Glycocalyx Research Articles

The Hyaluronan Pericellular Coat and Cold Atmospheric Plasma Treatment of Cells

In different tumors, high amounts of hyaluronan (HA) are correlated with tumor progression. Therefore, new tumor therapy strategies are targeting HA production and degradation. In plasma medicine research, antiproliferative and apoptosis-inducing effects on tumor cells were observed using cold atmospheric plasma (CAP) or plasma-activated media (PAM). Until now, the influence of PAM on the HA pericellular coat has not been the focus of research. PAM was generated by argon-plasma treatment of Dulbecco’s modified Eagle’s Medium via the kINPen®09 plasma jet. The HA expression on PAM-treated HaCaT cells was determined by flow cytometry and confocal laser scanning microscopy. Changes in the adhesion behavior of vital cells in PAM were observed by impedance measurement using the xCELLigence system. We found that PAM treatment impaired the HA pericellular coat of HaCaT cells. The time-dependent adhesion was impressively diminished. However, a disturbed HA coat alone was not the reason for the inhibition of cell adhesion because cells enzymatically treated with HAdase did not lose their adhesion capacity completely. Here, we showed for the first time that the plasma-activated medium (PAM) was able to influence the HA pericellular coat.

Evolutionary algorithms converge towards evolved biological photonic structures

Nature features a plethora of extraordinary photonic architectures that have been optimized through natural evolution in order to more efficiently reflect, absorb or scatter light. While numerical optimization is increasingly and successfully used in photonics, it has yet to replicate any of these complex naturally occurring structures. Using evolutionary algorithms inspired by natural evolution and performing particular optimizations (maximize reflection for a given wavelength, for a broad range of wavelength or maximize the scattering of light), we have retrieved the most stereotypical natural photonic structures. Whether those structures are Bragg mirrors, chirped dielectric mirrors or the gratings on top of Morpho butterfly wings, our results indicate how such regular structures might have spontaneously emerged in nature and to which precise optical or fabrication constraints they respond. Comparing algorithms show that recombination between individuals, inspired by sexual reproduction, confers a clear advantage that can be linked to the fact that photonic structures are fundamentally modular: each part of the structure has a role which can be understood almost independently from the rest. Such an in silico evolution also suggests original and elegant solutions to practical problems, as illustrated by the design of counter-intuitive anti-reflective coatings for solar cells.

Adhesive stem cell coatings for enhanced retention in the heart tissue.

Injection into the heart tissue is a direct route for optimally placing mesenchymal stem cells (MSC) to regulate local inflammation following a heart attack. The retention of MSCs at the injection site is severely limited by the fluid flows that rapidly wash cells away and minimize their capacity to modulate cardiac inflammation. To prevent this loss of MSCs and their function, antibody coatings were designed for the surface of MSCs to enhance their adhesion to the inflamed tissue. MSCs were biotinylated, and biotinylated antibodies against intercellular cell adhesion molecules were conjugated to the cell surface through an intermediate layer of streptavidin. MSC surfaces were modified with ~7,000 biotin/μm2 and ~23 antibodies/μm2. The heart tissue injection of antibody-coated MSCs offered a 3-fold increase of cell retention in an infarcted heart over the injection of uncoated MSCs. We supported the mechanism of adhesion through analysis of MSC adhesion to inflamed endothelial cells and also surfaces of purified adhesion molecules on glass under microfluidic shear flow.

High Fat Diet versus Disturbed Blood Flow Conditions: Implications for Endothelial Glycocalyx Integrity and Pre‐Atherosclerotic Inflammation

