bluuebunny/crossref_metadata_2025_split huggingface dataset:
Kidney alkalization increases macula densa NOS1β expression and improves transplanted renal graft function
Authors: Wang Lei 
Published: April 2020

Ischemia‐reperfusion injury (IRI) is a major hurdle for the survival of transplanted grafts. Currently, there are no effective strategies for prevention and treatment of IRI during kidney transplantation (KTX). Neuronal NO synthase beta (NOS1β) is the primary splice variant of NOS1 in macula densa (MD) and contributes to most of the MD‐NO generation, which blunts tubuloglomerular feedback (TGF) response and increase GFR, which is one of the important factors in evaluation of renal function. While TGF response modulated by NOS1β has been demonstrated to play an essential role in the long‐term control of hemodynamics, the role of NOS1 in the MD in the prevention of IRI and protection of transplanted graft function remains unknown.KTX was performed in C57BL/6 mice. The expression of NOS1 and graft function were evaluated. To test the response of NOS1 to the tubular pH, the mice were given bicarbonate or NH4Cl for two weeks. Blood, urine pH and expression of NOS1 were measured. KTX were performed after the treatment. Graft injury and function were evaluated again. To further investigate the role of the NOS1 in the protection of the graft function, we repeated the above experiments in kidney specific NOS1 knockout (KO) mice and compared the results with C57BL/6 mice.The control recipients showed moderate graft injury with plasma creatinine (Pcr) of 0.68 ± 0.13 mg/dl and reduced graft function indicated by 23% decrease in GFR at 5 days after KTX. NOS1β expression decreased by 60%. TGF response was measured by micropuncture which was 4.1 ± 0.3 mmHg for sham‐operated mice and 7.2 ± 0.9 mmHg for KTX mice. These data indicated that decreased NOS1β expression may contribute to the decrease of GFR and deterioration in graft function by enhanced TGF response. KO recipients showed more severe graft injury with the Pcr of 1.07±0.14mg/dl which demonstrated that deletion of NOS1 from macula densa significantly aggravated renal graft injury.The pH‐NOS activity response curve showed that NOS1 was sensitive to tubular pH with the optimal pH of 8.0. Bicarbonate treatment increased NOS1 expression in macula densa by 65% while NH4Cl intake decreased the NOS1 expression by 40%.KTX were performed in C57BL/6 and KO mice after two weeks treatment of bicarbonate or NH4Cl. Pcr levels were about 30% and 100% lower in bicarbonate‐treated group than H2O and NH4Cl groups, respectively, indicating that bicarbonate treatment significantly alleviated graft injury. However, there were no significant differences in the graft injury for the KO mice among different groups.In summary, NOS1β expression in the macula densa decreases after KTX, which enhances TGF responsiveness and contributes to the decrease in GFR and decline of graft function. Rescue of NOS1β in the macula densa by renal alkalization decreases kidney injury and improves transplanted graft functions.
The role of the rostral ventrolateral medulla in systolic blood pressure frequency content within the Mayer wave band
Authors: Badenhorst Cecilia , Squair Jordan W. , Gautier Matthieu , Soriano Jan Elaine , Courtine Gregoire , Phillips Aaron 
Published: April 2020

A primary supraspinal structure involved in sympathetically‐mediated blood pressure control is the rostral ventrolateral medulla (RVLM). The RVLM is where the cell‐bodies of descending sympatho‐excitatory axons responsible for blood pressure control are located. The frequency through which the sympathetic nervous system modulates blood pressure has been termed the Mayer wave. The precise origin of Mayer waves has never been clearly identified.Using a rat model, our objective was to assess the causal role of the RVLM in Mayer waves, which in rats occur in the low‐frequency range of 0.4–1.0 Hz. We tested this by assessing wavelet spectrograms in response to 1) optogenetic stimulation of the RVLM, 2) orthostatic activation of the RVLM and 3) surgical disruption of descending sympatho‐excitatory pathways. We optogenetically stimulated tyrosine hydroxylase (TH)ON neurons of the RVLM in a TH:Cre rat and found increased power in the Mayer wave band of the wavelet spectrogram. When we applied an orthostatic challenge to the rodents, in order to increase baroreflex‐mediated RVLM activity, the increased power in the wavelet spectrogram was again observed. In contrast, we did not find this effect in rats after a severe T3 spinal cord contusion, which disrupts descending sympatho‐excitatory pathways. In conclusion, THON neurons in the RVLM appear to be responsible for systolic blood pressure frequency content in the Mayer wave band.Support or Funding InformationCanadian Institutes of Health Research, Libin Cardiovascular Institute, Hotchkiss Brain Institute, Wings for Life, Campus Alberta Neuroscience.
Glutathione Peroxidase Influence on Cardiac Remodeling in Progeny of Dams Exposed to Engineered Nanomaterials during Gestation
Authors: Kunovac Amina , Hathaway Quincy A. , Taylor Andrew D. , Durr Andrya J. , Goldsmith William T. , Pinti Mark V. , Fink Garrett K. , Nurkiewicz Timothy R. , Hollander John M. 
Published: April 2020

