Role Of Blood Research Articles

Blood and imaging biomarkers in the long-term follow-up of bicuspid aortic valve patients

Abstract Background Bicuspid aortic valve (BAV) is a common congenital heart defect. Patients with BAV are at risk for long-term complications such as arrhythmias and reinterventions. Purpose This study aimed to investigate (1) sex differences in blood and imaging biomarkers and (2) the role of blood and echocardiographic biomarkers in the long-term risk prediction of patients with BAV. Methods In this multicenter prospective observational study, all included adult BAV patients underwent venous blood sampling and transthoracic echocardiography including speckle tracking at the time of inclusion. Analyzed blood biomarkers were: RDW, creatinine, CRP, TroponinT, NT-proBNP and TGF-beta. Sex differences were analyzed at baseline. Associations between blood & echocardiographic biomarkers and arrhythmia-free & intervention-free survival were determined by Cox proportional hazards models. Results A total of 182 patients were included (median age 34 (Interquartile range: 23-46) years, 55.5% male). Significant differences were found in blood biomarkers at baseline between men and women. CRP, NT-proBNP and RDW were higher in women while creatinine, TroponinT and TGF-beta were higher among men (figure 1). After a median follow-up time up of 6.9 (IQR: 6.5- 9.9) years, arrhythmia-free and event-free survival rates were respectively 81.0% and 73.1%. NT-proBNP was the only blood biomarker independently associated with both arrhythmia free and intervention free survival. On echocardiography an increase in LA size, LVEDD, LV mass index and E/E’ratio were associated with arrhythmia free survival. Intervention free survival was associated with increased LA size, LV mass index, aortic valve peak velocity and the presence of moderate or severe aortic regurgitation. No associations were found with ejection fraction and global longitudinal strain (figure 2). Conclusion Significant sex-related differences were observed in blood biomarkers between men and women with BAV at baseline. NT-proBNP was the only blood biomarker associated with arrhythmia-and intervention-free survival. The strongest associations of echocardiographic markers were those indicating diastolic function and LV mass index.Sex differences in blood biomarkersForest plot

Protective effects of KLF4 on blood–brain barrier and oxidative stress after cerebral ischemia–reperfusion in rats through the Nrf2/Trx1 pathway

PurposeTo investigate the role of KLF4 in CI/R injury and whether Nrf2/Trx1 axis acted as a downstream pathway of KLF4 to exert the protective role in blood–brain barrier destruction after CI/R. MethodsThe tMCAO rat model in vivo was constructed and received the intracerebroventricular injection of 5 μg/kg and 10 μg/kg rhKLF4 before operation. TTC, brain water content, neurological function, ELISA, behavioral tests, HE, TUNEL, and qRT-PCR were performed to detect the protective role of KLF4 on CIR. Double-fluorescence staining and western blot were performed to determine the localization and spatiotemporal expression in brain tissues. Furthermore, we also analyzed the effect of KLF4 on the blood–brain barrier (BBB) and related mechanisms in vivo and in vitro. Nrf2 inhibitor tretinoin was applied, which was intraperitoneally injected into CIR rat. Evans blue staining was conducted. In vitro OGD/R models of bEnd.3 cells were also established, and received KLF4 overexpressed transfection and 12.5 µM tretinoin incubation. The permeability of bEnd.3 cells was evaluated by TEER and FITC-dextran leakage. BBB-related factors and oxidative stress were also analyzed, respectively. The tubular ability of KLF4 on OGD/R bEnd3 cells was also evaluated. ResultsIn vivo study confirmed that KLF4 was expressed in astrocyte, and its content increased with time. KLF4 protected against brain injury caused by cerebral ischemia–reperfusion, reduced cerebral infarction area and oxidative stress levels, and promoted the recovery of behavioral ability in rats. Simultaneously, mechanism experiments confirmed that the repair effect of KLF4 on cerebral ischemia–reperfusion injury was closely related to the Nrf2/Trx1 pathway. KLF4 exerted the neuroprotective effect through upregulating Nrf2/Trx1 pathway. Consistent with in vivo animal study, in vitro study also confirmed the effect of KLF4 on the permeability of bEnd.3 cells after OGD/R injury through Nrf2/Trx1 pathway. ConclusionCollectively, KLF4 played neuroprotective role in CIR induced MCAO and OGD/R, and the beneficial effects of KLF4 was partly linked to Nrf2/Trx1 pathway.

