’s data base (7,687 of Juarez City and 5,279 of Mexico City) were screened for anthropometry and biochemical indicators. The significance of these factors to the HW phenotype was analyzed in groups segmented by university and sex. In both sexes, Mexico City students had higher abdominal obesity, triglycerides, glycemia and HW, but lower systolic blood pressure than Juarez students. In both populations the HW phenotype was associated with higher body weight, BMI, glucose, cholesterol, and blood pressure. Except for women in Juarez City, HW phenotype was associated to greater height. Simultaneous concurrence of hypertriglyceridemia and waist circumference increase risks factors for CVD in young adults from two regions of Mexico. It is necessary to evaluate other aspects to know the differences between these two populations to identify areas of intervention during their college stay.

Grant Funding Source: RV‐M is supported by PASPA, DGAPA, UNAM
Refractory wound healing and FoxO1/iNOS pathway in type 1 diabetes (1142.3)
Authors: Tie Lu , Li Xuejun
Published: June 2021

Aims/hypothesis Refractory wounds in diabetic patients constitute a serious complication that often leads to amputation with limited treatment regimens. The present study was designed to determine the role of FoxO1/iNOS pathway and effect of the soy isoflavone genistein on diabetic wound healing.

Methods Streptozotocin (STZ)‐induced type 1 diabetic mice with full‐thickness excisional wounds given intragastrically 1 and 3 mg/kg/day of iNOS selective inhibitor (L‐NIL) or received 0.2, 1 and 5 mg/kg/day of genistein via subcutaneous injection.

Results In diabetic wound tissues, the inducible nitric oxide synthase (iNOS) was activated, silent information regulator 1 (SIRT1) and forkhead box O transcription factor 1 (FoxO1) were elevated. Genistein administration prevented increased iNOS activity and attenuated diabetic cutaneous SIRT1 and FoxO1 levels in a dose‐dependent manner. Diabetic conditions significantly increased superoxide anion (O2.‐) production, nitrotyrosine formation, and decreased nitrite levels in wound tissues. L‐NIL and genistein treatments normalized the elevated O2.‐ production, nitrotyrosine formation and reversed the attenuated nitrite level. L‐NIL and genistein dose‐dependently rescued the delay of wound closure in diabetic mice. Five mg/kg/day of genistein treatment significantly increased the mean perfusion rate, and in vitro treatment with genistein protected against high glucose‐induced impairment of capillary tube formation in cultured endothelial cells.

Conclusions/interpretation FoxO1/iNOS signaling may contribute to the delayed wound healing in type 1 diabetes. Genistein could rescue the impaired wound healing and improved wound angiogenesis in type 1 diabetic mice, at least in part, by suppression of FoxO1, iNOS activity and oxidative stress.

Grant Funding Source: This work was supported by the NSFC and Beijing Higher Education Young Elite Teacher Project
Farnesol suppresses lipid accumulation in a HepaRG‐based model of hepatic steatosis (1142.8)
Authors: Pant Asmita , Kocarek Thomas
Published: June 2021

Non‐alcoholic fatty liver disease is one the most common metabolic syndromes characterized by accumulation of triglycerides (TG) in liver. Treatment with the endogenous isoprenoid farnesol lowers hepatic TG levels in rodents, and this effect appears to involve at least two nuclear receptors, peroxisome proliferator‐activated receptor α and farnesoid X receptor (FXR). Activation of constitutive androstane receptor (CAR) has also been shown to ameliorate hepatic steatosis in rodents. However, farnesol’s effects on human hepatic lipid metabolism are currently unknown. In this study, we evaluated the effect of farnesol treatment on human hepatic lipid accumulation in a cellular model of hepatic steatosis that was created by incubating the hepatocyte‐like HepaRG cell line with oleic acid. Incubation of differentiated HepaRG cells with oleic acid produced concentration‐ and time‐dependent increases in intracellular lipid droplets labeled with Oil Red O as well as in TG levels. This TG accumulation was suppressed by 32% when the oleate‐loaded HepaRG cells were co‐treated with farnesol. Farnesol treatment also caused concentration‐ and time‐dependent increases in the mRNA levels of small heterodimer partner and CYP2B6 in HepaRG cells, which are the targets of FXR and CAR, respectively. These results suggest that farnesol treatment can decrease fatty acid‐induced hepatic lipid accumulation in HepaRG cells, possibly by activating FXR and/or CAR.

