bluuebunny/crossref_metadata_2025_split huggingface dataset:
HlyA cause platelet activation and neutrophil/ monocyte interaction during urosepsis
Authors: Johnsen Nanna , Christensen Mette G. , Hamilton Aimi D.M. , Praetorius Helle 
Published: April 2020

The urinary tract is frequently a primary infection site in patients with severe sepsis or septic shock
1. Urinary tract infections are most commonly caused by E.coli sub‐strains that produce several virulence factors
4–
8, of which α‐haemolysin (HlyA) is the most abundant
9. Our previous data demonstrate that the biological effects of HlyA are intimately associated with extracellular ATP signalling secondarily to ATP release from cells through the HlyA‐pore. Our previous data support that HlyA is largely responsible for the septic symptoms observed in response to bacteraemia with uropathogenic E.coli. ATP and its degradation products are aggressive activators of immune cells
10 and platelets
11–
13, and prime candidates for promoting the intravascular coagulation observed during septic shock.We tested the effect of antagonising platelet P2Y receptors (P2Y12 and P2Y1) during sepsis with uropathogenic E.coli(ARD6, O6:K13:H1) in anaesthetised mice. In aggreement with the literature, infusion of the P2Y12 antagonist cangrelor (8.6 or 86mg/hour) did not change survival or any tested sepsis parameters. However, the P2Y1 inhibitor MRS2500 reduced platelet activation by pore‐forming bacterial toxins (70–80%) in vitro, measured as surface exposure of fibronectin, DC63 and P‐selectin. In vivo, infusion of MRS2500 (624μg/hour) increased the survival in mice exposed to uropathogenic E. coli. Moreover, infusion of MRS2500 partially prevented the 60% fall in platelet count otherwise seen in response to sepsis in control mice.Interestingly, targeting of P2Y2 receptors, which is highly expressed on neutrophils and monocytes
14 but not in platelets, showed that lack of P2Y2 receptors entirely prevented the fall in circulating platelets in response to sepsis. Moreover, P2Y2 receptor‐deficient mice die much earlier in response to intravenous application of uropathogenic E. coli compared to controls. By flow cytometry, we are able to detect increased complex formation between thrombocytes (CD42) and neutrophils (Ly6‐G) /monocyte (Ly6‐G/CD11B). These data could potentially suggest that complex formation between thrombocytes and neutrophils and/or monocytes are important for the observed reduction in free thrombocyte numbers during sepsis and that the complexes could dampen the septic reaction to the circulating bacteria and improve survival.Support or Funding InformationThis project is funded byThe Independent Research Foundation ‐ DenmarkThe Augustinus Foundation
Efficacy of the curcumin to ameliorate ovalbumin induced atopic dermatitis and progression of atopic march in mice
Authors: SHARMA SUKRITI , NAURA AMARJIT SINGH 
Published: April 2020

Atopic dermatitis (AD) is a chronic pruritic skin inflammatory condition which arises due to skin barrier disruption and immune dysregulation. Curcumin has been widely exploited in the treatment of various skin‐related pathologies due to its pharmacologic effects. The present study was designed to investigate the beneficial effect of curcumin in the amelioration of ovalbumin (OVA)‐induced atopic dermatitis and ensuing atopic march. AD was induced in female Balb/c mice by epicutaneous application of OVA at the back skin site for 1 week followed by a resting period of two‐weeks and same protocol was repeated thrice. Curcumin treatment was given either intraperitoneally or topically in two separate groups of animals. Curcumin was administered intraperitoneally at different doses during the last sensitization phase for a period of 7 days. Topical application of curcumin at a dose of 10 mg/kg was done for a period of 14 days during the second resting period. Skin tissues were examined for histologic changes, Th2 cytokines and the oxidative stress markers. Systemic curcumin administration ameliorated the onset of AD‐like lesions and significantly