bluuebunny/crossref_metadata_2025_split huggingface dataset:
Disturbance of post‐natal lung development in Norwich terriers
Authors: Anttila Marjukka , Dillard Kati , Vainio-Siukola Katri 
Published: June 2021

An arrest in the post‐natal lung development is a major cause of death in Norwich terrier puppies. Pedigree analysis indicates that the disorder is inherited. In dogs the lungs at term are in the saccular phase and the development into the alveolar stage occurs during the neonatal period.In the affected lungs the alveolar septae are poorly developed and the alveolar walls are thicker than normal. Immunohistochemical staining for vimentin, smooth muscle actin (SMA), CD31 and thrombomodulin was used to characterize the cells forming the alveolar parenchyme. There is a sequential and spatial pattern of SMA staining in normal canine lung examined 3 days, 3 weeks and 3 months after birth. The pattern appears to be similar in the affected puppies up to 3 weeks of age.Our canine model enables us to study a spontaneously occurring disease. The aim is to describe the genetic background of this congenital pulmonary disorder and to learn more about the molecular events involved in post‐natal lung development. In addition we wish to develop a DNA test for breeding purposes to eradicate the disease in Norwich terrier breed.
Effects of dopamine beta‐hydroxylase (DBH) inhibition on cocaine‐induced reinstatement in squirrel monkeys
Authors: Cooper Debra Andrea , Kimmel Heather , Howell Leonard 
Published: June 2021

Disulfiram (Antabuse) has shown promise in clinical trials as a pharmacotherapy for cocaine dependence. One of many biochemical actions of disulfiram is inhibition of dopamine beta‐hydroxylase (DBH), the enzyme that converts dopamine to norepinephrine (NE); consequently, reducing brain NE levels. It is believed that this mechanism is responsible for the therapeutic effects of disulfiram. In rats, disulfiram blocks cocaine‐primed reinstatement (an animal model of relapse), corroborating with clinical results. Furthermore, nepicastat, a selective DBH inhibitor also blocks cocaine‐primed reinstatement in rats, supporting the therapeutic use of DBH inhibitors. In this study, we attempted to replicate these results in non‐human primates. Squirrel monkeys trained to self‐administer cocaine under a second‐order schedule were pretreated with disulfiram or nepicastat prior to cocaine‐primed reinstatement sessions. Neither pretreatment altered cocaine‐primed reinstatement. Unexpectedly, nepicastat alone was sufficient to induce a modest reinstatement effect. The discrepancies found between squirrel monkeys and rats may be explained by differences in pharmacokinetic or metabolic profiles in the two species. Ongoing studies are investigating whether metabolic differences are present, which will ultimately be instrumental in influencing the translation of these therapies to a human population.Support: DA00517, DA12514, 5T32DA015040, UL1 RR025008, RR00165
Nitric Oxide Synthase Knockout Modulates Ca2+‐Sensing Receptor Expression and Signaling in Mesenteric Arteries
Authors: Awumey Emmanuel Modesto , Bridges Lakeesha E , Williams Cicely L , Diz Debra I 
Published: June 2021

Ca2+e‐induced relaxation of mesenteric artery is dependent on the perivascular nerve (PvN) Ca2+‐sensing receptor (CaSR). The CaSR stimulates a cannabinoid vasodilator pathway, with a small (~20%) nitric oxide (NO)‐dependent component. Wire myography of phenylephrine (PE)‐contracted mesenteric arteries was used to determine the source of NO contributing to CaSR‐mediated relaxation in NO synthase (NOS) gene knockout mice in combination with NOS inhibitors. . Endothelial NOS gene knockout (eNOS−/−) up‐regulates, whereas neuronal NOS gene knockout (nNOS−/−) down‐regulates CaSR protein expression. nNOS−/− reduced maximum Ca2+ e‐induced relaxation by ~ 34% with no change in EC50 values, consistent with down‐regulation of the receptor as the non‐selective NOS inhibitor, L‐N5‐(1‐Iminoethyl)‐Ornithine (10 μM) reduced Ca2+ e‐induced relaxation in C57BL/6 control mice by ~ 38% (p < 0.05), but not eNOS−/− mice. The nNOS inhibitor, 7‐Nitroindazole had no effect on arteries from nNOS−/− mice. Both NG‐Nitro‐L‐Arginine Methylester and NG‐Monomethyl‐L‐Arginine reduced Ca2+‐induced relaxation maxima by about 55%, 50% and 30% in C57BL/6, nNOS−/− and eNOS−/− mice, respectively. Thus eNOS down‐regulates the CaSR in mesenteric arteries and contributes to the NO‐mediated Ca2+‐induced relaxation, but nNOS is not a major contributor.Support: HL064761, HL059868, MD000175 & HL099139
Ca2+‐related proteins associated with intracellular stores in rat lymphatics
Authors: Nepiyushchikh Zhanna V , Bridenbaugh Eric A , Muthuchamy Mariappan , von der Weid Pierre-Yves , Zawieja David C 
Published: June 2021