Grant Funding Source: Supported by NIH grant HL050710
Compounds in Psoralea corylifolia as human MAOb natural inhibitors (1143.12)
Authors: Zarmouh Najla , Mazzio Elizabeth , Soliman Karam
Published: June 2021

Treatment for Parkinson’s disease (PD) involves the use of drugs that address the loss of dopaminergic function by restoring neurotransmitter homeostasis. However, the current drugs can be limiting, in that they do not address disease progression. A renewed interest in monoamine oxidase B inhibitors (MAObIs) for PD as independent therapeutics has emerged after recent clinical trials has shown efficacy of deprenyl and rasagiline monotherapy for early disease management. However, conventional MAOIs have food and drug counter‐interactions and side effects which limit its use. Previously, we screened 905 natural ethanol plant extracts on the enzymatic activity of recombinant human MAOb. In this study, we confirm and determine selectivity of MAOb / MAOa. The data show that extracts of Glycyrrhiza uralensis root have an IC50 of 0.04 mg/ml and a 266 fold MAOb/MAOa specificity; Phellodendron Amurense an IC50 of 0.02 mg/ml and a MAOb/MAOa specificity 45.29 fold; Psoralea Corylifolia (PC) an IC50 of 0.08 mg/ml and a 18.34 fold MAOb/MAOa specificity; Ferula Assa‐Foetida an IC50 of 0.24 mg/ml and a 30.23 fold MAOb/MAOa specificity. Further studies for PC active ingredients revealed that bakuchiol content within the plant may contribute to MAOI.

Grant Funding Source: Supported by NIMHD G12 MD007582 and P20 MD006738
Involvement of RAGE in isoflurane‐induced neuronal apoptosis (1143.3)
Authors: Koladia Mohit , Zhong Tiecheng , Sun Chengwen , Vetter Stefan
Published: June 2021

Anesthetics, including isoflurane, have been demonstrated to induce apoptotic neurodegeneration when administered during neurodevelopment. There is inadequate clinical data to conclude whether anesthetics induce neurotoxicity in the developing human brain. The mechanism behind anesthesia induced neurotoxic effects is still unclear. Isoflurane has been reported to cause neurotoxicity and neurocognitive impairments in neonatal rats exposed to this gas. Previous reports suggest elevated levels of S100B after anesthesia and brain trauma. The receptor for advanced glycation end products (RAGE) is the only known cell surface receptor for S100B and expressed in the brain. RAGE activation in neurons has also been shown to lead to apoptosis and neurodegeneration. Our research investigates the hypothesis that anesthesia leads to RAGE activation and subsequently to neuronal apoptosis. In preliminary studies, we investigated the expression of RAGE in different areas of brain using immunohistochemical methods and found that RAGE was highly expressed in hippocampal region of brain. Apoptotic cells were detected in brain sections by colorimetric and fluorometric immunohistochemistry using cleaved caspase‐3 as marker molecule. Immunofluorescence studies were used to investigate co‐localizes RAGE expression and apoptotic cells and to identify cell types based on neuronal marker molecules. On the basis of our results, we demonstrate that RAGE may be involved in the neuronal apoptosis induced by isoflurane. Further investigation in this direction may help to develop therapeutic agents targeting RAGE that may be route for potential treatments to attenuate the post‐anesthetic cognitive dysfunction in both young and aged patients.
The neuroprotective effects of fluoxetine on rat hippocampus, after chronic and acute 5‐fluorouracil treatment (1143.6)
Authors: Chambers Annabelle , Maqbool Ayesha , Wigmore Peter
Published: June 2021

5‐Fluorouracil (5FU) an anti cancer therapy, can cause long term cognitive decline. 5FU reduces cell proliferation in hippocampal neurogenesis which is associated with reduced spatial memory in rodents. Prior antidepressant treatment (Fluoxetine (FLX) prevents the behavioural/cellular effects of 5FU. This quantifies neural stem cell (NSC) number and proliferation after drug treatment.

Male rats were treated for 19 days with 10mg/kg/day FLX, via drinking water. Rats received 1 (acute) or 5 (chronic) injections of 35mg/kg 5FU or saline i.p., with or without continued FLX dosing. 1 day or 1 week post treatment, Ki67 (proliferation), Sox 2 (NSC’s) positive cells in the sub granular zone were counted.

