bluuebunny/crossref_metadata_2025 huggingface dataset:
Mechanism of activation of ERK and H-K-ATPase by isoproterenol in rat cortical collecting duct
Authors: Laroche-Joubert Nicolas , Marsy Sophie , Luriau Stéphanie , Imbert-Teboul Martine , Doucet Alain
Published: March 2015

Isoproterenol stimulates H-K-ATPase activity in rat cortical collecting duct β-intercalated cells through a PKA-dependent pathway. This study aimed at determining the signaling pathway underlying this effect. H-K-ATPase activity was determined in microdissected collecting ducts preincubated with or without specific inhibitors or antibodies against intracellular signaling proteins. Transient cell membrane permeabilization with streptolysin-O allowed intracellular access to antibodies. Isoproterenol increased phosphorylation of ERK in a PKA-dependent manner, and inhibition of the ERK phosphorylation prevented the stimulation of H-K-ATPase. Antibodies against the monomeric G protein Ras or the kinase Raf-1 curtailed the stimulation of H-K-ATPase by isoproterenol, whereas antibodies against the related proteins Rap-1 and B-Raf had no effect. Pertussis toxin and inhibition of tyrosine kinases with genistein also curtailed isoproterenol-induced stimulation of H-K-ATPase. It is proposed that activation of PKA by isoproterenol induces the phosphorylation of β-adrenergic receptors and the switch from Gsto Gicoupling. In turn, βγ-subunits released from Giwould activate a tyrosine kinase-Ras-Raf-1 pathway, leading to the activation of ERK1/2 and of H-K-ATPase.
Tubular reabsorption ofmyo-inositol vs. that ofd-glucose in rat kidney in vivo et situ
Authors: Silbernagl Stefan , Völker Katharina , Dantzler William H.
Published: March 2015

Filtered myo-inositol, an important renal intracellular organic osmolyte, is almost completely reabsorbed. To examine tubule sites and specificity and, thus possible mechanism of this reabsorption, we microinfused myo-[3H]inositol ord-[3H]glucose into early proximal (EP), late proximal (LP), or early distal tubule sections of superficial nephrons and into long loops of Henle (LLH) of juxtamedullary nephrons and papillary vasa recta in rats in vivo et situ and determined urinary fractional recovery of the3H label compared with comicroinfused [14C]inulin. To determine the extent to which the proximal convoluted tubule (PCT) alone contributes to myo-inositol reabsorption, we also microperfused this tubule segment between EP and LP puncture sites. We examined specificity of reabsorptive carrier(s) by adding high concentrations of other polyols and monosaccharides to the infusate. The results show that >60% of the physiological glomerular load of myo-inositol can be reabsorbed in the PCT and >90% in the short loop of Henle (SLH) by a saturable, phloridzin-sensitive process. myo-Inositol can also be reabsorbed in the ascending limb of LLH and can move from papillary vasa recta blood into ipsilateral tubular structures. Essentially no reabsorption occurred in nephron segments beyond the SLH or in collecting ducts. Specificity studies indicate that reabsorption probably occurs via a luminal Na+- myo-inositol cotransporter.
Molecular characterization of human and rat organic anion transporter OATP-D
Authors: Adachi Hisanobu , Suzuki Takehiro , Abe Michiaki , Asano Naoki , Mizutamari Hiroya , Tanemoto Masayuki , Nishio Toshiyuki , Onogawa Tohru , Toyohara Takafumi , Kasai Satoshi , Satoh Fumitoshi , Suzuki Masanori , Tokui Taro , Unno Michiaki , Shimosegawa Tooru , Matsuno Seiki , Ito Sadayoshi , Abe Takaaki
Published: March 2015

We have isolated and characterized a novel human and rat organic anion transporter subtype, OATP-D. The isolated cDNA from human brain encodes a polypeptide of 710 amino acids ( Mr76,534) with 12 predicted transmembrane domains. The rat clone encodes 710 amino acids ( Mr76,821) with 97.6% amino acid sequence homology with human OATP-D. Human and rat OATP-D have moderate amino acid sequence homology with LST-1/rlst-1, the rat oatp family, the prostaglandin transporter, and moat1/MOAT1/KIAA0880/OATP-B. Phylogenetic tree analysis revealed that OATP-D is branched in a different position from all known organic anion transporters. OATP-D transports prostaglandin E1( Km48.5 nM), prostaglandin E2( Km55.5 nM), and prostaglandin F2α, suggesting that, functionally, OATP-D encodes a protein that has similar characteristics to those of the prostaglandin transporter. Rat OATP-D also transports prostaglandins. The expression pattern of OATP-D mRNA was abundant mainly in the heart, testis, brain, and some cancer cells. Immunohistochemical analysis further revealed that rat OATP-D is widely expressed in the vascular, renal, and reproductive system at the protein level. These results suggest that OATP-D plays an important role in translocating prostaglandins in specialized tissues and cells.
The step response: a method to characterize mechanisms of renal blood flow autoregulation
Authors: Wronski T. , Seeliger E. , Persson P. B. , Forner C. , Fichtner C. , Scheller J. , Flemming B.
Published: March 2015