Changes in CSF sPDGFRβ level and their association with blood–brain barrier breakdown in Alzheimer’s disease with or without small cerebrovascular lesions

BackgroundCSF-soluble platelet-derived growth factor receptor beta (sPDGFRβ) is closely associated with pericyte damage. However, the changes in CSF sPDGFRβ levels and their role in blood–brain barrier (BBB) leakage at different stages of Alzheimer’s disease (AD), with or without cerebral small vessel disease (CSVD) burden, remain unclear.MethodsA total of 158 individuals from the China Aging and Neurodegenerative Disorder Initiative cohort were selected, including 27, 48, and 83 individuals with a clinical dementia rating (CDR) score of 0, 0.5, and 1–2, respectively. CSF total tau, phosphorylated tau181 (p-tau181), Aβ40, and Aβ42 were measured using the Simoa assay. Albumin and CSF sPDGFRβ were measured by commercial assay kits. CSVD burden was assessed by magnetic resonance imaging.ResultsCSF sPDGFRβ was the highest level in the CDR 0.5 group. CSF sPDGFRβ was significantly correlated with the CSF/serum albumin ratio (Q-alb) in the CDR 0–0.5 group (β = 0.314, p = 0.008) but not in the CDR 1–2 group (β = − 0.117, p = 0.317). In the CDR 0–0.5 group, CSF sPDGFRβ exhibited a significant mediating effect between Aβ42/Aβ40 levels and Q-alb (p = 0.038). Q-alb, rather than CSF sPDGFRβ, showed a significant difference between individuals with or without CSVD burden. Furthermore, in the CDR 0.5 group, CSF sPDGFRβ was higher in subjects with progressive mild cognitive impairment than in those with stable mild cognitive impairment subjects (p < 0.001). Meanwhile, CSF sPDGFRβ was significantly associated with yearly changes in MMSE scores in the CDR 0.5 group (β = − 0.400, p = 0.020) and CDR 0.5 (A+) subgroup (β = − 0.542, p = 0.019).ConclusionsWe provide evidence that increased CSF sPDGFRβ is associated with BBB leakage in the early cognitive impairment stage of AD, which may contribute to cognitive impairment in AD progression.

An Unseen Invasion: Vampirism as Contagion in Bram Stoker’s Dracula

As creatures that come back from the dead to haunt and hunt the living, vampires represent the fear of an uncontrollable, mysterious, foreign threat. Bram Stoker’s Gothic novel Dracula (1897) presents vampirism not only as a curse that resurrects the dead human body as an undead monster, but also as a metaphor for a contagious disease. Transgressing boundaries between life and death, human and animal, past and present, the vampire breaches Gothic purity by contaminating the individual. Arriving as an elusive, unseen disease to the English soil and subject, Dracula exposes his victims to his contaminated blood in his quest to invade Victorian England. Consuming the body by feeding on the life giving blood, contaminating the victim by causing illness and death, and transforming the human into a vampire, Dracula embodies a contagion upon Victorian society. Accordingly, the aim of this paper to explore the links between the Victorian vampire and infection, looking closely at the role of blood in the creation of the monstrous threat. To that end, this paper aims to analyse how vampirism as a disease and Dracula as an agent of contagion reflect Victorian anxieties regarding the individual identity, social changes, fears of degeneration, and invasion of a foreign Other bringing destruction within.

Surgical Management of Synucleinopathies.

Synucleinopathies represent a diverse set of pathologies with significant morbidity and mortality. In this review, we highlight the surgical management of three synucleinopathies: Parkinson’s disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). After examining underlying molecular mechanisms and the medical management of these diseases, we explore the role of deep brain stimulation (DBS) in the treatment of synuclein pathophysiology. Further, we examine the utility of focused ultrasound (FUS) in the treatment of synucleinopathies such as PD, including its role in blood–brain barrier (BBB) opening for the delivery of novel drug therapeutics and gene therapy vectors. We also discuss other recent advances in the surgical management of MSA and DLB. Together, we give a diverse overview of current techniques in the neurosurgical management of these pathologies.

