bluuebunny/crossref_metadata_2025 huggingface dataset:
Temporal gene expression profiling of the hypothalamus following trauma
Authors: Bowman Phillip Dalton , Bynum James A. , Craig Teresa , Meledeo M. Adam , Darlington Daniel N.
Published: June 2021

The hypothalamus plays an important role in homeostatic regulation. In order to identify biological processes relevant to the response to severe traumatic injury, the hypothalamic transcriptome was examined following polytrauma. Sprague‐Dawley rats (300–400g) were anesthetized with isoflurane and trauma induced by damaging the small intestine, left and medial liver lobes, right leg skeletal muscle, and a right femur break. Rats had 40% of their blood volume removed in 30–60 min with no resuscitation. All rats survived the 240 min experimental period. Brains were removed at 0.5, 1, 2, 3, and 4 hours post injury and placed in RNAlater.™ Hypothalami were dissected from 3 mm coronal sections (4 per time point) and RNA extracted. Following labeling and hybridization to rat whole genome microarrays (Agilent), genes statistically altered in their expression compared to sham were determined with BRB Arraytools. Within 30 min, 350 genes were upregulated >2‐fold and 660 were downregulated >2. By reference to Ingenuity Pathways Analysis program, canonical pathway activation included genes reported to be involved in hepatic stellate cell activation as well as glucocorticoid receptor, Il–10, acute phase response, and G‐protein coupled receptor signaling. Understanding the hypothalamic response to polytrauma may provide new targets for improving survival. Project funding provided by US Army Medical Research and Material Command
VE‐cadherin Signaling Induces EB3 Phosphorylation to Suppress Microtubule Growth and Stabilize Adherens Junctions
Authors: Geyer Melissa , Huang Fei , Daneshjou Nazila , Garcia Alexander , Kreutz Barry , Mehta Dolly , Malik Asrar B. , Komarova Yulia
Published: June 2021

End Binding (EB) proteins, microtubule (MT) plus‐end factors, promote MT growth in cells. Here we demonstrate an unexpected role of Vascular Endothelial (VE)‐cadherin homotypic adhesion in suppressing MT growth by phosphorylation of EB3 on serine 162. In endothelial monolayers, MTs displayed short episodes of growth; however, disruption of VE‐cadherin‐mediated adhesion by either extracellular Ca2+“switch” or treatment with a specific anti‐adhesion peptide resulted in persistent MT growth. MT growth was secondary to increased cytosolic [Ca2+] and EB3 dephosphorylation resulting from the loss of VE‐cadherinmediated adhesion. Inhibition of calcineurin (CaN), the Ca2+‐dependent phosphatase, prevented EB3 dephosphorylation and MT growth. Expression of a phospho‐defective S162A EB3 mutant promoted MT growth in confluent monolayers and destabilized adherens junctions (AJs). Thus, VE‐cadherin outside‐in signaling regulates cytosolic Ca2+homeostasis and EB3 phosphorylation, which in turn stabilizes AJs. These results reveal a novel function of VE‐cadherin homotypic interaction in modulating endothelial AJ barrier function through the tuning of MT dynamics. This work was supported by NIH grants R01 HL103922 and Giles F. Filley Memorial Award to Y. K.; R01 HL 45638 to A.B. M.; T32 HL07829‐17 support to M. G. and T32 HL07829‐18 support to N. D.
Dietary iodole 3, 3′‐diindolylmethane protects cultured human alveolar epithelial cells from oxidative stress induced cell death
Authors: galam lakshmi , Prasanna TP , younus Asfiya , Hodgkins Amanda , Gurukumar KR , Cox Ruan R , RajanBabu  , Jutaro F , Lockey Richard F , Kolliputi Narasaiah
Published: June 2021

Dietary indoles including indole‐3‐carbinol (I3C) and its derivative, diindolylmethane (DIM) are found in cruciferous vegetables have been known for their anti cancer and anti‐inflammatory properties. Oxidative stress and alveolar epithelial cell death play a key role in the pathogenesis of several pulmonary diseases. In this report, we investigated the protective ability of DIM to ameliorate oxidative stress and alveolar epithelial cell death in vitro. Human type II alveolar epithelial cells (A549) were treated with the oxidative stress inducer hydrogen peroxide (H2O2), in the presence and absence of DIM. In another set of experiments, we first induced apoptosis by starvation in epithelial cells and then post treated with DIM to determine the reversal of epithelial cell apoptosis. The results showed that the compound DIM inhibited the H2O2 induced the alveolar epithelial cell necrosis and apoptosis. This property of DIM ensured the protection of alveolar epithelial cells from oxidative stress induced cell death. In addition DIM also reverses cell death and protects alveolar epithelial cells from starvation induced cell damage and cell death. Taken together our data demonstrated that the natural dietary compound DIM attenuates oxidative stress induced epithelial cell death. This work is supported by R01 HL105932 (NIH) and AmericanHeart Association National Scientist Development Grant 09SDG2260957
The effects of high and low intensity exercise on heat shock protein accumulation in rat skeletal muscle and vasculature
Authors: Kowalchuk Hana , Melling C.W. James , Noble Earl G
Published: June 2021