Response of renal vasculature to changes in renal perfusion pressure (RPP) involves mechanisms with different frequency characteristics. Autoregulation of renal blood flow (RBF) is mediated by the rapid myogenic response, by the slower tubuloglomerular feedback (TGF) mechanism, and, possibly, by an even slower third mechanism. To evaluate the individual contribution of these mechanisms to RBF autoregulation, we analyzed the response of RBF to a step increase in RPP. In anesthetized rats, the suprarenal aorta was occluded for 30 s, and then the occlusion was released to induce a step increase in RPP. Three dampened oscillations were observed; their oscillation periods ranged from 9.5 to 13 s, from 34.2 to 38.6 s, and from 100.5 to 132.2 s, respectively. The two faster oscillations correspond with previously reported data on the myogenic mechanism and the TGF. In accordance, after furosemide, the amplitude of the intermediate oscillation was significantly reduced. Inhibition of nitric oxide synthesis by Nω-nitro-l-arginine methyl ester significantly increased the amplitude of the 10-s oscillation. It is concluded that the parameters of the dampened oscillations induced by the step increase in RPP reflect properties of autoregulatory mechanisms. The oscillation period characterizes the individual mechanism, the dampening is a measure for the stability of the regulation, and the square of the amplitudes characterizes the power of the respective mechanism. In addition to the myogenic response and the TGF, a third rather slow mechanism of RBF autoregulation exists.
Contrasting roles for STAT4 and STAT6 signal transduction pathways in murine renal ischemia-reperfusion injury
Authors: Yokota Naoko , Burne-Taney Melissa , Racusen Lorraine , Rabb Hamid
Published: March 2015

Recent data support a modulatory role for CD4 T cells in experimental renal ischemia-reperfusion injury (IRI). CD4 T cells can functionally differentiate to either a Th1 (IFN-γ producing) or the counterbalancing Th2 (IL-4) phenotype. The enzymes signal transducers and activators of transcription (STAT) 4 and STAT6 regulate Th1 or Th2 differentiation and cytokine production, respectively. We therefore hypothesized that mice that were STAT4 deficient would be protected from renal IRI and that STAT6-deficient mice would have a more severe course. Intracellular cytokine staining of splenocytes from STAT4–/– or STAT6–/– exhibited distinct IFN-γ and IL-4 cytokine expression profiles. STAT6–/– had markedly worse renal function and tubular injury postischemia compared with wild type. STAT4–/– had only mildly improved function. Renal phagocyte infiltration and ICAM-1 upregulation were similar in STAT4–/–, STAT6–/–, and wild type. To evaluate if the mechanism of the marked worsening in the STAT6–/– mice could be due to IL-4 deficiency, IL-4-deficient mice were studied and had similar postischemic phenotype to STAT6–/– mice. These data demonstrate that the STAT6 pathway has a major protective role in renal IRI. IL-4 deficiency is a likely mechanism underlying the STAT6 effect. A “yin-yang” role for inflammation is emerging in renal IRI, similar to recent observations in atherosclerosis.
Effects of pathophysiological concentrations of albumin on NHE3 activity and cell proliferation in primary cultures of human proximal tubule cells
Authors: Lee E. M. , Pollock C. A. , Drumm K. , Barden J. A. , Poronnik P.
Published: March 2015

The progression of renal disease correlates strongly with hypertension and the degree of proteinuria, suggesting a link between excessive Na+reabsorption and exposure of the proximal tubule to protein. The present study investigated the effects of albumin on cell growth and Na+uptake in primary cultures of human proximal tubule cells (PTC). Albumin (1.0 mg/ml) increased cell proliferation to 134.1 ± 11.8% ( P < 0.001) of control levels with no change in levels of apoptosis. Exposure to 0.1 and 1.0 mg/ml albumin increased total22Na+uptake to 119.1 ± 6.3% ( P = 0.005) and 115.6 ± 5.3% ( P < 0.006) of control levels, respectively, because of an increase in Na+/H+exchanger isoform 3 (NHE3) activity. This was associated with an increase in NHE3 mRNA to 161.1 ± 15.1% ( P < 0.005) of control levels in response to 0.1 mg/ml albumin. Using confocal microscopy with a novel antibody raised against the predicted extracellular NH2terminus of human NHE3, we observed in nonpermeabilized cells that exposure of PTC to albumin (0.1 and 1.0 mg/ml) increased NHE3 at the cell surface to 115.4 ± 2.7% ( P < 0.0005) and 122.4 ± 3.7% ( P < 0.0001) of control levels, respectively. This effect was paralleled by significant increases in NHE3 in the subplasmalemmal region as measured in permeabilized cells. These albumin-induced increases in expression and activity of NHE3 in PTC suggest a possible mechanism for Na+retention in response to proteinuria.
Oxidative and nitrosative stress in acute renal ischemia
Authors: Noiri Eisei , Nakao Akihide , Uchida Koji , Tsukahara Hirokazu , Ohno Minoru , Fujita Toshiro , Brodsky Sergey , Goligorsky Michael S.
Published: March 2015

First Published July 12, 2001; 10.1152/ajprenal.0071.2001.—Generation of reactive oxygen species and nitric oxide in hypoxia-reperfusion injury may form a cytotoxic metabolite, peroxynitrite, which is capable of causing lipid peroxidation and DNA damage. This study was designed to examine the contribution of oxidative and nitrosative stress to the renal damage in ischemic acute renal failure (iARF). iARF was initiated in rats by 45-min renal artery clamping. This resulted in lipid peroxidation, DNA damage, and nitrotyrosine modification confirmed both by Western and immunohistochemical analyses. Three groups of animals were randomly treated with an inhibitor of inducible nitric oxide synthase (NOS),l- N6-(1-iminoethyl)lysine (l-Nil), cell-permeable lecithinized superoxide dismutase (SOD), or both. Each treatment resulted in amelioration of renal dysfunction, as well as reduced nitrotyrosine formation, lipid peroxidation, and DNA damage, thus suggesting that peroxynitrite rather than superoxide anion is responsible for lipid peroxidation and DNA damage. Therefore, in a separate series of experiments, a scavenger of peroxynitrite, ebselen, was administered before the reperfusion period. This treatment resulted in a comparable degree of amelioration of iARF. In conclusion, the present study provides the first attempt to elucidate the role of peroxynitrite in initiation of the cascade of lipid peroxidation and DNA damage to ischemic kidneys. The results demonstrate that l-Nil , lecithinized SOD, and ebselen treatments improve renal function due to their suppression of peroxynitrite production or its scavenging, consequently preventing lipid peroxidation and oxidative DNA damage.
Food restriction prevents age-related polyuria by vasopressin-dependent recruitment of aquaporin-2
Authors: Combet S. , Teillet L. , Geelen G. , Pitrat B. , Gobin R. , Nielsen S. , Trinh-Trang-Tan M.-M. , Corman B. , Verbavatz J.-M.
Published: March 2015