Role of NADPH Oxidases in Blood-Brain Barrier Disruption and Ischemic Stroke.

NADPH oxidases (Nox) are one of the main sources of reactive oxygen species (ROS) in the central nervous system (CNS). While these enzymes have been shown to be involved in physiological regulation of cerebral vascular tone, excessive ROS produced by Nox1-5 play a critical role in blood–brain barrier (BBB) dysfunction in numerous neuropathologies. Nox-derived ROS have been implicated in mediating matrix metalloprotease (MMP) activation, downregulation of junctional complexes between adjacent brain endothelial cells and brain endothelial cell apoptosis, leading to brain microvascular endothelial barrier dysfunction and consequently, increases in BBB permeability. In this review, we will highlight recent findings on the role played by these enzymes in BBB disruption induced by ischemic stroke.

Targeting the blood–brain barrier disruption in hypertension by ALK5/TGF-Β type I receptor inhibitor SB-431542 and dynamin inhibitor dynasore

IntroductionIn this study, we aimed to target two molecules, transforming growth factor-beta (TGF-β) and dynamin to explore their roles in blood–brain barrier (BBB) disruption in hypertension. MethodsFor this purpose, angiotensin (ANG) II-induced hypertensive mice were treated with SB-431542, an inhibitor of the ALK5/TGF-β type I receptor, and dynasore, an inhibitor of dynamin. Albumin-Alexa fluor 594 was used to assess BBB permeability. The alterations in the expression of claudin-5, caveolin (Cav)-1, glucose transporter (Glut)-1, and SMAD4 in the cerebral cortex and the hippocampus were evaluated by quantification of immunofluorescence staining intensity. ResultsANG II infusion increased BBB permeability to albumin-Alexa fluor 594 which was reduced by SB-431542 (P < 0.01), but not by dynasore. In hypertensive animals treated with dynasore, claudin-5 immunofluorescence intensity increased in the cerebral cortex and hippocampus while it decreased in the cerebral cortex of SB-431542 treated hypertensive mice (P < 0.01). Both dynasore and SB-431542 prevented the increased Cav-1 immunofluorescence intensity in the cerebral cortex and hippocampus of hypertensive animals (P < 0.01). SB-431542 and dynasore decreased Glut-1 immunofluorescence intensity in the cerebral cortex and hippocampus of mice receiving ANG II (P < 0.01). SB-431542 increased SMAD4 immunofluorescence intensity in the cerebral cortex of hypertensive animals, while in the hippocampus a significant decrease was noted by both SB-431542 and dynasore (P < 0.01). ConclusionOur data suggest that inhibition of the TGFβ type I receptor prevents BBB disruption under hypertensive conditions. These results emphasize the therapeutic potential of targeting TGFβ signaling as a novel treatment modality to protect the brain of hypertensive patients.

European Stroke Organisation (ESO) guideline on screening for subclinical atrial fibrillation after stroke or transient ischaemic attack of undetermined origin.

We aimed to provide practical recommendations for the screening of subclinical atrial fibrillation (AF) in patients with ischaemic stroke or transient ischaemic attack (TIA) of undetermined origin. These guidelines are based on the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) methodology. Five relevant Population, Intervention, Comparator, Outcome questions were defined by a multidisciplinary module working group (MWG). Longer duration of cardiac rhythm monitoring increases the detection of subclinical AF, but the optimal monitoring length is yet to be defined. We advise longer monitoring to increase the rate of anticoagulation, but whether longer monitoring improves clinical outcomes needs to be addressed. AF detection does not differ from in- or out-patient ECG-monitoring with similar monitoring duration, so we consider it reasonable to initiate in-hospital monitoring as soon as possible and continue with outpatient monitoring for more than 48 h. Although insertable loop recorders (ILR) increase AF detection based on their longer monitoring duration, comparison with non-implantable ECG devices for similar monitoring time is lacking. We suggest the use of implantable devices, if feasible, for AF detection instead of non-implantable devices to increase the detection of subclinical AF. There is weak evidence of a useful role for blood, ECG and brain imaging biomarkers for the identification of patients at high risk of AF. In patients with patent foramen ovale, we found insufficient evidence from RCT, but prolonged cardiac monitoring in patients >55 years is advisable for subclinical AF detection. To conclude, in adult patients with ischaemic stroke or TIA of undetermined origin, we recommend longer duration of cardiac rhythm monitoring of more than 48 h and if feasible with IRL to increase the detection of subclinical AF.

