bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of Low‐to‐Moderate Ethanol Consumption on Colonic Cell Proliferation, Differentiation and Apoptosis in Young Adult and Middle‐Aged Rats
Authors: Voutour Luciano , Murase Yuko , Wells Nicole , Quigley Jacqueline , Pascua Jeremy , Pinkowski Natalie , Almaiman Lama , Brasser Susan M. , Hong Mee Young 
Published: April 2020

Excessive alcohol consumption is a risk factor associated with colorectal cancer; however, some epidemiological studies have reported that moderate alcohol consumption may not contribute additional risk for developing colorectal cancer while others suggest that moderate alcohol consumption may provide a protective effect that reduces colorectal cancer risk. Prior experimental data have indicated the importance of proliferation, differentiation, and apoptosis as parameters to consider when evaluating colonic cell growth and tumorigenesis. Chemopreventative agents are reported to decrease cell proliferation and to increase differentiation and apoptosis. The present study examined whether chronic low‐to‐moderate ethanol consumption had an effect on these parameters of colonic cell growth. Twenty‐four nondeprived young adult (109 days old) Wistar rats (n=12/group) and twenty‐four nondeprived middle‐aged (420 days old) Wistar rats (n=12/group) were allowed to voluntarily consume a 20% v/v ethanol solution on alternate days for 13 weeks (45 ethanol drinking sessions total) or were given access only to water (non ethanol‐exposed control). The intermittent access 20% ethanol drinking paradigm utilized results in mean blood alcohol levels of ~30–50 mg/dl in the outbred Wistar strain when measured 30–120 min into a standard drinking session. Following the final experimental session, colon tissues were collected for quantitative immunohistochemical analyses of cell proliferation, differentiation and apoptosis using Ki‐67, goblet cell and TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling), respectively. Ethanol treatment resulted in a lower cell proliferation index in both young (P = 0.05) and older (P < 0.001) rats, as well as a lower proliferative zone (P < 0.01 for both ages). Older rats had a shorter colon, but there was no significant difference for proliferation index between young and old rats. Cell differentiation between ethanol‐treated animals and controls was not significant for young rats, but older rats had a lower level of differentiation in the ethanol group. For apoptosis, there was no significant effect of ethanol treatment for either young or old rats. Younger rats had higher values than older rats for both differentiation and apoptosis (P < 0.05). These findings suggest that moderate consumption of ethanol improves at least one notable parameter (cell proliferation) in colonic tumorigenesis regardless of age, however, reduced cell differentiation among older animals.Support or Funding InformationSupport Contributed By: NIH AA023291
Long‐lasting Effects of Early‐life Western Diet and Exercise on Adult Gut Microbiome Composition in Mice
Authors: McNamara Monica P. , Cadney Marcell D. , Singleton Jennifer M. , Ruegger Paul M. , Borneman James , Garland Theodore 
Published: April 2020

Increases in availability of energy‐dense foods and simultaneous reductions in physical activity of Western industrialized societies has created an environment that promotes obesity, but alterations in the gut microbiome may also play a role. As with other organismal characteristics, the microbiome may be influenced by factors experienced early in life. We previously demonstrated that early‐life wheel access for 3 weeks, starting at weaning, followed by an 8‐weeks washout period, increases adult wheel running in both selectively bred High Runner (HR) and non‐selected Control (C) lines of mice. We extended these studies by examining effects of early‐life treatments on the gut microbiome. Mice were given early‐life wheel access and/or Western diet from weaning until sexual maturation at 6 weeks of age, then housed individually without wheels and on standard diet until 14 weeks of age, when fecal samples were taken. We profiled the gut microbiome by extracting DNA from fecal samples using MoBio Laboratories PowerSoil DNA Isolation kit, amplified ITS rRNA, sequenced the libraries using an Illumina MiSeq, and chose OTUs using Usearch. For bacterial richness (number of OTUs and Chao1 index) we found a 3‐way interaction of linetype, juvenile Western diet, and juvenile wheel access. These results constitute one of the first reports of juvenile exercise and/or diet having long‐lasting effects on the adult gut microbiome after a substantial washout period (equivalent to ~6 human years).Support or Funding InformationSupported in part by NIH grant R21HD084856
Human microglia respond to malaria‐induced extracellular vesicles
Authors: Mbagwu Smart , Lannes Nils , Walch Micheal , Filgueira Luis , Mantel Pierre-Yves 
Published: April 2020