Vulnerable populations, like the developing fetus, may be more susceptible to sustained detriments elicited by inhaled engineered nanomaterials (ENMs) entering the bloodstream and prompting systemic effects. Previous studies have demonstrated diminished mitochondrial bioenergetics in progeny following maternal exposure to a highly utilized ENM, nano‐titanium dioxide (nano‐TiO2). Cardiac dysfunction and reduced ROS scavenging ability were observed with decreased mitochondria phospholipid hydroperoxide glutathione peroxidase (mPHGPx) expression. This study aimed to elucidate whether overexpression of mPHGPx could provide sufficient antioxidant protection to attenuate cardiac dysfunction in pregnant dams and progeny following gestational ENM inhalation exposure. Wild‐type (WT) and transgenic (mPHGPx) female mice were bred with transgenic and WT males, respectively, to delineate maternal vs progeny protection. Pregnant dams were exposed to nano‐TiO2 (6 hrs/day for 6 days) with a primary particle size of 21nm and mass concentration of 12mg/m3 or sham air. Conventional (M‐Mode) and speckle tracking stress‐strain echocardiographic parameters were assessed in pregnant dams, as well as in fetal (gestational day 14) and adult (11 wks) offspring. Following inhalation exposure to nano‐TiO2 during gestation, WT dams exhibited signs of concentric remodeling marked by significantly decreased (left ventricular) LV diastolic diameter (12%) and volume (26%), and significant increases in both LV posterior wall thickness (23%) and relative wall thickness (39%), which were preserved in mPHGPx pregnant dams. Fetal offspring of mPHGPx dams had significantly enhanced cardiac output (87%) and stroke volume (43%) compared to fetal offspring of WT dams. Stress‐strain analysis revealed that fetal WT offspring of nano‐TiO2–exposed mPHGPx dams had significantly higher short‐axis systolic circumferential strain rate (~3 fold) and displacement (~2 fold) than WT offspring of WT dams. Adult WT offspring of transgenic dams presented with a significantly lower ejection fraction (6% and 8%) and fractional shortening (18% and 22%) than sham WT and nano‐TiO2 transgenic offspring, respectively. This may suggest that in offspring lacking the transgene, enhanced maternal antioxidant defense alone may not provide sufficient protection into adulthood. This study highlights the beneficial role of enhanced antioxidant defense (during fetal development and adulthood) on cardiac dysfunction following gestational ENM exposure.Support or Funding InformationSupported by: R01 HL‐128485 (JMH), CFO for the Ohio Valley Whipkey Trust (JMH), R01‐ES015022 (TRN), AHA‐17PRE33660333 (QAH).
Changes in cerebral expression of ERK 1/2 related to commercial sweetener consumption in BALB/c mice
Authors: Contreras-Chavez Gerson Gabriel , Estrada José A. , Contreras Irazú 
Published: April 2020

