Ultrastructural morphometry of the blood-air barrier in pulmonary sarcoidosis.

Stereologic techniques were utilized in an electron microscopic study of biopsy samples obtained from the lungs of seven patients with pulmonary sarcoidosis. Relative fractional volumes of alveolar septal components and the arithmetic mean thickness and harmonic mean thickness of alveolocapillary membranes (blood-air barrier) were compared with values for normal lungs. Based on morphometric analysis, increases in the arithmetic mean thickness and the harmonic mean thickness of the alveolocapillary membranes appeared too small to account for the reduction in gas transfer present; however, there was a quantitative relative increase in interstitial tissue in alveolar septa, which does not take part in gas exchange, at the expense of the capillary bed, which is critical to this function.     

Arterial micropuncture. An advance in blood-gas analysis

Worldwide, uncountable arterial macropunctures and cannulations are performed for blood-gas analysis. The associated complications are accepted as inevitable. 8,000 arterial micropunctures performed under controlled conditions show that this sampling method is not only simpler but also less traumatic and less susceptible to errors of method. As these advantages are not countered by any disadvantages, microsampling is suggested for general use.     

肺心病呼吸衰竭血气分析指标与预后关系分析

目的：研究肺心病呼吸衰竭患的血气指标变化对预后的影响。方法：分析患血气分析指标的变化与预后的关系。结果：（1）血气分析指标中的pH值越低，死亡率越高。（2）血气分析指标中的二氧化碳分压越高，死亡率也相应升高；（3）血气分析指标中的氧分压高低对预后无明显影响；（4）酸中毒总死亡率大于碱中毒，三重性酸碱平紊乱死亡率最高。结论：（1）pH值对预后有绝对影响，两呈负相关系；（2）二氧化碳对预后同样有影响，两呈正相关系；（3）同一患出现二种或以上酸碱平衡紊乱时，预后差、死亡率高。     

Signaling pathways involved in OxPAPC-induced pulmonary endothelial barrier protection

Increased tissue or serum levels of oxidized phospholipids have been detected in a variety of chronic and acute pathological conditions such as hyperlipidemia, atherosclerosis, heart attack, cell apoptosis, acute inflammation and injury. We have recently described signaling cascades activated by oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (OxPAPC)in the human pulmonary artery endothelial cells (EC) and reported potent barrier-protective effects of OxPAPC, which were mediated by small GTPases Rac and Cdc42. In this study we have further characterized signal transduction pathways involved in the OxPAPC-mediated endothelial barrier protection. Inhibitors of small GTPases, protein kinase A (PKA), protein kinase C (PKC), Src family kinases and general inhibitors of tyrosine kinases attenuated OxPAPC-induced barrier-protective response and EC cytoskeletal remodeling. In contrast, small GTPase Rho, Rho kinase, Erk-1,2 MAP kinase and p38 MAP kinase and PI3-kinase were not involved in the barrier-protective effects of OxPAPC. Inhibitors of PKA, PKC, tyrosine kinases and small GTPase inhibitor toxin B suppressed OxPAPC-induced Rac activation and decreased phosphorylation of focal adhesion kinase (FAK) and paxillin. Barrier-protective effects of OxPAPC were not reproduced by platelet activating factor (PAF), which at high concentrations induced barrier dysfunction, but were partially attenuated by PAF receptor antagonist A85783. These results demonstrate for the first time upstream signaling cascades involved in the OxPAPC-induced Rac activation, cytoskeletal remodeling and barrier regulation and suggest PAF receptor-independent mechanisms of OxPAPC-mediated endothelial barrier protection.     

Electron microscopic studies on the alveolar-capillary barrier in the patients of chronic pulmonary edema

Electron microscopic studies on the alveolar-capillary barrier were carried out in 13 patients of chronic pulmonary edema and/or congestion resulting from heart disease of various etiologies. The characteristic findings are tremendous proliferation of type II granular pneumocyte and irregular thickening of alveolar epithelial and capillary basement membrane. These ultrastructural changes correlated to the duration of heart failure and mean pulmonary arterial wedge pressure despite of disease groups and the age of the patients. In particular, lamination of capillary basement membrane with fragmentation was more specific finding which was observed only in patients with mean pulmonary arterial wedge pressure above 35 mmHg and duration of heart failure over 6 hears. There was no apparent relationship existed between the ultrastructural changes and pulmonary arterial pressure. In addition to these characteristic changes the remaining ultrastructural changes of the alveolar-capillary barrier were difficult to make a correlation to clinical course and cardio-pulmonary hemodynamics.     