First published August 8, 2001; 10.1152/ajprenal.00139.2001.—The mechanisms underlying the prevention of age-related polyuria by chronic food restriction were investigated in female WAG/Rij rats. The decreased osmolality of renal papilla observed in senescent rats was not corrected by food restriction. A reduced urea content in the inner medulla of senescent rats, fed ad libitum or food-restricted, was suggested by the marked decrease in expression of UT-A1 and UT-B1 urea transporters. Aquaporin-2 (AQP2) downregulation in the inner medulla of senescent rats was partially prevented by food restriction. Both AQP2 and the phosphorylated form of AQP2 (p-AQP2), the presence of which was diffuse within the cytoplasm of collecting duct principal cells in normally fed senescent rats, were preferentially targeted at the apical region of the cells in food-restricted senescent animals. Plasma vasopressin (AVP) was similar in 10- and 30-mo-old rats fed ad libitum, but was doubled in food-restricted 30-mo-old rats. This study indicates that 1) kidney aging is associated with a marked decrease in AQP2, UT-A1, and UT-B1 expression in the inner medulla and a reduced papillary osmolality; and 2) the prevention of age-related polyuria by chronic food restriction occurs through an improved recruitment of AQP2 and p-AQP2 to the apical membrane in inner medulla principal cells, permitted by increased plasma AVP concentration.
In rat inner medullary collecting duct, NH 4 + uptake by the Na,K-ATPase is increased during hypokalemia
Authors: Wall Susan M. , Fischer Michael P. , Kim Gheun-Ho , Nguyen Bich-May , Hassell Kathryn A.
Published: March 2015

In rat terminal inner medullary collecting duct (tIMCD), the Na,K-ATPase mediates NH[Formula: see text] uptake, which increases secretion of net H+equivalents. K+and NH[Formula: see text]compete for a common binding site on the Na,K-ATPase. Therefore, NH[Formula: see text] uptake should increase during hypokalemia because interstitial K+concentration is reduced. We asked whether upregulation of the Na,K-ATPase during hypokalemia also increases basolateral NH[Formula: see text] uptake. To induce hypokalemia, rats ate a diet with a low K+content. In tIMCD tubules from rats given 3 days of dietary K+restriction, Na,K-ATPase β1-subunit (NK-β1) protein expression increased although NK-α1protein expression and Na,K-ATPase activity were unchanged relative to K+-replete controls. However, after 7 days of K+restriction, both NK-α1and NK-β1subunit protein expression and Na,K-ATPase activity increased. The magnitude of Na,K-ATPase-mediated NH[Formula: see text]uptake across the basolateral membrane ( J [Formula: see text]) was determined in tIMCD tubules perfused in vitro from rats after 3 days of a normal or a K+-restricted diet. J [Formula: see text] was the same in tubules from rats on either diet when measured at the same extracellular K+concentration. However, in either treatment group, increasing K+concentration from 10 to 30 mM reduced J [Formula: see text] >60%. In conclusion, with 3 days of K+restriction, NH[Formula: see text] uptake by Na,K-ATPase is increased in the tIMCD primarily from the reduced interstitial K+concentration.
Expression of the urate transporter/channel is developmentally regulated in human kidneys
Authors: Hyink Deborah P. , Rappoport Joshua Z. , Wilson Patricia D. , Abramson Ruth G.
Published: March 2015

First published August 15, 2001; 10.1152/ajprenal.00352. 2001.—Recombinant protein prepared from cDNA cloned from rat kidney and its human homolog function as urate transporter/channels in lipid bilayers. Using the antibody (anti-uricase) that detected the rat cDNA clone, we now demonstrate that normal human kidneys contain an immunoreactive protein of identical size to that in rat kidney (36–37 kDa), presumably the human urate transporter/channel (hUAT). The amount of hUAT in kidney homogenates increases progressively from 13 wk of gestation to the early postnatal period. During gestation, hUAT expression is confined to the cytoplasm of proximal tubules of Stage III and/or IV nephrons. However, at 1 yr of age hUAT is primarily located subapically and within brush borders of proximal tubules. Xenopus laevisoocytes and differentiated A6 cells injected with cRNA and transfected with cDNA of hUAT, respectively, demonstrated a similar pattern: hUAT is not detected in oocytes but is abundantly expressed in cytoplasm and plasma membranes of A6 cells. These data imply that different developmental factors regulate the initiation of cytoplasmic hUAT expression and subsequent insertion into human proximal tubule brush-border membranes.
Heat-induced force suppression and HSP20 phosphorylation in swine carotid media
Authors: O'Connor Matthew J. , Rembold Christopher M.
Published: March 2015