Calcium plays essential roles in the contraction of striated and smooth muscle types. Changes in cytoplasmic concentration of Ca2+ depend in general on the calcium channels and pumps at the cytoplasmic and endoplasmic/sarcoplasmic reticulum membrane. As we have shown previously, lymphatic muscle has a unique combination of the striated and smooth muscle contractile and regulatory proteins (Muthuchamy et al., 2003). However, very little is known about the composition of the Ca2+ ‐channels and pumping structures in the intracellular stores of lymphatic muscle. Our immunohistochemistry data with confocal microcopy from whole isolated vessels preparation, frozen sections and lymphatic muscle cell culture demonstrates the presence of inositol 1,4,5‐triphosphate receptor type 3 (IP3R3) channels, Ca2+ transporting ATPase type 2a1 (SERCA2a1) and Ca2+ transporting ATPase type 2a2 (SERCA2a2) at intracellular stores of rat mesenteric lymphatic vessels, cervical lymphatics and thoracic duct.Supported by NIH HL096552
Sensitization of Rats with Low dose of Angiotensin II through Multiple Signaling Pathways in Lamina Terminalis
Authors: Zhang Zhongming , Xue Baojian , Guo Fang , Beltz Terry , Johnson Alan K 
Published: June 2021

A one wk infusion of a non‐pressor dose of ANGII (induction phase; I) followed by a 1 wk rest (delay period; D) results in a sensitized hypertensive response produced by a subsequent 2 wk infusion of ANGII (expresson period; E). Several brain areas including structures along the lamina terminalis (LT) participate in the regulation of blood pressure. The present study investigated the effects of the non‐pressor sensitizing dose of ANGII (10ng/min/kg) on candidate second messengers in LT that are implicated in sensitization during E. Following a 1 week infusion of either vehicle or ANGII during I LT tissue was collected from rats at the end of D. The dissected tissue was prepared for western blot or for real time PCR.Western blot analyses of LT tissue indicated that phosphorylated cAMP responsive element binding protein (CREB), phosphorylated ERK, and phosphorylated phospholipase C gamma (PLCã) increased 1.5‐, 1.4‐, 1.4 and 1.6 fold respectively (p<0.05). Brain derived neurotrophic factor (BDNF) increased 1.5‐fold. These findings were consistent with real time PCR data showing that BDNF mRNA levels increased 1.6 fold and c‐fos is increased 1.6 fold. These data suggested that the induction by ANGII activates gene expression through PKA, MAPKs and PLC pathways. BDNF may be directly upregulated through the activation of PKA pathway, which further activates TrkB so that AngII‐induced signaling is amplified.
SLC2A2 gene is upregulated in diabetes and dowregulated in insulin treatment: participation of HNFs (1α, 3β and 4α)
Authors: Silva Aline David , Freitas Helayne Soares , Furuya Daniela Tomie , Poletto Ana Cláudia , Silva Robinson Sabino , Okamoto Maristela Mitiko , Machado Ubiratan Fabres 
Published: June 2021

