bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dendritic Cell Produced IL‐12 Augments CD8+ T cell Activation through the Production of the Chemokines CCL1 and CCL17
Authors: Henry Curtis James , Grayson Jason M , Ornelles David A , Brzoza Kristina L , Hiltbold Elizabeth M 
Published: June 2021

Dendritic cells (DC) shape immune responses by producing cytokines such as Interleukin‐12 (IL‐12). IL‐12 is important in the clearance of the intracellular bacterial pathogen Listeria monocytogenes (Lm) and activates CD8+ T cells (CTL), which are required for sterilizing immunity. Our initial studies on the role of IL‐12 on CTL function determined that IL‐12p40 signals were transduced primarily during the first 48 hours of priming. However, these studies did not address the individual contributions of IL‐12 or IL‐23 nor how they affect the physical interactions of priming.To address these questions wild‐type, IL‐12p35−/−, or IL‐12p40−/− DC were infected with Lm and then used to prime CTL. IL‐12p40−/− mice immunized with p35−/− DC or p40−/− exhibited a reduced ability to clear Listeria compared to mice immunized with wild‐type DC. We found that CTL primed in vitro by p35−/− or p40−/− DC produced less IFN‐γ with the greatest reduction seen with the p40−/− DC, whereas proliferation was not affected. CTL primed in the absence of IL‐12 engaged in shorter duration interactions with DC which correlated with the absence of the chemokines CCL1 and CCL17. Neutralization of these chemokines resulted in decreased IFN‐γ production by CTL. Thus, CTL priming by DC is influenced more by the activity of IL‐12 than IL‐23 and in part through CCL1 and CCL17 whose production requires IL‐12. 1RO1AI057770‐01A1 and 1F31AI073245‐01A1
Modulation of IFN‐γ‐induced gene expression by TLR2 signaling in macrophages is mediated by an NF‐κB‐dependent mechanism
Authors: Benson Sarah , Grabenstein Jens , Ernst Joel 
Published: June 2021

Mycobacterium tuberculosis (Mtb) uses multiple mechanisms to avoid destruction by the immune system. We have previously shown that Mtb can inhibit selected macrophage responses to IFN‐γ using TLR2‐dependent and –independent mechanisms. To address the role of TLR2 signaling in mediating this inhibition, we used microarray analysis to identify IFN‐γ‐inducible genes whose expression decreased when macrophages were treated with the specific TLR2/1 ligand Pam3CSK4 (Pam). Four of the genes identified—Ciita, Klrk1, Dnase1l3, and Cxcl11—were further characterized. Ciita and Klrk1 induction by IFN‐γ was rapidly inhibited by Pam while Dnase1l3 and Cxcl11 required prolonged Pam pretreatment prior to IFN‐γ. Histone H4 acetylation at the Ciita and Klrk1 promoters was reduced in macrophages treated simultaneously with Pam and IFN‐γ compared to those treated with IFN‐γ alone. This effect was dependent on histone deacetylase activity. Treatment with the NF‐κB inhibitor parthenolide restored IFN‐γ‐induced gene expression and histone H4 acetylation at the promoters of selected Pam‐inhibited genes including Klrk1 and Cxcl11. These results indicate that TLR2‐dependent inhibition of IFN‐γ‐induced gene expression by Mtb depends on the length of exposure to a TLR2 agonist, is mediated by a mechanism affecting chromatin remodeling, and involves multiple signaling pathways including NF‐κB.NIH‐R01‐AI46097
Effect of PBMC shipment and storage conditions on ELISpot activity
Authors: Dryga Sergey A , Moskowitz Keith A , Moshkoff Dmitry A , Jeffrey Lisa , Manak Mark 
Published: June 2021