In vascular smooth muscle, cyclic nucleotide-dependent phosphorylation of heat shock protein 20 (HSP20) on serine-16 (Ser16) has been suggested to cause force suppression, i.e., reduced force with only minimal myosin regulatory light chain (MRLC) dephosphorylation. We hypothesized that heat pretreatment also suppresses force by increasing HSP20 phosphorylation. After heat pretreatment of swine carotid artery at 44.5°C for 4 h and reduction to 37°C for 1 h, Ser16-HSP20 phosphorylation was increased and histamine-induced increases in contractile force were suppressed. Subsequent addition of nitroglycerin induced additive force suppression. Heat and nitroglycerin induced a similar relation between Ser16-HSP20 phosphorylation and force. Heat pretreatment induced a small, but significant, increase in total HSP20 immunostaining. These results demonstrate that vascular smooth muscle responds to thermal stress by increasing Ser16-HSP20 phosphorylation in addition to a possible small increase in total HSP20 concentration. The resulting heat-induced reduction in force should be considered “force suppression” because histamine-induced increases in MRLC phosphorylation were not significantly altered by heat pretreatment. These processes may bring about a resistance to contractile agonists, which could have clinical significance in conditions such as hyperthermia and/or sepsis with vasodilatory shock.
Propranolol prevents epinephrine from limiting insulin-stimulated muscle glucose uptake during contraction
Authors: Hunt Desmond G. , Ding Zhenping , Ivy John L.
Published: March 2015

β-Blockade results in rapid glucose clearance and premature fatigue during exercise. To investigate the cause of this increased glucose clearance, we studied the acute effects of propranolol on insulin-stimulated muscle glucose uptake during contraction in the presence of epinephrine with an isolated rat muscle preparation. Glucose uptake increased in both fast- (epitrochlearis) and slow-twitch (soleus) muscle during insulin or contraction stimulation. In the presence of 24 nM epinephrine, glucose uptake during contraction was completely suppressed when insulin was present. This suppression of glucose uptake by epinephrine was accompanied by a decrease in insulin receptor substrate (IRS)-1-phosphatidylinositol 3 (PI3)-kinase activity. Propranolol had no direct effect on insulin-stimulated glucose uptake during contraction. However, epinephrine was ineffective in attenuating insulin-stimulated glucose uptake during contraction in the presence of propranolol. This ineffectiveness of epinephrine to suppress insulin-stimulated glucose uptake during contraction occurred in conjunction with its inability to completely suppress IRS-1-PI3-kinase activity. Results of this study indicate that the effectiveness of epinephrine to inhibit insulin-stimulated glucose uptake during contraction is severely diminished in muscle exposed to propranolol. Thus the increase in glucose clearance and premature fatigue associated with β-blockade could result from the inability of epinephrine to attenuate insulin-stimulated muscle glucose uptake.
Modulation of cholinergic responsiveness through the β-adrenoceptor signal transmission pathway in bovine trachealis
Authors: Brichetto Lorenzo , Song Pingfang , Crimi Emanuele , Rehder Kai , Brusasco Vito
Published: March 2015

The effects of pharmacological stimulation at different levels of the β-adrenoceptor (AR) pathway, including the receptor, the receptor-coupled Gs protein, and adenylyl cyclase, were studied by simultaneous measurements of acetylcholine (ACh) release and isometric force evoked by electric stimulation in isolated bovine trachealis. The β-AR agonists isoproterenol (10-6 and 10-5 M) and salbutamol (10-7 to 10-5 M) significantly attenuated both ACh release and contractile force. Forskolin, at 10-6 M, significantly increased ACh release without effect on contractile force, whereas at 10-5 M it increased ACh release but significantly decreased force. Activation of Gs protein by cholera toxin (10 μg/ml) significantly attenuated both ACh release and contractile force, but its effect on ACh release was abolished by calcium-activated potassium (KCa)-channel blocker iberiotoxin (10-7 M). The KCa-channel opener NS-1619 (10-4 M) attenuated significantly both ACh release and contractile force. It is concluded that β-AR agonists attenuate cholinergic neurotransmission in isolated bovine trachealis model by a mechanism not involving cAMP but KCa channels.
Distribution and mitogen response of peripheral blood lymphocytes after exertional heat injury
Authors: DuBose David A. , Wenger C. Bruce , Flinn Scott D. , Judy Thomas A. , Dubovtsev Alexandre I. , Morehouse David H.
Published: March 2015

To determine whether immune disturbances during exertional heat injury (EHI) could be distinguished from those due to exercise (E), peripheral lymphocyte subset distributions and phytohemagglutinin-stimulated CD69 mitogen responses as discriminated by flow cytometry were studied in military recruits [18.7 ± 0.3 (SE) yr old] training in warm weather. An E group (3 men and 3 women) ran 1.75–2 miles. During similar E, 11 recruits (10 men and 1 woman) presented with suspected EHI. EHI (40.4 ± 0.3°C) vs. E (38.6 ± 0.2°C) body temperature was significantly elevated ( P < 0.05). Heat illness was largely classified as EHI, not heatstroke, because central nervous system manifestations were generally mild. Blood was collected at E completion or EHI onset (0 h) and 2 and 24 h later. At 0 h (EHI vs. E), suppressor, natural killer, and total lymphocyte counts were significantly elevated, helper and B lymphocyte counts remained similar, and the helper-to-suppressor ratio was significantly depressed. By 2 h, immune cell dynamics between groups were similar. From 0 to 24 h, T lymphocyte subsets revealed significantly reduced phytohemagglutinin responses (percent CD69 and mean CD69 fluorescent intensity) in EHI vs. E. Thus immune cell dynamics with EHI were distinguishable from E. Because heat stress as reported in exercise or heatstroke is associated with similar immune cell disturbances, these findings in EHI contributed to the suggestion that heat stress of varying severity shares a common pathophysiological process influencing the immune system.
Modeling of impact of gas molecular diffusion on nitric oxide expired profile
Authors: Van Muylem Alain , Noël Claire , Paiva Manuel
Published: March 2015