improved the dermal destruction in mice upon OVA exposure. Curcumin reduced the levels of various inflammatory mediators such as Th2 promoting cytokines (TSLP and IL‐33), Th2 cytokines (IL‐4, IL‐5, IL‐13 and IL‐31) and also downregulated the activation/expression of Stat‐6 and GATA‐3. Next, when curcumin preparation was applied topically, it shows a remarkable efficacy in restoration of the histo‐architecture of the skin suggesting that the phytochemical based ointment(s) can offer a newer treatment option for AD and associated atopic march. Finally, curcumin blocked the OVA initiated progression of AD towards asthma as evident by the reduced number of BALF inflammatory cells and suppressed expression of Th2 cytokines in the lung tissue upon aerosolized exposure of the allergen to the AD mice. Overall, our findings demonstrate for the first time that curcumin has the efficacy to ameliorate AD and halt the progression of associated atopic march.Support or Funding InformationFinancial support from Department of Biotechnology, Government of India [BT/PR17968/MED/122/33/2016]; University Grants Commission, New Delhi, India [UGC‐SAP] and [UGC‐BSR‐SRF]
Nonselective Inhibition of PDE4 Induces Gastroparesis in Mice
Authors: McDonough Will , Aragon Ileana , Rich Justin , Murphy James M. , Saleh Lina Abou , Boyd Abigail , Koloteva Anna , Richter Wito 
Published: April 2020

PurposeNon/PAN‐selective inhibitors of Type 4 phosphodiesterases (PDE4s), a group of isoenzymes that hydrolyze the second messenger cAMP, exert promising therapeutic benefits, such as anti‐inflammatory as well as cognition‐ and memory‐enhancing effects. However, these drugs also induce several side effects, in particular emesis and nausea, curbing their clinical use. The PDE4 family consists of four subtypes/genes (PDE4A to D). As each plays unique and non‐overlapping roles in the body, targeting individual PDE4 subtypes is a promising approach to separate the therapeutic benefits from the side effects of current PDE4 inhibitors. To this end, this study aimed to identify the specific PDE4s involved in emesis and nausea. While mice are anatomically unable to vomit, PDE4 inhibition in mice induces gastroparesis, an abnormal retention of food in the stomach, which is a well‐known cause of nausea and vomiting in humans and was thus used as a correlate.MethodsGastroparesis in knock‐out mice deficient in individual PDE4 subtypes and in mice treated with PAN‐PDE4 inhibitors was compared to their respective controls. We used a long term model, in which food accumulated in the stomach of mice fed ad libitum for 3 days, as well as an acute model, in which fasted mice were given a FITC‐labeled food bolus by oral gavage, euthanized 30 min later, and FITC/food‐retention in the stomach was then measured.ResultsTreatment of wildtype mice with PAN‐PDE4 inhibitors increased stomach sizes to twice that of controls without changing their food intake or the weight of their intestines, suggesting a specific effect of PDE4 inhibition in restricting gastric emptying. Structurally distinct PDE4 inhibitors all induced gastroparesis, indicating it is a class effect. Conversely, YM976, an inhibitor which does not cross the blood‐brain‐barrier, did not induce gastroparesis suggesting that PDE4 inhibitor‐induced gastroparesis results from altered neuronal regulation, rather than from PDE4 inhibition in the stomach/periphery. Ablation of PDE4A, B, C or D in the respective KO mice did not induce, nor did it protect the animals from PAN‐PDE4 inhibitor‐induced gastroparesis, implying that it results from the concurrent inhibition of multiple PDE4s.ConclusionsSelective ablation of individual PDE4 subtypes does not induce gastroparesis in mice. Thus, potentially, any of the four PDE4 subtypes may be targeted individually for therapeutic benefits without inducing nausea or emesis. In addition, PDE4 inhibitors that do not cross the blood‐brain barrier may also be free of these side effects.Support or Funding InformationSupported by grants from the NIH (HL76125, HL141473, HL066299).
