bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genotyping the FCGR3a polymorphism using melt curve analysis
Authors: Burchard Paul R. , Malhotra Saurabh , Kaur Prabhjot , Tsongalis Gregory J. 
Published: June 2021

BackgroundFCGR3a (FcγRIIIA or CD16) plays an important role in immunoinflammatory processes. Recently a polymorphism in this gene has been shown to influence response to rituximab (anti CD20) therapy. Here, we evaluate a new method to genotype this polymorphism.Methods69 samples of genomic DNA extracted from either human blood or FFPE tissue were tested for the FCGR3a polymorphism using a real time PCR followed by melt curve analysis or by a Taqman allelic discrimination assay using the AB 7500 FAST real time PCR instrument. The Taqman assay consisted of the following PCR conditions: 60C for 1:00; 95C for 10:00; 40 cycles of 92C for 15 sec, 60C for 1:00; 60C for 1:00. The PCR/melt curve assay was performed using the following conditions: 95C for 5:00; 40 cycles of 95C for 20 sec, 56C for 20 sec, 72C for 30 sec followed by a continuous melt form 72C for 10 sec then 1% continuous temperature increase until 97C for 10 sec.ResultsThe TaqMan assay identified 29/69 (42%) cases as homozygous wild type F/F, 11/69 (16%) cases as homozygous V/V and 29/69 (42%) cases as heterozygous V/F. The real time PCR followed by melt curve analysis correctly identified 68/69 (98.6%) of the previous TaqMan results, thus demonstrating a high interassay concordance. The one discordant case was identified as the V/F genotype by the TaqMan assay, but as the F/F genotype by the melt curve assay.ConclusionsBoth the TaqMan allelic discrimination and the melt curve assays gave concordant results. The assay utilizes standard technologies in a molecular diagnostic laboratory and could be used for clinical assessment of the FCGR3a polymorphism.
Decreased eNOS relaxations and relation with hsp90 in hypercholesterolemic rabbit aorta
Authors: Ozsarlak‐Sozer Gonen , Arun Mehmet , Sevin Gulnur , Ertuna Elif , Yilmaz Zeynep , Ulasan Sema , Yetik‐Anacak Gunay 
Published: June 2021

Hsp90 activates eNOS in vasculature. we investigated basal and agonist evoked eNOS dependent relaxations and the role of hsp90 on these endothelial relaxations in hypercholesterolemia, since it is not studied well.Rabbits were fed with a regular/high cholesterol diet (%2) for 6 weeks. Hypercholesterolemia almost totally abolished agonist evoked eNOS dependent KCl‐ACh relaxations (93,9 % inhibition), however inhibited phenylephrine‐ACh relaxations only by 32,8 % in aorta in krebs buffer with indomethacine suggesting increased EDHF responses in hypercholesterolemic aorta.Basal eNOS dependent LNA‐induced further contraction and papaverin‐induced relaxations were not changed in hypercholesterolemic aorta.Hsp90 inhibitor radicicol (10 μg/ml) for 15 hrs decreased ACh relaxation after pre‐contraction with KCl (11,22±1,18 vs 7,33±1,58 *p<0,01 student t test, n=5) however did not cause any further inhibition in hypercholesterolemic aorta, suggesting that hsp90 inhibitors do not cause any further inhibition since hsp90 dependent eNOS activation may be already disrupted in hypercholesterolemia.We concluded that although basal eNOS dependent relaxations were not affected, since the agonist evoked eNOS relaxations were decreased which seems mainly depend on hsp90, a compansotory increase in EDHF relaxations may develop in hypercholesterolemic aorta. (Supported by TUBA GEBÝP‐2010 and FABAL).
Pro‐osteolytic effects of chronic morphine administration in a model of breast cancer‐induced bone pain.
Authors: Symons‐Liguori Ashley M , Sayers Nicole Marie , Vanderah Todd W 
Published: June 2021