Claudin-17 Deficiency in Mice Results in Kidney Injury Due to Electrolyte Imbalance and Oxidative Stress.

The multi-gene claudin (CLDN) family of tight junction proteins have isoform-specific roles in blood–tissue barrier regulation. CLDN17, a putative anion pore-forming CLDN based on its structural characterization, is assumed to regulate anion balance across the blood-tissue barriers. However, our knowledge about CLDN17 in physiology and pathology is limited. The current study investigated how Cldn17 deficiency in mice affects blood electrolytes and kidney structure. Cldn17−/− mice revealed no breeding abnormalities, but the newborn pups exhibited delayed growth. Adult Cldn17−/− mice displayed electrolyte imbalance, oxidative stress, and injury to the kidneys. Ingenuity pathway analysis followed by RNA-sequencing revealed hyperactivation of signaling pathways and downregulation of SOD1 expression in kidneys associated with inflammation and reactive oxygen species generation, demonstrating the importance of Cldn17 in the maintenance of electrolytes and reactive oxygen species across the blood-tissue barrier.

RNA-Seq Profiling of Neutrophil-Derived Microvesicles in Alzheimer's Disease Patients Identifies a miRNA Signature That May Impact Blood-Brain Barrier Integrity.

(1) Background: Systemic infection is associated with increased neuroinflammation and accelerated cognitive decline in AD patients. Activated neutrophils produce neutrophil-derived microvesicles (NMV), which are internalised by human brain microvascular endothelial cells and increase their permeability in vitro, suggesting that NMV play a role in blood–brain barrier (BBB) integrity during infection. The current study investigated whether microRNA content of NMV from AD patients is significantly different compared to healthy controls and could impact cerebrovascular integrity. (2) Methods: Neutrophils isolated from peripheral blood samples of five AD and five healthy control donors without systemic infection were stimulated to produce NMV. MicroRNAs isolated from NMV were analysed by RNA-Seq, and online bioinformatic tools were used to identify significantly differentially expressed microRNAs in the NMV. Target and pathway analyses were performed to predict the impact of the candidate microRNAs on vascular integrity. (3) Results: There was no significant difference in either the number of neutrophils (p = 0.309) or the number of NMV (p = 0.3434) isolated from AD donors compared to control. However, 158 microRNAs were significantly dysregulated in AD NMV compared to controls, some of which were associated with BBB dysfunction, including miR-210, miR-20b-5p and miR-126-5p. Pathway analysis revealed numerous significantly affected pathways involved in regulating vascular integrity, including the TGFβ and PDGFB pathways, as well as Hippo, IL-2 and DNA damage signalling. (4) Conclusions: NMV from AD patients contain miRNAs that may alter the integrity of the BBB and represent a novel neutrophil-mediated mechanism for BBB dysfunction in AD and the accelerated cognitive decline seen as a result of a systemic infection.

The arginine deaminase system plays distinct roles in Borrelia burgdorferi and Borrelia hermsii.

Borrelia species are amino acid auxotrophs that utilize di- and tri- peptides obtained through their oligopeptide transport system to supply amino acids for replicative growth during their enzootic cycles. However, Borrelia species from both the Lyme disease (LD) and relapsing fever (RF) groups harbor an amino acid transport and catabolism system, the Arginine Deiminase System (ADI), that could potentially augment intracellular L-arginine required for growth. RF spirochetes contain a “complete”, four gene ADI (arcA, B, D, and C) while LD spirochetes harbor arcA, B, and sometimes D but lack arcC (encoding carbamate kinase). In this study, we evaluated the role of the ADI system in bacterial survival and virulence and discovered important differences in RF and LD ADIs. Both in vitro and in a murine model of infection, B. hermsii cells significantly reduced extracellular L-arginine levels and that reduction was dependent on arginine deiminase expression. Conversely, B. burgdorferi did not reduce the concentration of L-arginine during in vitro growth experiments nor during infection of the mammalian host, suggesting a fundamental difference in the ability to directly utilize L-arginine compared to B. hermsii. Further experiments using a panel of mutants generated in both B. burgdorferi and B. hermsii, identified important differences in growth characteristics and ADI transcription and protein expression. We also found that the ADI system plays a key role in blood and spleen colonization in RF spirochetes. In this study we have identified divergent metabolic strategies in two closely related human pathogens, that ultimately impacts the host-pathogen interface during infection.