Serine/theonine kinase 4 is a potential biomarker for assessment of β‐catenin‐mediated colon cancer prognosis
Authors: Hsu Tai-I , Lu Pei-Jung , Hsiao Micheal 
Published: April 2020

Colon cancer has highly incident rate in world, and the surgical therapy can cure early stage colorectal cancer patients. But the most difficult is to predict patients’ disease prognosis. STK4 is a one of the Hippo pathway component and was shown to have important role to control the organ size and cell to cell contact inhibition ability and was considered a potential tumor suppressor. However, little is known about the relationship between the expression of STK4 and patient survival. The aim of this study was to investigate prognostic value of STK4 in colon cancer. STK4 expression in normal and tumor tissues from 140 colon cancer patients was examined by immunohistochemistry. The relationship between STK4 expression and clinicopathological factors as well as patient survival was analyzed. Human colon cancer cell lines (H3347and DLD‐1) were used to analyze the effect of STK4 knockdown and overexpression on invasion/migration in vitro. IHC analysis showed STK4 was significantly downregulated in tumor tissues compared to their corresponding non‐tumor part (P< 0.01) and was significantly associated with tumor size (P = 0.003), distal metastasis (P = 0.03), disease recurrence and poor survival (P< 0.005). Univariate and multivariate analysis indicated loss of STK4 expression predicted significantly poor outcome. Knockdown of STK4 promoted, whereas overexpression of STK4 inhibited invasion and migration of human colon cancer cells in vitro. STK4 is a prognosis maker to predict patient survival in colorectal cancer. Loss of STK4 expressed in tissue is correlated with poor prognosis and survival.Support or Funding InformationThis research was supported by Academia Sinica [AS‐SUMMIT‐108].
Myoendothelial Junctions of Mature Coronary Vessels Express Notch Signaling Proteins
Authors: McCallinhart Patricia E. , Wani Elsa , Biwer Lauren A. , Wolpe Abigail G. , Isakson Brant E. , Lilly Brenda , Trask Aaron J. 
Published: April 2020

The Notch signaling pathway, via cell‐cell communication, is critical during normal vascular development and homeostasis, and it is known to regulate vascular smooth muscle (VSMC) phenotype, including proliferation; however, few studies have addressed the role of Notch signaling in the mature vasculature. The current study tested the hypothesis that Notch signaling is present and active at mature myoendothelial junctions (MEJs), signaling hubs that occur at breaks in the internal elastic lamina (IEL). In mature, intact normal adult mouse coronary resistance microvessels (CRMs) embedded within heart sections, we observed Notch1, Notch2, Notch3, and Jagged1 expression within the fenestrae of the CRM IEL where MEJs are known to occur. Immunofluorescence clearly shows bridging from the VSMCs to endothelial cells (ECs). Immunofluorescence in larger mouse left anterior descending (LAD) coronary arteries demonstrate the expression Notch2 and Notch3 protein immunofluorescence within fenestrae of the IEL. We found that, on average, 39% and 33% of IEL fenestrae were positive for Notch2 and Notch3 expression, respectively. We also confirmed the presence of Notch3 expression in the MEJ by immunogold electron microscopy. In normal mouse femoral arteries, we found that, on average, 46%, 51%, and 58% of IEL fenestrae were positive for Notch1, Notch3, and Jagged1 expression, respectively. To determine whether Notch signaling components were localized to the MEJ, we co‐cultured primary human coronary ECs (hcECs) and primary human coronary VSMCs (hcVSMCs) across transwell inserts, and we utilized fluorescent microscopy to immunostain for Pai‐1, Jagged1, Notch1, and Notch2. PAI‐1 immunofluorescence was observed on the EC side of the transwells and within the transwell pores, indicating the formation of in vitro MEJs. We also identified distinct staining of Notch1, Notch2, and Jagged1 in the pores of the co‐culture transwells, which co‐localized with PAI‐1 within the pores. To assess whether Notch signaling was active at the MEJ, we co‐cultured hcECs and hcVSMCs on transwells to create in vitro MEJs. Smooth muscle cells were first transfected with a Notch‐sensor luciferase plasmid followed by transwell co‐culture of the transfected cells with endothelial cells. Notch activity was stimulated by 3‐fold across the MEJ in hcVSMC (1.00 ± 0.15 vs. 3.02 ± 0.43, p<0.001), which was completely abrogated by the non‐selective Notch γ‐secretase inhibitor, DAPT (1.15 ± 0.16, p<0.01). In this study, we demonstrate for the first time that Notch receptors and ligands are expressed within and are active at mature coronary MEJs, demonstrating a previously unrecognized mode of Notch signaling regulation between the endothelium and smooth muscle.Support or Funding InformationFunding: EW was supported by NIH R25HD086885. NIH R00HL116769, R21EB026518, and S10OD023438 to AJT.