Background and ObjectiveMicroglia are the chief immune cells of the brain and have been reported to be activated in severe malaria. Their activation may drive towards neuroinflammation in cerebral malaria. Malaria‐infected red blood cell derived‐extracellular vesicles (MiREVs) are produced during the blood stage of malaria infection. They mediate intercellular communication and immune regulation among other functions. During cerebral malaria, the breakdown of the blood‐brain barrier can promote the migration of substances such as MiREVs from the periphery into the brain targeting cells such as microglia. Microglia and extracellular vesicles interactions in different pathological conditions have been reported to induce neuroinflammation. Unlike in astrocytes, microglia‐extracellular vesicle interaction has not yet been described in malaria infection. Therefore, in this study, we aimed at investigating the uptake of malaria‐infected red blood cell derived‐extracellular vesicles (MiREVs) by human microglia cells and their cytokine response.MethodHuman blood monocyte‐derived microglia were generated from buffy coats of anonymous healthy donors using Ficoll‐Paque density gradient centrifugation. The malaria infected red blood cell derived‐EVs (MiREVs) were isolated from the Plasmodium falciparum cultures. They were purified by ultracentrifugation and labelled with PKH67 green fluorescent dye. The internalization of MiREVs by MoMi was observed after 4hours of co‐incubation on coverslips placed in a 24‐well plate at 37°C using confocal microscopy. Cytokine‐gene expression was investigated using rt‐qPCR following the stimulation of the MoMi cells with supernatants from the parasite cultures at 2,4 and 24h respectively.Results and ConclusionMiREVs were internalized by the microglia and accumulated in the perinuclear region. MiREVs‐treated cells increased gene expression of the inflammatory cytokine TNFα and reduced gene expression of the immune suppressive IL‐10. Overall, the results indicate that MiREVs may act on microglia which would contribute to enhanced inflammation in cerebral malaria.
Deletion of NEDD4‐2 Abolishes the Inhibitory Effect of High Sodium (HS) Intake on the Basolateral Kir4.1/Kir5.1 of the DCT and Thiazide‐Sensitive Na‐Cl Cotransport (NCC)
Authors: Zhang Dandan , Duan Xin-Peng , Wu Peng , Xiao Yu , Gao Zhong-Xiuzi , Wang Wen-Hui , Lin Dao-Hong 
Published: April 2020