Non‐nutritive sweeteners are food additives that provide sweet taste with low energy content. Consumption of these compounds has become a common practice in the general population to reduce calorie intake from the diet. Nonetheless, there is evidence suggesting consumption of non‐nutritive sweeteners can induce changes in feeding behavior, altering energy intake, body weight and adiposity.Previous studies have revealed that intake of sucralose and steviol glycosides lead to changes in the JAK/STAT3 signaling pathway in the brain of mice, altering food intake patterns, body weight and adiposity. However, the effect of these compounds on other signaling pathways, such as MAPK and ERK 1/2 in particular, which are also involved in the regulation of food intake in the central nervous system, has not been studied.The main objective of this project is to determine the effects of frequent intake of sucrose, sucralose or steviol glycosides on the expression of total and phosphorylated ERK 1/2 protein in the brain of BALB/c mice.To this end, 8 week‐old BALB/c male and female mice were supplemented with sucrose, sucralose or steviol glycosides for 6 weeks in their daily drinking water. Subsequently, total brain protein extracts were used to analyze the expression of total and phosphorylated ERK 1/2 proteins by western blot.Our preliminary results show that expression of pERK was reduced in male mice supplemented steviol glycosides compared to non‐supplemented controls, while sucrose and sucralose groups has increased expression of pERK. In contrast, female animals supplemented with sucrose had decreased expression of pERK, which was increased in the steviol glycosides group compared to non‐supplemented controls. No differences were observed in total ERK expression among groups.In conclusion, our preliminary results suggest that chronic consumption of non‐nutritive sweeteners may induce sex‐specific alterations in the ERK signaling pathway, with possible effects on feeding behavior and body weight in vivo.
Prenatal chronic stimulation of the endocannabinoid system affects the respiratory motor outputs of juvenile male and female rats
Authors: Patrone Luis Gustavo A. , Karlen-Amaranre Marlusa , Bicego Kenia C. , Zoccal Daniel B. , Gargaglioni Luciane H. 
Published: April 2020

The use of drugs of abuse during pregnancy can severely affect the development of physiological systems including the central respiratory network, mainly because prenatal period is extremely susceptible to external and pharmacological interventions that may cause postnatal consequences to the offspring. The endocannabinoid system is already present in the central nervous system in early stages of embryonic development, including in areas responsible for respiratory control. Nevertheless, the effect of external cannabinoids on the central cardiorespiratory network development as well as in the chemosensitivity and the future consequences during juvenile period is still unclear. Here, we evaluate the effects of prenatal exposure to cannabinoid on the respiratory motor outputs of juvenile (P27–28) male and female rats by using a decerebrated, arterially perfused in situ preparation. To this end, osmotic pumps were implanted subcutaneously in pregnant rats at embryonic day 0 and delivered vehicle (VEH) or CB1 receptor agonist (WIN 55212‐2, 0.5 mg/Kg/day) for 21 days. In situ preparations of juvenile rats were used for recording the respiratory motor and sympathetic outputs during normocapnia (5% CO2), hypercapnia (8% CO2) and stimulation of peripheral chemoreceptors with potassium cyanide (KCN; 25 μg in 50 μL). Our data showed that prenatal WIN exposure resulted in a tonic increase in phrenic burst amplitude in male animals, which was sustained during the KCN and CO2 challenge, while WIN‐treated females exhibited elevated phrenic burst frequency. The activity of the vagus nerve was also higher in male WIN‐treated animals, either under basal condition, KCN or hypercapnic exposure, although no effects were observed in the female group. The frequency of late‐expiratory events in abdominal activity, which indicate active expiration, was significantly higher (37%) for male treated animals during KCN stimulus, with no significant effects in the female group. In both sexes, prenatal WIN treatment did not modify the levels of thoracic sympathetic activity. These results suggest that prenatal chronic stimulation of endocannabinoid system promotes alterations in the respiratory network development affecting the activity of nerves responsible for the ventilatory motor drive during juvenile period. These effects seem to be potentiated in males.Support or Funding InformationFAPESP and CNPq
Renal Protection against Post Ischemia‐reperfusion Injury in Fatty Acid Amide Hydrolase Knockout Mice
Authors: Chen Chaoling , Wang Weili , Hu Gaizun , Xie Dengpiao , Ritter Joseph , Li Ningjun 
Published: April 2020