INTRODUCTION: Cellular immune response is often evaluated on frozen Peripheral Blood Mononuclear Cells (PBMC). Here, we evaluate the effect of shipping and storage conditions on PBMC viability and functional response using ELISpot assay.METHODS: Cryopreserved human PBMC were evaluated after overnight placement on dry ice and subsequent storage at −80oC, −20°C, in LN2 and on dry ice for up to 6 months. Cell count and viability was measured at time of thaw and after 24 hour rest. ELISpot assays were performed using huIFN‐γ ELISpot Kit (SeraCare) after stimulation with PHA, CMV and CEF peptide pools.RESULTS: Cells stored overnight on dry ice showed no significant decrease in viability at thaw and post rest. Cells stored at −20°C were not suitable for ELISpot assay. Cells stored on dry ice, at −80°C or in LN2 for 6 month demonstrated 92–93% viability upon thaw and 80–86% after 24 hr rest. The control cells remained 90% viable both post thaw and post rest. ELISpot responses of cells stored at −80°C or in LN2 for 6 month, or cells stored on dry ice for 4 weeks were comparable to control.CONCLUSIONS: The PBMC shipped in dry ice can be stored at −80°C for up to 6 months or on dry ice for 4 weeks with no appreciable loss of viability or functionality. Therefore, the use of PBMC for ELISpot proficiency testing is possible even in laboratories which lack ultra‐cold storage facilities.SUPPORT: This work was supported by NIAID contract NO1‐AI‐85341.
Associated Factors in diabetes prevalence in Mexican elderly people
Authors: Ruán Ma del Carmen Morales , Levy Teresa Shamah , Hernández Salvador Villalpando , Aguilar Alejandra Jimenez 
Published: June 2021

The prevalence of type 2 diabetes (TTD) is very high in Mexico especially in elderly population and its complications represent high expenditures in health care and reduction in their quality of life. The objective of this investigation is to assess some risk factors associated to TTD in a probabilistic sample of Mexican 60 yr. an over from the Mexican National Health and Nutrition Survey 2006 (ENSANUT 2006).A health questionnaire including information about previous diagnostic of TTD made by a doctor or other health personnel, weight, height, waist circumference, blood pressure measurements and capillary blood cholesterol determinations in a sub sample 1568 individuals were performed. The overall prevalence of TTD by previous diagnostic was 17.8% (women 19.6 %, men 5.4 %). In a logistic regression model the risk for TTD increased with Factors waist circumference > 90 cm (OR 1.6, CI95% 0.9, 2.6), to live in an urban community (OR 3.0, CI95% 2.0, 4.6) and cholesterol ¡Ý 240 mg/dL (OR 2.8 CI95% 1.4, 5.8).Nutritional intervention to maintain ideal weight, a waist circumference under 90 cm and adequate blood cholesterol concentrations, are essential in this population to guarantee a good quality of life in this elderly population.
Subependymal giant cell astrocytoma incidentally found in an elderly patient: Report of an autopsy case
Authors: Takei Hidehiro , Powell Suzanne Z 
Published: June 2021

Background: Subependymal giant cell astrocytoma (SEGA) is a benign, slowly growing tumor typically occurring in the first two decades of life. SEGA usually occurs in the setting of tuberous sclerosis complex (TSC), however there are several reported cases in which patients with solitary SEGA had no other stigmata of TSC. These lesions are generally considered to be forme fruste of TSC. We report such a case of SEGA in the oldest patient reported to date.Case report: The patient was a 75‐year‐old woman who had a history of radical vulvectomy for malignant melanoma and died of widespread systemic metastasis, which was confirmed at autopsy. Postmortem examination of the brain revealed a single intraventricular nodule in the left lateral ventricle, measuring 2.1×1.0×0.8cm. Histologically, the tumor was composed of interlacing bundles of spindle shaped tumor cells with thin delicate processes admixed with relatively large atypical cells with abundant glassy eosinophilic cytoplasm, as seen in SEGA. Ultrastructural study supported the diagnosis. Autopsy study showed no manifestations of TSC systemically or intracranially.Discussion: This is a very rare case of solitary SEGA incidentally found at autopsy in an elderly patient lacking any other findings of TSC. This is the second reported case of SEGA found in an elderly patient. This autopsy case showed that SEGA can occur outside the setting of TSC.
Myofibroblastoma presenting as a cystic lesion with a mural nodule in the brain parenchyma
Authors: Gaurav Gupta , Meera Hameed , Catrambone Jeffrey , Eun‐Sook Cho 
Published: June 2021