IntroductionEndothelial glycocalyx (GCX) shedding plays a role in endothelial dysfunction and increases vessel wall permeability to inflammatory cells contributing to atherogenesis. We sought to determine whether a high fat diet (HFD) or disturbed blood flow conditions, both well‐known atherogenic risk factors, would more detrimentally contribute to pre‐atherosclerotic loss of GCX integrity and vascular inflammation.Materials and MethodsC57BL/6‐background apolipoprotein E knockout (ApoE‐KO) mice were fed either a HFD or chow diet, and/or underwent a ligation of the left carotid artery (LCA), inducing disturbed flow conditions. After one week, mice were sacrificed and LCAs and right carotid arteries (RCAs) were preserved to compare GCX coverage and thickness and inflammatory macrophage accumulation in carotid arterial walls amongst and between cohorts.Results and DiscussionEndothelial GCX damage occurred in LCAs of HFD fed mice when compared to the control. More significant GCX damage occurred in the LCAs of mice exposed to disturbed flow by partial LCA ligation, also compared to control. Flow simulations conducted through SimVascular adequately characterized flow patterns, pressure, and wall shear stresses in non‐ligated vs. ligated mice. They confirmed the notion that the LCA partial ligation surgery does indeed reduce wall shear stress and increase pressure, characteristics of disturbed flow. No difference in macrophage accumulation in carotid arterial walls was observed when comparing the LCAs of control mice to the LCAs of HFD fed mice. However, macrophage infiltration in vessel walls showed a 20‐fold increase in LCAs exposed to disturbed flow following ligation, when compared to control LCAs.ConclusionsThis study was the first to demonstrate that disturbed flow contributes more detrimentally to pre‐atherosclerotic loss of GCX integrity and vascular inflammation. The study concluded that disturbed flow conditions induced by partial ligation, compared to HFD conditions, compromised the integrity of the endothelial GCX to a greater extent. We anticipate more rapid atherogenesis as a consequence of the severity of this flow induced GCX damage.Support or Funding InformationWe are pleased to acknowledge that this work was funded by the National Institute of Health (NIH) K01‐ HL125499 awarded to E Ebong, NIH R21‐DA042583 awarded to S Sridhar, and Air Force Office of Scientific Research (AFOSR) FA2386‐17‐1‐4042 awarded to S Sridhar.

Branch‐Chain Interpenetration of the Mesothelial Glycocalyx with Naturally‐Occurring Biopolymers

The mesothelial surface layer of the lung, heart, bowel, and liver is a dynamic epithelial tissue that regulates a variety of physiologic processes including the healing of visceral organ injuries. A major barrier to addressing these injuries is the slippery and nonadhesive surface of visceral organs, which consists of a carbohydrate‐rich glycocalyx. The glycocalyx is difficult to study, however, because traditional fixation methods destroy this carbohydrate layer. To define the mesothelial glycocalyx, we utilized a high pressure (2100 bar) and rapid freezing technique of the pleura. Examining the glycocalyx with scanning electron microscopy, we demonstrated that the pleural glycocalyx is more than 100‐times thicker than previously realized (more than 12 um) and is notable for its branch‐chain polysaccharide structure. The unique branch‐chain structure of the mesothelial glycocalyx is reminiscent of the plant structural heteropolysaccharide called pectin. Pectin has a branch‐chain polysaccharide structure that appears to mirror the glycocalyx. When 35 mm circular pectin films were applied to porcine mesothelium, the pectin films appeared to fuse with the mesothelial surface. Our hypothesis is that pectin adhesion to mesothelium is via branch‐chain entanglement. Quantitative adhesive strength testing demonstrated that pectin was significantly more adhesive to the mesothelial surface than linear‐chain polysaccharide polymers (p<0.0001). The work of adhesion between pectin and mesothelial glycocalyx was measured to be 80 to 90 N*sec while the work of adhesion measured between linear‐chain polymers and mesothelium was 10 to 20 N*sec. Transillumination stereomicroscopy of the interaction between two branch‐chain polymers demonstrated entanglement of the polymer chains, while stereomicroscopy of the branch‐chain polymer with the linear‐chain polymer or two linear‐chain polymers demonstrated no entanglement. We conclude that the effective binding of pectin to the mesothelial glycocalyx is the result of reciprocal branch‐chain interactions between these structural polysaccharide polymers. We anticipate that the adhesivity between branch‐chain polymers will provide important experimental and therapeutic opportunities in the study of the mesothelium.

Flow-regulated endothelial glycocalyx determines metastatic cancer cell activity.