Opposing Roles of Vascular Endothelial Growth Factor C in Metastatic Dissemination and Resistance to Radio/Chemotherapy: Discussion of Mechanisms and Therapeutic Strategies.

Many cancers can be cured by combining surgery with healthy margins, radiation therapy and chemotherapies. However, when the pathology becomes metastatic, cancers can be incurable. The best situation involves "chronicization" of the pathology even for several years. However, most of the time, patients die within a few months. To disseminate throughout the body, cancer cells must enter the vascular network and seed in another organ. However, during the initiation of cancer processes, the tumor is avascular. Later, the production of angiogenic factors causes tumor neovascularization and subsequent growth and spread, and the presence of blood and/or lymphatic vessels is associated with high grade tumors. Moreover, during tumor development, cancer cells enter lymphatic vessels and disseminate via the lymphatic network. Hence, blood and lymphatic vessels are considered as main routes of metastatic dissemination and cancer aggressiveness. Therefore, anti-angiogenic drugs entered in the therapeutic arsenal from 2004. Despite undeniable effects however, they are far from curative and only prolong survival by a few months.Recently, the concepts of angio/lymphangiogenesis were revisited by analyzing the role of blood and lymphatic vessels at the initiation steps of tumor development. During this period, cancer cells enter lymphatic vessels and activate immune cells within lymph nodes to initiate an antitumor immune response. Moreover, the presence of blood vessels at the proximity of the initial nodule allows immune cells to reach the tumor and eliminate cancer cells. Therefore, blood and lymphatic networks have a beneficial role during a defined time window. Considering only their detrimental effects is a concern. Hence, administration of anti-angio/lymphangiogenic therapies should be revisited to avoid the destruction of networks involved in antitumor immune response. This review mainly focuses on one of the main drivers of lymphangiogenesis, the VEGFC and its beneficial and pejorative roles according to the grade of aggressive tumors.

The role of blood lnc-ZFAS1 in acute ischemic stroke: correlation with neurological impairment, inflammation, and survival profiles.

BackgroundLong non‐coding RNA zinc finger antisense 1 (lnc‐ZFAS1) has been reported to inhibit neuronal damage in acute ischemic stroke (AIS). However, the role of lnc‐ZFAS1 in AIS patients remains unclear. Therefore, we assessed the relationship of lnc‐ZFAS1 with neurological impairment, inflammation, and prognosis in AIS patients.MethodsTotally, 241 AIS patients and 120 controls were enrolled. lnc‐ZFAS1 in peripheral blood mononuclear cells was evaluated using reverse transcription‐quantitative polymerase chain reaction. Besides, a 3‐year follow‐up was conducted to assess recurrence‐free survival (RFS) and overall survival (OS) in AIS patients.Resultslnc‐ZFAS1 was reduced in AIS patients compared to that in controls (Z = −10.693, p < 0.001). In AIS patients, lnc‐ZFAS1 was negatively correlated with National Institutes of Health Stroke Scale score (r s = −0.335, p < 0.001), C‐reactive protein (r s = −0.284, p < 0.001), tumor necrosis factor‐alpha (r s = −0.293, p < 0.001), interleukin‐1β (r s = −0.149, p = 0.021), and interleukin‐6 (r s = −0.161, p = 0.012), but not underlying diseases (all p > 0.05). Besides, lnc‐ZFAS1 was divided into high and low levels based on the median expression in AIS patients. Indeed, high lnc‐ZFAS1 predicted better RFS (χ 2 = 6.222, p = 0.013); the 1‐year, 2‐year, and 3‐year RFS rates were 94.2%, 88.3%, and 85.5%, respectively, in patients with high lnc‐ZFAS1, then 87.5%, 79.2%, and 71.6%, respectively, in those with low lnc‐ZFAS1. However, lnc‐ZFAS1 was not correlated with OS (χ 2 = 1.404, p = 0.236); the 1‐year, 2‐year, and 3‐year OS rates were 98.3%, 95.8%, and 94.0%, respectively, in patients with high lnc‐ZFAS1, then 96.7%, 93.9%, and 89.6%, respectively, in those with low lnc‐ZFAS1.ConclusionLower lnc‐ZFAS1 expression is connected with increased neurological impairment and inflammation as well as worse RFS in AIS patients.