An 18‐year‐old female with no past medical history presented with left sided generalized tonic clonic seizures. Imaging showed a large cystic lesion within the right frontal lobe, just superior to the sylvian fissure. It measured 3.3cm × 4.0cm × 4.1cm. Within this large cystic lesion, there was a homogeneously enhancing lobulated nodule measuring 1.5cm × 1.0cm × 0.9cm. There was edema in the surrounding frontal white matter.At operation the lesion had no attachment to the dura and was completely intraparenchymal. The mural nodule was resected en bloc. Histologically the tumor showed alternating zones of spindle or ovoid cells that were separated by thick collagen bundles. Reticulin fibers are abundant and frequently wrapped around individual cells. Immunocytochemistry showed diffuse positivity for vimentin and focal positivity for bcl 2 and α‐smooth muscle actin. Tumor cells are negative for CD34, desmin, HHF35, S‐100, and EMA. This immunocytochemical profile is consistent with a myofibroblastoma.Myofibroblastoma is a benign mesenchymal tumor, which typically arises in the breast, less frequently in the lymph nodes and in other soft tissue. There are three reported cases of intracranial myofibroblastoma, all of which was attached to the meninges. Current case is unique in that it presented as a mural nodule in a large cyst within the brain parenchyme.
Endogenous Cholinergic Anti‐Inflammatory Pathway in Hepatic Ischemia‐Reperfusion (IR)
Authors: Crockett Elahe T , Dowlatshahi Shadi , Kolamal Nageswari , Galligan James J 
Published: June 2021

We recently showed anti‐inflammatory role of cholinergic pathway (Ch.Path) in liver IR by pharmacological stimulation of acetylcholine receptor. Here we tested whether endogenous Ch.Path (e.Ch.Path) activation would exert a similar protective effect, and studied liver gene expression modulation by Ch.Path activation.MethodsMice treated with CCK‐8 or nicotine, i.p. prior to ischemia (CCK‐8 stimulates acetylcholine release via Ch.Path). A 2nd group was subjected to bilateral vagotomy prior to CCK‐8, while a 3rd group subjected to electrical vagal stimulation for 20min, followed by liver IR (90min I : 3h R). Plasma ALT and cytokines levels and liver histopathology were assessed.ResultsCCK‐8‐ or nicotine treatment significantly reduced liver injury (87%, and %69 respectively) and cytokine production, as compared to saline‐treated mice. CCK‐8 protective effect was absent in vagotomized mice, indicating e.Ch.Path role. Electrical vagal nerve stimulation confirmed the observation as ALT levels and liver injury were significantly decreased in this group. Gene array data showed significant modulation of several pathways and associated genes.Conclusionse.Ch.Path activation via humoral or electrical stimuli provides a protective effect on liver IR injury, which could offer therapeutic means to selectively act on targeted genes for treatment of IR inflammatory diseases. Support: NIH HL075475.
A simple method for estimation of agonist activity at GPCRs: characterization of responses elicited by M2 and M3 muscarinic receptors
Authors: Tran John A , Chang Alex , Matsui Minoru , Ehlert Frederick J 
Published: June 2021