ObjectiveInvestigate the role of HNFs (1á, 3â, 4á) at SLC2A2 (GLUT2 gene) in liver of diabetic and insulin treated diabetic rats.Methods3 month old Wistar rats, allocated into non diabetic (C), diabetic (D), diabetic treated with saline (DS) or insulin (DI) for 1, 4 and 6 days (d). The mRNA of GLUT2 and HNFs was analyzed by RT‐PCR. The HNFs binding activity into SLC2A2 promoter was analyzed by EMSA.ResultsIn comparison to C, D rats showed: hyperglycemia, polyuria, glycosuria without ketonuria, less gain of weight; increase of: I) GLUT2 (P<0,0001), HNF1á (P<0,05), HNF3â (P<0,01) and HNF4á (P<0,05) mRNA expression, II) bindig activity of HNF1á (P<0,05), HNF3â (P<0,01) and HNF4á (P<0,05) into SLC2A2. In comparison to DS, DI rats showed: reduction of glycemia, increased of body weight; decrease of: I) mRNA expression: GLUT2 after 4 and 6d (P< 0,001), HNF1á after 6d (P<0,05), HNF3â after 4 and 6d (P<0,001) and HNF4á after 6d (P<0,05), II) binding activity into SLC2A2: HNF1á after 4 and 6d (P<0,01), HNF3â after 1, 4 (P<0,05) and 6d (P<0,01) and HNF4á after 6d (P<0,05).ConclusionThe SLC2A2 overexpression is related to expression and activity of HNFs (1á, 3â, 4á). Furthermore, insulin therapy‐induced regulation of SLC2A2 expression also involves HNFs. Considering the importance of the GLUT2, we hope that the knowledge of the SLC2A2 transcriptional regulation may contribute to control gene expression.FAPESP 2009/03871‐7
Endothelin‐1‐induced inhibition of NKCC2 activity is blunted in thick ascending limbs from angiotensin II‐hypertensive rats
Authors: Ramseyer Vanesa Daniela , Garvin Jeffrey Lawrence 
Published: June 2021

Thick ascending limbs (THALs) absorb 25% of the filtered NaCl. Endothelin‐1 (ET‐1) inhibits NaCl absorption in THALs by increasing NO and reducing Na/K/2Cl cotransporter (NKCC2) activity via ETB receptors. THAL NaCl absorption is enhanced while NO production is reduced in angiotensin II (Ang II)‐hypertensive rats. Thus, we hypothesized that inhibition of NKCC2 by ET‐1 is blunted in Ang II‐induced hypertension. Rats were infused with vehicle or Ang II 200 ng/kg/min. At day 5 we measured: 1) mean arterial blood pressure (MABP) in anesthetized rats; 2) NKCC2 activity in perfused THALs by fluorescence microscopy and; 3) NOS3 and ETB receptor expression in THALs by Western blot. Ang II infusion increased MABP from 84 ± 5 mmHg to 98 ± 1 mmHg (p< 0.01). In vehicle‐treated rats, 1 nM ET‐1 reduced NKCC2 activity from 1.65 ± 0.15 to 0.84 ± 0.14 AU/s (Δ= ‐50 ± 7 %, p<0.01). In contrast, in Ang II‐hypertensive rats, ET‐1 did not decrease NKCC2 activity (from 1.46 ± 0.24 to 1.21 ± 0.22 AU/s). Ang II treatment reduced NOS3 expression by 35 ± 13% (p<0.03) but did not decrease ETB receptor expression (Δ= 10 ± 10%). We conclude that inhibition of NKCC2 activity by ET‐1 is blunted in Ang II‐induced hypertension and this is likely due to a reduction in NOS3 expression rather than ETB receptors. Impaired ET‐1‐induced inhibition of NKCC2 activity could be the cause of the enhanced NaCl absorption seen in this model of hypertension.Sources of funding: NIH HL 028982 to J.L.G. and AHA 11PRE7510005 to V.D.R
COX‐2 inhibition exacerbates SOD1 dowregulation and the progression of renal oxidative stress in response to UUO
Authors: Nilsson Line , Frøkiær Jørgen , Nørregaard Rikke 
Published: June 2021