Acute 5FU causes a significant decline in cell proliferation 1 day later while 1 week animals returned to control levels. Pre‐treatment with FLX prevented the decrease caused by 5FU at one day. Chronic 5FU causes prolonged decline in proliferation, also prevented by FLX. Preliminary results indicate that NSC levels are little affected but their proliferative activity is altered.

FLX appears to change the proliferative behaviour of NSC under 5FU treatment. NSC location, number and proliferation are influenced by both compound; use of variations in acute/chronic effects aim to deduce specific mechanisms behind FLX’s neuroprotection.

Grant Funding Source: Supported by School of Life Sciences, University of Nottingham, and AstraZeneca UK
A novel triple reuptake inhibitor with rapid antidepressant properties identified by virtual screening (1144.1)
Authors: Talbot Jeffery , Geffert Laura , Wolters Nicholas , Amos Mary , Dallman Elizabeth , Munro Caitlin , Tolle Erica , Larkey Jessica , Tanda Gianluigi , Katz Jonathan , Indarte Martin , Madura Jeffry , Surratt Christopher
Published: June 2021

A continuing challenge in treating depression is therapeutic delay and resistance to available treatments. Hence, the growing interest in developing antidepressants (AD) with rapid onset (hours to days) and novel mechanisms of action. For example, the glutamatergic antagonist ketamine has rapid AD efficacy but is limited by poor bioavailability, short duration of effect, and potential for abuse. Here we report MI‐4, identified through virtual screening targeting the human dopamine transporter, inhibited the human monoamine transporters of dopamine (DA), norepinephrine (NE), and serotonin (5‐HT) in vitro. In vivo administration of MI‐4 in mice induced robust, dose‐dependent AD‐like behaviors in acute models of learned helplessness (tail suspension test and forced swim test). Moreover, 21‐day administration of MI‐4 reduced drinking latency times comparable to fluoxetine in the novelty‐induced hypophagia test, which requires chronic treatment to produce AD effects. Interestingly, like ketamine, MI‐4 also exhibits NMDA receptor antagonism and produced rapid (3‐day) and chronic (21‐day) AD‐like effects in the social avoidance test following 10 days of social defeat, by increasing social interaction scores 2‐fold vs. vehicle controls while improving resiliency to near 70%. Unlike ketamine, MI‐4 exhibited minimal abuse liability in behavioral and neurological models (conditioned place preference and DA in vivo microdialysis). These data suggest MI‐4 is a novel, multiple mechanism antidepressant with rapid and chronic properties.
Correlation of beta‐arrestin1 levels in mononuclear leukocytes with depression scores for women with premenstrual dysphoric disorder (1144.2)
Authors: Nayyar Tultul , Alam Farzana , Richie William , Ansah Twum , Bailey Rahn
Published: June 2021

OBJECTIVE: Depression is very common in reproductive women with premenstrual dysphoric disorder (PMDD), a severe form of premenstrual syndrome (PMS). This study examined whether a measurement for beta‐arrestin1 levels in peripheral blood mononuclear leukocytes (PBMC) could aid in the diagnosis of PMDD and support the results from questionnaires. DESIGN AND METHODS: 18‐42 years old non‐pregnant women at the luteal phase with the symptoms of PMS/PMDD, but not taking any antidepressants, were studied. The protein levels of beta‐arrestin1 in the PBMC were determined by ELISA and compared with the Hamilton Depression Rating Scale scores. The different criteria for Axis 1 mental disorders were compared. RESULTS: The criteria for Axis 1 mental disorders were significantly higher and beta arrestin1 protein levels in mononuclear leukocytes were significantly lower in women with depression (PMDD) as compared to no‐depression (PMS) group. The reduction in beta arrestin1 protein levels was significantly correlated with the severity of depressive symptoms. CONCLUSION: These findings support the implication of beta‐arrestin1 protein in the pathophysiology of depression in women with PMDD. Beta‐arrestin1 measurements in women with PMDD may potentially serve for biochemical diagnostic purposes and might be useful as evidence‐based support for questionnaires.