De novo Lipogenesis in Astrocytes Promotes the Repair of Blood–Brain Barrier after Transient Cerebral Ischemia Through Interleukin-33

Astrocytes experience significant metabolic shifts in the “sensitive period” of neurological function recovery following cerebral ischemia. However, the changes in astrocyte lipid metabolism and their implications for neurological recovery remain unknown. In the present study, we employed a mouse middle cerebral artery occlusion model to investigate the changes in de novo lipogenesis and interleukin-33 (IL-33) production in astrocytes and elucidate their role in blood–brain barrier (BBB) repair in the subacute phase of cerebral ischemia. Neurological behavior evaluation was used to assess functional changes in mice. Pharmacological inhibition and astrocyte-specific downregulation of fatty acid synthase (FASN) were used to evaluate the role of de novo lipogenesis in brain injury. Intracerebroventricular administration of recombinant IL-33 was performed to study the contribution of IL-33 to BBB disruption. Extravasation of Evans blue dye, dextran and IgG were used to assess BBB integrity. Western blotting of tight junction proteins ZO-1, Occludin, and Claudin-5 were performed at defined time points to evaluate changes in BBB. It was found that de novo lipogenesis was activated, and IL-33 production increased in astrocytes at the subacute stage of cerebral ischemia injury. Inhibition of lipogenesis in astrocytes decreased IL-33 production in the peri-infarct area, deteriorated BBB damage and interfered with neurological recovery. In addition, supplementation of IL-33 alleviated BBB destruction and improved neurological recovery worsened by lipogenesis inhibition. These findings indicate that astrocyte lipogenesis increases the production of IL-33 in the peri-infarct area, which promotes BBB repair in the subacute phase of cerebral ischemia injury and improves long-term functional recovery.

The Role of Blood in Regulating Body Heat in Bovines

The Role of Blood in Regulating Body Heat in Bovines

Role of Intermediate Filaments in Blood-Brain Barrier in Health and Disease.

The blood–brain barrier (BBB) is a highly selective cellular monolayer unique to the microvasculature of the central nervous system (CNS), and it mediates the communication of the CNS with the rest of the body by regulating the passage of molecules into the CNS microenvironment. Limitation of passage of substances through the BBB is mainly due to tight junctions (TJ) and adherens junctions (AJ) between brain microvascular endothelial cells. The importance of actin filaments and microtubules in establishing and maintaining TJs and AJs has been indicated; however, recent studies have shown that intermediate filaments are also important in the formation and function of cell–cell junctions. The most common intermediate filament protein in endothelial cells is vimentin. Vimentin plays a role in blood–brain barrier permeability in both cell–cell and cell–matrix interactions by affecting the actin and microtubule reorganization and by binding directly to VE-cadherin or integrin proteins. The BBB permeability increases due to the formation of stress fibers and the disruption of VE–cadherin interactions between two neighboring cells in various diseases, disrupting the fiber network of intermediate filament vimentin in different ways. Intermediate filaments may be long ignored key targets in regulation of BBB permeability in health and disease.

Establishment of a Mass-Spectrometry-Based Method for the Identification of the In Vivo Whole Blood and Plasma ADP-Ribosylomes.