Many common cancers have a high propensity for bone metastasis; for breast cancer, bone metastasis affects 70% of advanced patients and is the most common site for secondary tumor relapse. Tumor manifestation disrupts bone homeostasis and results in debilitating chronic skeletal pain that decreases patient functional status, quality of life and survival. Morphine remains the golden standard for managing cancer‐induced bone pain; however, opioids including morphine are associated with a host of side effects that complicate chronic dosing schedules. Furthermore, it has been posited that morphine may accelerate bone wasting in patients; in models of secondary bone metastasis, morphine exacerbates fracture rate, markers of osteolysis, and pain behaviors.We sought to determine whether morphine exacerbates bone loss in the 66.1‐Balb/C model of breast cancer‐induced bone metastasis (BCBM). Sham and tumor‐inoculated animals received a constant morphine infusion for 7 days (20 mg/kg/day). Morphine produced initial analgesia but failed to manage behaviors indicative of cancer‐induced bone pain over time; additionally, morphine treatment increased TRACP 5b in sera and RANKL in femur extrudate, suggesting that morphine enhances osteolysis in BCBM. In vitro studies evaluated the ability of morphine to induce osteoclastogenesis in the presence of permissive levels of RANKL. Morphine did not appear to affect tumor burden in vivo or in vitro viability of tumor cells. These data suggest that morphine worsens bone loss in the 66.1‐Balb/C model and may act directly on osteoclast cells; however, the exact mechanism of morphine‐induced bone loss is unknown and remains a barrier to improving pain relief in suffering BCBM patients.
Altered protein phosphatase 1 (PP1) activity and expression contribute to the differential regulation of proximal tubule NHE3 before and after onset of hypertension in SHR
Authors: Crajoinas Renato Oliveira , Pessoa Thaissa Dantas , Malnic Gerhard , Girardi Adriana Castello Costa 
Published: June 2021

We have previously demonstrated that NHE3 activity is higher in the proximal tubule of young pre‐hypertensive SHR (Y‐SHR) compared to adult hypertensive SHR (A‐SHR). This fall in NHE3 activity with blood pressure increase was associated with a higher ratio of phosphorylated NHE3 at serine 552 to total NHE3 (P‐NHE3/ total). The present study was undertaken to test the hypothesis that the increased levels of phosphorylated NHE3 at the PKA consensus site serine 552 was due to an increase of PKA activity, or a decrease of PP1 activity, or both. To confirm that PKA‐mediated phosphorylation of NHE3 regulation is altered in SHR, we examined the effect of 6MB‐cAMP (an cAMP analog that specifically activates PKA) on both phosphorylation levels and activity of NHE3 in young and adult SHR. We found that the 6MB‐cAMP‐induced increase in the levels of NHE3 phosphorylation at the serine 552 was much higher in the renal cortex from Y‐SHR than from A‐SHR (179 ± 14 vs. 36 ± 4%, P < 0.001). Likewise, the inhibitory effect of 6MB‐cAMP on NHE3‐mediated bicarbonate reabsorption was much more pronounced in proximal tubule from Y‐SHR than from A‐SHR (65 ± 10 vs. 23 ± 9%, P < 0.01). PKA activity remained unchanged between Y‐SHR and A‐SHR. On the other hand, Y‐SHR exhibited higher PP1 activity than A‐SHR (1640 ± 107 vs. 940 ± 119 pM/μg, P < 0.01). Furthermore, PP1α expression was decreased in the renal cortex of A‐SHR compared to Y‐SHR (32 ± 8 %, P < 0.01). We conclude that the differential regulation of proximal tubule NHE3 activity and phosphorylation levels before and after onset of hypertension in SHR is, at least in part, due to changes in the activity and expression of PP1.Supported by FAPESP
HIF‐1alpha activation improves mitochondrial function and morphology in the subtotal nephrectomy model of CKD
Authors: Singh Prabhleen , Pham Hai , Finley Joseph , Perkins Guy , Petrosyan Susanna , Murphy Anne , Patel Hemal 
Published: June 2021