Grant Funding Source: U54 MD 007593 (NIMHD)
Developmental differences in the effect of chronic corticosterone treatment in pre‐ and post‐pubescent female juvenile rats (1144.6)
Authors: Nickle Tyler , Middlemas David
Published: June 2021

The aim of this study was to determine the effect of chronic corticosterone (CORT) administration on pre‐pubescent and post‐pubescent female rats in order to develop a rat model of depression in these age groups. Rats were administered daily injections of CORT (40 mg kg‐1) for twenty‐one days. Depressive‐like behavior was assessed using the modified forced swim test (FST) and the sucrose preference test (SPT). Immunohistochemistry was used to determine changes in mitogenesis and survival of newborn neurons in the dentate gyrus of the hippocampus. Chronic CORT treatment produced significant changes in the FST for post‐pubescent rats, but not pre‐pubescent rats. Conversely, CORT produced significant changes in the SPT for pre‐pubescent rats, but not post‐pubescent rats. Ongoing investigation will determine the effect of CORT on mitogenesis and survival of newborn neurons in the dentate gyrus of pre‐pubescent and post‐pubescent female rats. These results suggest that CORT may induce depressive‐like behavior in female pre‐pubescent and post‐pubescent rats; however, the physiological effect is manifested differently at different stages of rat development.

Grant Funding Source: Supported by AT Still University of Health Sciences grant and the Warner‐Fermaturo Fund
PDE11A negatively regulates lithium responsivity in mice possibly due to an interaction with AKT/PKB (1144.8)
Authors: Kelly Michy , Pathak Geetanjali , Hartman Alexander
Published: June 2021

Lithium is a first line mood stabilizer for patients with bipolar disorder, but for unknown reasons approximately half of all patients do not respond to lithium. Lithium responsiveness in patients with bipolar disorder has been genetically associated with single nucleotide polymorphisms (SNPs) in Phosphodiesterase 11A (PDE11A), a unique enzyme that breaks down cAMP and cGMP only in the hippocampus. These PDE11A SNPs are intronic; therefore, the objective of this study was to determine if decreasing PDE11A expression was sufficient to increase lithium responsivity in mice. We measured PDE11A expression by in situ hybridization and Western blot in the hippocampus of inbred mouse strains known to respond well (C57BL/6J), moderately (129S6) or poorly (Balb/cJ) to lithium. PDE11A mRNA and protein levels were lowest in C57BL/6J mice, the strain most responsive to lithium, and highest in Balb/cJ mice, the least responsive strain. Consistent with these observations, PDE11A KO mice given 0.4% lithium chow for 3 weeks exhibited greater lithium responsivity relative to wild‐type littermates in tail suspension, an anti‐depressant predictive assay, and in amphetamine hyperlocomotion, an anti‐manic predictive assay. Lithium does not directly affect PDE11A catalytic activity; however, in vitro experiments suggest that PDE11A may influence lithium responsiveness due to an interaction with AKT, an enzyme that is required for lithium’s behavioral effects. AKT is predicted to phosphorylate PDE11A at several residues, including serines 117 and 162. Phosphomimic mutations of serines 117 and 162 shift the subcellular distribution of PDE11A away from a punctate, membrane enriched compartmentalization towards a more diffuse, cytoplasmic localization. Together, these findings suggest that PDE11A negatively regulates lithium responsiveness by inhibiting the activation of AKT but that AKT can feed back and release this brake by phosphorylating PDE11A, thereby removing PDE11A from the membrane.

Grant Funding Source: Research supported by a PhRMA Foundation and the University of South Carolina
The effect of nitric oxide on blockade of the norepinephrine transporter, uptake‐1, by methylphenidate (1145.6)
Authors: Simaan Joseph
Published: June 2021

We previously showed that fresh synthesis of nitric oxide (NO) mediates transport of substrates across uptake ‐1, norepinephrine (NE), tyramine (T), mephentermine, ephedrine, imipramine and atomoxetine, and blockade by cocaine. Synthesis of NO was blocked by Nw‐nitro‐L‐arginine (L‐NNA) and the rise in arterial pressure it induces was restored to starting level by an infusion of nitroglycerin (NG), a NO donor. In this study we explored the role of NO synthesis on the blockade of uptake‐1 by methylphenidate (MP), a drug used in the treatment of attention deficit disorder, in Sprague Dawley rats in which mean arterial pressure was measured. NE (0.05, 0.1, 0.2 ug) with direct action on adrenergic receptors but is transported across uptake‐1 and sequestered in sympathetic nerve terminals, showed potentiation of its pressor effect after MP (1.52+/‐0.4 mg/kg) by 34%‐56% and not changed further after MP/L‐NNA/NG. The pressor effect of T (0,025, 0.05, 0.1 mg) which acts indirectly by entering across uptake‐1 to release NE, was reduced after MP by 78%‐85% and restored by 424% ‐650% after MP/L‐NNA/NG. The pressor effect of methoxamine, not a substrate of uptake‐1, was not affected by pretreatment with MP or MP/L‐NNA/NG. It is concluded that the blocking effect of MP on uptake‐1 is confirmed and this blockade is significantly dependent on synthesis of NO. Supported by the American University of Beirut.
Dihydromyricetin attenuates high glucose‐induced oxidative stress in human umbilical vascular endothelial cells (1146.2)
Authors: Zhang Ting , Mi Mantian , Liang Xinyu , Shi Linying
Published: June 2021

