bluuebunny/crossref_metadata_2025_split huggingface dataset:
Kefir exhibits anti‐proliferative and pro‐apoptotic effects on colon adenocarcinoma cells with no significant effects on cell migration and invasion (647.24)
Authors: Rizk Sandra , El Khoury Nathalie , El‐Hayek Stephany , Tarras Omayr , El‐sibai Mirvat , El‐Sabban Marwan 
Published: June 2021

Background: Previous studies showed that kefir, a fermented milk product, exhibited anti‐tumoral activity in vivo; yet its mechanism of action remains elusive. Recent studies have focused on the mechanism of action of kefir on cancer cells in vitro. The current study aims at examining the effect of kefir on cell survival, proliferation and motility of colorectal cancer (CRC) cells.Methods: Kefir’s anti‐cancer potential was tested on CRC cell lines, Caco‐2 and HT‐29, through cytotoxicity, proliferation, and apoptotic assays. The expression of certain genes involved in proliferation and apoptosis was assessed using RT‐PCR and western blot analysis to determine the possible mechanism of action of kefir. To assess the effect of kefir on cancer metastasis, wound healing and time‐lapse movies, in addition to collagen‐based invasion assay, were used.Results: Results have shown that cell‐free fractions of kefir exhibit an anti‐proliferative effect on Caco‐2 and HT‐29 cells. Analysis of DNA content by flow cytometry revealed the ability of kefir to induce cell cycle arrest at the G1 phase. Kefir was also found to induce apoptosis, as seen by cell death ELISA. Results from RT‐PCR showed that kefir decreases the expression of TGF‐α and TGF‐β1 in HT‐29 cells. Western blot results revealed an upregulation in Bax:Bcl‐2 ratio, confirming the pro‐apoptotic effect of kefir, and an increase in p53 independent‐p21 expression upon kefir treatment. MMP expression was not altered by kefir treatment. Furthermore, results from time‐lapse motility movies, wound‐healing, and invasion assays showed no effect on the motility of colorectal as well as breast (MCF‐7 and MB‐MDA‐231) cancer cells upon kefir treatment.Conclusion: Our data suggest that kefir is able to inhibit the proliferation and induce apoptosis in HT‐29 and Caco‐2 CRC cells, yet it does not exhibit a significant effect on the motility and invasion of these cells in vitro.Grant Funding Source: Supported by SRDC, Lebanese American University
Novel insights in post‐infectious irritable bowel syndrome in experimental giardiasis (650.15)
Authors: Halliez Marie , Le Goff Laetitia , Francois Arnaud , Colasse Elodie , Gargala Gilles , Villena Isabelle , Favennec Loic , Lapointe Tamia , Altier Christophe , Buret Andre 
Published: June 2021

Giardia duodenalis infections can lead to post‐infectious irritable bowel syndrome (PI‐IBS), a disorder characterized by chronic visceral pain. Using a new model of post‐giardiasis IBS, this study aimed to reveal novel mechanisms responsible for PI‐IBS. Weaning rats were gavaged with live Giardia trophozoites (assemblages A or B). Visceral hypersensitivity was assessed 50 days post‐infection (DPI) by measuring blood pressure changes in response to jejunal/rectal balloon distension. Spinal cord expression of c‐fos was used as a marker of nociceptive signaling. Caco‐2 monolayers were used to assess non‐invasive E. Coli HB101 translocation, as well as the integrity of epithelial barrier structure and function at early time points. c‐fos activation was observed during the acute phase. While the infection was cleared by 21DPI, jejunal and rectal hypersensitivity was observed 50 DPI with either assemblage along with mast cell infiltration. In vitro challenge with Giardia also facilitated the paracellular translocation of E. coli, increased epithelial permeability, and disrupted the tight junctional (TJ) proteins occludin and claudin‐4. These data show that Giardia‐induced IBS is associated with TJ proteins disruption, translocation of a commensal E. coli and c‐fos activation during the acute phase of the infection, as well as mast cell infiltration and visceral hypersensitivity. Findings in this model will help identify the mechanisms of Giardia leading to post‐giardiasis‐IBS.Grant Funding Source: Supported by the French ministry of secondary education and research, NSERC‐CREATE
Sensitization of sodium channels by cystathionine β‐synthetase activation in colon primary sensory neurons contributes to visceral hypersensitivity in adult rats with neonatal maternal deprivation (651.20)
Authors: Xu Guangyin 
Published: June 2021