HIF‐1α activation in subtotal nephrectomy (STN) lowers elevated renal oxygen consumption, ameliorates hypoxia and improves GFR. HIF‐1α also has significant effects on mitochondrial metabolism. Since the kidney derives most of its energy from oxidative metabolism, we examined the regulation of mitochondrial oxidative metabolism by HIF‐1α in early STN kidney model of CKD.Mitochondrial oxygen consumption, ATP generation, ROS production and morphology were examined in 1‐week STN rats and DMOG (HIF‐1α inducer) treated STN rats and compared with controls. Mitochondrial oxygen consumption rates were elevated in STN. HIF‐1 α activation lowered the mitochondrial oxygen consumption in both STN and controls. Mitochondrial ATP generation was modestly elevated and ROS (superoxide) production was significantly elevated in STN. HIF‐1 α activation significantly lowered both ATP and ROS production. All results were significant (p<0.05). Mitochondrial morphology was significantly altered in early STN with cortical mitochondria showing an elongated phenotype (fusion) and medullary mitochondrial showing a rounded phenotype (fission). Mitochondrial lengths, volume density and cristae abundance were increased in STN cortical mitochondria compared to control and STN medullary mitochondria. HIF‐1 α activation increased mitochondrial number and volume density in the STN cortical mitochondria and mitochondrial lengths in the control cortical and medullary mitochondria.In conclusion, there are significant alterations in mitochondrial function and morphology and function in early STN kidney. Several of these are ameliorated by HIF‐1α activation, which may be important in the preservation renal function and morphology and to slow the progression of disease.
The Kv1.3 Blocker PAP‐1 Reduces Infarction and Neurological Deficit in a Rat Model of Reperfusion Stroke
Authors: Chen Yi‐Je , Singh Vikrant , Garing April , Wulff Heike 
Published: June 2021

Activated microglia significantly contribute to the secondary inflammatory damage in ischemic stroke and therefore constitute attractive targets for post‐infarct intervention. Microglia express the voltage‐gated Kv1.3 and the calcium‐activated KCa3.1 channel and both channels are involved in microglia mediated neuronal killing, oxidative burst and inflammatory cytokine production. We previously found that KCa3.1 blockade with TRAM‐34 reduces neurological deficit and infarct area in a rat model of ischemic stroke even if initiated 12 hours after reperfusion. We now investigated whether Kv1.3 blockade would have similar effects and whether combined blockade of both microglial K+ channels would offer additional benefits. We subjected male Wistar rats to 90 min of middle cerebral artery occlusion (MCAO) and administered either vehicle or the Kv1.3 blocker PAP‐1 (40 mg/kg i.p. BID) for 7 days starting 12 h after reperfusion. PAP‐1 significantly reduced infarct area and improved neurological deficit in the infarcted hemisphere on day‐7. However, combined blockade of both Kv1.3 and KCa3.1 with PAP‐1 and TRAM‐34 did not further reduce infarct area compared to treatment with either TRAM‐34 or PAP‐1 alone suggesting that blockade of one microglial K+ channel is sufficient to improve outcomes of ischemic stroke.Supported by RO1 GM076063 from the National Institute of Health.
hERG1 agonist NS1643 inhibits proliferation of ER‐neg breast cancer cells by activating a senescence program
Authors: Gentile Saverio , Lansu Katherine , Ha Geun‐Hyoung , Breuer Eun‐Kyoung 
Published: June 2021