BackgroundWe have previously demonstrated that the deletion of NEDD4‐2 increased the basolateral Kir4.1/Kir5.1 channel activity in the distal convoluted tubule (DCT), suggesting the role of NEDD4‐2 in the regulation of basolateral Kir4.1/Kir5.1. Also, it has been demonstrated that dietary sodium intake regulated the basolateral Kir4.1/Kir5.1 channel activity in the DCT such that low sodium (LS) intake stimulated whereas HS intake inhibited the Kir4.1/Kir5.1 channels in the DCT. The HS‐induced inhibition of basolateral Kir4.1/Kir5.1 channels in the DCT was essential for the HS‐induced inhibition of NCC expression/activity because HS failed to inhibit NCC in the kidney‐specific Kir4.1 knockout mice. The aims of the present study are: 1) To explore whether the effect of HS on the basolateral Kir4.1/Kir5.1 in the DCT is compromised in the absence of Nedd4‐2; 2) To examine whether the deletion of Nedd4‐2 abolishes the effect of HS on NCC activity.MethodsWe have used the patch‐clamp methods to measure the basolateral K (Kir4.1/Kir5.1) channel activity of the DCT and DCT membrane potential, and employed renal clearance method to measure thiazide‐sensitive renal sodium excretion in Nedd4l+/+ (WT) mice and renal tubule‐specific Nedd4l−/− mice (Ks‐NEDD4‐2 KO).ResultsWe confirmed the previous finding that HS inhibited the basolateral Kir4.1/Kir5.1 channel activity in the DCT and depolarized the DCT membrane and that LS intake increased the basolateral K conductance and hyperpolarized DCT membrane. The deletion of NEDD4‐2 increased the basolateral Kir4.1/Kir5.1 channel activity in the DCT and hyperpolarized DCT membrane. Moreover, the effect of either HS or LS on the basolateral Kir4.1/Kir5.1 channel activity was absent in Ks‐NEDD4‐2 KO mice. Renal clearance experiments showed that LS increased while HS decreased thiazide‐sensitive urinary sodium excretion, indicating the stimulation of NCC and the inhibition of NCC, respectively. The deletion of Nedd4‐2 not only augmented the thiazide‐induced natriuresis but also abolished the inhibitory effect of HS on the thiazide‐induced natriuresis. Furthermore, LS intake failed to further increase thiazide‐sensitive renal sodium excretion. Thus, the effect of HS and LS intake on NCC activity was abolished in Ks‐NEDD4‐2 KO mice.ConclusionNEDD4‐2 plays an important role in mediating the inhibitory effect of HS on the basolateral Kir4.1/Kir5.1 channel activity and NCC activity/expression.Support or Funding InformationDK115366
Pharmacological characterization of a novel, orally available GluN2B antagonist JNJ‐63612445
Authors: Balana Bartosz , Schoellerman Jeffrey , Lord Brian , Wyatt Ryan , Matta Jose , Neff Robert , Welty Natalie , Bhattacharya Anindya , Bonaventure Pascal , Lovenberg Tim , Letavic Michael 
Published: April 2020

IntroductionBlockade of GluN2B (NR2B)‐containing NMDA receptors has been proposed as a therapy for a number of neurological and psychiatric diseases including mood disorders. A number of GluN2B antagonists have been identified and developed over the last three decades. These compounds, however, have significant limitations in regard to their selectivity or drug‐like properties (e.g. oral bioavailability).MethodsThe pharmacology of JNJ‐63612445 was explored using in‐vitro assays (calcium mobilization, competitive radioligand binding, electrophysiology), ex‐vivo slice autoradiography and in‐vivo electrophysiology (synaptic transmission and plasticity) and behavior.ResultsJNJ‐63612445 is a high affinity (pKi=8.0), potent (pIC50=7.8) and selective negative allosteric modulator of GluN2B receptors. Upon oral dosing, the compound occupied GluN2B receptors in rat hippocampus in time‐ and concentration‐dependent manner, reaching 50% occupancy at the plasma concentration of 443 ng/ml. JNJ‐63612445 at 1 μM blocked 54% of NMDA receptor‐mediated EPSC in rat neonatal slices similar to other GluN2B antagonists. The compounds at the dose 10 mg/kg s.c. inhibited in vivo long‐term depression measured in the CA1 of anesthetized rats without effects on the basal synaptic transmission. Given orally at the dose 30 mg/kg in mice, the compound significantly reduces immobility time in a tail‐suspension test, while occupying approximately 89% of the GluN2B receptors in the hippocampus.ConclusionsJNJ‐63612445 represents an example of new generation of potent, selective, and orally available GluN2B antagonists.Support or Funding InformationFunding informationAuthors are full time employees of Janssen Research & Development, LLC. The research was funded by the company.
SPARC: Minimally Invasive Recording of Neural Activity During Natural Voiding in Anaesthetized Rats
Authors: Payne Sophie C. , Osborne Peregrine B. , Keast Janet R. , Fallon James B. 
Published: April 2020