Cancer metastasis and secondary tumor initiation largely depend on circulating tumor cell (CTC) and vascular endothelial cell (EC) interactions by incompletely understood mechanisms. Endothelial glycocalyx (GCX) dysfunction may play a significant role in this process. GCX structure depends on vascular flow patterns, which are irregular in tumor environments. This work presents evidence that disturbed flow (DF) induces GCX degradation, leading to CTC homing to the endothelium, a first step in secondary tumor formation. A 2-fold greater attachment of CTCs to human ECs was found to occur under DF conditions, compared to uniform flow (UF) conditions. These results corresponded to an approximately 50% decrease in wheat germ agglutinin (WGA)-labeled components of the GCX under DF conditions, vs UF conditions, with undifferentiated levels of CTC-recruiting E-selectin under DF vs UF conditions. Confirming the role of the GCX, neuraminidase induced the degradation of WGA-labeled GCX under UF cell culture conditions or in Balb/C miceand led to an over 2-fold increase in CTC attachment to ECs or Balb/C mouse lungs, respectively, compared to untreated conditions. These experiments confirm that flow-induced GCX degradation can enable metastatic CTC arrest. This work, therefore, provides new insight into pathways of secondary tumor formation.

Porin' through the wax

Microbiology Mycobacterium tuberculosis has a distinctive physiology that allows it to persist in the human body, including a wax-like cell coat that is largely impermeable and can resist attack by host immune response effectors. Wang et al. identified a simple molecule that effectively crosses this cell coat and kills M. tuberculosis cells. Whole-genome sequencing of mutants resistant to this molecule, 3,3- bis -di(methylsulfonyl)propionamide, showed scattered mutations in a protein called PPE51, and these mutants resulted in a spectrum of nutrient-related growth defects. Experiments suggest that PE/PPE family proteins are small molecule–selective channels analogous to outer membrane porins, which allow M. tuberculosis to take up nutrients while maintaining an otherwise impermeable barrier. Science , this issue p. [1147][1] [1]: /lookup/doi/10.1126/science.aav5912

PE/PPE proteins mediate nutrient transport across the outer membrane of Mycobacterium tuberculosis.

Mycobacterium tuberculosis has an unusual outer membrane that lacks canonical porin proteins for the transport of small solutes to the periplasm. We discovered that 3,3-bis-di(methylsulfonyl)propionamide (3bMP1) inhibits the growth of M. tuberculosis, and resistance to this compound is conferred by mutation within a member of the proline-proline-glutamate (PPE) family, PPE51. Deletion of PPE51 rendered M. tuberculosis cells unable to replicate on propionamide, glucose, or glycerol. Growth was restored upon loss of the mycobacterial cell wall component phthiocerol dimycocerosate. Mutants in other proline-glutamate (PE)/PPE clusters, responsive to magnesium and phosphate, also showed a phthiocerol dimycocerosate-dependent growth compromise upon limitation of the corresponding substrate. Phthiocerol dimycocerosate determined the low permeability of the mycobacterial outer membrane, and the PE/PPE proteins apparently act as solute-specific channels.

The Fabrication of Micro-/Nano-Structural Antirefraction Coatings Based on the PDMS Film for Solar Cells Application

A simple and low-cost methodology to fabricate micro/nanostructure anti-refraction coatings (ARCs) of solar cells is introduced. The nanostructure ARCs are generated by O2 Plasma surface modification technique for a flexible polydimethylsiloxane (PDMS) polymeric film, the microstructure ARCs are obtained by soft imprint lithography, and the micro-nano complex structure PDMS ARCs are fabricated by soft imprint lithography technique combined with surface modification technique. The measured results show that these PDMS ARCs can decrease reflection loss of light from the solar cells and increase the solar cells photovoltaic conversion efficiency.

Targeted Intravenous Nanoparticle Delivery: Role of Flow and Endothelial Glycocalyx Integrity.

Therapies for atherosclerotic cardiovascular disease should target early disease stages and specific vascular sites where disease occurs. Endothelial glycocalyx (GCX) degradation compromises endothelial barrier function and increases vascular permeability. This initiates pro-atherosclerotic lipids and inflammatory cells to penetrate vessel walls, and at the same time this can be leveraged for targeted drug delivery. In prior cell culture studies, GCX degradation significantly increased endothelial cell uptake of nanoparticle vehicles that are designed for drug delivery, compared to the effects of intact GCX. The present study assessed if the cell culture findings translate to selective nanoparticle uptake in animal vessels. In mice, the left carotid artery (LCA) was partially ligated to disturb blood flow, which induces GCX degradation, endothelial dysfunction, and atherosclerosis. After ligation, the LCA vessel wall exhibited a loss of continuity of the GCX layer on the intima. 10-nm gold nanospheres (GNS) coated withpolyethylene glycol (PEG) were delivered intravenously. GCX degradation in the ligated LCA correlated to increased GNS infiltration of the ligated LCA wall. This suggests that GCX dysfunction, which coincides with atherosclerosis, can indeed be targeted for enhanced drug delivery, offering a new approach in cardiovascular disease therapy.