Present descriptions of nitric oxide (NO) transport in the lungs use two compartment models: airway compartment without mixing and alveolar compartment with perfect mixing. These models neglect NO molecular diffusion in the airways. To assess the impact of axial diffusion on expired NO profile, we solved a transport equation that incorporated diffusion, convection, and NO sources in the symmetrical Weibel model of the lung. When NO parameters computed from experimental data with the two compartment models are used in our model as NO sources, simulated end-expired NO is 29–45 and 64–78% of experimental values at expiratory flows of 50 and 2,000 ml/s, respectively. These lower values are because of NO axial diffusion: During expiration, NO back diffusion (opposed to convection) prevents some NO from being expired, so a two- to fivefold increase of airway NO excretion is necessary to simulate end-expired NO consistent with experimental data. We conclude that, insofar as a significant amount of NO is produced in small airways, models neglecting NO axial diffusion underestimate excretion in the airways.
Mesenteric microvascular inflammatory responses to systemic hypoxia are mediated by PAF and LTB4
Authors: Casillan Alfred J. , Gonzalez Norberto C. , Johnson Jennifer S. , Steiner Dawn R. S. , Wood John G.
Published: March 2015

Systemic hypoxia produces a rapid microvascular inflammatory response characterized by increased reactive oxygen species (ROS) levels, leukocyte-endothelial adherence and emigration, and increased vascular permeability. The lipid inflammatory mediator leukotriene B4 (LTB4) is involved in the early hypoxia-induced responses (ROS generation and leukocyte adherence). Whether other lipid inflammatory mediators participate in this phenomenon is not known. The objective of these experiments was to study the role of platelet-activating factor (PAF) in the microvascular inflammatory response to hypoxia and its potential interactions with LTB4 in this response. Intravital microscopy was used to examine mesenteric venules of anesthetized rats. We found that WEB-2086, a PAF receptor antagonist, completely prevented the increase in ROS levels and leukocyte adherence during a brief reduction in inspired Po 2 to anesthetized rats; administration of either WEB-2086 or the LTB4 antagonist LTB4-DMA attenuated leukocyte emigration and the increase in vascular permeability to the same extent during prolonged systemic hypoxia in conscious rats. Furthermore, no additive effect was observed in either response when both antagonists were administered simultaneously. This study demonstrates a role for PAF in the rapid microvascular inflammatory response to hypoxia, as well as contributions of PAF and LTB4 to the slowly developing responses observed during sustained hypoxia. The incomplete blockade of the hypoxia-induced increases in vascular permeability and leukocyte emigration by combined administration of both antagonists indicates that factors in addition to LTB4 and PAF participate in these phenomena.
Sound transmission in the lung as a function of lung volume
Authors: Bergstresser T. , Ofengeim D. , Vyshedskiy A. , Shane J. , Murphy R.
Published: March 2015

We were interested in how the transmission of sound through the lung was affected by varying air content in intact humans as a method of monitoring tissue properties noninvasively. To study this, we developed a method of measuring transthoracic sound transit time accurately. We introduced a “coded” sound at the mouth and measured the transit time at multiple microphones placed over the chest wall by using a 16-channel lung sound analyzer (Stethographics). We used a microphone placed over the neck near the trachea as our reference and utilized cross-correlation analysis to calculate the transit times. The use of the coded sound, composed of a mix of frequencies from 130 to 150 Hz, greatly reduced the ambiguity of the cross-correlation function. The measured transit time varied from 1 ms at the central locations to 5 ms at the lung bases. Our results also indicated that transit time at all locations decreased with increasing lung volume. We found that these results can be described in terms of a model in which sound transmission through the lung is treated as a combination of free-space propagation through the trachea and a propagation through a two-phase system in the parenchyma.
Assessment of brachial artery blood flow across the cardiac cycle: retrograde flows during cycle ergometry
Authors: Green Daniel , Cheetham Craig , Reed Chris , Dembo Lawrie , O'Driscoll Gerry
Published: March 2015

We describe a novel software system that utilizes automated algorithms to perform edge detection and wall tracking of high-resolution B-mode arterial ultrasound images, combined with synchronized Doppler waveform envelope analysis, to calculate conduit arterial blood flow (BF) across the cardiac cycle. Furthermore, we describe changes in brachial arterial BF to the resting forearm during incremental cycle ergometry in eight subjects. During exercise, peak BF during the cardiac cycle increased at each workload ( P < 0.001), because of increased velocity in the presence of unaltered cross-sectional area. In contrast, mean BF calculated across each cardiac cycle decreased at lower workloads before increasing at 100 and 160 W ( P< 0.001). Differences in the pattern of peak and mean cardiac cycle flows were due to the influence of retrograde diastolic flow, which had a larger impact on mean flows at lower workloads. In conclusion, BF can be measured with high temporal resolution across the cardiac cycle in humans. Resting brachial arterial flow, including retrograde flow, increases during lower limb exercise.
Mechanical properties of rat soleus after long-term spinal cord transection
Authors: Talmadge Robert J. , Roy Roland R. , Caiozzo Vincent J. , Edgerton V. Reggie
Published: March 2015