Aims: This study was designed to determine roles for sodium channels and endogenous H2S signaling in a rat model of chronic visceral hypersensitivityinduced by neonatal maternal deprivation (NMD). Methods: Neonatal male rats were exposed to a 3 hour period of daily maternal separation. Colon‐specific DRG neurons were acutely dissociated for measuring excitability and sodium channel currents under whole‐cell patch clamp configurations. Expression of NaV1.8 was analyzed by western blot. Results: NMD significantly increased AWR scores. TTX‐resistant sodium current density was greatly enhanced in colon DRG neurons, activation curves showed a leftward shift in NMD rats. Furthermore, NMD remarkably enhanced expression of NaV1.8 at protein levels. AOAA, an inhibitor of CBS, significantly suppressed the expression of NaV1.8. AOAA treatment also reduced the TTX‐R sodium current density, and reversed hyperexcitability of colon‐specific DRG neurons in NMD rats. Conversely, addition of NaHS, a donor of H2S, greatly enhanced TTX‐R sodium current density, leftshifted activation curve and enhanced excitability of colon DRG neurons in age‐matched healthy rats. Conclusion: Our data suggest that NMD enhances TTX‐resistant sodium activity of colon DRG neurons, which is most likely mediated by CBS‐H2S signaling, thus contributing to chronic visceral hypersensitivity.Grant Funding Source: Supported by National Natural Science Foundation of China
UM206, a frizzled‐receptor antagonist attenuates adverse remodeling and cardiac function deterioration following myocardial infarction (652.10)
Authors: Hermans Kevin , Daskalopoulos Evangelos , Janssen Ben , Blankesteijn W. Matthijs 
Published: June 2021

Activation of the Wnt/frizzled signaling pathway has a key role in wound healing and cardiac remodeling following myocardial infarction (MI). We have previously shown that Wnt/frizzled signaling blockade with UM206 immediately after MI suppresses adverse remodeling and prevents heart failure (HF) development in a mouse MI model. Here, we investigated the role of UM206 after short‐time early treatment and long‐term late treatment, in order to unveil the optimal anti‐HF regimen.Swiss male mice were subjected to MI by left anterior descending coronary artery permanent ligation. Two different regimens were followed: a) short‐time early treatment: 2 weeks of UM206 treatment followed by 3 weeks without treatment and b) long‐term late treatment: 3 weeks without treatment followed by 5 weeks of UM206 treatment. UM206 was administered via an osmotic mini‐pump (6µg/kg/day) and controls were administered normal saline.Early UM206 treatment led to improved end diastolic volume (178±6 mm3) compared to saline (214±20 mm3, p<0.01) or to late UM206 treatment (278±24 mm3, p<0.01). Both effects were less pronounced compared to the full UM206 treatment (132±30 mm3). Ejection fraction was improved in early (24.4±0.1%) and late (23.2±2.6%) treatment compared to saline (16.5±0.2, p<0.01 and 11.0±3.2%, p<0.05 respectively) but full treatment was the optimal (31.4±0.4%), p<0.001).Both short‐time early and long‐term late UM206‐treatment regimens demonstrate a cardioprotective effect by halting the progression to HF. Nevertheless, full UM206 treatment immediately after MI is the one eliciting the most beneficial effects for the injured myocardium.Grant Funding Source: Supported by the Dutch Heart foundation (2010B196) and Cyttron (FES0908)
CaMKIIδ splice variants exert differential effects on heart failure development and myocardial ischemia/reperfusion injury (652.14)
Authors: Gray Charles , Xiang Sunny , Mishra Shikha , Heller Brown Joan 
Published: June 2021

Mice that lack CaMKIIδ (KO) are significantly protected from the development of myocardial damage following ischemia reperfusion (I/R). Cardioprotection in KO animals was associated with the loss of NF‐κB‐mediated inflammatory responses. CaMKIIδ is alternatively spliced in the heart to generate the CaMKIIδB and δC subtypes. In vitro data also suggest that δC is deleterious while δB may be cardioprotective. Furthermore, mice expressing δC rapidly and spontaneously progress to heart failure and death while those expressing δB have a milder phenotype. We postulated that CaMKIIδB and δC play opposing roles in response to I/R injury. Mice expressing CaMKIIδB and δC were subjected to global ischemia in the Langendorff mode for 25 minutes followed by reperfusion for various times. CaMKII activation and substrate phosphorylation were assessed by western blot. Subcellular fractionation was performed to determine compartment‐specific CaMKII activation. Infarct size was used as a measure of myocardial injury. I/R led to activation of CaMKIIδC but not δB in the cytosolic compartment. IκB‐Kinase (IKK) phosphorylation was increased in mice expressing CaMKIIδC but not in those expressing δB. Mice expressing δC showed large infarcts compared to WT, while mice expressing CaMKIIδB show greatly diminished infarct formation following I/R. The observation that pharmacological inhibition of IKK mitigated the deleterious effects of δC overexpression suggests that selective activation of CaMKIIδC in the cytosol and subsequent activation of inflammatory signaling contributes to I/R injury.Grant Funding Source: Supported by NIH Grant HL080101
Electrically monitoring effects of cisplatin on transendothelial invasion of human ovarian cancer cells (655.11)
Authors: Lo Jun‐Chih , Lo Chun‐Min 
Published: June 2021