Renal damage and oxidative stress resulting from unilateral ureteral obstruction (UUO) may be aggravated by reactive oxygen species (ROS). Deleterious effects of ROS are attenuated by antioxidant enzymes. We investigated the role of COX‐2 in the progression of renal oxidative stress induced by UUO in mice.Mice were subjected to UUO and treated with the selective COX‐2 inhibitor parecoxib for 7 days. Using immunoblot analysis, we examined the regulation of antioxidant enzymes, catalase, superoxid dismutase 1 and 2 (SOD1 and −2) in renal cortex and inner medulla. Immunohistochemistry was performed with oxidative stress markers.COX‐1 and −2 protein were increased in response to UUO. SOD1 and −2 protein were increased in IM in parecoxib treated sham operated mice. Parecoxib treated UUO mice had more reduced SOD1 protein level in IM and cortex than untreated UUO mice, whereas SOD2 and cortical catalase protein levels were unchanged.COX‐2 inhibition increased the renal SOD1 and −2, consistent with a stress response to increased ROS production. UUO reduced SOD1 and catalase, and inhibited increased SOD1 and −2 seen with COX‐2 inhibition.In conclusion, suppression of renal antioxidant enzymes by UUO contributes to the progression of renal oxidative stress, a process exacerbated by COX‐2 inhibition.
Analysis and expression pattern of slc12a genes
Authors: Di Fulvio Mauricio 
Published: June 2021

Mammalian cells exhibit a non‐equilibrium distribution of Cl− across their plasma membranes. This distribution plays a key role in cell volume regulation and growth, salt transport, K+ scavenging, hormone secretion and GABA signaling. The [Cl−]i results from the functional balance between Cl− transporters and channels expressed in the cellˈs plasma membrane. The family of solute carriers slc12a is the most relevant one involved in the regulation of the [Cl−]i. This family includes Na+K+2Cl−, Na+Cl− and K+Cl− co‐transporters: NKCCs, NCCs and KCCs, respectively. Two NKCC, one NCC and four KCC genes encode for NKCC1, NKCC2, NCC and KCC1‐4 isoforms. NKCC2 and NCC are considered expressed exclusively in the kidney, whereas NKCC1 is ubiquitously distributed. Similarly, KCC2 is considered a neuron‐specific isoform whilst KCC1, 3 and 4 appear to have less restricted expression patterns. To determine molecular identity, classify known slc12a members and establish patterns of expression we searched and analyzed nucleotide/protein sequences data posted in GenBank, EST and GEO databases. We found that at least two NKCC1 splice variants exist in humans, but different NKCC1s exist in rodents. Some NKCC2 variants were cloned from human kidneys, but were not found in rodents or vice versa. Moreover, at least ten KCC3 splice variants have been cloned from human tissues whilst rodents express a few ones. Further, NKCC2 or NCC mRNAs are detected outside the kidney. Therefore, splice variants of slc12a members may be species‐specific and/or non‐restricted to particular tissues. The molecular classification of slc12a variants and their pattern of expression will aid in the interpretation/extrapolation of functional/pharmacological data.This work has been supported by the American Diabetes Association (Grant JF‐1001730 to MDiF).
Effects of Oxytocin on Gastric‐Projecting Neurons in a Rat Model of Functional Dyspepsia
Authors: Babic Tanja , Browning Kirsteen N , Travagli Renato Alberto 
Published: June 2021

Emotional stress triggers functional dyspepsia (FD) and gastric dysmotility. Oxytocin (OXY) modulates gastric motility and has a recognized antistressor role. We have shown that although OXY receptors are expressed in the dorsal vagal complex, OXY perfusion does not modulate the critical GABAergic inputs to identified gastric‐projecting preganglionic vagal (DMV) neurons. Here we tested the hypothesis that FD induces an abnormal response to OXY in the gastric‐related vagal circuits.Following induction of FD, we did whole‐cell patch‐clamp recordings from gastric‐projecting DMV neurons from FD and control rats. In control rats, OXY had no effect on GABA currents in any of the neurons tested unless the neurons were pretreated with the stress hormone corticotrophin releasing factor (CRF). Following CRF, OXY altered GABAcurrents in 6/15 neurons. Conversely, in FD animals, OXY altered GABA currents in 10/25 neurons, independently of CRF. In FD rats, pretreatment of neurons with H89, a protein kinase A (PKA) inhibitor, blocked the effects of OXY on GABA currents in 7/8 neurons tested.These data show that FD induces an abnormal response to OXY in the gastric‐related vagal circuits similar to that obtained by the stress‐related hormone CRF. We also suggest that that the effects are mediated by activation of cAMP/PKA pathway.Supported by: NIH DK55330
Severe sustained hypoxia elicits long‐lasting phrenic motor facilitation (pMF) by an adenosinergic mechanism
Authors: Nichols Nicole L , Devinney Michael J , Mitchell Gordon S 
Published: June 2021