Lipoatrophy‐associated insulin resistance and hepatic steatosis are attenuated by intake of diet rich in omega 3 fatty acids.
Authors: Festuccia William T. , Moustaid-Moussa Naima , Moreira Rafael 
Published: April 2020

We evaluated in this study glucose homeostasis and progression of nonalcoholic fatty liver disease (NAFLD) and hepatomegaly in severe lipoatrophic mice and their modulation by intake of a diet rich in omega 3 (n‐3) fatty acids. Severe lipoatrophic mice induced by adipocyte‐specific PPARγ deletion (A‐PPARγ KO) and littermate controls (A‐PPARγ WT) were evaluated for glucose homeostasis and liver mass, proteomics, lipidomics, inflammation and fibrosis after intake of a chow diet. Lipoatrophic mice were heavier than controls due to increased food intake and energy efficiency, severely glucose intolerant and hyperinsulinemic, and developed NAFLD characterized by increased liver glycogen, triacylglycerol and diacylglycerol contents, DNA and mitotic index, apoptosis, inflammation, steatosis score and fibrosis. This was associated with activation of de novo lipogenesis and liver enrichment with monounsaturated in detriment of polyunsaturated fatty acids including n‐3 fatty acids. Administration of a high‐fat diet rich in n‐3 fatty acids (HFO) inhibited de novo lipogenesis, enriched liver with n‐3 fatty acids, reduced hepatic steatosis, apoptosis, inflammation and steatosis and improved glucose homeostasis in lipoatrophic mice. In conclusion, diet enrichment with n‐3 fatty acids is an effective strategy to improve glucose homeostasis and reduce liver steatosis and inflammation without affecting hepatomegaly in severe lipoatrophic mice.Support or Funding InformationFunded by FAPESP (2015/19530‐5).
Quantitatively defining spatial patterns of myoendothelial junctions in the arterial wall
Authors: Ruddiman Claire A. , Corliss Bruce A. , Peirce Shayn M. , Isakson Brant E. 