We investigated a parameter termed intrinsic relative activity (RAi), which provides a measure of agonist activity in functional assays. The RAi value is equivalent to the product of observed affinity and intrinsic efficacy of the agonist‐receptor‐G protein complex. To demonstrate that the validity of the RAi analysis, we measured the observed affinity and intrinsic efficacy values of selected agonists using Furchgott's method of partial receptor inactivation and compared the product of these estimates with the corresponding RAi estimate. We found good agreement between RAi and the product of affinity and efficacy in experiments on M3 muscarinic receptor stimulation of contraction of mouse ileum. Since observed affinity and efficacy are parameters inherent to the agonist‐receptor‐G protein complex, the RAi value is expected to be the same for various biological systems utilizing the same receptor‐G protein signaling pathway. We tested this hypothesis by comparing RAi values of muscarinic agonists for inhibiting the electrically paced guinea pig atrium and for inhibiting forskolin stimulated cAMP accumulation in CHO cells expressing the M2 receptor. Our results showed good agreement between the RAi estimates of selected agonists (oxotremorine‐M, oxotremorine, S‐aceclidine, arecoline, bethanechol, pilocarpine and McN‐A‐343) across the two assays.Supported by NIH grant 69829
Parathyroid hormone acts as a pharmacological chaperone on the parathyroid hormone receptor
Authors: Klenk Christoph , Lohse Martin J. 
Published: June 2021

Parathyroid hormone receptor (PTH1R) belongs to class B G‐protein‐coupled receptors and is the major regulator of extracellular calcium homeostasis. Binding of the native ligand parathyroid hormone (PTH) or biologically active peptide fragments of PTH to the receptor lead to the activation of Gs and Gq/G11 pathways with subsequent stimulation of adenylyl cyclase and phospholipase C. Activation of the PTH1R is followed by the recruitment of beta‐arrestin and subsequently to an efficient internalization of the receptor from the cell surface within several minutes.Here we show that long‐term stimulation of the PTH1R over several hours with PTH resulted in increased cellular protein levels and enhanced membrane distribution of the receptor. This increase of receptor levels was accompanied by an accumulation of high molecular mass moieties of the PTH1R. Analysis of the maturation steps during biosynthesis of the PTH1R revealed that the increased molecular mass was caused by a differential glycosylation pattern of the receptor. Blocking the export from the endoplasmic reticulum of the newly synthesized receptor by Brefeldin A completely inhibited the PTH induced mass changes. We therefore propose a ligand‐mediated stabilization of immature receptors in the endoplasmic reticulum which could eventually lead to a more efficient maturation of the PTH1R.
Insulin Resistance Induced by Low Salt Intake is Linked with RAS Activity and Inflammatory Signaling in Tissues
Authors: Coelho Michella Soares , Negri Heloisa B. , Oliveira Ivone B. , Shimizu Maria HM. , Hirata Aparecida E. , Casarini Dulce E. , Prada Patricia O. , Dolnikoff Miriam S. , Saad Mario JA. , Heimann Joel C. 
Published: June 2021

Low salt diet (LS) impairs insulin (INS) signaling and increases angiotensin II (AII) that may enhance JNK and IRS1ser307phosphorylation (PHOS), and inflammatory pathways (IP) with consequent reduction in INS action.Objective: To evaluate if losartan (LOS) improves INS resistance induced by LS by decreasing JNK and IRS1ser307activity.Methods: Twelve‐week‐old rats were fed a LS (0.15% NaCl) or normal salt diet (NS: 1.3%) since weaning. Groups were studied: LS+LOS (30 mg/kg/day), LS+vehicle, NS+LOS and NS+vehicle. Blood glucose (GLU), plasma INS and TBARS (oxidative stress marker), HOMA index, and tissue AII (immunohistochemistry and HPLC) were evaluated. The steps of INS signaling and IP were determined by immunoblotting.Results (mean±SEM, P<0.05, n = 8): INS, HOMA index and TBARS were higher on LS (INS: 712±60) than on NS (INS: 257±40 pmol/l). GLU and TBARS were lower and INS was higher on LS+LOS (TBARS: 4±0.5) than on LS (TBARS: 6±0.6 nmol/ml). Liver and muscle (M) AII were higher on LS (10±1) compared with NS (2±0.9 cell/mm2). Liver IkBα PHOS was lower on LS (20±2) than on NS (100±±4 %) suggesting an activation of IP on LS. Insulin receptor, PI3K and Akt PHOS (90±4) were higher and JNK (120±3) and IRS1ser307 (100±6 %) PHOS were lower in the liver of LS+LOS compared to LS. Similar results were observed in M.Conclusions: INS resistance induced by LS is associated with dysregulation of INS signaling, activated IP, and higher tissue AII and TBARS. LOS improves INS signaling by decreasing JNK and IRS1ser307 PHOS, with consequent reduction in GLU in LS.FAPESP
Chronic aortic denervation plus high sodium diet. A model of salt‐sensitive normotension in rat
Authors: Rodríguez‐Martínez Manuel , Rodríguez‐Pérez Adriana S , Brito‐Orta Dolores , López‐Rodríguez Juan F 
Published: June 2021