Function of the hERG1 potassium channel has been well characterized in the heart where it plays an important role in contraction. However, hERG1 channel is expressed in cancers of different hystogenesis. Although inhibiting hERG1 activity can lead to apoptosis, this approach is not likely to be successful in cancer therapy because of the association between hERG1 blocking agents and cardiac ventricular fibrillation. Nevertheless, the hERG1 agonist NS1643 has been found to protect against cardiac arrhythmia but nothing is known about the effects of this drug in cancer cells.The most successful breast cancer treatment relies on inhibition of the estrogen (E2) receptor (ER) in mediating the effect of E2 on cell proliferation (Tamoxifen). However, this approach cannot be employed for about 30% of breast tumors as they do not express ERs (ERneg).Our work demonstrates that application of the hERG1 activator NS1643 to ERneg breast cancer cells strongly inhibits proliferation by causing cell cycle arrest in G1 phase. In addition, cells treated with NS1643 lost invading and migrating capability.Interestingly, NS1643 treatment did not cause apoptosis but expression level of the tumor suppressor proteins p21 and p16INK4 increased significantly.In conclusion, these data suggest that hyper stimulation of hERG1 activity in ERneg breast cancer cells can arrest cell proliferation by activating a senescence program.
Ion Channel Phosphorylopathy: a link between genomic variation and human disease.
Authors: Gentile Saverio 
Published: June 2021

Many genomic variations that change amino acids in cytoplasmic domains of ion channels have been found to be associated with several diseases, including cardiac arrhythmias and neurodegeneration. However, a mechanism linking these mutations to ion channel malfunction has not been clearly established and in some cases is totally unknown.Our present work gives an overview on a possible mechanism by which disease‐associated mutations in cytoplasmic domains of several ion channels can affect current activity by creating or disrupting phosphorylation sites for specific kinases. We call these events “Ion Channel Phosphorylopathies”. Understanding Ion Channel Phosphorylopathies offers the opportunity to find a mechanism linking genomic variations to human disease and is crucial to the process of designing an effective pharmacological strategy.
Lack of CFTR in a cystic fibrosis pig model disrupts airway smooth muscle development
Authors: Stoltz David A. , Gansemer Nick D. , Rector Michael V. , Bouzek Drake , Moninger Thomas O. , Meyerholz David K. 
Published: June 2021

To better understand the pathogenesis of cystic fibrosis (CF) lung disease, we recently generated CFTR‐targeted pigs. CF pigs develop lung disease typical of human CF. An unexpected finding at birth, prior to the onset of infection and inflammation, was that CF tracheas have a reduced lumen area, irregularly developed airway cartilage, and airway smooth muscle (ASM) with prominent appearing bundles. We hypothesized that these abnormal‐appearing ASM bundles were due to differences in ASM proliferation. To test this hypothesis, tracheas were harvested at birth for histological examination. Despite a reduction in tracheal ASM area in CF, we found that ASM cell density was increased in CF (CF: 0.52±0.06 vs. non‐CF: 0.39±0.02 cells/100 μm2). To investigate if cell proliferation was increased in CF, we stained for the proliferation marker Ki‐67 and found reduced Ki‐67 positive cells in CF (CF: 0.04%±0.01 vs. non‐CF: 0.1%±0.01). This decrease was associated with a reduction in CF ASM cell area (197±19 vs. 259±14 μm2). Finally, electron microscopic studies revealed that at birth CF ASM has a contracted appearance, likely accounting for the reduced cell area. These data suggest that CFTR is important in ASM development and might explain, in part, the increased incidence of airway hyper‐responsiveness in human CF.This work was supported, in part, by the American Asthma Foundation (Sandler).
Anoctamin‐1 is expressed and functional in the Pulmonary Vascular Endothelium
Authors: Allawzi Ayed Mamdouh , Vang Alexander , Li Anlong , Choudhary Gaurav 
Published: June 2021