Published: April 2020

The regulation of vasoconstriction and vasodilatory pathways is essential in resistance arteries, and can be modulated by heterocellular communication between the endothelial and smooth muscle cells. This can occur via direct endothelial‐smooth muscle cell contact at myoendothelial junctions (MEJs) through fenestrations in the internal elastic lamina (IEL). While it has been demonstrated that MEJs do not occupy every fenestration, their spatial distribution throughout the endothelium has not yet been defined and could reveal localization tendencies to cellular landmarks. We hypothesized that the MEJ and IEL fenestration spatial distribution would exhibit a non‐random distribution pattern throughout the endothelium with respect to various cellular landmarks, including the interendothelial junctions. In order to quantify this relationship, the spatial distributions of both holes and MEJs with respect to various cellular landmarks were examined. Third order mesenteric resistance arteries isolated from C57Bl6 male mice were prepared en face to view the endothelium and detect the IEL, nuclei, myoendothelial junctions, and interendothelial junctions (e.g., claudin‐5). A Matlab based analysis pipeline was developed to calculate the proximity of IEL fenestrations to cellular landmarks, such as interendothelial junctions. En face views of the IEL often have patches of varying signal intensity, posing a challenge for detecting fenestrations through basic thresholding methods. To overcome this, parameters within Image Processing Toolbox documentation were optimized for each image prior to thresholding. This allowed for accurate assignment of XY coordinates to fenestration boundaries (bwboundaries documentation) and calculation of center points. En face views of claudin‐5 were similarly thresholded and their boundaries also defined by XY coordinate matrix. Next, Euclidean distance was calculated between each fenestration center point and all claudin‐5 XY coordinates and plotted as a probability distribution. Monte Carlo simulations were also performed as a negative control. In the simulation, the fenestrations are replaced by randomly generated circles that mimic the number and radii of fenestrations in the original image. This is compared to the original distribution to assess if identified patterns are random. Our analysis of >100 simulations indicate that IEL fenestrations appear to be randomly distributed across the IEL. However, we find the MEJs within the IEL fenestrations may be strategically placed near interendothelial junctions. Preliminary data indicates that this particular MEJ localization pattern may be opportunistic because of the cytoskeletal components at interendothelial junction that can be recruited to support the signaling microdomains within MEJs.Support or Funding InformationThis work supported by 1F31HL149228‐01 (CAR) and HL088554 (BEI).
Brainstem catecholaminergic neurons mediate cognitive impairment in heart failure rats through regulation of hippocampal synaptic‐related gene expression.
Authors: Toledo Camilo , Martínez Milka , Diaz Hugo , Schwartz Karla , Pereyra Katherin , Diaz-Jara Esteban , Del Rio Rodrigo 
Published: April 2020

Heart failure (HF) is common in elder population and it is highly associated with cognitive impairment (CI). Several studies suggest that autonomic imbalance contribute to age‐related CI. We previously showed that catecholaminergic neurons (C1) from the rostral ventrolateral medulla (RVLM) displayed chronic activation in HF. In addition, we also shown that HF rats showed CI. Whether RVLM‐C1 neurons contribute to CI in the setting of HF is completely unknown. Therefore, we aimed to study the contribution of RVLM‐C1 neurons on cognitive function in HF. Adult male Sprague‐Dawley rats underwent volume overload to induce HF. Anti‐dopamine β‐hydroxylase‐saporin (DβH–SAP: 5ng/150nl) was used to selectively destroy RVLM‐C1 neurons. Morris water maze (MWM) test was used to assess learning and memory performance. Micropunches containing the hippocampus were dissected to measure the expression of several synaptic‐related genes using a custom 12‐gene array by qRT‐PCR. HF rats displayed learning and memory impairment compared to control animals. Indeed, HF showed an increased traveled distance to the target platform along 5 trial/days during MWM test (15.1±0.5 vs. 30.7±2.1 m; Control vs. HF, respectively; p<.05). In addition, when the platform was changed daily to a different quadrant (i.e. to evaluate reference memory) HF rats showed an increase in the number of trials required to reach the platform compared to control rats (4.2±0.1 ± vs. 8.2±1.2 trials; Control vs. HF, respectively; p<.05). Partial ablation of RVLM‐C1 neurons in HF rats decreased traveled distance to reach the target platform compared to HF rats without treatment (18.1±1 vs 30.7±2.1 m; HFDβH–SAP vs. HF, respectively; p<.05). Also, during memory testing HF untreated rats needed almost twice the number of trials to accomplish the task when compared to HF+DβH–SAP rats (4.4±0.4 vs. 8.2±1.2 trials; HFDβH–SAP vs. HF, respectively; p<.05). Finally, compared to control rats, HF rats display (Ctrl vs. HF, in AU) decreased levels of PSD‐95 (1.0±0.2 vs. 0.4±0.3), Hommer1 (1.0±0.2 vs. 0.6±0.3), CaMKIV (1.0±0.2 vs. 0.5±0.3), Piccolo (1.0±0.3 vs. 0.4±0.2), Erc2 (1.0±0.3 vs. 0.2±0.1), RIM (1.0±0.5 vs. 0.3±0.2), Neuroligin 3 (1.0±0.4 vs. 0.5±0.3), Neuroxin 1 (1.0±0.3 vs. 0.6±0.4) and Synaptotagmin‐1 (1.0±0.3 vs. 0.6±0.1). Importantly, DβH–SAP treatment in HF significantly restored the expression levels of Hommer1, Piccolo, Erc2, RIM and Synaptotagmin‐1 to levels observed in control rats. Taken together, our results show that catecholaminergic RVLM neurons contribute to CI in HF rats. In addition, our results suggest that RVLM‐C1 may regulate hippocampal synaptic transmission in the setting of HF through the regulation of gene expression in the hippocampus.Support or Funding InformationSupported by FONDECYT 1180172 and 3190659 and the basal Center of Excellence in Aging and Regeneration (AFB 170005).