Cunea russae n. sp. (Amoebozoa, Dactylopodida), another cryptic species of Cunea Kudryavtsev and Pawlowski, 2015, inhabits a continental brackish-water biotope

The genus Cunea Kudryavtsev and Pawlowski, 2015 (Amoebozoa, Dactylopodida) was initially described from the oceanic benthos: C. profundata, from over 5 km depth in the Atlantic Ocean, and C. thuwala from the Red Sea benthos at ca. 60 m depth. Both species are identical to each other in morphology (including cell coat ultrastructure), but differ significantly in the gene sequence data, including barcoding loci of small subunit ribosomal RNA and cytochrome oxidase subunit 1 gene, as well as actin. This paper describes the third species of Cunea, C. russae n. sp. isolated from a brackish water habitat without a direct connection to the ocean, a small spring of brackish water (19‰) emerging from a 246 m deep hole in the earth. This species is morphologically identical to the previous two amoebae, but differs from them significantly in the gene sequence data and ecological preferences. In particular, this species has the broadest salinity tolerance range, being able to reproduce well already at 2.5‰. It is also capable of resisting cold temperatures, like C. profundata. The data obtained suggest that the genus Cunea may comprise a significant taxonomic diversity represented by morphologically identical, but quickly diverging species with significant ecological plasticity.

The role of Sigma-1 receptor agonist in hepatic ischemiareperfusion injury.

Sigma-1 receptor (Sig-1R) is a ligand-operated protein that modulates activity of various proteins. It is expressed within the endoplasmic reticulum membranes of multiple organs. We examined the role of Sig-1R in hepatic ischemia-reperfusion injury (IRI). We studied IRI indicators in Sig-1R-/- mice and compared them with wild-type controls. In addition, we assessed the influence of Sig-1R agonist, fluvoxamine, on IRI in both types of animals. We found that Sig-1R-/- mice exhibited significantly decreased liver damage after hepatic IRI as compared to wild-type mice. This effect was manifested by decreased serum levels of alanine aminotransferase (AST), aspartate aminotransferase (ALT), myeloperoxidase (MPO), and supernatant level of lactate dehydrogenase (LDH), decreased endothelial glycocalyx shedding indexed by decreased serum levels of heparan sulfate and syndecan-1, slightly improved liver histology and reduced metalloproteinase-9 expression. Furthermore, in comparison to Sig-1R-/- mice, fluvoxamine significantly increased serum levels of AST, ALT, MPO, and LDH in wildtype animals in a dose-dependent manner at 6 h after IRI. Our findings demonstrate that the absence of Sig-1R provides a protective effect during hepatic IRI. Sig-1R-mediated signaling pathways may play distinct roles in IRI in different organs. The dynamic interaction between Sig-1R and other signaling molecules in different organs needs to be examined further.

Fabrication of transparent, durable and self-cleaning superhydrophobic coatings for solar cells

A superhydrophobic coating with high transparency and ultrahigh adhesive force is prepared for application on the glass covers of solar cells, which also exhibits excellent thermal stability and strong acid–base corrosion resistance.

HALL EFFECT ON HEAT TRANSPORT OF MAGNETIZED Cu-ENGINE OIL OVER A ROTATING SLIPPING DISK WITH CONVECTIVE HEATING IN A POROUS SPACE

The purpose of this article is mainly to explore the effect of Hall currents and slip conditions on the unsteady magneto-hydrodynamic flow and heat transfer of electrically conducting copper-engine oil (non-Newtonian Casson nanoliquid) over a rotating porous disk with hydrodynamic slip and convective heating in a porous space of Darcy resistance. The Casson model is used to describe the rheology of the non-Newtonian fluid in this study. Non-Newtonian Casson nano-liquid is engineered by dispersing copper (Cu) nanoparticles in engine oil (EO). The disk is subjected to convective boundary conditions and the effect of viscous and Joule dissipations are taken into account in this analysis. The present physical model is analyzed under the influence of strong Lorentz force to investigate the effect of the Hall current on the flow behavior. An exact solution is established for the governing momentum equation by employing the Laplace transform method. The MATLAB built-in function pedpe is used to numerically solve the nonlinear energy equation. The impact of significant parameters on the flow fields is analyzed by plotting graphs and tables and discussed in detail. The computations demonstrate that the Hall current generates substantial enhancement in the velocity component profiles and significantly elevates the temperature in the flow domain. The disk temperature is substantially reduced in the presence of suction. A comparative study of EO Casson and Cu-EO Casson nanoliquids is presented and interpreted. The inferences obtained from our investigation may effectively be used in disk cleaners, rotor-systemic manufacturing of magnetic nano-polymer coatings, fuel cell coatings, print ink, self-cleaning ceramics, cosmetics, etc.