Exercise is a stressor that activates cytoprotective heat shock proteins (Hsps) in skeletal muscle (SM). Though exercise intensity influences this response, the localization of Hsps to skeletal myofibers versus the vasculature following various exercise loads has yet to be examined. The hypothesis tested was that induction of Hsp70 and 90 in skeletal muscle and blood vessels (BVs) is intensity‐dependent, with greater increases following high‐intensity (HIEX) than low‐intensity exercise (LOEX). Rats (n=10 per group) were sacrificed 24hr after a single exercise bout of 1hr treadmill running (2%incline) at either LOEX (15m/min) or HIEX (30m/min). A control group (CON) was similarly handled yet underwent no exercise. Immunofluorescent localization of Hsp70 and 90 was conducted on the white portion of the vastus muscle (WV). Hsp70 positive fibres were absent in WV of both CON and LOEX but found in HIEX (~5% of all fibres). Hsp70 was not detected in CON BV, was observed mainly in large BV's in LOEX and had a greater distribution to most BV's of HIEX. Hsp90 was equally robust in larger BVs in both HI and LOEX and greater than in CON. These observations confirm that in skeletal myofibers and vasculature Hsp70 is induced post exercise and exercise intensity determines the extent and localization of the response. Hsp90 is found solely in the vasculature and is elevated by exercise independent of intensity.
Analysis of the osteogenic effects of TGF‐beta in a calvarial defect model
Authors: SOTO JUAN MANUEL SOLIS , MORENO VANESSA SOUSA , NAJERA ROSA ISELA SANCHEZ , CEPEDA MARIA ARGELIA AKEMI NAKAGOSHI , CEPEDA SERGIO EDUARDO NAKAGOSHI , MEESTER IRENE
Published: June 2021

TGF‐beta is involved in several cellular processes, including acting as a growth factor.

Objectives

The aim of this study was to investigate the effect of TGF‐beta on bone regeneration and osteocalcin production.

Methods

Calvarial osteotomy defects were created in adult male Sprague‐Dawley rats, they received TGF‐beta in an absorbable collagen sponge. After 6 weeks, morphometric analysis of histological sections was performed to evaluate the osteoid and bone growth, and the immunoreactive cells to osteocalcin.

Results

TGF‐beta induced a significative increase (p<0.01) of osteoid and bone growth in comparison to controls. Similarly, tissues treated with TGF‐beta displayed a higher number (p<0.01) of immunoreactive cells to osteocalcin.

Conclusion

The osteoinductive capacity of TGF‐beta is demonstrated, in terms of the production of osteocalcin (mineralization factor) and bone growth. This work was supported by PAICYT UANL.
Early Interference with p44/42 Mitogen‐Activated Protein Kinase (MAPK) Signaling in Hypothalamic Paraventricular Nucleus (PVN) Attenuates Angiotensin II‐Induced Hypertension in Rats
Authors: Yu Yang , Xue Bao-Jian , Zhang Zhi-Hua , Wei Shun-Guang , Beltz Terry G , Guo Fang , Johnson Alan Kim , Felder Robert B
Published: June 2021

Brain p44/42 MAPK signaling can upregulate angiotensin (ANG) II type‐1 receptors (AT1‐R) and proinflammatory cytokines in the PVN, a critical cardiovascular and autonomic center. We tested the hypothesis that brain p44/42 MAPK signaling contributes to the development of ANG II‐induced hypertension. Rats with blood pressure (BP) measured by telemetry received ANG II (120 ng/kg/min, SC) for two weeks. Pooled small interfering RNAs (siRNAs) for p44/42 (28 ng), a control siRNA or vehicle (artificial CSF) was microinjected bilaterally into the PVN early (at the onset of the ANG II infusion) or 1 week later. Compared with vehicle or control siRNA, the early (but not the later) p44/42 siRNA treatment significantly attenuated the rise in BP and the response of BP to ganglionic blockade. Real time PCR revealed a significant reduction in AT1‐R mRNA in the PVN at 1 week and 2 weeks after the early p44/42 siRNA treatment, and at 2 weeks after the later p44/42 siRNA treatment, compared with the vehicle and control siRNA treatments. There were no differences among groups in mRNA for the pro‐inflammatory cytokines. These results suggest that the full expression of ANG II‐induced hypertension depends upon p44/42 MAPK‐mediated upregulation of AT1‐R in the PVN. MAPK signaling in the PVN may be a novel target for early intervention in the progression of ANG II‐dependent hypertension. Supported by NIH RO1 HL096671 and Dept of Veterans Affairs.
Evidence that Leptin Signaling in POMC Neurons Regulates HDL‐c
Authors: Marino Joseph S , Dowling Abigail , Faulkner Latrice , Hill Jennifer W
Published: June 2021

Mice lacking leptin or its receptor have elevated serum HDL‐c due to dysfunctional hepatic ApoA‐1 and ApoA‐2 regulation. Data from our lab suggests the brain's pro‐opiomelanocortin (POMC) neurons, a major target of leptin, are involved in mediating the affects of leptin on HDL‐c.

Using cre‐Lox genetic recombination, we bred mice that lack leptin receptors (LepR) specifically on POMC neurons (LepRPOMC). We measured serum HDL‐c, LDL/vLDL, total cholesterol, triglycerides, and ApoA‐1 from LepRPOMC and littermate controls (LepRflox/flox).

Serum HDL‐c and ApoA‐1 levels were significantly elevated in LepRPOMC compared to control mice. LepRPOMC mice also have elevated triglycerides, LDL/vLDL ratio, and exhibit obesity. Thus, the elevated HDL‐c and ApoA‐1 levels do not appear to act in a cardio‐protective manner. Instead, their elevation may be due to dysfunctional hepatic regulation of HDL‐c associated lipoproteins as demonstrated in mice lacking leptin or its receptor.