To assess whether progressive arterial baroreceptor denervation results in salt‐sensitive changes in blood pressure, mean arterial pressure (MAP) was measured in six groups of conscious male Wistar rats before and ∼30 days after bilateral: a) sham denervation (SH), b) sino‐aortic denervation (SD) and c) aortic denervation (AD). During the last 21 days of post‐denervation period three groups received high sodium diet (HNa = 8%) and the other three groups low sodium diet (LNa = 0.04%). The remaining baroreflex control of ERSNA was examined in the anesthetized state as %Δ ERSNA vs MAP. The modelling indicated that there were no effects of denervation nor diet on MAP ∼ 30 days after denervation. However, the modelling of Δ MAP (MAP after – MAP before) indicated effect of both denervation and diet but not interaction, the highest and more lasting Δ MAP being observed in AD + HNa group (21± 3 mmHg). The modelling of remaining baroreflex control of ERSNA indicated effect of denervation but not of diet.Conclusion:AD + HNa is a novel experimental model of salt‐sensitive normotension, though AD does not impair the remaining baroreflex control of ERSNA.Supported by: SMRF06‐UASLP

Range %
Slope %/mmHg
X to ½ range mmHg
Lower Plateau %
Gmax

SH
119± 7
−.085±.007
130±5
−85±6
−2.5±0.1

AD
125± 9
−.065±.005*
128±4
−82±5
−2.0±0.2

SD
61± 6*
−.059±.005*
153±7*
−66±6*
−.9±.1*

p<0.05 vs SH.

Conclusion:AD + HNa is a novel experimental model of salt‐sensitive normotension, though AD does not impair the remaining baroreflex control of ERSNA.Supported by: SMRF06‐UASLP
Hypovolemia affects cortical activity patterns associated with the cardiovascular response to moderate lower body negative pressure (LBNP)
Authors: Usselman Charlotte Willemina , Wong Savio W , Kimmerly Derek S , Rothwell Amanda , Shoemaker J Kevin 
Published: June 2021

We tested the hypothesis that hypovolemia (HV; reduced blood volume) modifies the cortical autonomic network (CAN) response to moderate (−35 mmHg) LBNP. CAN activity was assessed using functional magnetic resonance imaging (fMRI) with blood oxygen level‐dependent (BOLD) contrast in young healthy subjects (n = 9). Testing was repeated under normovolemia (NV) and HV (spironolactone: 100mg/day for 3 days; randomized). BOLD data were collected at baseline and during 3 repeated 60s bouts of LBNP (separated by 90s rest periods). Random effects analysis revealed BOLD signal changes (p < 0.005, uncorrected) in the CAN during both the onset (baroreceptor unloading) and offset (baroreceptor loading) phases of LBNP. In both NV and HV, LBNP onset induced CAN BOLD signal changes in the thalamus, hippocampus, putamen, mid‐cingulate cortex and posterior insular cortex. In addition, LBNP onset in HV also elicited BOLD signal changes in the caudate nucleus, and anterior and posterior cingulate cortex. Changes in these additional areas were observed again during HV offset, although HV offset did not elicit a change in hippocampal activation. Compared to NV, these results suggest that HV was associated with more widespread CAN activation during the dynamic phases of baroreceptor manipulation.Supported by the Canadian Space Agency.
Glycerol‐induced fluid shift attenuates the vestibulosympathetic reflex in humans
Authors: Dyckman Damian J , Ray Chester A 
Published: June 2021