Renal fibrosis is a typical characteristic of progressive chronic kidney disease (CKD). Current treatment options to slow renal fibrosis are still very limited. Endocannabinoids known as anandamide and 2‐arachidonoylglycerol play a critical role in regulating renal physiology via cannabinoid receptors (CB1 and CB2). Fatty acid amide hydrolase (FAAH) is one of the major enzymes to metabolize endocannabinoids. Studies have shown that endocannabinoid system is present in the kidney and that CB1 and CB2 are involved in CKD. However, the role of FAAH in CKD remains unclear. The present study tested the hypothesis that FAAH pathway participates in the kidney damage in post ischemia‐reperfusion (PIR) model, which produces chronic renal injury and fibrosis, using FAAH knockout mice. Male wild type C57BL6 (WT) and Faah−/− mice were subjected to PIR by unilateral ischemia‐reperfusion and contralateral nephrectomy 10 days after ischemia‐reperfusion. Compared with WT mice, Faah−/− mice showed significantly reduced BUN (mean: 22.0 ± 1.16, 102.0± 9.50, and 61± 4.35 mg/dL in WT‐shame, WT‐PIR, and Faah−/−‐PIR mice, respectively, p<0.05) and plasma creatinine level (mean: 0.18 ± 0.02, 1.18 ± 0.06, and, 0.54± 0.05 mg/dL in WT‐shame, WT‐PIR, and Faah−/−‐PIR mice, respectively, p<0.05) in response to PIR. Western blot analysis showed that increased expression of fibrotic protein alpha‐smooth muscle actin in the kidneys were significantly reduced in Faah−/− mice, whereas remained high in WT mice under PIR (1.85‐fold versus 2.46‐fold, p<0.05). Collagen staining by Sirius Red showed the same pattern as alpha‐smooth muscle actin expression in the kidneys from different groups of mice. Histological evaluation by Periodic acid–Schiff (PAS) staining showed that Faah−/− mice exhibited less tissue damage especially in tubular structure in response to PIR compared with WT mice. These results suggest that inhibition of FAAH may preserve renal function and prevent renal fibrosis and that FAAH may serve as a target for developing novel and effective treatment for CKD.Support or Funding InformationNIH grants R01DK54927, R01DK107991, R01HL145163
Comparative Anticoagulant Effects of Parenteral Sulodexide as Measured by Activated Clotting Time. Relevance to Dosing in Various Indications
Authors: Daravath Bharathi , Fareed Jawed , Iqbal Omer , Kouta Ahmed , Hoppensteadt Debra , Siddiqui Fakiha , Lewis Mary , Lewis Bruce 
Published: April 2020

IntroductionSulodexide is a mucosal tissue derived glycosaminoglycan of mammalian origin which is clinically used in oral (capsule) and parenteral (ampules) forms for various clinical indications. The biologic potency of sulodexide is expressed in lipoproteinlipase releasing units (LSU). As a parenteral drug sulodexide produces sizable anticoagulant effects in various laboratory assays including the activated clotting time which is routinely used during surgical procedures. In comparison to heparin the USP referenced potency of sulodexide is reported to be 95 – 105 U/mg by anti‐Xa assay and 30 – 40 U/mg by anti‐IIa assay. The purpose of this study is to determine the whole blood anticoagulant effect of parenteral preparations of sulodexide and its neutralization by protamine sulfate.Materials and MethodsParenteral formulations (ampules) containing 600 LSU/2 ml were diluted with saline to make 10 and 100 LSU/ml working solutions. Protamine sulfate was obtained from the Loyola hospital pharmacy and diluted to 1.00 – 0.12 mg/ml. Anticoagulant effects of sulodexide over the concentration range of 1.0 – 0.25 LSU/ml were measured on the Hemachrom ACT instrument in freshly drawn normal human blood (n=5–10). Protamine neutralization studies were carried out at a fixed sulodexide level (1.0 LSU/ml) and variable protamine sulfate levels (50–12.5 μg/ml).ResultsSulodexide produced a concentration‐dependent anticoagulant effect in normal human whole blood over a concentration range of 0.25 – 1.0 LSU/ml in the range of (238 ± 24 to 401 ± 30 seconds) in comparison to saline control (136 ± 12 seconds). Protamine sulfate supplementation at a concentration range of 50 – 12.5 μg/ml produced varying levels of neutralization of the anticoagulant effects of sulodexide (1 LSU/ml). 50 μg/ml protamine completely neutralized sulodexide at 1 LSU/ml whereas at lower concentrations it partially neutralized the anticoagulant effects.DiscussionThe results demonstrate that sulodexide in parenteral formulation produces a strong anticoagulant effect as measured by whole blood ACT. At a concentration of 1.0 LSU/ml, sulodexide produces an anticoagulant effect that is comparable to that observed with surgical anticoagulation levels of heparin (>3 μg/ml). The anticoagulant effects of sulodexide are neutralized by protamine sulfate which can be used as an antidote for sulodexide.ConclusionParenteral (ampules) sulodexide can be used for anticoagulation in dose dependent fashion and protamine sulfate can be used as an antidote for this agent thus in the event of a shortage of unfractionated heparin sulodexide can be used as a substitute anticoagulant drug for the indications in which heparin is considered.
Using Exogenous βHB as an Epigenetic Modifier to Mitigate the Symptoms of Kabuki Syndrome
Authors: Moss Sara , Soliven Maricel , Poff Angela , D’Agostino Dominic 
Published: April 2020