The effects of a complete spinal cord transection (ST) on the mechanical properties of the rat soleus were assessed 3 and 6 mo post-ST and compared with age-matched controls. Maximal tetanic force was reduced by ∼44 and ∼25% at 3 and 6 mo post-ST, respectively. Similarly, maximum twitch force was reduced by ∼29% in 3-mo and ∼17% in 6-mo ST rats. ST resulted in faster twitch properties as evidenced by shorter time to peak tension (∼45%) and half-relaxation time (∼55%) at both time points. Maximum shortening velocity was significantly increased in ST rats whether measured by extrapolation from the force-velocity curve (approximately twofold at both time points) or by slack-test measurements (over twofold at both time points). A significant reduction in fatigue resistance of the soleus was observed at 3 (∼25%) and 6 mo (∼45%) post-ST. For the majority of the speed-related properties, no significant differences were detected between 3- and 6-mo ST rats. However, the fatigue resistance of the soleus was significantly lower in 6- vs. 3-mo ST rats. These data suggest that, between 3 and 6 mo post-ST, force-related properties tended to recover, speed-related properties plateaued, and fatigue-related properties continued to decline. Thus some specific functional properties of the rat soleus related to contractile force, speed, and fatigue adapted independently after ST.
Computational model of airway narrowing: mature vs. immature rabbit
Authors: Lambert R. K. , Ramchandani R. , Shen X. , Gunst S. J. , Tepper R. S.
Published: March 2015

Immature rabbits have greater maximal airway narrowing and greater maximal fold increases in airway resistance during bronchoconstriction than mature animals. We have previously demonstrated that excised immature rabbit lungs have more distensible airways, a lower shear modulus, and structural differences in the relative composition and thickness of anatomically similar airways. In the present study, we incorporated anatomic and physiological data for mature and immature rabbits into a computational model of airway narrowing. We then investigated the relative importance of maturational differences in these factors as determinants of the greater airway narrowing that occurs in the immature animal. The immature model demonstrated greater sensitivity to agonist, as well as a greater maximal fold increase in airway resistance. Exchanging values for airway compliance between the mature and immature models resulted in the mature model exhibiting a greater maximal airway response than the immature model. In contrast, exchanging the shear moduli or the composition of the airway wall relative to the airway size produced relatively small changes in airway reactivity. Our results strongly suggest that the mechanical properties of the airway, i.e., greater compliance of the immature airway, can be an important factor contributing to the greater airway narrowing of the immature animal.
Heat acclimation increases skin vasodilation and sweating but not cardiac baroreflex responses in heat-stressed humans
Authors: Yamazaki Fumio , Hamasaki Kunshige
Published: March 2015

In the present study, to test the hypothesis that exercise-heat acclimation increases orthostatic tolerance via the improvement of cardiac baroreflex control in heated humans, we examined cardiac baroreflex and thermoregulatory responses, including cutaneous vasomotor and sudomotor responses, during whole body heating before and after a 6-day exercise-heat acclimation program [4 bouts of 20-min exercise at 50% peak rate of oxygen uptake separated by 10-min rest in the heat (36°C; 50% relative humidity)]. Ten healthy young volunteers participated in the study. On the test days before and after the heat acclimation program, subjects underwent whole body heat stress produced by a hot water-perfused suit during supine rest for 45 min and 75° head-up tilt (HUT) for 6 min. The sensitivity of the arterial baroreflex control of heart rate (HR) was calculated from the spontaneous changes in beat-to-beat arterial pressure and HR. The HUT induced a presyncopal sign in seven subjects in the preacclimation test and in six subjects in the postacclimation test, and the tilting time did not differ significantly between the pre- (241 ± 33 s) and postacclimation (283 ± 24 s) tests. Heat acclimation did not change the slope in the HR-esophageal temperature (Tes) relation and the cardiac baroreflex sensitivity during heating. Heat acclimation decreased ( P < 0.05) the Tes thresholds for cutaneous vasodilation in the forearm and dorsal hand and for sweating in the forearm and chest. These findings suggest that short-term heat acclimation does not alter the spontaneous baroreflex control of HR during heat stress, although it induces adaptive change of the heat dissipation response in nonglabrous skin.
Invited Review: HXB/BXH rat recombinant inbred strain platform: a newly enhanced tool for cardiovascular, behavioral, and developmental genetics and genomics
Authors: Printz Morton P. , Jirout Martin , Jaworski Rebecca , Alemayehu Adamu , Kren Vladimir
Published: March 2015

This review deals with the largest set of rat recombinant inbred (RI) strains and summarizes past and recent accomplishments with this platform for genetic mapping and analyses of divergent and complex traits. This strain, derived by crossing the spontaneously hypertensive rat, SHR/Ola, with a Brown Norway congenic, BN-Lx, carrying polydactyly-luxate syndrome, is referred to as HXB/BXH. The RI strain set has been used for linkage and association studies to identify quantitative trait loci for numerous cardiovascular phenotypes, including arterial pressure, stress-elicited heart rate, and pressor response, and metabolic traits, including insulin resistance, dyslipidemia and glucose handling, and left ventricular hypertrophy. The strain's utility has been enhanced with development of a new framework marker-based map and strain distribution patterns of polymorphic markers. Quantitative trait loci for behavioral traits mapped include loci for startle motor response and habituation, anxiety and locomotion traits associated with elevated plus maze, and conditioned taste aversion. The polydactyly-luxate syndrome Lx mutation has allowed the study of alleles important to limb development and malformation phenotypes as well as teratogens. The RI strains have guided development of numerous congenic strains to test locus assignments and to study the effect of genetic background. Although these strains were originally developed to aid in studies of rat genetic hypertension and morphogenetic abnormalities, this rodent platform has been shown to be equally powerful for a wide spectrum of traits and endophenotypes. These strains provide a ready and available vehicle for many physiological and pharmacological studies.
Effects of 14 days of head-down tilt bed rest on cutaneous vasoconstrictor responses in humans
Authors: Wilson Thad E. , Shibasaki Manabu , Cui Jian , Levine Benjamin D. , Crandall Craig G.
Published: March 2015