Increased resistance of the gastric mucosal barrier to barrier breakers in the rat.

The effects of two exposures of the gastric mucosa to either 10 mM taurocholic acid (TcA) or 20% ethanol, both in 150 mM HCl, on transmucosal potential difference (PD) and net fluxes of H+, Na+, and K+ ions have been tested in the rat. The interval between exposures was 30 min. The results demonstrated that the first exposure of the gastric mucosa, either to TcA or to ethanol, reduced the net fluxes of H+, Na+, K+ and the change in transmucosal PD induced by the second exposure, indicating an increased resistance of the mucosa to the barrier breaking effects of TcA or ethanol.     

Heat exchnage in the black skipjack, and the blood-gas relationship of warm-bodied fishes

The black skipjack, Euthynnus lineatus, uses a centrally located vascular heat exchanger to maintain core body temperatures warmer than ambient sea water. The heat exchanger is composed of the dorsal aorta, the posterior cardinal vein, and a large vertical rete. The dorsal aorta is embedded in the posterior cardinal vein and is completely bathed in venous blood. Skipjack hemoglobin appears similar to that of the bluefin tuna in that oxygen capacity is unaffected by changing temperature. Temperature-insensitive hemoglobin may function in warm-bodied fishes to prevent the premature dissociation of oxygen from hemoglobin as blood is warmed en route to the muscles.     

The effect of dextran solution on pulmonary plasma-lymph barrier in awake sheep

We investigated the effect of dextran solution on lung lymph flow in awake sheep with chronic lung lymph fistulas. Ten percent of dextran (molecular weight 40,000) solution or normal saline were infused at 1000 ml/h for 2 h through left atrial catheter. We measured pulmonary arterial pressure (Ppa), left atrial pressure (Pla), aortic pressure (Psa), cardiac output (CO), oncotic pressures of both plasma (IImv) and lung lymph (IIpmv), lung lymph flow rate (Qlym), and lymph-to-plasma ratio of total protein (L/P). Infusion of dextran solution caused significant increases in Ppa, Pla and CO and decrease in plasma-lung lymph oncotic pressure gradient (IImv-IIpmv), without changes in L/P. Infusion of normal saline caused significant increases in Ppa and Pla and no changes in IImv-IIpmv and CO, and slight decrease in L/P. The calculated filtration coefficients increased by 2.2 fold after dextran infusion and 1.7 fold after normal saline infusion. Moreover, an apparent increase in protein transport across microvessels as evidenced by the normal L/P despite increases in hydrostatic pressure occurred after dextran solution infusion. These results suggest that dextran solution may increase permeability of microvascular wall, as well as effective net pressure gradient across microvessels, both of which result in a large net fluid filtration from microvessels to perimicrovascular compartment.     

Soluble adenylyl cyclase reveals the significance of cAMP compartmentation on pulmonary microvascular endothelial cell barrier

Subtle elevations in cAMP localized to the plasma membrane intensely strengthen endothelial barrier function. Paradoxically, pathogenic bacteria insert adenylyl cyclases (ACs) into eukaryotic cells generating a time-dependent cytosolic cAMP-increase that disrupts rather than strengthens the endothelial barrier. These findings bring into question whether membrane versus cytosolic AC activity dominates in control of cell adhesion. To address this problem, a mammalian forskolin-sensitive soluble AC (sACI/II) was expressed in pulmonary microvascular endothelial cells. Forskolin stimulated this sACI/II construct generating a small cytosolic cAMP-pool that was not regulated by phosphodiesterases or Galphas. Whereas forskolin simultaneously activated the sACI/II construct and endogenous transmembrane ACs, the modest sACI/II activity overwhelmed the barrier protective effects of plasma membrane activity to induce endothelial gap formation. Retargeting sACI/II to the plasma membrane retained AC activity but protected the endothelial cell barrier. These findings demonstrate for the first time that the intracellular location of cAMP synthesis critically determines its physiological outcome.     

Cimetidine and the gastric mucosal barrier.

The histamine H2-receptor antagonist, cimetidine, when given topically or parenterally, did not affect net luminal Na+ gain or net luminal H+ loss in vagally-denervated pouches of antrectomised dogs. Cimetidine did not affect disruption of the mucosal barrier by 5 mmol/l or 20 mmol/l sodium taurocholate.     