Background and ObjectiveThe expression and functional role of Anoctamin‐1 (ANO1), a Ca2+ activated Cl channel in the endothelial cells (EC) is unclear. We sought to confirm the presence of ANO1 in the pulmonary vascular EC (PVEC) and the effect of ANO1 activation on EC membrane potential (Vm).MethodsWhole lung lysates and primary isolated rat lung EC (RLEC) were isolated from 8 week old Sprague‐Dawley rats. ANO1 expression was evaluated in whole lung, commercially available primary rat lung microvascular EC (RLMVEC) and human pulmonary artery EC (HPAEC) lysates by immunoblot, in RLEC and RLMVEC by RT‐PCR, and in RLEC by immunofluorescence (IF). Changes in RLMVEC Vm was assessed using DiBAC4(3) and fluorescent live cell imaging.ResultsImmunoblotting confirmed the presence of ANO1 in whole lung lysates, RLMVEC, and HPAEC. RT‐PCR validated ANO1 expression in RLMVEC and RLEC. IF confirmed the expression of ANO1 on RLEC. Activation of ANO1 with 10μM Eact, a ANO1 opener, resulted in a hyperpolarization of Vm, which was attenuated in presence of a low chloride solution.ConclusionANO1 is expressed in the PVEC and can alter the resting Vm.
Smooth muscle microRNAs play a crucial role in regulation of myogenic tone in small mesenteric arteries
Authors: Bhattachariya Anirban , Nordström Ina , Swärd Karl , Hellstrand Per , Albinsson Sebastian 
Published: June 2021

Myogenic, pressure‐induced, tone is an intrinsic mechanism for the regulation of blood pressure and blood flow to the organs. Small non‐coding microRNAs (miRNAs) have been implicated in the regulation of smooth muscle differentiation and vascular contractility. However, the role of miRNAs in pressure‐induced contractile responses has not been investigated previously.The aim of this study was to determine the role of miRNAs in myogenic tone using a pressure myograph and smooth muscle specific, tamoxifen‐inducible knockout of Dicer, an enzyme essential for the biogenesis of miRNAs. The results from these mice were then compared to those of the miR143/145−/− and miR143/145+/− mice. In Dicer KO, myogenic tone was abolished 5 weeks post tamoxifen treatment, even though KCl‐induced contraction was only moderately reduced. A similar phenotype was observed in miR‐143/145−/− mice. Interestingly, myogenic tone was also abolished in miR‐143/145+/− mice, which exhibit normal KCl‐induced contraction. This result indicates that that myogenic constriction is highly sensitive to a reduction of these specific miRNAs. Furthermore, we found that the L‐type calcium channel opener Bay K 8644 partially rescued myogenic tone in Dicer KO and 143/145+/− but not in 143/145−/−. Our results suggest that miRNAs play a crucial role in the regulation of myogenic tone, possibly via regulation of L‐type channel activity
Increased expression of AT2 receptors in the nucleus of the solitary tract improves baroreflex function in renovascular hypertensive rats.
Authors: Blanch Graziela T , Freiria‐Oliveira Andre H , Rigatto Katya , Li Hongwei , Colombari Eduardo , Sumners Colin , Colombari Debora S A 
Published: June 2021

Renovascular hypertensive (2K1C) rats exhibit impaired baroreflexes. Here we studied the effects of increased AT2R expression in the nucleus of the solitary tract (NTS) on baroreflex function of 2K1C rats. The left renal artery was partially obstructed in male Holtzman rats (150–180g) using a silver clip (0.2 mm width). Three weeks later, rats were microinjected in the NTS with either AAV2‐CBA‐AT2R or AAV2‐CBA‐eGFP to elicit chronic increases in AT2R or GFP (control) expression respectively. Thirty days later, femoral artery and vein were cannulated to measure mean arterial pressure (MAP) and heart rate (HR) and perform drug infusions, respectively. The next day, phenylephrine (5 μg/kg, iv) and sodium nitroprusside (30 μg/kg, iv) were used to elicit respective pressor and depressor responses in conscious rats. Reflex HR responses were analyzed to each 10 mmHg change in MAP. Baseline MAP in 2K1C‐AT2R rats (173 ± 7 mmHg) was lower than in 2K1C‐GFP rats (199 ± 9 mmHg; p <0.05). 2K1C‐GFP rats exhibited a significant impairment of the baroreflex (slope: −0.95 ± 0.05, vs. sham‐GFP −1.37 ± 0.06, p<0.05), which was improved in the 2K1C‐AT2R rats (slope: −1.44 ± 0.07). Baroreflex in 2K1C‐AT2R was similar to sham‐AT2R rats (slope: −1.56 ± 0.06) or sham‐GFP. The results suggest that AT2R over expression in the NTS of 2K1C rats improves baroreflex and this may contribute to the reduction in blood pressure observed in these animals.Supported by:PNPD/CAPES, NIH and FAPESP/PRONEX.
Desensitization of cough during exercise in anesthetized rabbits
Authors: Poussel Mathias , Bosser Gilles , Tiotiu Angelica , Chalon Bernard , Chenuel Bruno 
Published: June 2021