Lipopolysaccharide Stress Induces Cryptic Exon Splice Variants of the Human Glucocorticoid Receptor.

Glucocorticoids are widely used in the treatment of numerous inflammatory conditions, including sepsis. Unfortunately, patient response to glucocorticoid therapy can be inconsistent. Variations in the human glucocorticoid receptor (hGR) may contribute to the differential patient response. We screened for hGR variants in the buffy coats of burn patients and peripheral blood mononuclear cells (PBMCs) treated with lipopolysaccharide. Three novel splice variants containing cryptic exons were upregulated in the PBMCs after lipopolysaccharide exposure at 3 and 13 h with the greatest observed expression at 3 h. Luciferase assays revealed that two of the isoforms had no significant activity in comparison with the reference hGR when stimulated with hydrocortisone. The third isoform had an augmented response that was greater than the reference hGR at a high cortisol dose. This shows that PBMCs are able to produce variant hGR isoforms in response to stress. Furthermore, lipopolysaccharide stress appears to induce these hGR variants, potentially by influencing mRNA splicing. In the future, identifying hGR expression profiles may be a key component in individually tailoring a patient's treatment to sepsis and injury.

Electrospun Nitrogen-doped TiO2 Nanofibrous Thin Film for Photovoltaic Application

This research study aims at fabrication of fine size nitrogen doped TiO2 nanofiber using electrospinning method and evaluation of the performance of TiO2 in a photovoltaic cell under visible light irradiation. Undoped and N doped TiO2 nanoparticles were synthesized by sol gel method where titanium isopropoxide was used as the source of TiO2 and ammonium nitrate was used as the source of N dopant. TiO2/PVA composite material was prepared by stabilizing TiO2 particle in to 10 wt % of PVA (aq) solution in order to prepare thin film that can be coated on photovoltaic (PV) cells. Coating of solid thin film PV cells by TiO2/PVA nanofibers was conducted using electrospinning and doctor blade method. In both systems, doping the TiO2 with nitrogen improved its optical properties which it successfully lowered the band gap energy from 3.14 to 2.76 eV and shifted its optical response to the visible light region. The presence of O-H stretching vibration, O-H bending and vibration of the N-Ti bond contributed to an increased performance of the PV cells. The electrospun N-doped TiO2 produced better power output than doctor blade method coated PV cells with power of 0.040 and 0.026 mW, respectively.

Endothelial barrier reinforcement relies on flow-regulated glycocalyx, a potential therapeutic target.

The onset of many disease processes depends on the function of the endothelial cell (EC) glycocalyx (GCX) which acts as a flow-dependent barrier to cellular infiltration and molecular transport across the blood vessel wall. This review aims to examine these processes with the potential end goal of implementing GCX repair to restore EC barrier function and slow the progression of disease. Cell and mouse studies were employed to examine the state of EC GCX in healthy versus disruptive flow conditions. Correlations of observations of the GCX with a number of EC functions were sought with an emphasis on studies of trans-endothelial barrier integrity against vessel wall infiltration of cells and molecules from the circulation. To demonstrate the importance of GCX as a regulator of trans-endothelial infiltration, assays were performed using ECs with an intact GCX and compared to assays of ECs with an experimentally degraded GCX. Studies were also conducted of ECs in which a degraded GCX was repaired. In healthy flow conditions, the EC GCX was found to be thick and substantially covered the endothelial surface. GCX expression dropped significantly in complex flow conditions and coincided with a disease-like cellular and molecular accumulation in the endothelium or within the blood vessel wall. Therapeutic repair of the GCX abolished this accumulation. Regenerating the degraded GCX reverses EC barrier dysfunction and may attenuate the progression of vascular disease.