Cisplatin is a platinum based chemotherapy drug used to treat various types of cancers, including sarcomas, some carcinomas, lymphomas and germ cell tumors. In this study human ovarian cancer cell lines, OVCA429 and ALST, were challenged with different concentrations of cisplatin (1E‐6M, 1E‐5M, 5E‐5M and 1E‐4M) and the effects on cancer cell morphology and micromotion were monitored and analyzed by electric cell‐substrate impedance sensing (ECIS) and microscopy. Moreover, the transendothelial invasion of the ovarian cancer cells to human umbilical vein endothelial cells (HUVECs) was examined. Our measurements showed that 1E‐4M of cisplatin caused a decrease of junctional resistance between cells and the reduction in micromotion. In addition, 1E‐6M and 1E‐5M of cisplatin were not toxic to the integrity of HUVEC layers but effectively inhibited transendothelial invasion of ovarian cancer cells. Thus, we report a strategic approach for using ECIS to analyze potential drugs for their abilities to suppress metastasis, particularly at the level of transendothelial invasion.Grant Funding Source: Supported by the National Science Council in Taiwan
The soluble epoxide hydrolase as a novel target for chronic neuropathic pain (659.15)
Authors: Wagner Karen , Inceoglu Bora , Hammock Bruce D. 
Published: June 2021

Neuropathic pain remains an unmet clinical need. In addition to nerve injury cases of neuropathy diabetes is increasing worldwide and diabetic neuropathy, the most frequent secondary condition to diabetes, will increase with it. Recent investigations reveal lipids as signaling mediators and their functional significance in nociception is receiving attention. Notably, there is evidence of antihyperalgesia mediated by epoxy fatty acids (EpFAs), metabolites generated by cytochrome P450 epoxygenase in an arm of the arachidonic acid cascade. These endogenous lipid mediators are subsequently degraded by the soluble epoxide hydrolase (sEH) enzyme to their mostly inactive diols. Metabolomic studies show that administration of sEH inhibitors (sEHI) stabilize EpFA to diol ratios in vivo and lead to decreased inflammation and pain related behavior in inflammatory pain models. Unexpectedly, the sEHI are also active against a model of type I diabetic neuropathy where the role of inflammation is minimal. The experiments here in rat and mouse demonstrate that potent sEHI mediate antihyperalgesia in both the sciatic nerve ligation (SNL) neuropathic pain and diabetic neuropathic pain (DNP) models. While mechanical withdrawal thresholds were used to assess both the SNL and DNP modeled pain, there are limitations to using acute stimulus reflex responses to assess chronic neuropathy. Therefore, the conditioned place preference (CPP) assay was used to evaluate the tonic nature of chronic pain and to address these limitations. Here, the CPP assay alongside withdrawal thresholds demonstrates the dose dependent antihyperalgesic efficacy of sEHI in the murine DNP model. Importantly, the CPP effect was absent both in control mice and in sEH knockout mice indicating the sEHI is not positively reinforcing or rewarding. Overall, these experiments indicate that inhibiting the sEH attenuates neuropathic pain and offers an alternative to current therapies to meet this clinical need.Grant Funding Source: Supported by R01 ES002710, P42 ES004699, R21AR062866
Role of cAMP signaling in mucin production in response to IL‐13 in human bronchial epithelial cells (660.12)
Authors: Al‐Sawalha Nour , Senny Christina , Hernandez Adrian , Valdez Daniel , Pokkunuri Indira , Thanawala Vaidehi , Bond Richard , Knoll Brian 
Published: June 2021