This study tested the hypothesis that head-down tilt bed rest (HDBR) reduces adrenergic and nonadrenergic cutaneous vasoconstrictor responsiveness. Additionally, an exercise countermeasure group was included to identify whether exercise during bed rest might counteract any vasoconstrictor deficits that arose during HDBR. Twenty-two subjects underwent 14 days of strict 6° HDBR. Eight of these 22 subjects did not exercise during HDBR, while 14 of these subjects exercised on a supine cycle ergometer for 90 min a day at 75% of pre-bed rest heart rate maximum. To assess α-adrenergic vasoconstrictor responsiveness, intradermal microdialysis was used to locally administer norepinephrine (NE), while forearm skin blood flow (SkBF; laser-Doppler flowmetry) was monitored over microdialysis membranes. Nonlinear regression modeling was used to identify the effective drug concentration that caused 50% of the cutaneous vasoconstrictor response (EC50) and minimum values from the SkBF-NE dose-response curves. In addition, the effects of HDBR on nonadrenergic cutaneous vasoconstriction were assessed via the venoarteriolar response of the forearm and leg. HDBR did not alter EC50 or the magnitude of cutaneous vasoconstriction to exogenous NE administration regardless of whether the subjects exercised during HDBR. Moreover, HDBR did not alter the forearm venoarteriolar response in either the control or exercise groups during HDBR. However, HDBR significantly reduced the magnitude of cutaneous vasoconstriction due to the venoarteriolar response in the leg, and this response was similarly reduced in the exercise group. These data suggest that HDBR does not alter cutaneous vasoconstrictor responses to exogenous NE administration, whereas cutaneous vasoconstriction of the leg due to the venoarteriolar response is reduced after HDBR. It remains unclear whether attenuated venoarteriolar responses in the lower limbs contribute to reduced orthostatic tolerance after bed rest and spaceflight.
Spinoreticular neurons that receive group III input are inhibited by MLR stimulation
Authors: Degtyarenko Alexandr M. , Kaufman Marc P.
Published: March 2015

In decerebrate paralyzed cats, we examined the responses of 18 spinoreticular neurons to electrical stimulation of the mesencephalic locomotor region. The activity of each of the spinoreticular neurons was recorded extracellularly from laminae IV through VI of the L7 and S1 spinal cord. In addition, each of the 18 spinoreticular neurons received group III afferent input from the tibial nerve. Spinoreticular projections were established for each of 18 neurons by antidromic invasion of the ventro lateral medulla at the P11 though P14 levels. The onset latencies and current thresholds for antidromic invasion from the ventro lateral medulla averaged 15.0 ± 3.8 ms and 117 ± 11 μA, respectively. Electrical stimulation of the mesencephalic locomotor region attenuated the spontaneous activity or the responses of each of the spinoreticular neurons to tibial nerve stimulation at currents that recruited group III afferents. Our data support the notion that thin-fiber muscle afferent input to the ventrolateral medulla is gated by a central command to exercise.
Detection of local lung air content by electrical impedance tomography compared with electron beam CT
Authors: Frerichs Inéz , Hinz José , Herrmann Peter , Weisser Gerald , Hahn Günter , Dudykevych Taras , Quintel Michael , Hellige Gerhard
Published: March 2015

The aim of the study was to validate the ability of electrical impedance tomography (EIT) to detect local changes in air content, resulting from modified ventilator settings, by comparing EIT findings with electron beam computed tomography (EBCT) scans obtained under identical steady-state conditions. The experiments were carried out on six anesthetized supine pigs ventilated with five tidal volumes (Vt) at three positive end-expiratory pressure (PEEP) levels. The lung air content changes were determined both by EIT (Goe-MF1 system) and EBCT (Imatron C-150XP scanner) in six regions of interest, located in the ventral, middle, and dorsal areas of each lung, with respect to the reference air content at the lowest Vt and PEEP, as a change in either local electrical impedance or lung tissue density. An increase in local air content with Vt and PEEP was identified by both methods at all regions studied. A good correlation between the changes in lung air content determined by EIT and EBCT was revealed. Mean correlation coefficients in the ventral, middle, and dorsal regions were 0.81, 0.87, and 0.93, respectively. The study confirms that EIT is a suitable, noninvasive method for detecting regional changes in air content and monitoring local effects of artificial ventilation.
Pulmonary hypertension in TNF-α-overexpressing mice is associated with decreased VEGF gene expression
Authors: Fujita Masaki , Mason Robert J. , Cool Carleyne , Shannon John M. , Hara Nobuyuki , Fagan Karen A.
Published: March 2015