Microfiber Optic Arrays as Top Coatings for Front-Contact Solar Cells toward Mitigation of Shading Loss

Microfiber optic array structures are fabricated and employed as an optical structure overlaying a front-contact silicon solar cell. The arrays are synthesized through light-induced self-writing in a photo-crosslinking acrylate resin, which produces periodically spaced, high-aspect-ratio, and vertically aligned tapered microfibers deposited on a transparent substrate. The structure is then positioned over and sealed onto the solar cell surface. Their fiber optic properties enable collection of non-normal incident light, allowing the structure to mitigate shading loss through the redirection of incident light away from contacts and toward the solar cell. Angle-averaged external quantum efficiency increases nominally by 1.61%, resulting in increases in short-circuit current density up to 1.13 mA/cm2. This work demonstrates a new approach to enhance light collection and conversion using a scalable, straightforward, light-based additive manufacturing process.

Glycocalyx breakdown is increased in African children with cerebral and uncomplicated falciparum malaria.

Cerebral malaria (CM) from Plasmodium falciparum infection is associated with endothelial dysfunction and parasite sequestration. The glycocalyx (GCX), a carbohydrate-rich layer lining the endothelium, is crucial in vascular homeostasis. To evaluate the role of its loss in the pathogenesis of pediatric CM, we measured GCX degradation in Tanzanian children with World Health Organization-defined CM (n = 55), uncomplicated malaria (UM; n = 20), and healthy controls (HCs; n = 25). Urine GCX breakdown products [glycosaminoglycans (GAGs)] were quantified using dimethylmethylene blue (DMMB) and liquid chromatography-tandem mass spectrometry assays. DMMB-GAG and mass spectrometry (MS)-GAG (g/mol creatinine) were increased in CM and UM compared with HCs (P < 0.001), with no differences in DMMB-GAG and MS-GAG between CM and UM children or between those with and without a fatal outcome. In CM survivors, urinary GCX DMMB-GAG normalized by d 3. After adjusting for disease severity, DMMB-GAG was significantly associated with parasitemia [partial correlation coefficient (Pcorr) = 0.34; P = 0.01] and plasma TNF (Pcorr = 0.26; P = 0.04) and inversely with plasma and urine NO oxidation products [Pcorr = -0.31 (P = 0.01) and Pcorr = -0.26 (P = 0.03), respectively]. GCX breakdown is increased in children with falciparum malaria, with similar elevations in CM and UM. Endothelial GCX degradation may impair endothelial NO production, exacerbate adhesion-molecule expression, exposure, and parasite sequestration, and contribute to malaria pathogenesis.-Yeo, T. W., Bush, P. A., Chen, Y., Young, S. P., Zhang, H., Millington, D. S., Granger, D. L., Mwaikambo, E. D., Anstey, N. M., Weinberg, J. B. Glycocalyx breakdown is increased in African children with cerebral and uncomplicated falciparum malaria.

The protective effect of hydroxyethyl starch solution on the glycocalyx layer in an acute hemorrhage mouse model

PurposeFluid therapy focused on glycocalyx (GCX) protection in hemorrhagic shock is a current focus of research. Hydroxyethyl starch (HES) solution is commonly used for fluid resuscitation; however, its effects on the GCX remain unclear. The primary aim of this study was to explore the protective effect of HES130 in maintaining GCX thickness and reducing plasma syndecan-1 expression.MethodsAn acute hemorrhage murine model with the dorsal skin chambers was used to measure GCX thickness and to evaluate vascular permeability. Groups of mice were treated with normal saline (NS), albumin (NS-A), HES130 (NS-V), or no exsanguination or infusion (C). We measured syndecan-1 plasma concentrations, performed blood gas analysis, and analyzed the 7-day cumulative mortality.ResultsGCX thickness in NS mice was significantly reduced compared to that in group C, but no other groups showed a difference compared to group C. The plasma concentration of syndecan-1 was significantly higher in NS mice than in group C. There were no significant differences in the fluorescence intensity of dextran in the interstitial space. HES70 leakage was suppressed in NS-V mice compared to those in other groups. HES70 was localized to the inner vessel wall in C, NS, and NS-A mice, but not in group NS-V. Blood gas analysis indicated that pH and lactate showed the greatest improvements in NS-V mice. The 7-day cumulative mortality rate was the highest in group NS.ConclusionResuscitation with HES130 protected the GCX and suppressed vascular permeability of HES70 during early stages of acute massive hemorrhage.