BackgroundControlling urination with bioelectric neuromodulation devices is an emerging treatment strategy for bladder incontinence/retention as it can be rapidly adjusted to respond to changing patient needs, known as closed‐looped control. However, true closed‐loop bioelectric neuromodulation requires sensing of neural activity to allow accurate and timely delivery of the stimulation to maximally affect the end‐organ. As such, developing a neural recording feedback system that is highly specific for different neural fibre types is a crucial first step towards closed‐loop control.ObjectiveTo develop and validate a minimally invasive recording technology that differentiates between afferent and efferent neural activity, as well as distinguishing between fast and slow fibre types.MethodsA custom‐designed, two pair platinum electrode array was implanted onto the pelvic nerve of urethane‐anaesthetised Sprague Dawley rats (male; 8–10 weeks; n=6) with catheterised bladders. Evoked compound action potentials (ECAPs) were generated to confirm placement of the electrode array and cystometry and neural activity recorded during bladder filling and voiding.ResultsECAPs show recruitment of slow and fast conduction neural populations. Our recording technology distinguished between afferent and efferent activity during the micturition reflex. Within these neural types, distinctive patterns of fast and slow conducting fibres were identified and their firing patterns were consistent with results reported using invasive single‐ and multi‐unit recording techniques.ConclusionClosed‐loop bioelectric neuromodulation requires the ability to stimulate or inhibit neural activity, as well as precisely sense ongoing neural activity. We have completed the first step in developing closed loop technology by validating technology that precisely records multiple distinct populations of neural activity using a minimally invasive technique. Future studies will investigate the effectiveness of this recording technique in awake, freely moving rats.Support or Funding InformationResearch reported in this publication was supported by the Office Of The Director, National Institutes Of Health of the National Institutes of Health under Award Number OT2OD023872.
Supplemental arginine improves intestinal morphology, but not expression of enzymes related with synthesis of polyamines in heat stress pigs
Authors: Morales Adriana , González Fernanda , Bernal Hugo , Camacho Lucero , Htoo John K. , González-Vega Caroline , Cervantes Miguel 
Published: April 2020

Animals exposed to high ambient temperature suffer heat stress (HS); this condition reduces feed intake and damages their intestinal epithelia, affecting nutrient digestion and absorption. Polyamines are involved in the restoration of intestinal epithelia by stimulating the proliferation of enterocytes. Because arginine is a precursor for the synthesis of polyamines, we examined the effect of supplying extra arginine in the diet on the expression of enzymes involved in polyamine synthesis and the intestinal morphology of HS pigs. This experiment was conducted when natural ambient temperature (AT) fluctuated daily from 28.2 to 38.7 °C. Twenty pigs (25.3±4.4 kg BW) were assigned to one of two treatments: control wheat‐soybean meal diet, and the control diet added with 0.20% free arginine. Pigs were fed at 0700 and 1900‐h, same amount (900 g) each time. At the end of a 21‐d trial, six pigs per treatment were sacrificed to collect intestinal mucosa samples and 5‐cm fragments from duodenum, jejunum, and ileum to analyze the intestinal morphology, and mRNA expression of spermidine/spermine N1‐acetyltransferase (SSAT), ornithine decarboxylase (ODC), and argininosuccinate synthase (ASS). Extra dietary arginine increased the villi height in duodenum, jejunum, and ileum, and the villi height:crypt depth ratio in duodenum and jejunum (P<0.01). However, no effect of extra arginine was observed on the expression of polyamine‐related enzymes (P>0.10). These results indicate that supplementing extra free arginine restores the morphology of the small intestine, but there is no clear explanation for the lack of effect of extra arginine on expression of enzymes involved in the polyamine synthesis.Support or Funding InformationCONACYT, México, CB‐2016‐01‐287052. Evonik Nutrition & Care GmbH.
The Role of SGS1 Helicase in Mismatch Repair During Single Strand Annealing
Authors: Afreen Faria 
Published: April 2020