The ‘cough center’ exhibits plasticity at the sensor and integration levels leading to modulation of the reflex in its strength or in its pattern. Little is known about interactions between cough and human activities, especially during exercise. The present study was designed to determine whether exercise, mimiked by electrically‐induced muscular contractions, is able to change the occurrence and/or the strenght of cough following a mechanical stimulation of the trachea in anesthetized rabbits.Thirteen anesthetized, tracheotomized rabbits were studied allowing 329 tracheal stimulations : 208 at rest and 121 during exercise.During exercise, the incidence of the cough reflex (CR) decreased and the expiration reflex (ER) increased (p < 0.0001). Whereas the sensitivity of the CR and the ER are both decreased during exercise compared to the sensitivity of the CR at rest (p < 0.02), the strength of the expulsive response is unchanged.These results indicate that adjustments occurring during exercise likely down‐regulates tracheal defensive reflexes in anesthetized rabbits.
Types of tracheal submucosal glands: implications of literature controversy
Authors: Barros Mirna Duarte , Landim Amanda P , Dobre Ana Clara M , Andreotti Joao Duvilio Biazzi , Curcio Daniella Franco , dos Anjos Duarte Ana Lúcia 
Published: June 2021

In what concerns the morphological description of tissues and organs there are some conceptual discrepancies in the classical literature used as the knowledge foundation of basic sciences in health and medical colleges. Such controversies may create conflicts and jeopardize the relationship of trust between students and faculty members. This paper compares basic histology books and atlases with respect to the main information on tracheal subucosal glands. We analyzed 23 books and 21 atlases (from which 17 were virtual atlases), focusing information regarding presence and classification of tracheal submucosal glands. All references agreed that exocrine glands were present in the tracheal submucosa, however we found controversy about its types: presence of “seromucous glands” were referred by 39.1% of the references; presence of “mucous and serous glands”, by 26.1%; “mucous glands with serous components”, 17.4%; presence of both “mucous” and “seromucous”, 4.3%; “mixed glands” 4.3%; mucous, serous and mixed glands, 2.2%; mucous only, 2.2%; serous only, 2.2%. No information on gland type could be found in 2.2% of the references.CONCLUSIONThere is no consensus in what concerns the classification of the glands present in the trachea submucosa. The lack of consistence in basic information and terms may be a source of problems in standardization of histological terminology. Furthermore, such controversy may affect student's confidence in the subject and the staff, since the reliability of the theoretical basic knowledge is questioned.
Comparison between Student Desired Grades and Actual Final Grades in Undergraduate Human Anatomy
Authors: Dunham Stacey 
Published: June 2021

Anat A215, Basic Human Anatomy, is a large enrollment (400+) lecture/lab course offered each fall and spring semester at Indiana University. Each semester students complete a short survey in which they indicate name, major, standing in school, previous anatomy coursework, and the lowest grade they would want for the course.In this retrospective study the information provided by students in these surveys was compared to their final course grade earned in Anat A215, as recorded in the instructor grade files. The first goal of this study was to identify how accurately students are predicting their own grades. Second, this study intended to identify any associations between ability to accurately predict course grades and the variables included in the survey, including demographic information and students’ reported desired grade and corresponding rationale for earning that grade (to maintain GPA, to raise GPA, to get into specific undergraduate or graduate programs). Results show that of 350 students, 40% scored lower than their stated “lowest” desired grade. Further analysis will look for trends impacting both successful and failing grade predictions by students.Grant Funding Source: None