Objective To investigate the effects of Dihydromyricetin on high glucose‐induced cell damage in human umbilical vascular endothelial cells (HUVECs).

Methods HUVECs were cultured in 5.56 mmol/L glucose as normal control, 5.56mmol/L glucose plus 27.8mmol/L mannitol as an osmotic control, or 33.36 mmol/L high glucose for 48 hours in the absence and presence of Dihydromyricetin, cell viability and apoptosis were detected by CCK‐8 assays and annexin V‐FITC/PI double staining using flow cytometry, respectively. In addition, the intracellular reactive oxygen species (ROS), malondialdehyde (MDA) content and the activity of superoxide dismutase (SOD) and catalase (CAT) in the culture supernatants were measured.

Results High concentration of glucose significantly decreased cell viability and the activity of SOD and CAT, while increased the apoptotic rate, the intracellular ROS and the MDA content in HUVECs (P<0.05). Dihydromyricetin pretreatment significantly inhibited the decrease in cell viability and the increase in cell apoptosis induced by high glucose. The ROS and MDA levels were reduced while the SOD and CAT levels were increased after pretreatment with Dihydromyricetin following high glucose stimulation.

Conclusion Dihydromyricetin attenuated high glucose‐induced oxidative stress in HUVECs.

Grant Funding Source: Supported by The National Natural Science Foundation of China (81102129)
Leukocyte GPR120 differentiates hypolipidemic vs. anti‐inflammatory effects of long chain n‐3 and n‐6 polyunsaturated fatty acids in inducing atherosclroprotection in LDLrKo mice (1146.7)
Authors: Shewale Swapnil , Boudyguina Elena , Bi Xin , Zhu Xuewei , Brown Amanda , Mishra Nila , Amexander‐Miller Martha , Oh Da Young , Olefsky Jerrold , Parks John
Published: June 2021

Background: Whether anti‐inflammatory effects of leukocyte GPR120 impart atheroprotection in‐vivo was unknown. We determined contribution of leukocyte GPR120 in n‐3 vs.n‐6 polyunsaturated fatty acid (PUFA) induced atheroprotection. Methods: Leukocyte GPR120 wildtype (WT) or knockout (KO) mice in the LDLr‐/‐ C57BL/6 background were generated by bone marrow transplantion and were fed one of the four atherogenic diets for 16 weeks (palm (+ control), fish(‐ control), echium (n‐3 PUFA enriched) or borage oil (n‐6 PUFA enriched), (PO, FO, EO or BO)). Plasma lipids‐lipoproteins, circulating and splenic leukocytosis, monocyte recruitment to aortic root and aortic atherosclerosis were analyzed. Results Compared to PO, mice fed BO and EO had significantly lower plasma total cholesterol, VLDL cholesterol, aortic root intimal area and aortic Cholesteryl Esters (CE) levels comparable to the FO group. EO and FO also had lower plasma triglyceride concentrations compare to PO and BO. Lack of leukocyte GPR120 did not significantly affect plasma lipids or atherosclerosis severity within any diet group. However, KO BM recipients, fed FO, EO and BO had increased circulating and splenic neutrophilia and splenic monocytosis characterized by higher proportion of Ly6Chi monocytes leading to increased monocyte recruitment into the aortic root atherosclerotic lesions. Summary: n‐3 as well as n‐6 PUFAs may activate leukocyte GRP120 in vivo and possess GRP120‐independent and ‐dependent atheroprotective properties. Leukocyte GPR120 has minimal effect on plasma lipid‐ lipoprotein concentrations and aortic atherosclerosis however; PUFA induced attenuation of neutrophilia, splenic monocytosis and pro‐inflamatory monocyte (Ly6chi) phenotype is leukocyte GPR120 dependent. Conclusion: n‐3 and n‐6 PUFA induced VLDL cholesterol lowering is primary atheroprotective mechanism in LDLrKO mice.