Tumor necrosis factor-α (TNF-α) transgenic mice have previously been found to have characteristics consistent with emphysema and severe pulmonary hypertension. Lungs demonstrated alveolar enlargement as well as interstitial thickening due to chronic inflammation and perivascular fibrosis. In the present report, we sought to determine potential mechanisms leading to development of pulmonary hypertension in TNF-α transgenic mice. To determine whether sustained vasoconstriction was an important component of this pulmonary hypertension, nitric oxide was administered and hemodynamics were measured. Nitric oxide (25 ppm) failed to normalize right ventricular pressure in transgene-positive mice, suggesting that the pulmonary hypertension was not due to sustained vasoconstriction. Structural analysis of the pulmonary arteries found adventitial thickening and a trend toward medial hypertrophy in pulmonary arteries of transgene-positive mice, suggesting that vascular remodeling had occurred. Echocardiographic measurement of the percent fractional shortening of the left ventricle as a measurement of ventricular function in vivo revealed that left ventricular dysfunction was not contributing to pulmonary hypertension. We examined expression of genes known to be important in regulation of vascular tone and structure. Messenger RNA expression of vascular endothelial growth factor and its receptor flk-1 was reduced compared with transgene-negative littermates at all ages. Endothelial and inducible nitric oxide synthase mRNA levels were similar in both groups. Endothelin-1 mRNA was also decreased in TNF-α transgenic mice. Interestingly, female transgenic mice had decreased survival rate compared with male transgenic mice. We conclude that chronic overexpression of TNF-α is associated with decreased vascular endothelial growth factor and flk-1 gene expression, pulmonary vascular remodeling, and severe pulmonary hypertension, although the precise mechanism is unknown.
Regional brain gray matter loss in heart failure
Authors: Woo Mary A. , Macey Paul M. , Fonarow Gregg C. , Hamilton Michele A. , Harper Ronald M.
Published: March 2015

Heart failure (HF) patients exhibit enhanced sympathetic tone, aberrant responses to blood pressure challenges, and sleep-related breathing disorders, suggesting that the syndrome is accompanied by central neural deficits. We assessed regional gray matter volumes over the entire brain in nine HF patients (51 ± 10 yr; left ventricular ejection fraction 0.27 ± 0.06; six men) and 27 healthy controls (46 ± 12 yr; 22 men) using T1-weighted magnetic resonance imaging to evaluate potential neural damage. Regional volumes were evaluated by using voxel-based morphometry while controlling for age, gender, and handedness. HF patients showed significant and largely lateralized gray matter loss in autonomic and respiratory-related areas as well as regions not classically associated with such control, including the insula and basal ganglia, right cingulate gyrus, parahippocampal/fusiform gyrus, dorsal midbrain extending to the posterior and medial thalamus, ventral and superior lateral frontal cortex, bilateral cerebellar quadrangular lobules and right fastigial and neighboring nuclei, and bilateral deep parietal and lateral parietal-occipital cortex. Areas of gray matter loss may contribute to inappropriate cognitive, autonomic, and breathing regulation in HF.
Tracking of airway and tissue mechanics during TLC maneuvers in mice
Authors: Hantos Zoltán , Collins Rachel A. , Turner Debra J. , Jánosi Tibor Z. , Sly Peter D.
Published: March 2015

A tracking impedance estimation technique was developed to follow the changes in total respiratory impedance (Zrs) during slow total lung capacity maneuvers in six anesthetized and mechanically ventilated BALB/c mice. Zrs was measured with the wave-tube technique and pseudorandom forced oscillations at nine frequencies between 4 and 38 Hz during inflation from a transrespiratory pressure of 0-20 cmH2O and subsequent deflation, each lasting for ∼20 s. Zrs was averaged for 0.125 s and fitted by a model featuring airway resistance (Raw) and inertance, and tissue damping and elastance ( H). Lower airway conductance (Glaw) was linearly related to volume above functional residual capacity (V) between 0 and 75-95% maximum V, with a mean slope of dGlaw/dV = 13.6 ± 4.6 cmH2O-1 · s-1. The interdependence of Raw and H was characterized by two distinct and closely linear relationships for the low- and high-volume regions, separated at ∼40% maximum V. Comparison of Raw with the highest-frequency resistance of the total respiratory system revealed a marked volume-dependent contribution of tissue resistance to total respiratory system resistance, resulting in the overestimation of Raw by 19 ± 8 and 163 ± 40% at functional residual capacity and total lung capacity, respectively, whereas the lowest frequency reactance was proportional to H; these findings indicate that single-frequency resistance values may become inappropriate as surrogates of Raw when tissue impedance is changing.
One- and two-year change in body composition as measured by DXA in a population-based cohort of older men and women
Authors: Visser Marjolein , Pahor Marco , Tylavsky Frances , Kritchevsky Stephen B. , Cauley Jane A. , Newman Anne B. , Blunt Barbara A. , Harris Tamara B.
Published: March 2015

Changing body composition has been suggested as a pathway to explain age-related functional decline. No data are available on the expected changes in body composition as measured by dual-energy X-ray absorptiometry (DXA) in a population-based cohort of older persons. Body composition data at baseline, 1-yr follow-up, and 2-yr follow-up was measured by DXA in 2,040 well-functioning black and white men and women aged 70–79 yr, participants of the Health, Aging, and Body Composition Study. After 2 yr, a small decline in total body mass was observed (men: −0.3%, women: −0.4%). Among men, fat-free mass and appendicular lean soft tissue mass (ALST) decreased by −1.1 and −0.8%, respectively, which was masked by a simultaneous increase in total fat mass (+2.0%). Among women, a decline in fat-free mass was observed after 2 yr only (−0.6%) with no change in ALST and body fat mass. After 2 yr, the decline in ALST was greater in blacks than whites. Change in total body mass was associated with change in ALST ( r = +0.58 to +0.70; P < 0.0001). Among participants who lost total body mass, men lost relatively more ALST than women, and blacks lost relatively more ALST than whites. In conclusion, the mean change in body composition after a 1- to 2-yr follow-up was 1–2% with a high interindividual variability. Loss of ALST was greater in men compared with women, and greater in blacks compared with whites, suggesting that men and blacks may be more prone to muscle loss.