Asthma, a chronic airway inflammatory disease, is associated with mucus hypersecretion by airway epithelium. Accumulated mucus leads to airway obstruction and morbidity. We have shown that β2AR signaling has pro‐inflammatory effects in cultured normal human bronchial epithelial cells (NHBECs) in response to IL‐13(1). Our objective here was to elucidate the role of cAMP signaling in mucin production.NHBECs growing at air‐liquid interface (ALI) were subjected to various treatments, and levels of MUC5AC transcripts, intracellular mucin, cAMP levels, pCREB and pSTAT6 were measured on day 14 ALI.Treatment with forskolin+IBMX (to raise global intracellular cAMP levels) increased MUC5AC transcripts and intracellular mucin in the presence or absence of epinephrine in response to IL‐13 pCREB and pSTA6 were also elevated only in the presence of epinephrine, IL‐13 and forskolin+IBMX. A compartmentalized increase in cAMP caused by treatment with roflumilast, a PDE4 inhibitor, did not affect MUC5AC expression but reduced intracellular mucin only in the presence of epinephrine+IL‐13. Roflumilast did not affect the levels of cAMP, pCREB or pSTAT6.Our data provide evidence for the different influences of global and compartmentalized increases in cAMP on mucin production in human airway epithelium in response to IL‐13.1.Pokkunuri et al, BPS, 2011 Winter MeetingGrant Funding Source: Supported by NIH 1RO1A179236 awarded to RAB and BJK
Modulation by tetrodotoxin of action potential discharge in guinea pig vagal tracheal nodose afferent A‐delta cough receptors (660.15)
Authors: Herbstsomer Adam , Meeker Sonya , Undem Bradley 
Published: June 2021

We used the isolated vagally innervated guinea pig trachea/bronchus preparation to characterize the inhibitory effect of tetrodotoxin (TTX) on action potential discharge evoked by electrical (concentric stimulating electrode), and chemical (citric acid) activation of capsaicin‐insensitive nodose A‐delta fibers. We previously described these nerves and bona fide “cough receptors” (Canning et al. J. Physiol, 2004). We have also previously noted at the mRNA, protein, and electrophysiological level that the cell bodies of these nerves express TTX‐sensitive (mainly NaV 1.7) and TTX‐insensitive (both NaV 1.8 and NaV 1.9) channels (J. Physiol. 2008). Here we evaluated 18 nodose fibers with mean CVs of 5.1 ± 0.5 m/s. The voltage threshold of the terminals was 13 ± 1.5 V. The peak following Hz (>95% success for 5s) averaged 30 ± 2 Hz. All fibers responsed to citric acid with an average peak AP discharge of 35 ±6 Hz. Over 95% (17/18) of the fibers were completely silenced with 1 µM TTX. At 0.1 µM, TTX increased the voltage threshold to 24 ± 4 V, reduced the acid response to 10 ± 2 Hz, but had no effect on peak following Hz (studied at 2x V threshold). Jugular neurons project capsaicin‐sensitive A‐delta fibers with CVs the same as the nodose fibers (CV = 6.2 ± 1.4 m/s) and C‐fibers (CV = 0.6 ±0.1 m/s) to the trachea/bronchi. In contrast to nodose A‐delta fibers, only about 50% of jugular A‐delta (5/11) or C‐fibers (5/9) fibers were silenced with 1 µM TTX. These data indicate that although the TTX‐resistant NaVs (1.8 and 1.9) are expressed in nodose tracheal A‐delta cough receptors , they cannot alone support action potential discharge in healthy animals; this is in contrast to the jugular A‐delta and C‐fibers in the trachea/bronchi where TTX‐resistant channels may play a more important role.Grant Funding Source: NIH, Bethesda Maryland, USA
Alveolar type II cell‐localized p120‐catenin is an essential regulator of lung barrier function and innate immunity (660.8)
Authors: Chignalia Andreia , Vogel Stephen , Reynolds Albert , Malik Asrar , Liu Yuru 
Published: June 2021