Moderate acute intermittent hypoxia (AIH; 3, 5‐min episodes) elicits serotonin‐dependent phrenic long‐term facilitation (pLTF); this form of pLTF is enhanced by spinal adenosine 2A (A2A) receptor inhibition. Severe AIH also elicits pLTF, but via an adenosine A2A receptor dependent (serotonin independent) mechanism. This shift may reflect greater spinal adenosine release/formation during severe AIH. Moderate sustained hypoxia (SH; 9–20‐min) fails to elicit pMF, demonstrating pMF pattern‐sensitivity. However, since severe SH may increase extracellular adenosine levels, we hypothesized that severe SH would elicit pMF via an A2A receptor dependent mechanism. Severe SH (PaO2=25–35; 25 min) was studied in anesthetized, paralyzed and ventilated rats +/− spinal A2A receptor inhibition (MSX‐3, i.t., C4). Severe SH elicits pMF (53%±12% at 60 min post‐SH) vs. time controls (10%±3%; p<0.05, both n=8). MSX‐3 pre‐treatment attenuated severe SH induced pMF (10%±5%; p<0.05, n=7). Thus, severe SH and AIH both elicit spinal A2A‐dependent pMF, unlike moderate SH and AIH (Devinney et al., ibid), indicating that adenosine‐dependent pMF is not pattern sensitive. Understanding mechanisms of pattern sensitivity in respiratory plasticity may enable us to “harness” plasticity as a treatment for respiratory insufficiency.[Supported by NIH HL080209 and T32 007654 and the Francis Families Foundation]
Spinal A2A receptor inhibition reveals phrenic motor facilitation following moderate sustained hypoxia
Authors: Devinney Michael J , Nichols Nicole L , Mitchell Gordon S 
Published: June 2021

Moderate acute intermittent hypoxia (AIH; 3, 5‐min episodes) elicits serotonin‐dependent phrenic long‐term facilitation (pLTF); this form of phrenic motor facilitation (pMF) is enhanced by spinal adenosine 2A (A2A) receptor inhibition. In contrast, moderate sustained hypoxia (SH; 9–20‐min) fails to elicit pMF, demonstrating pLTF pattern‐sensitivity. Since longer moderate hypoxic episodes may increase extracellular adenosine levels and negate pLTF, we hypothesized that A2A receptor blockade reveals pMF after moderate SH. pMF was studied in anesthetized, paralyzed, and ventilated rats ± spinal A2A receptor inhibition (MSX‐3, C4 i.t.). Although, moderate SH (25 min, PaO2 45–55) caused no significant pMF (n=9, 14±5%) vs. time controls (n=8, 10±3%; P >0.05) 60 min post SH, MSX‐3 pre‐treatment revealed significant pMF (n=8, 39±5% %, p<0.05 vs. SH). Thus, A2A receptor activation during moderate SH negates pMF. Since spinal A2A receptor activation and severe SH (25 min, PaO2 25–35) elicit adenosine‐dependent pMF (Nichols et al., ibid), pLTF pattern‐sensitivity may arise from emergent properties of parallel signaling networks giving rise to pMF; inhibitory interactions between these competing mechanisms negates pMF after moderate SH. Understanding mechanisms of pattern‐sensitivity in respiratory plasticity may enable us to better harness plasticity as a therapeutic tool.Supported by NIH HL080209, T32 007654 and the Francis Families Foundation.
Reduced lung glutamate (GLU) production is the cause of decreased systemic glutamate availability in hyperdynamic sepsis
Authors: Ten Have Gabriella A , Engelen Marielle P , Wolfe Robert R , Deutz Nicolaas E 
Published: June 2021