In humans, interspersed repeated sequences constitute at least 35% of the genome. Single Strand Annealing (SSA) is a repair pathway which occurs when a double strand break is produced between homologous repeats. The product of SSA is a single copy of one of the homologous repeats. As such SSA is a highly mutagenic repair pathway. If an SSA event occurs and consequently deletes important genes such as BRAC2, which itself contains repeats, the result can be catastrophic.To study mismatch correction (MMC) during SSA, a yeast strain was created to have 7 mismatches (6 base‐pair substitutions and 1‐bp insertion/deletion), resulting in a 3% divergence between the repeats. Previous results show that strains with 3% divergence between the repeats, designated as the F‐A strain, reduced SSA two‐fold compared to the identical strains, designated as the A‐A strain. This reduction in SSA is due to heteroduplex rejection which occurs when the mismatch repair complex, MSH2‐MSH6, recognizes the mismatches in the DNA‐duplex and stimulates the unwinding of the DNA via SGS1 helicase, resulting in cell death. In the absence of heteroduplex, MSH2‐MSH3 will bind branched SSA intermediate, and recruit RAD1‐RAD10 to remove the non‐homologous tails.Sgs1Δ results in a very efficient SSA repair, with 2 folds increased viability, in both F‐A and A‐A strains, compared to WT strains as SGS1 is required for heteroduplex rejection. Sequencing analysis of sgs1Δ F‐A strain shows an asymmetric gradient of correction favoring the F fragment, similar to the WT strain, suggesting that SGS1 has minimal effects on MMC outcomes. In both sgs1Δ and WT strains tailless strains, there is a uniform correction of MMs, suggesting that the non‐homologous tails influence MMC. All together current data suggests that MMC is more likely to occur during replication, once the SSA repair is completed. Future directions include knocking‐out MLH1, a mismatch repair protein shown to interact with SGS1, and investigating if sectored colonies, cells with unrepaired mismatches is due to replication.Support or Funding InformationBrandeis Division of Science Summer Research Fellowship
Blood Pressure Maintained in Response to a Hypotensive Stimulus During Mild Acute Inflammation
Authors: Schroeder Elizabeth C. , Hilgenkamp Thessa I. M. , Lefferts Wesley K. , Fernhall Bo 
Published: April 2020

IntroductionSevere acute inflammation such as sepsis disrupts autonomic nervous system function and jeopardizes blood pressure regulation. Severe experimental inflammation (endotoxin) blunts the vascular baroreflex to increase muscle sympathetic nerve activity in response to reductions in blood pressure and abolishes baroreflex control of heart rate. Whether mild acute inflammation alters the ability to maintain blood pressure and peripherally vasoconstrict during a hypotensive challenge is unknown.PurposeDetermine whether mild acute inflammation alters the ability to maintain blood pressure and peripherally vasoconstrict during lower body negative pressure (LBNP).MethodsThirteen young, healthy adults (26 ± 4.5 yrs; male n=5; 21.3 ± 2.0 kg/m2) completed assessments before (baseline) and 24h after the induction of acute inflammation via typhoid vaccination. Measures were completed after 10 min rest (rest) and after 2 minutes of equilibration to −20 mmHg of LBNP. Measures included brachial blood flow via ultrasonography and brachial blood pressure and heart rate via an automatic blood pressure cuff. Conductance was calculated as blood flow/mean arterial pressure (MAP). ELISAs were performed on serum blood samples from baseline and 24h to assess inflammatory markers C‐reactive protein (CRP) and interleukin‐6 (IL‐6).ResultsAcute inflammation was evident at 24h by increases in IL‐6 (0.98 ± 0.55 to 1.78 ± 0.68 pg/mL, p<0.01) and CRP (0.42 ± 0.98 to 1.30 ± 1.44 mg/L, p=0.02). Acute inflammation did not alter the blood flow, conductance, diameter, or MAP response to LBNP (see table
, p for interactions >0.05). Blood flow, conductance, and velocity were lower during LBNP compared to rest (p<0.01) whereas brachial diameter and MAP were maintained (p>0.05). Heart rate was elevated at 24h and increased during LBNP at baseline (p<0.01) but did not change during LBNP at 24h (interaction p<0.01).ConclusionThe ability of young adults to respond to a hypotensive challenge and maintain blood pressure was preserved during mild acute inflammation, however, the heart rate response was altered. Our data may suggest that mild acute inflammation alters baroreflex control of the heart rate response to a hypotensive challenge.
Hemodynamic response to LBNP at baseline and during acute inflammation.