Osmotic flows across the blood-joint barrier.

The effective osmotic pressure across the blood-joint barrier is a key factor controlling synovial fluid volume and joint effusions. The effect of plasma colloid osmotic pressure (COP) on trans-synovial flow was studied directly in rabbit knees expanded by intra-articular fluid infusion. The synovial microcirculation was perfused with blood of varying COP. Absorption of fluid from the joint cavity was a linear function of COP; but albumin COP was only 78% effective across the blood-joint interface. Hyperosmolar solutions of small solutes (e.g., glucose) generated transient osmotic flows across the blood-joint barrier, but far less effectively than albumin. The hydraulic permeability of synovium increased at pathological intra-articular pressures--a phenomenon of potential importance to effusion kinetics.     

Immunological function of the blood-cerebrospinal fluid barrier.

Because the brain lacks a true lymphatic system, it is unclear how peripheral lymphocytes recognize foreign antigens present in the central nervous system. This report demonstrates that the choroid plexus, which constitutes the blood-cerebrospinal fluid barrier, is able to present foreign antigen to, and stimulate the proliferation of, peripheral helper T lymphocytes through an Ia-dependent, major histocompatibility complex-restricted mechanism. Furthermore, in vivo, choroid plexus epithelial cells have access to, and are capable of taking up, virus-sized particles injected elsewhere into the cerebrospinal fluid. Thus these data suggest that the blood-cerebrospinal fluid barrier may play a role in immunological communication between the central nervous system and periphery, a function relevant to the initiation of immunological responses to central nervous system infections and autoimmune processes and for the surveillance of tumor cells in the cerebrospinal fluid.     

Ca(2+)-inhibitable adenylyl cyclase modulates pulmonary artery endothelial cell cAMP content and barrier function.

Maintenance by the endothelium of a semi-permeable barrier is critically important in the exchange of oxygen and carbon dioxide in the lung. Intracellular free Ca2+ ([Ca2+]i) and cAMP are principal determinants of endothelial cell barrier function through their mutually opposing actions on endothelial retraction. However, details of the mechanisms of this antagonism are lacking. The recent discovery that certain adenylyl cyclases (EC 4.6.1.1) could be acutely inhibited by Ca2+ in the intracellular concentration range provided one possible mechanism whereby elevated [Ca2+]i could decrease cAMP content. This possibility was explored in pulmonary artery endothelial cells. The results indicate that a type VI Ca(2+)-inhibitable adenylyl cyclase exists in pulmonary artery endothelial cells and is modulated by physiological changes in [Ca2+]i. Furthermore, the results suggest the inverse relationship between [Ca2+]i and cAMP that is established by Ca(2+)-inhibitable adenylyl cyclase plays a critical role in modulating pulmonary artery endothelial cell permeability. These data provide evidence that susceptibility to inhibition of adenylyl cyclase by Ca2+ can be exploited in modulating a central physiological process.     

Expanding the definition of the blood-brain barrier to protein.

Tight junctions between cerebral endothelial cells and the near absence of pinocytosis and vesicular transport of blood-borne protein into and across these cells are believed to constitute the mammalian blood-brain barrier. In the present investigation evidence is provided to indicate that the capillary endothelium of the mouse brain pinocytosis the enzymatic tracer horseradish peroxidase (EC 1.11.1.17) from cerebral blood under normal conditions. This protein and the internalized endothelial surface membrane associated with it are directed, for the most part, to acid hydrolase-positive lysosomes for degradation. Although peroxidase was never seen in the perivascular clefts, the lysosomes of pericytes were peroxidase-positive. Pericytes are macrophage-like cells located on the abluminal surfaces of cerebral microvasculature; these cells may serve as the first line of defense once the blood-brain barrier is breached. The definition of the blood-brain barrier should be expanded to include consideration of the lysosomal system of organelles in endothelial cells and pericytes.     

A blood-gas nomogram of the chick fetus: blood flow distribution between the chorioallantois and fetus

This paper presents equations for quantifying the relationships between the O2 and CO2 concentrations and tensions in the blood of the 18-day chick fetus. A blood-gas nomogram showing these relationships is presented. Starting with the reported chorioallantoic artery and vein gas tensions and using the blood-gas equations, the range of embryonic arterial and venous gas tensions as well as the distribution of the cardiac output and the degree of mixing between the chorioallantoic and embryonic circulations are explored. It is concluded that at least 65% of the blood in the chorioallantoic artery consists of blood of embryonic mixed venous composition. A model of the blood flow distribution is proposed in which chorioallantoic and embryonic flows are equal, with 70% of the blood returning from the tissues of the embryo going to the chorioallantois and vice versa.     