We believe this is the first evidence for the contribution of specific leptin responsive neurons to the regulation of HDL‐c. Studies are ongoing to examine the mechanisms for leptin‐mediated regulation of HDL‐c.
Acid‐Base Balance in Camels (Camelus dromedarius): Effects of Exercise and Dehydration
Authors: Okab Aly Bassunny , Abdoun Khalid A. , Samara Emad M. , Al-Haidary Ahmed A.
Published: June 2021

There is a lack of information pertaining to the reference levels of acid‐base parameters and the effects of dehydration and exercise on the acid‐base balance in camels. The effect of dehydration and exercise on acid‐base balance was investigated using five (2‐years old) male dromedary camels. The camels were dehydrated for 7 days or exercised for 2 hours at 6km/h. Respiratory rate and blood gases were determined immediately before, immediately after and during the first 48 hours after rehydration and exercise. Despite that dehydration and exercise induced (P<0.05) elevation of respiration rate, blood pH did not affected significantly (P≥0.05) by dehydration or exercise. Never the less, pCO2, tCO2, HCO3 and BE were increased (P<0.05) as influenced by dehydration and exercise. Forty eight hours post rehydration or exercise approved sufficient (P≥0.05) to retain the blood acid‐base parameters to their pre‐dehydration or exercise levels. This study demonstrates that blood pH remains within the normal range in dehydrated and /or exercised camels. However, the primary challenge to dehydrated or exercised camels is the mild respiratory alkalosis that was induced by reduction of blood pCO2 which in turn a result of an accelerated respiratory rate. We acknowledged the financial support of this project by King Abdul‐Aziz City for Science and Technology (KACST), Saudi Arabia under the grant SGP‐14‐14.
Infrared thermal imaging of human skin temperature during combined simulated hemorrhage and thermal stress
Authors: Moralez Gilbert , Cox Paul , Lucas Rebekah A.I. , Ganio Matthew S. , Pearson James , Crandall Craig G. , Cooke William H.
Published: June 2021

Digital infrared thermographic imaging (DITI) may be useful as a remote diagnostic tool for assessment of hemorrhagic injury. Under normal ambient temperatures DITI tracks reductions in central blood volume during simulated hemorrhage from changes in forehead skin temperature. This study tested the hypotheses that (1) DITI can detect changes in forehead skin temperature during simulated hemorrhage under elevated ambient conditions, and (2) changes in skin temperature detected with DITI correlate with changes in skin temperature using thermocouples. Ten healthy subjects were instrumented for ECG, beat‐by‐beat arterial pressure (Finometer), forehead skin temperature (thermocouple and DITI) and rested in the supine position for 60 min in a heated chamber (40 °C). Data were collected during continuous lower body negative pressure (LBNP) until pre‐syncope. DITI tracked the progressive reduction in forehead skin temperature during LBNP (baseline 35.9 ± 0.3 °C vs. pre‐syncope 35.3 ± 0.3 °C, p = 0.001). Correlations between DITI and thermocouple measurements throughout the protocol were high (r2 = 0.88), and did not differ at baseline (p = 0.53) or during LBNP (p ≥ 0.20). These data suggest that DITI may be an appropriate surrogate for remote, or non‐contact assessment of skin temperature in elevated ambient conditions and may prove to be a useful tool in the development of future technology for remote triage.
MicroRNA‐16 regulates ENaC expression in alveolar epithelial cells
Authors: parthasarathy Prasanna Tamarapu , Yunus Asfiya , Ramanathan Gurukumar Kollongod , Huynh Bao , Fukumoto Jutaro , Lockey Richard F , Kolliputi Narasaiah
Published: June 2021

Objective

MicroRNAs (miRNAs) have emerged as a novel class of gene regulators which play a critical role in complex diseases like acute lung injury (ALI). Our miRNA profiling studies in animal models of ALI revealed that miR‐146a, mir‐16 and miR‐155 were dysregulated. Mir16 targets Serotonin transporter (SERT) which is involved in serotonin uptake. Elevated levels of serotonin in ALI are known to regulate epithelial sodium channel (ENac) expression. The aim of this study is to investigate the functional effects of mir16 on SERT and ENaC in human alveolar epithelial cells (A549) in vitro.

Methods

A549 cells were commercially obtained and cultured according to the manufacturer instructions. Cells were transfected with plasmid containing miR16 expressing or control vector by Lipofectamine. q PCR and immunoblot analysis were performed to assess the effects of miR‐16 on SERT, ENaC and other downstream signalling molecules.

Results

A549 cells transfected with miR16 decreased the expression levels of SERT when compared to controls. Dose dependant regulation of SERT was observed in untransfected A549 cells treated with serotonin. Increased expression of ENaC β was observed in miR16 transfected cells compared to controls.

Conclusion

miR16 over expression altered SERT and ENaC expression in human alveolar epithelial cells.

Research supported by:RO1HL105932(NIH) and 09SDG2260957(AHA)
Measurements of Dynamic [Cl‐]i responses in stably C1C2 expressed cells with quantum dots(QD) chloride nanobiosensor, Cl‐QD
Authors: Mao Hua , Wang Yuchi , Wang Ling , Wong Lid B
Published: June 2021

C1C‐2 channel is one of the drug targets for the treatments of chloride channelopathies and imbalanced fluid secretions. Herein we describe the results of the [Cl−]i in a stably expressed ClC‐2 transfected HEK293F and T84 human cell lines measured by Cl‐ QD™. We have evaluated the role of C1C‐2 on [Cl−]i homeostasis with different putative chloride channel inhibitors and activators in both HEK293F/HEK293FC1C2 and T84/T84C1C2. Cl‐QDs were loaded into the cells using a cell‐penetrating TAT peptide protocol. [Cl−]i responses were measured using a photon counting fluorescence microscope. Five experiments were performed per condition. All four tested chloride channel inhibitors, DIDS, DCPIB, DPC and glibenclamide, caused dose‐dependent increases of [Cl−]i in C1C‐2 expressed cells compared to their respective wild types. Conversely, both tested chloride channel activators, lubriprostone and genistein, caused a lesser decrease of [Cl−]i in the C1C‐2 expressed cells compared to their respective wild types. These data demonstrate that cells stably expressed with ClC‐2 channels enhanced the responses of many putative chloride inhibitors. These data support the hypothesis that C1C‐2 channels could be a potential drug target for the treatment of diarrhea, an alternate option to the conventional cAMP‐ and Ca‐dependent chloride pathways. (Supported by NIDDK R44‐DK084600)
Cardiac glycogen influences protective responses of newly born rats to hypoxia
Authors: Fewell James Edwin , Zhang Chunfen
Published: June 2021