Maintenance of alveolar epithelial barrier (AEB) integrity is crucial for normal lung gas exchange function, fluid balance, and regulation of immune response. The mechanisms whereby AEB is kept are not clear but adherens junctions (AJ) are central. P120‐catenin (p120) is an important component of AJ that stabilizes E‐cadherin. The alveolar epithelium is composed of two types of cells: flat type I cells comprising ~95% of the gas‐exchange surface, and cuboidal type II cells (ATII), which secrete surfactant and play a role in modulating lung immunity. Here we addressed if p120 contributes to ATII regulation of AEB and immunity by developing an inducible ATII‐specific p120 knockout mouse (P120EKO). P120EKO lungs displayed persistently leaky alveolar barrier and chronic lung inflammation. The inflammatory phenotype included infiltration of inflammatory cells, increased MPO and NFκB activity, and enhanced ICAM1and MIP2 expression. p120EKO ATII displayed E‐cadherin instability and decreased transepithelial resistance. p120EKO lungs showed barrier leakiness as shown by increased 125I‐BSA permeability in vivo. P120EKO lungs had increased surfactant proteins (SpB, SpC and SpD) and TLR4 expression. These findings indicate that while ATII constitute only 5% of alveolar surface area, ATII‐localized p120 plays a critical role in maintenance of alveolar epithelial barrier and homeostasis and modulate lung immune response.Grant Funding Source: Supported by CAPES fellowship (AZC), NIH grants HL105947‐01 (YL), HL07829‐16 (AM), HL090152 (AM)
The effect of aging on microvascular density and angiogenesis (665.4)
Authors: Sweat Richard , Chedister Lee , Sloas David , Stewart Scott , Murfee Walter 
Published: June 2021

Age related pathologies are commonly associated with vessel loss or excess vessel growth. An important question is whether or not angiogenesis, defined as the growth of new vessels from existing ones, is altered during aging. Angiogenesis in aged populations is commonly thought to be impaired. However, much of this supporting evidence is based on local changes in angio‐regulator molecules. The objective of this study was to quantitatively compare microvascular network remodeling metrics during angiogenesis in adult versus aged rat strains. Mesenteric tissues from adult (9 months) and aged (24 months) male Fischer 344 rats were harvested according to 4 experimental groups: 1) Adult Unstimulated (n=4 rats, 16 tissues), 2) Aged Unstimulated (n=4 rats, 16 tissues), 3) Adult Stimulated (n=4 rats, 16 tissues), 4) Aged Stimulated (n=4 rats, 16 tissues). For stimulated groups, tissues were harvested 3 days post compound 48/80‐induced mast cell degranulation stimulation. Unstimulated aged microvascular networks displayed larger mean vascular area per tissue area compared to the unstimulated adult networks. Following angiogenic stimulation, both adult and aged networks displayed similar increases in vascularized area and vessel length density compared to their respective unstimulated control groups, indicating that both adult and aged networks are capable of undergoing angiogenesis. However, capillary sprouting was significantly lower in stimulated aged networks compared with stimulated adult networks, supporting the hypothesis of impaired angiogenesis during aging. Interestingly, our results provide metric specific evidence for both comparable and impaired angiogenesis during aging and offer a possible explanation for conflicting results from the literature.Grant Funding Source: Supported by Tulane Center for Aging and NIH 5‐P20GM103629‐02
A novel low pHo‐induced Cl‐ conductance in nodose ganglion neurons is neuroprotective during simulated ischemia: implication for vagal afferent activity in heart failure (667.9)
Authors: Wang Runping , Lu Yongjun , Cicha Michael , Chapleau Mark , Benson Christopher , Abboud Francois 
Published: June 2021

We found that vagal afferent neurons isolated from nodose ganglia exhibit a large and prolonged low pHo‐conditioned Cl‐ current (25±5 pA/pF, n=60), when exposed to 3 brief pulse of low pHo (6.0) solution delivered at 20sec intervals. The current was similar to the one carried by the “Swell‐Activated Cl‐ Channel” (SAC) during hypoosmolarity. The activation pathways of this Cl‐ conductance are through NADPH oxidase (NOX)‐dependent production of ROS (dihydroethidine fluorescence). Since a similar Cl‐ conductance was found to be protective during ischemic preconditioning in cardiomyocytes (Draz et al 1999) we tested the hypothesis that the activation of this current may also be protective in cultured neurons exposed to “simulated ischemia” (low glucose, hypoxia and pH5.0). We measured the LDH release as an indication of cell injury. Without low pHo preconditioning the supernatant LDH concentration averaged 82±4 μU/ml and 441±62 μU/ml after exposure to 1 and 3 hours of simulated ischemia. In contrast, supernatant LDH was significantly reduced to 58±10 μU/ml and 227±25 μU/ml at 1 and 3 hours when neurons were preconditioned by activating the Cl‐ current before simulated ischemia. The protective effect of preconditioning was partially reversed by SAC channel inhibitor tamoxifen (10μM) (393±99 μU/ml at 3hours).Conclusions and Clinical Implications: A low pHo‐induced activation of a Cl‐ conductance in nodose neurons is NOX mediated and provides a putative autocrine protective mechanism for preservation of cardiac vagal afferents during ischemia or heart failure.Grant Funding Source: Supported by NIH HL 14388