Alveolar macrophage inducible nitric oxide synthase-dependent pulmonary microvascular endothelial cell septic barrier dysfunction

Inducible nitric oxide (NO) synthase (iNOS) from neutrophils and alveolar macrophages (AM) contributes to the pathophysiology of murine septic acute lung injury (ALI). It is not known if AM iNOS has a direct effect on septic pulmonary microvascular endothelial cell (PMVEC) permeability. We hypothesized that AM iNOS mediates PMVEC permeability in vitro under septic conditions through NO and peroxynitrite. 100,000 confluent PMVEC on cell-culture inserts were co-incubated with iNOS+/+ vs. iNOS-/- AM, in various ratios of AM to PMVEC. PMVEC injury was assessed by trans-PMVEC Evans Blue-labelled albumin flux in the presence or absence of cytomix (equimolar TNF-alpha, IL-1beta and IFN-gamma). Cytomix stimulation dose-dependently increased trans-PMVEC EB-albumin flux, which was exaggerated (1.4+/-0.1% vs. 0.4+/-0.1% in unstimulated PMVEC, p<0.05) in the presence of iNOS+/+, but not iNOS-/-, AM in the upper compartment. Similarly, iNOS+/+, but not iNOS-/-, AM in the lower compartment also enhanced septic trans-PMVEC albumin leak. The mechanism of iNOS-dependent septic PMVEC permeability was pursued through pharmacologic studies with inhibitors of NOS, and scavengers of NO, superoxide, and peroxynitrite, and treatment of PMVEC with the NO donor, DETA-NONOate. Septic iNOS+/+ AM-dependent trans-PMVEC albumin leak was significantly attenuated by pharmacologic iNOS inhibition (L-NAME and 1400W), and scavenging of either NO (oxyhemoglobin), superoxide (PEG-SOD), or peroxynitrite (FeTPPS). Exogenous NO (DETA-NONOate) had no effect on PMVEC permeability. These data are consistent with a direct role of AM iNOS in septic PMVEC barrier dysfunction, which is likely mediated, in part, through peroxynitrite.     

Alveolar macrophage inducible nitric oxide synthase-dependent pulmonary microvascular endothelial cell septic barrier dysfunction

Inducible nitric oxide (NO) synthase (iNOS) from neutrophils and alveolar macrophages (AM) contributes to the pathophysiology of murine septic acute lung injury (ALI). It is not known if AM iNOS has a direct effect on septic pulmonary microvascular endothelial cell (PMVEC) permeability. We hypothesized that AM iNOS mediates PMVEC permeability in vitro under septic conditions through NO and peroxynitrite. 100,000 confluent PMVEC on cell-culture inserts were co-incubated with iNOS+/+ vs. iNOS−/− AM, in various ratios of AM to PMVEC. PMVEC injury was assessed by trans-PMVEC Evans Blue-labelled albumin flux in the presence or absence of cytomix (equimolar TNF-α, IL-1β and IFN-γ). Cytomix stimulation dose-dependently increased trans-PMVEC EB-albumin flux, which was exaggerated (1.4 ± 0.1% vs. 0.4 ± 0.1% in unstimulated PMVEC, p < 0.05) in the presence of iNOS+/+, but not iNOS−/−, AM in the upper compartment. Similarly, iNOS+/+, but not iNOS−/−, AM in the lower compartment also enhanced septic trans-PMVEC albumin leak. The mechanism of iNOS-dependent septic PMVEC permeability was pursued through pharmacologic studies with inhibitors of NOS, and scavengers of NO, superoxide, and peroxynitrite, and treatment of PMVEC with the NO donor, DETA-NONOate. Septic iNOS+/+ AM-dependent trans-PMVEC albumin leak was significantly attenuated by pharmacologic iNOS inhibition (L-NAME and 1400W), and scavenging of either NO (oxyhemoglobin), superoxide (PEG-SOD), or peroxynitrite (FeTPPS). Exogenous NO (DETA-NONOate) had no effect on PMVEC permeability. These data are consistent with a direct role of AM iNOS in septic PMVEC barrier dysfunction, which is likely mediated, in part, through peroxynitrite.