Baseline

24H

p‐value

Rest

LBNP

Rest

LBNP

Visit

LBNP

Interaction

Brachial blood flow, mL/min

47.0 ± 14.4

40.8 ± 11.8

45.4 ± 14.2

42.0 ± 16

0.94

0.04

0.56

Conductance, mL/min/mmHg

52.2 ± 15.5

44.7 ± 11.9

49.8 ± 14.3

46.5 ± 16.6

0.93

0.03

0.46

Brachial diameter, mm

3.81 ± 0.85

3.83 ± 0.87

3.89 ± 0.84

3.81 ± 0.81

0.46

0.62

0.12

Mean velocity, cm/s

7.5 ± 3.3

6.4 ± 2.9

6.8 ± 3

6.4 ± 2.6

0.35

0.04

0.44

MAP, mmHg

90 ± 9

91 ± 8

91 ± 6

90 ± 7

0.81

0.87

0.23

Heart rate, bpm

52 ± 11

57 ± 11

56 ± 12

57 ± 12

0.08

<0.01

<0.01

Data mean ± standard deviation. MAP, mean arterial pressure.
Human Fetuses Accrue Osmotically Inactive Sodium Stores in Anticipation of Birth
Authors: Segar Jeffrey L. , Grobe Connie , Grobe Justin L. 
Published: April 2020

BackgroundSodium (Na) is stored in the skin and muscle in a non‐osmotic manner. It has been postulated that in adults, these stores protect from water loss and regulate immune function. In post‐weanling rats, osmotically inactive Na pools are mobilized to support growth. The presence of and role for such stores earlier in development have not been explored.ObjectiveDetermine if osmotically inactive Na pools exist in human infants.MethodsWe utilized literature providing human fetal body composition (Ziegler, EE, 1976) and water compartment distribution (Friis‐Hansen B, 1983) data to calculate changes in total body stores of Na, potassium (K) and chloride (Cl) during fetal development (24–40 weeks gestation). Total body mineral content was estimated by two methods, with calculations based upon 1) body water compartments (BW) and 2) reported mineral content per 100 g fat free mass (MC). Differences between these calculations were interpreted to represent osmotically inactive stores. The content of Na, K and Cl within water compartments were estimated to be 140, 4, and 110 mEq/L (extracellular) and 4, 150, and 10 mEq/L (intracellular), respectively.ResultsEstimates of total body K were nearly identical at each gestational time point (<10% variance; (Kmc−Kbw)/Kmc). Estimated total body Cl by MC exceeded that using BW compartments at all time points, with a maximum difference at 40 weeks gestation of 40 mEq (23% variance). In contrast, total body Na by MC greatly exceeded that calculated based upon BW compartments, with differences increasing from 17 mEq at 24 weeks gestation (variance of 25%) up to 100 mEq at 40 weeks gestation (variance of 35%).ConclusionThe disproportionate differences in total body mineral content between the two methods coupled with increasing variances at advancing gestational ages suggest that Na, and to a lesser extent Cl, are stored in osmotically inactive pools within the fetus. These pools increase in size with advancing gestational age. As breastmilk is relatively Na‐deficient (10 mEq/L), we speculate the fetal osmotically inactive Na pool is vital to maintain a sufficient total body Na content that is necessary for optimal postnatal growth.