Influence of heat acclimation on spontaneous wheel‐run exercise in mice
Authors: Masuda Yuta , Marui Shuri , Kato Issei , Nagashima Kei 
Published: April 2020

Introduction We previously found that, in mice, running distance on a running wheel went down in heat in mice, during which body core temperature became lower than that in no‐heat control. These results may suggest that mice determine exercise intensity depending on thermal input from the body surface. In the present study, we hypothesized that heat acclimation disinhibit such suppression of spontaneous running in heat.MethodsTwenty two male C57BL/6J3 mice (age, 7 w) were used, which were divided to two groups: the heat acclimation and control groups (HA group and CON groups, respectively). They had surgery of intra‐abdominal placement of the IC‐chip sensor device (nanotag, Kissei, Nagano) for the measurement of abdominal temperature (Tabd) and counts of movement under inhalation anesthesia with sevoflurane and were allowed to recover for 10 d at least. They were individually housed at an ambient temperature (Ta) of 33°C and 25°C in the HA and CON groups, respectively for 2 w. Before and after the acclimation period, each mouse had two different trials on two‐separated days: exposures at 33°C and 25°C at 19:00–1:00 in a dark phase. In the trials, each mouse was moved to a semi‐closed chamber with a running wheel inside, where Tabd, running distance on the wheel, and metabolism by indirect calorimetry. After the experiments, mice were killed by overdose injection of pentobarbital, and the dorsal root ganglia (DRG, Th 5‐12) was obtained to assess protein expression of TRPV1 and V4.ResultsBefore the acclimation period, the running distance was smaller in the in the 33°C trial than in the 25°C trial in both groups (P<0.05; e.g. 8.98 ± 0.86 and 13.68 ± 1.18 km, respectively, in the HA group). However, after the acclimation period, the distance in the HA group at 33°C trial became greater than the value before the acclimation period (P<0.05; 14.73 ± 1.32 km; post‐25°C, 14.50 ± 0.66 km).VO2 in the HA groups was higher in pre‐33°C trial and lower in post‐33°C trial than any other trial. Before the acclimation period, Tabd was lower in the 33°C trial than in the 25°C trial. After the acclimation period, Tabd was greater in the 33°C trial than in the 25°C trial. (pre‐33°C, 37.52 ± 0.28; pre‐25°C, 37.99 ± 0.30°C; post‐33°C, 37.93 ± 0.23; post‐25°C, 37.82 ± 0.30 °C). The expression TRPV1 on DRG tended to be lower in the HA group compared with the CON
group.ConclusionsWe found that, in mice, heat acclimation increases running distance on the wheel and decrease metabolic heat production. Based on physiological and biochemical analyses, decrease in thermal input from the skin and improve metabolic efficiency may be involved in the
mechanism.