Failure to autoresuscitate from apnea by gasping has been suggested to play a role in sudden infant death. Little is known, however, about the factors that influence the ability of gasping to sustain life during acute hypoxia in the newborn. The present experiments were carried out on newly born rat pups to investigate the influence of cardiac glycogen on the time to last gasp (TLG) during a single hypoxic exposure, and on the ability to autoresuscitate (AR) from primary apnea during repeated hypoxic exposures. Heart glycogen was decreased in one to two day‐old pups by isoproterenol‐induced transient tachycardia. Both the TLG (Veh 2318±239s vs ISO 1990±315s; n=22) and the number of successful AR (Veh 29±12 vs ISO 20±12; n = 10) were significantly decreased (p<0.05) in newly born rat pups which had decreased cardiac glycogen. Thus, our data provide evidence that cardiac glycogen supports cardiac function and plays a role in sustaining life during hypoxia in newly born rat pups.
Off Target therapeutic effects of commonly used drugs to treat acute mountain sickness
Authors: Lisk Christina , Irwin David , Connor Ian
Published: June 2021

Introduction

Acute mountain sickness is a syndrome that effects ~60% of individuals ascending to high altitude, yet the etiology is still unclear. It has been proposed that reactive oxygen species (ROS) formed from acute high altitude exposure contribute to brain vascular leak and development of AMS. NF‐E2‐related factor 2 (Nrf2) is a transcription factor that regulates expression of greater than 90% of antioxidant genes.

Hypothesis

We postulated that some drugs currently used to treat AMS symptoms could induce Nrf2 activation and that by increasing anti‐oxidant status would decrease vascular damage from oxidative stress in hypoxic conditions.

Methods

A MCF7 cell line stably transfected with a luciferase gene driven by an antioxidant response element, the known Nrf2 promoter, was used to screen. Drugs that showed favorable Nrf2 activation were further tested in brain microvascular endothelial cells to determine if they attenuated hypoxia induced ROS production and permeability. Finally, compounds were tested for their ability to induce Nrf2 and decrease brain leak (measured by Evans blue dye) in male Spraque Dawly rats exposed to high altitude (48 h, 19,000 ft).

Results

Of 9 drugs tested, Nifedipine and Methazolamide showed strong NRF2 activation. Cells in hypoxic conditions increased hydrogen peroxide production by two fold from normoxic. Pre‐treatment with Methazolamide, Nifedipine, and Protandim decreased ROS production back to normoxic concentrations. This response was blocked by siRNA against Nrf2. Resistance was decreased after 24 hr hypoxic exposure, which was restored to normoxic values with pre‐treatments of Protandim, Methazolamide, and Nifedipine. Only drugs that showed Nrf2 activation in‐vitro, decreased brain vascular leak, in‐vivo.

Conclusion

Nrf2 activators are effective at reducing brain vascular leak from acute high altitude exposures.
The role of myostatin on skeletal muscle energy metabolism during rest and exercise
Authors: Reisz-Porszasz Suzanne , Abraham Andrea , Kovanecz Istvan , Lew Henry , Constantinescu Silvana , Porszasz Janos
Published: June 2021

Myostatin (Mst) regulates skeletal muscle mass and adipogenesis, but its role in regulating metabolism and physical performance is unknown. We investigated three mouse models: Mst overexpressing transgenic [TG]; Mst knock‐out [KO]; wild type [WT]. Male (1 yr) mice were fed a regular (R), low carb (LC), high carb (HC) or high fat (HF) diet for 6 wks. Maximal (MS) and critical (CS) running speed and ‘work capacity’ (W’) were determined using a treadmill at 5% incline. Results were allometrically scaled for body mass. Calorific intake was not different among diets. Compared to R, body mass was lower all animals in LC (5–8%; P<0.05), but in HC was 9% lower only in KO (P<0.05). During HF body mass was 41% greater in TG than WT (P<0.05). MS was faster among TG animals, especially in LC diet (TG=50±2; WT=40±1; KO=35±1 m/min; P<0.05), and was slower in KO. Also, CS was slower in TG than WT (P<0.05), but only during HC and HF diets. W’ was independent of diet but significantly greater in TG (TG=41±10 m; WT=25±5 m; KO=18±5; P=0.012). The results are consistent with a metabolic regulatory role of Mst, where overexpression results in a greater reliance on â‐oxidation and a reduced glycogenolytic rate.

Supported by U54RR022776, SC1GM089648‐01A1.
Comparison of the cardiovascular responses evoked by activation of NMDA receptors in the right and left insular cortex
Authors: Marins Fernanda Ribeiro , Xavier Carlos Henrique , Vaz Gisele Cristiane , Fontes Marco Antonio Peliky
Published: June 2021