RATIONALEPreviously, we described in patients in the early phase of septic shock that reduced plasma GLU relates to mortality. As the mechanism is unclear, we studied GLU organ net balances (NB) in a Pseudomonas aeroginosa (PM) sepsis induced pig model.METHODSArterial GLU concentrations and transorgan GLU net balances were studied in 12 pigs with sepsis and 8 controls. GLU concentrations were determined with LC‐MS. Data are expressed as (mean (SE) control vs PM) in μM (conc) and nmol/kg bw/min (NB), statistics with Student t‐test.RESULTSIn PM pigs, arterial GLU concentration (81.3(6.8) vs 38.0(3.7)) and lung GLU net release (7472(1457) vs 1354(1204)) were reduced (resp. 53%, p<0.0001 and 82%, p=0.0088), while Liver net release was unchanged. Gut (195(49) vs 24(34)) and muscle (843(57) vs 376(73)) GLU net uptake were decreased (resp. 88%, p=0.009 and 55%, p=0.0002).CONCLUSIONThe reduced lung GLU production plays a major role in the reduced systemic GLU availability in sepsis.
Age‐related changes in the skeletal muscle DNA methylome and transcriptome
Authors: Thalacker-Mercer Anna Elizabeth , Day Kenneth , Cui Xiangqin , Waite Lindsay , Absher Devin , Merritt Edward , Bamman Marcas 
Published: June 2021

Advances in technology may assist with unraveling mechanisms linked to characteristic skeletal muscle (SkM) aging. The objective of this study was to identify age‐related differences in the SkM DNA methylome and transcriptome. DNA and RNA were isolated from resting state, old and young male, human SkM. Human Methylation BeadChips and Agilent microarrays were used to identify age‐related differences in SkM DNA methylation (DM) and gene expression (GE), respectively. Differentially methylated (1596) and expressed genes (1179) were identified between old and young SkM (False Discovery Rate of p<0.05). 52 genes were common between the male, aging DM and GE. Gene networks and functional annotation were determined with Ingenuity Pathways Analysis. Overrepresented functional categories related to the top gene network, molecular and cellular functions, and diseases/disorders included cellular development, growth, proliferation, and morphology, gene expression, cell cycle, inflammation, and neurological disease. Interestingly, 50% (26) of the common genes were associated with neurological diseases/disorders. Of these 26, 12 were related to neuromuscular disorders. These data support the hypothesis that age‐related changes in GE and perhaps phenotypes stem from changes in DM and highlight the need to understand the role of DM in neuromuscular aging.Support: 5R01AG017896, VA Merit, 5UL1 RR025777, UAB Center for Aging, P30DK056336
Mechanical Properties of Incisors in Rodents
Authors: Druzinsky Robert E , Naveh Gili , Weiner Steve , Brumfeld Vlad , Klein Ophir D. , Charles Cyril 
Published: June 2021

Rodents sharpen their incisors by grinding the lingual and buccal surfaces of opposing teeth. Normally the buccal surface is covered with enamel, whereas the lingual surface is not. While grinding enamel against dentin, the dentin wears and a sharp tip is created.We studied lower incisors in mice carrying mutations in sprouty genes, the protein products of which are antagonists of receptor‐tyrosine kinase signaling. Sprouty loss‐of‐function mutants (Spry2+/−;Spry4−/−) have phenotypic features that mimic the ancestral condition in rodents, which include second incisors and enamel on the lingual surfaces of the incisors. We found that in some individuals the phenotype was asymmetrical, with enamel on the lingual surface of one lower incisor but not on the other.We compared the properties of mutated and regular lower incisors from sprouty mutated mice. Teeth were axially loaded and scanned within a micro‐CT (Xradia). Normal incisors, without lingual enamel, bent lingually primarily at the worn and sharp part near the tip, whereas the teeth with enamel on both sides bent at a more apical location, with the result that the entire tooth bent. Much smaller excursions of the loading device were required to fracture the incisors with enamel on both sides. Our results demonstrate that enamel deposited on the lingual surface lowers the extent to which teeth bend and elevates the brittleness of the teeth.