BackgroundKabuki Syndrome is a rare genetic disorder that leads to many developmental abnormalities, and is caused by a heterozygous mutation in either KMT2D (Type 1) or KDM6A (Type 2) that leads to a loss of function. Both genes play a role in gene regulation via histone modification. KMT2D is a gene that codes for the protein kmt2d, a lysine methyltransferase responsible for methylating H3K4. The KDM6A gene encodes the protein kdm6a, a demethylase responsible for demethylating H3K27. Both gene functions contribute to the opening of the chromatin. When either of these two proteins are deficient, craniofacial, skeletal, mental, and dermatologic development are severely impacted. Other developmental processes are impacted, but to varying degrees across patients. The endogenous ketone metabolite beta‐hydroxybutyrate (βHB) is a known epigenetic modifier, functioning as a class I and IIa histone deacetylase inhibitor. Previous research has shown that the ketogenic diet increases H3K4 acetylation and methylation rates, making βHB a potential therapeutic intervention for Kabuki syndrome. Additionally, H3K4 is a beta‐hydroxybutyrylation site, further underlying the potential for βHB to affect the epigenetic modifications observed in the disease. However, there is no current research on beta‐hydroxybutyrylation in Kabuki, nor on the effects of continuous exogenous βHB supplementation in the disorder.MethodsKmt2d+/βGeo (Type 1 Kabuki syndrome model) mice were fed standard diet with or without 15% BHB‐MCT supplementation for 12–15 weeks. Blood ketones, glucose, and body weight were monitored during the treatment period. Open field, Elevated Plus Maze, Novel Object Recognition, and Grip Strength behavioral tests were performed at the end of treatment. Various tissues were harvested, including the brain, for ongoing ex vivo analysis of markers associated with learning, memory, and development.ConclusionsThe 15% BHB‐MCT supplementation is sufficient to raise blood ketone levels in Kmt2d+/βGeo mice. Behavioral and molecular analysis is ongoing.Support or Funding InformationDisruptive Nutrition, Florida high Tech Corridor
Effects of Vacuolar H+‐ATPase Inhibition and the a1 and a3 Isoforms on Secretion and Activation of Cathepsins in MDA‐MB231 Breast Cancer Cells
Authors: Hinton Ayana , El Sayed Mohamed , Henry Abhijeet , Swickard Jamie 
Published: April 2020

V‐ATPases are multi‐subunit H+ pumps that acidify intracellular compartments and are essential for bone degradation and synaptic signaling. They are at the cell surface of the highly metastatic breast cancer cell line MDA‐MB231 and may contribute to its metastatic phenotype. We hypothesize that V‐ATPases create a low pH in a localized environment which promotes tumor cell invasion by activating secreted cathepsins. Studies indicate that cathepsins secreted by cancer cells cleave extracellular matrix proteins and activate other proteases, facilitating tumor cell invasion. Our lab is investigating a connection between cell surface V‐ATPases, activation of secreted cathepsins and the metastatic phenotype of MDA‐MB231 cells. Previous data from our lab indicates that inhibition of V‐ATPases by a general inhibitor reduced the activity of secreted cathepsin B, but not cathepsin L. However, although there was a trend, the difference in activity for secreted cathepsin B after V‐ATPase inhibition was not statistically significant. Our current data supports the conclusion that there is a significant difference in the effects of V‐ATPase inhibition on secreted cathepsins. Inhibiting V‐ATPases does not appear to affect the activity of secreted cathepsin L, however it reduces the activity of secreted cathepsins B and D To investigate the role of V‐ATPases in the activation of cathepsins further we are using CRISPR/Cas9 to knockout the gene for two of the four V‐ATPase “a” subunit isoforms, a1 and a3. Published data indicates that the a3 and a4 isoforms are found in V‐ATPases at the cell surface of MDA‐MB231 cells. The a1 isoform is found in intracellular compartments. We hypothesize that the a3 isoform knockout cells will have reduced activation of secreted cathepsins B and D while not effecting activation of cathepsin L. We predict that the a1 isoform knockout will affected the activity of intracellular cathepsins, but not affect the activity of secreted cathepsin B, D or L. Separate abstracts submitted by our lab are focused on the a1 and a3 isoform knockouts.Support or Funding InformationDenison University Anderson AssistantshipDenison University Research Fund