In humans, stimulation of right insular cortex (R IC) increases the cardiac sympathetic outflow, whereas the left IC (L IC) seems to drive parasympathetic effects. Unilateral damage of IC unbalances the autonomic control, either to sympathetic or to parasympathetic prevalence, changing the cardiac performance. In rats, electrical stimulation of posterior IC showed that rostral portion drives tachycardia whereas the caudal IC drives bradycardia. We compared the cardiovascular responses evoked by stimulation of NMDA receptors into L or R IC (n=6 each). Under urethane (1.4g/kg) anesthesia, adult male Wistar rats were implanted with ECG electrodes, catheters in a femoral artery and vein. Left renal nerve was isolated to record sympathetic activity (RSNA). NMDA (0.2mM) was injected (100nL) unilaterally (0 mm posterior; 5.8 mm lateral; 7 mm ventral). NMDA into IC increased RSNA (Δ: IC L 20±6 vs. IC R 24±4 %; P<0.05 vs. baseline) and HR (Δ: IC L 28±8 vs. IC R 38±7 bpm; P<0.05 vs. baseline) without changing MAP (Δ: IC L 6±2 vs. IC R 7±4 mmHg). There were no differences in the magnitude of tachycardic and RSNA responses when comparing L and R sides of IC. No arrhythmias were detected in the recordings. We conclude that unilateral stimulation of L or R IC causes equipotent sympathoexcitatory responses. Further experiments are needed to better understand the role of this area in the autonomic control. Support: CNPq, Fapemig and Capes.
Fluoxetine Augments the Hypercapnic Ventilatory Response in CO2‐insensitive Brown Norway (BN) Rats
Authors: Puissant Madeleine M , Mouradian Gary Charles , Echert Ashley , Forster Hubert Vincent , Hodges Matthew R
Published: June 2021

The Brown Norway (BN) rat has a severely reduced hypercapnic ventilatory response (HCVR), potentially due to dysfunctional mechanisms of CO2/H+ chemoreception. BN rats also show reduced brain serotonin (5‐HT), dopamine, and norepinephrine, despite equal numbers of 5‐HT and catecholaminergic neurons. Here, we tested if daily treatment with a selective serotonin reuptake inhibitor (SSRI) fluoxetine (10 mg/kg/day) would augment the blunted HCVR in BN rats. We measured ventilation during room air (RA) and 7% CO2 breathing in young (age 3 weeks; n=10) and older (age 7 weeks; n=10) BN rats 2 days before and at multiple time points (3, 9, 14 and 21 days) during daily treatment with either fluoxetine (n=6, 6) or saline (n=4, 4). We found no differences in RA breathing or the HCVR (expressed as % room air breathing) prior to treatment, and found no effects of saline treatment at any time points during the experiment. Fluoxetine injections in young and old BN rats increased the HCVR by day 3 of treatment and remained elevated (30–60%) throughout treatment. The older BN group was also studied after discontinuing treatment, where the HCVR remained elevated up to day 3 of withdrawal, after which the HCVR was not different from the saline injected group. Thus, daily SSRI treatment reversibly augments the HCVR in CO2‐insensitive BN rats at multiple ages, further suggesting inherent 5‐HT system dysfunction in the BN rat. Supported by NHLBI HL 097033.
AN ANATOMICAL MODEL OF HEAD IN WAX MADE BY ZUMBO (1700): OBSERVATION AND PHOTOGRAPHICAL TRI‐DIMENSIONAL RECONSTRUCTION
Authors: Le Floch-Prigent Patrice Pierre , Jean-François UHL
Published: June 2021

A composite head in wax which was mounted on a real skeleton, was studied. It was photographed with a numerical camera, in rotation every 5° all along 360° and with three incidences in relationship with the sample: horizontal, at 25° and at 50°. 72 photographs were shot in each incidence, i.e.: a total of 216 was obtained. This wax was realized in the year 1700 by Gaetano Zumbo. It belonged to the Orfila museum collection closed in december 2009. The Quick Time Virtual Reality software permited the tri‐dimensional photographic reconstruction. The introduction on the internet permitted to spread the knowledge of this sample to the general public. he photographic data completed previous personal works (Le Floch‐Prigent, 1999; Le Floch‐Prigent, Guerini, Chevrot, 2006) radiographical ones and CT‐scanographical ones which proved the composite nature of the sample and the youth and female nature of the underneath head skeleton with an intra‐cranial capacity of 1350cc3.
AS160: a new Na,K‐ATPase partner that regulates the trafficking of the sodium pump in response to energy depletion and renal ischemia
Authors: Alves Daiane S , Thulin Gunilla , Loffing Johannes , Kashgarian Michael , Caplan Michael J
Published: June 2021

The Na,K‐ATPase appears to be located in two different cellular pools. The major sodium pump pool is located at the plasma membrane, while the other pool resides in cytoplasmic vesicular compartments which can translocate to the plasma membrane in response to physiological stimuli. We have identified AS160 as a new Na,K‐ATPase partner. We also demonstrated that the knockdown of AS160 increases the level of cellular Na,K‐ATPase activity, suggesting that AS160 modulates the activity of the enzyme. In the present study we characterized aspects of the physiological role of the interaction between AS160 and Na,K‐ATPase. We find that AS160 is required for the intracellular accumulation of the Na, K‐ATPase that occurs in response to energy depletion in cultured epithelial cells. In intact animals ischemic kidney injury has been shown to induce internalization of the Na,K‐ATPase from the plasma membranes of renal epithelial cells. We assessed the distribution of the Na,K‐ATPase following renal ischemia and reperfusion in kidneys from wild type and AS160 knockout mice. We observed a substantial decrease in the injury‐induced cytoplasmic accumulation of the Na,K‐ATPase in the renal epithelial cells of the AS160 knockout mice as compared to those of the wild type controls. We propose that AS160 plays a key role in regulating the redistribution of the Na,K‐ATPase that occurs in response to energy depletion and renal ischemia.
Relationship between inulin clearance and serum creatinine continuous plot using spline cubic interpolation
Authors: Morais Alceni do Carmo , Razvickas Clara Versolato , Patrícia Semedo , Oliveira-Sales Elizabeth B , Durão Marcelino , Batista Marcelo Costa , Monte Julio Cesar
Published: June 2021

To analyze the relationship among serum creatinine (SC) and inulin clearance (GRF) in a mathematical model at Acute Kidney Injury (AKI) ongoing. Correlate it with morphological changes and molecular markers of cellular process.