Classically, the glycerol dehydration test (GDT) has been used to test for the presence of Ménière's disease and can cause acute alterations in vestibular reflexes in both normal and pathological states. The vestibulosympathetic reflex (VSR) elicits increases in muscle sympathetic nerve activity (MSNA) and peripheral vasoconstriction. We hypothesized that the GDT would attenuate the VSR through an acute fluid shift of the inner ear. Nine male subjects (27±1 years) performed head‐down rotation (HDR), which engages the VSR, before and after administration of either the GDT or saline. MSNA (microneurography), arterial blood pressure, and leg blood flow (venous occlusion plethysmography) were measured during HDR. Subjects performed 80‐degree head‐up tilt for up to 30 minutes after the post‐tests. Before drug administration, HDR significantly increased MSNA in burst frequency (Δ5±1 bursts/min, Δ8±1 bursts/min; p<0.01) and total activity (Δ44±13% Δ77±17%; p<0.01), and decreased calf vascular conductance (Δ15±6%, Δ20±3%; p<0.01), in both the saline and glycerol trials, respectively. Post‐saline injection, HDR still significantly increased MSNA (Δ6±2 bursts/min, Δ83±20% total activity; p<0.01) and decreased calf vascular conductance (Δ21±4%, p<0.01), which was not significantly different from pretesting. In contrast, post‐GDT resulted in an attenuation of MSNA (Δ3±1 bursts/min, Δ22±3% total activity) and reduction in calf vascular conductance (Δ7±4%) during HDR. These results suggest that a fluid shift of the inner ear via glycerol dehydration attenuates the VSR. These data provide support that dynamic fluid shifts can have a significant effect on the VSR.Research supported by NIH DC006459, HL077670, & M01RR10732 and NSBRI CA00404
Isoflurane Preconditioning Delays Opening of Mitochondrial Permeability Transition Pore via Protein Kinase C Signaling Pathway
Authors: Pravdic Danijel , Sedlic Filip , Mueller Samantha J , Bosnjak Zeljko J 
Published: June 2021

Isoflurane protects the myocardium from ischemia and reperfusion (I/R) injury via protein kinase C (PKC) signaling pathway. It has been suggested that the mitochondrial permeability transition pore (mPTP) is the end effector of I/R injury, whereas PKC has a protective effect by blocking mPTP opening. Here we investigated whether anesthetic preconditioning (APC) with isoflurane delays mPTP opening and whether the PKC plays a role in isoflurane‐induced delay in mPTP opening. Isoflurane preconditioned rat cardiomyocytes were loaded with fluorescent probe tetramethylrhodamine ethyl ester (TMRE) and subjected to a simulated I/R injury by laser‐induced ROS production. Using the laser‐scanning confocal microscopy, we measured the time to 50% decrease in TMRE fluorescence, which coincides with depolarization of the mitochondria and represents an opening of mPTP. To confirm the role of mPTP, we used cyclosporine A (CsA). The role of PKC was tested using the non‐specific PKC inhibitor chelerythrine (CHEL). APC with isoflurane prolonged the mean depolarization time of the control fluorescence of 216±8 s, to 261±16 s. A similar time delay of 279±22 s was found in cells pretreated with CsA. Isoflurane‐induced delay in an mPTP opening was abrogated to 219±7 s in cells treated with CHEL. In conclusion, our results indicate that APC with isoflurane delays the opening of mPTP in a PKC‐dependent manner.Supported by NIH (HL 034708 and GM 066730)