Animals(rats) were submitted to AKI by administration of cisplatin(CIS) (0.6 mg/kg i.p). Animals were followed until day 8. At days 1,2,3,5,6,8 animals were sacrificed. Blood and urine were collect for functional analysis, and kidneys for histopathological, immunohistochemistry, Real Time PCR analyses

GFR interpolation graph shows the minimum point between day 2 (cis group 0.24 ml/min vs. control 1.11ml/min) when SC is still at half way from the maximum point day 2 (cis group 0.70mg/dl vs. control 0.50mg/dl). This suggests that the deterioration finish in the second day after onset of AKI. In the stabilization period observed increased of mRNA expression of markers for proliferation(Cdc2), apoptosis(Bcl2) and regeneration (Pax2 and vimentin).

This study has provided insights, into timing, of cellular process during AKI. Recognizing the end of deterioration and onset of recovery is important and it affects medical decision making.
The influence of controlled breathing on cerebrovascular control
Authors: Barrera Chelsea R. , Rickards Caroline A. , Cooke William H.
Published: June 2021

High frequency breathing (HF; >0.2 Hz) may limit the influence of respiration at the low frequency (LF; 0.07–0.2 Hz) where cerebrovascular control is assessed. We tested the hypotheses that: 1) LF breathing will increase, and HF breathing will decrease LF coherence (COH) between mean arterial pressure (MAP) and mean middle cerebral artery velocity (MCAvmean ); and 2) reductions of COH expected when subjects breathe at 0.25 Hz are due, in part, to reduced CO2 associated with faster breathing. We recorded ECG, finger arterial pressure (Finometer), MCAvmean (transcranial Doppler), respiratory rate, and end‐tidal CO2 in 20 volunteers (28 ± 2 yrs). Four 7‐min breathing protocols were followed: 1) spontaneous breathing (SB); 2) paced breathing at 0.1 Hz; 3) paced breathing at 0.25 Hz; and 4) paced breathing at 0.25 Hz with CO2 control (5% CO2 bled in continuously; 0.25 Hz C). LF COH was assessed between MCAvmean and MAP. End‐tidal CO2 was lower during 0.25 Hz breathing than all other conditions (P<0.05). COH was lower during 0.25 Hz breathing than both 0.1 Hz breathing and SB (P≤0.008). COH was indistinguishable during 0.25 Hz breathing with controlled and uncontrolled CO2 (P =0.299). Controlled breathing in the HF limits contributing influences of respiration on cerebrovascular control despite moderate reductions in end‐tidal CO2.
Adeno‐associated viral delivery of TrkB receptor enhances functional recovery after cervical spinal hemisection
Authors: Gransee Heather M , Zhan Wen-Zhi , Sieck Gary C , Mantilla Carlos B
Published: June 2021

Unilateral C2 spinal cord hemisection (C2SH) removes premotor drive to phrenic motoneurons, paralyzing the ipsilateral diaphragm muscle (DIAm). In previous studies, we found that intrathecal brain‐derived neurotrophic factor (BDNF) increases the proportion of rats displaying recovery of ipsilateral DIAm activity over time. We hypothesized that enhancing expression of the tropomyosin‐related kinase receptor subtype B (TrkB) in phrenic motoneurons will promote functional recovery ipsilateral to C2SH. Adult rats were injected intrapleurally with AAV7 expressing TrkB.FL‐FLAG, and 3 weeks later C2SH was performed. A separate group of C2SH animals served as controls. Bilateral DIAm EMG activity was chronically monitored to assess recovery up to 2 weeks after C2SH. In AAV‐treated rats, delivery of TrkB.FL‐FLAG to phrenic motoneurons was evident by the presence of FLAG protein in the phrenic motor nucleus. At 7‐days post‐C2SH, the proportion of rats displaying ipsilateral DIAm EMG activity was 13% and 40% in the untreated control and AAV‐treated groups, respectively. At 14‐days post‐C2SH, all AAV‐treated rats displayed functional recovery compared to 40% of C2SH rats. We conclude that targeted delivery of TrkB.FL to phrenic motoneurons enhances functional recovery of DIAm respiratory activity following upper cervical spinal cord injury.

Supported by NIH grant HL96750 and Paralyzed Veterans of America.
The effect of DAMGO on the preBötzinger Complex (preBC) in young and adult rabbits
Authors: Stucke Astrid G , Zuperku Edward J , Geissinger Nicole M , Hopp Francis A , Prkic Ivana , Stuth Eckehard A
Published: June 2021

In vitro studies suggest the preBC as the site of opioid‐induced respiratory depression. This study evaluated the effect of injections of the mu‐opioid agonist [D‐Ala2, N‐MePhe4, Gly‐ol]‐enkephalin (DAMGO) into the preBC in young (13–24 days) and adult rabbits, which were decerebrate, tracheotomized, ventilated and vagus‐intact. The preBC was identified by injection of D,L‐homocysteic acid (1mM). DAMGO (100μM) was injected into the preBC and 1mm rostral and 1mm caudal on both sides of the brainstem. When bradypnea or apnea ensued, the opioid antagonist naloxone (1mM) was injected into the same locations. In 9 young rabbits, DAMGO caused significant bradypnea or apnea. Local naloxone reversed this depression. In 3/6 adult rabbits DAMGO injection into the preBC caused apnea, which was reversible with local naloxone. In the other three animals, DAMGO had no effect. In conclusion, opioid injection into the preBC causes respiratory depression in young but not in all adult rabbits. It remains to be seen whether this effect is still relevant at clinical opioid concentrations. Supported by FAER grant MRTG‐BS‐02/15/2010‐Stucke.
Hematocrit and Plasma Chemistry Values in Adult Collared Scops Owls (Otus lettia) and Crested Serpent Eagles (Spilornis cheela hoya)
Authors: Hsu Tien-Huan , Chan Fang-Tse
Published: June 2021

It is about the hematocrit and plasma chemistry values of adult captive collared scops owls (Otus lettia) and crested serpent eagles (Spilornis cheela hoya). In addition, the values are also differ from gender. On 37 adult captive collared scops owls and 39 adult captive crested serpent eagles, measuring their hematocrit (HCT) and plasma chemistry values for uric acid (UA), plasma urea nitrogen (BUN), total protein (TP), albumin (ALB), glucose (GLU), cholesterol (CHO), Triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), total bilirubin (TBIL), creatine (CRE), creatine phosphokinase (CPK), amylase (AMY), calcium (CA), ionic phosphorous (IP), sodium (NA), potassium (K), and chloride ions (CL). Measuring for UA, GLU, and CPK on collared scops owl, the concentrations have obvious diversity in sex. Males have evidently higher concentrations of UA and GLU (p < 0.05) than females, however, males have evidently lower concentration of CPK than females. Even so, to crested serpent eagles, there is evidently higher CPK concentration (p < 0.05) on males. Above all, it shows that raptors have varies hematocrit and plasma chemistry values due to species, body size, and sex. All these results would be helpful reference of plasma values for adult captive collared scops owls and crested serpent eagles. Also, it benefits the diagnostic tools on veterinary care of both captive species and individuals.
Using Facebook as a discussion forum for teaching Medical Physiology
Authors: Guadagnoli Jutta A.
Published: June 2021

Social networking technologies are an integral part of the lives of many students (Thompson, 2007). The users of social networking are growing in number, including both individual users and institutions. Upon matriculation all students are enrolled in Blackboard, our platform for the distribution of information and content about coursework. Learning theory has recognized that the process of learning includes not only cognitive function, but participation in a social realm (Lave and Wenger, 1991), which may go outside the traditional classroom to enhance the community of practice (CoP). While Blackboard is user friendly in many realms, its discussion interface is lacking. In an effort to enhance the CoP among our 135 students, Facebook was utilized as a mechanism to engender exchange of ideas and questions between students, peer teaching, and instructor involvement within our medical physiology class. Joining the “I Love Physiology” page was completely voluntary. Students were surveyed as to how useful the page was in enhancing their understanding of the material. Sixty seven percent of students enrolled in the Facebook page. Of these 48% posted or commented, while 83% read through the comments and posts as a review for their exam. Of the respondents, 84% found “I Love Physiology” to be useful. Facebook definitely enhanced the CoP so much so that students have formally requested its use other courses.
Up‐Regulation of Replication Factor C‐40 (RFC40) in estrogen positive as well as negative breast cancer
Authors: Gupte Rakhee
Published: June 2021

Background

Currently most therapies for breast cancer are targeted against growth factor and endocrine receptors. Identification of non receptor based molecules that will target receptor negative breast cancers is of paramount importance. Replication Factor C 40 (RFC40) protein required for DNA replication and it influences the cell cycle from DNA replication to cell division. RFC40 is over‐expressed in choriocarcinoma, acute and chronic myeloid leukemia, nasopharyngeal cancer and glioblastomas, suggesting that it may be directly responsible for unrestricted DNA replication and cell proliferation. However, its role in breast cancer has not been explored.

Methods

We determined the expression of RFC40 message and protein by qRT‐PCR and Western blot analyses in normal, estrogen positive and negative breast cancer cell lines and performed immunohistochemistry for RFC40 protein in patient breast tumor microarrays (BTMAs). We performed fluorescent in situ hybridization analyses with BTMAs to determine amplification of the RFC40 gene copy numbers in breast cancer.

Results

We found that RFC40 message and protein is significantly up‐regulated in all the breast cancer cell lines and in the patient BTMAs. We observed RFC40 gene amplification and polysomy of chromosome 7 in BTMAs

Conclusions

RFC40 is up‐regulated in breast cancer accompanied by either gene amplification or polysomy of chromosome 7.
Dysfunctional DNA double‐strand break repair is present in a subset of primary t‐aml myeloblasts
Authors: DeJesus Rigoberto , Jacoby Meagan , Shao Jin , Koboldt Daniel , Walter Matthew
Published: June 2021

We hypothesized that dysregulation of double‐strand DNA break (DSB) repair occurs in treatment related acute myeloid leukemia (t‐AML), a tumor characterized by chromosomal abnormalities, and may result from acquired mutations in homology‐directed repair (HDR) or non‐homologous end joining (NHEJ) pathway genes. We used next‐generation sequencing technology to identify acquired mutations in 6/24 t‐AML patients in 3 genes (RAD51L3, EME1, TP53) involved in HDR/NHEJ and DNA damage response. We used array comparative genomic hybridization array (aCGH) to identify 104 copy number alterations (CNAs) in 30 t‐AML samples, of which 40 CNAs were detected only by aCGH and not by cytogenetics, with many CNAs containing DNA repair genes. Finally, a functional assay was used to evaluate the induction and kinetics of histone H2AX (pH2AX) phosphorylation, a well established marker for DSB, following irradiation (IR) of primary tumor cells. Compared to normal human CD34+ hematopoietic progenitor cells (controls), 11/15 t‐AML patients had abnormal pH2AX levels at baseline or post IR, consistent with dysfunctional DSB repair. 10/11 abnormal t‐AML samples had a mutation or CNA in a DNA repair gene compared to 1/4 t‐AML samples with no pH2AX abnormality (p<0.05). This study confirms that DNA repair genes are mutated in t‐AML and suggests that dysfunctional DSB repair is present in t‐AML tumor cells. T32‐HL007088, K12 HL087107.
Design of a Biological Sensor for Potassium Ions using a DNA Aptamer and Calorimetry
Authors: Diegelman-Parente Amy
Published: June 2021

DNA quadruplexes are an unusually stable potassium‐binding structural motif that has the potential for wide‐reaching biological implications. While significant work has been performed towards the general development of these motifs as potassium sensors, little work has been done to rationally design these motifs with predictable, and wide‐ranging potassium binding constants. This project aims to correlate the [K+] required for quadruplex formation with the stability associated with an alternative hairpin structure. In an effort to facilitate such a rational design of ligandbinding oligonucleotides, we used nearest‐neighbor parameters to derive a set of DNA quadruplexes whose binding constants were determined by potassium titration experiments and circular dichroism. In addition, HT‐ITC (High‐Throughput Isothermal Titration Calorimetry) and HT‐DSC (High‐Throughput Differential Scanning Calorimetry) were used to provide a more complete picture of the ways in which alternative secondary structure formation can be used to tune the affinity of RNA and DNA sensors of small molecules. Our findings demonstrate that free energy values for both of these processes (hairpin to quadruplex formation and stability of hairpin structure) are linearly correlated, such that a DNA quadruplex‐forming sequence can be rationally designed to selectively bind K+ at any desired KD.
mTOR complex 2 (mTORC2) regulation of hepatic gene expression
Authors: Boylan Joan M , Lamming Dudley W , Neretti Nicola , Ferreira Jonathan , Sabatini David M , Gruppuso Philip A
Published: June 2021

mTORC2 has been connected to various disorders, including diabetes and cancer. It is required for insulin‐mediated suppression of hepatic glucose production. Our aim was to examine the role of mTORC2 in the regulation of liver metabolism. Control mice and RKO mice (liver‐specific knockout of Rictor, an mTORC2 constituent) were fasted overnight, fed for 3 hr, then sacrificed. Liver RNA (n = 4 per group) was analyzed using Affymetrix Genechip Mouse Gene 1.0 ST arrays. Gene Set Enrichment Analysis (GSEA) revealed upregulation of three categories of genes in the RKO mice: ribosomal proteins, TCA cycle/oxidative phosphorylation, and components of the 26S proteosome. GSEA results were consistent using both KEGG and Reactome datasets. Ingenuity Pathway Analysis (IPA) showed gene function categories relating to carbohydrate and lipid metabolism. In addition, three transcription factors were significantly altered in the RKO mice. FoxO1 was activated relative to the controls while CTNNB1 and NR1L3 were inactivated. Our results indicate that mTORC2 has broad effects on hepatic gene expression. mTORC2 is linked, in particular, to carbohydrate, lipid and mitochondrial energy metabolism. It also appears to regulate genes involved in ribosome and proteosome biogenesis. Supported by grants from NIH/NIA (DWL), the American Federation of Aging Research (DMS), Howard Hughes Medical Institute (DMS), and NIH/NICHD (PAG).
Structure of the Carnitine Palmitoyltransferase‐1A Gene in Rats, Mice and Humans
Authors: Cook George A. , Park Edwards A.
Published: June 2021

Carnitine Palmitoyltransferase‐1A (CPT1A) is expressed in all tissues of mammals except skeletal muscle and adipose tissue. CPT1A is the key regulatory enzyme of long‐chain fatty acid oxidation, and it is especially important in liver and kidney where it is important for controlling ketogenesis. When inhibited by its physiological inhibitor, malonyl‐CoA, CPT1A is also important for directing fatty acids into pathways of esterification resulting in increased secretion of lipoproteins. Since knowledge of the structure and control of the CPT1A gene will be important for understanding diabetes, altered thyroid states, and other aberrant energy states, we examined gene sequence data, genomic structure and mRNA expression data in rats, mice and human cell lines. We initially used RACE reactions to produce the full‐length rat cDNA and discovered that the initially published sequence for the rat cDNA did not include the most upstream exons. In experiments reported here, we used real‐time PCR to examine the structure of genes encoding CPT1A in these three species to see which exons were expressed in each species. The human CPT1A gene expresses 2 alternative exons (1a and 1b) upstream of the exon containing the start site for transcription. We found that the rat gene expresses a similar but larger exon (340 bp vs. 163 bp) that is approximately the same distance upstream (28 kbp) of the ATG. Furthermore, we found that rat gene expresses 3 additional intervening exons, all of which are different, both in size and location, from the human exon 1b. The mouse liver also expresses an initial exon in the same position as the human and rat gene, and it has 89% similarity with the rat exon 1a. The mouse also has sequences that are approx. 90% similar to the two additional rat exons, but they were not found to be expressed in mature mouse mRNA. In conclusion, the rat liver expresses at least 4 CPT1A mRNA species, whereas there are only 2 species expressed in humans. In mouse liver, 2 species of mRNA are expressed, but they are substantially different from the human mRNA species.
Aerobic exercise training during pregnancy improves mitochondrial function in human placenta
Authors: Ramírez‐Vélez Robinson
Published: June 2021

This was a randomized, controlled clinical trial of 20 healthy primigravid women between 16 and 20 weeks’ gestation. The pregnant women who were assigned to the exercise group received a supervised aerobic exercise program of 60 min sessions, three times a week. The effect of aerobic exercise training on mitochondrial function from human placenta was studied during the second half of pregnancy. Respiratory control showed a tendency to diminish in placental mitochondria from exercised women, as compared with control subjects. Mitochondrial membrane potential was not significantly affected by training. Mitochondrial O2‐levels and H2O2 production rate were decreased by 6% and 26% respectively in placental mitochondria from exercised women, as compared with control subjects (p<0.05). Activity of complex I‐III of the mitochondrial respiratory chain was increased by 29% in human placental mitochondria from exercised women, as compared with control subjects (p<0.05). Also, significant increases of 45% and 21%, (p<0.05) were observed in complexes II‐III and in complex IV respectively in placental mitochondria from exercised women, as compared with the controls. Aerobic training improves mitochondrial function in human placenta by increasing mitochondrial respiratory complexes activity and decreasing O2‐ and H2O2 production.