Blood and plasma proteins are heavily investigated as biomarkers for different diseases. However, the post-translational modification states of these proteins are rarely analyzed since blood contains many enzymes that rapidly remove these modifications after sampling. In contrast to the well-described role of protein ADP-ribosylation in cells and organs, its role in blood remains mostly uncharacterized. Here, we discovered that plasma phosphodiesterases and/or ADP-ribosylhydrolases rapidly demodify in vitro ADP-ribosylated proteins. Thus, to identify the in vivo whole blood and plasma ADP-ribosylomes, we established a mass-spectrometry-based workflow that was applied to blood samples collected from LPS-treated pigs (Sus scrofa domesticus), which serves as a model for human systemic inflammatory response syndrome. These analyses identified 60 ADP-ribosylated proteins, 17 of which were ADP-ribosylated plasma proteins. This new protocol provides an important step forward for the rapidly developing field of ADP-ribosylation and defines the blood and plasma ADP-ribosylomes under both healthy and disease conditions.

Macrophage-Mediated Tissue Vascularization: Similarities and Differences Between Cornea and Skin.

Macrophages are critical mediators of tissue vascularization both in health and disease. In multiple tissues, macrophages have been identified as important regulators of both blood and lymphatic vessel growth, specifically following tissue injury and in pathological inflammatory responses. In development, macrophages have also been implicated in limiting vascular growth. Hence, macrophages provide an important therapeutic target to modulate tissue vascularization in the clinic. However, the molecular mechanisms how macrophages mediate tissue vascularization are still not entirely resolved. Furthermore, mechanisms might also vary among different tissues. Here we review the role of macrophages in tissue vascularization with a focus on their role in blood and lymphatic vessel formation in the barrier tissues cornea and skin. Comparing mechanisms of macrophage-mediated hem- and lymphangiogenesis in the angiogenically privileged cornea and the physiologically vascularized skin provides an opportunity to highlight similarities but also tissue-specific differences, and to understand how macrophage-mediated hem- and lymphangiogenesis can be exploited for the treatment of disease, including corneal wound healing after injury, graft rejection after corneal transplantation or pathological vascularization of the skin.

Electrochemical Detection of Insulin with a Novel Protein-Based Artificial Receptor

In this paper, we have developed a new electrochemical method to detect insulin with a protein-based artificial receptor. Diabetes is one of the most serious diseases all over the world. For the treatment of diabetes and maintenance of the patient's quality of life, the control of blood glucose level is important. Usually the blood glucose level in human body is regulated by insulin. Therefore, it is very important to know the status of both glucose and insulin in blood for the treatment of diabetes. Currently, it has become possible to measure the blood glucose level by patients themselves with bioelectrochemical sensors, however, there is no convenient method to measure insulin by patients themselves. In recent years, antibody-based and aptamer-based electrochemical insulin sensors have been developed. Although these sensors are highly sensitive, they may also detect physiologically inactive insulin such as partially digested or mutation insulin because the insulin recognition molecules of these sensors specifically recognize a part of insulin. However, it is desired to detect only active insulin for accurate evaluation of insulin which is functional in blood. Therefore, it is necessary to develop a novel sensor based on a new target recognition mechanism for the accurate evaluation of insulin. In our strategy, we utilized an insulin receptor, because the recognition by the insulin receptor is one of the essential processes for insulin to exhibit its functional role in blood. It has been reported that two domains in the insulin receptor, αCT segment and L1CR domain, recognize an insulin molecule by sandwiching format. However, the insulin receptor is typically buried in a cellular membrane and thus it is difficult to use the receptor with a native format for electrochemical detection. Hence, we have developed a novel artificial insulin receptor by connecting these two domains with a linker. We considered that this new artificial insulin receptor alters its structure in response to insulin recognition, and thus electrochemical properties of an electrode modified with the artificial receptor should reflect the structural change. In order to modify the electrode, we utilized three-time-repeats of a gold-binding peptide (GBP) that is known to assemble on a gold surface spontaneously. These domains were genetically fused and resulting artificial insulin receptors were termed as 3GαL and αL3G depending on their sequence of the domains, respectively. The novel artificial receptors were expressed by introducing plasmids encoding each recombinant gene to animal cells, then purified utilizing histidine-tag affinity. The expression and purification of each receptor was confirmed by SDS-PAGE. The ability of self-assembling on a gold surface was evaluated by QCM-D. The artificial receptor modified electrode was prepared by placing each protein solution on the gold surface of electrodes. The electrochemical response of the modified electrode to insulin was evaluated using electrochemical impedance spectroscopy (EIS). EIS measurement was performed in buffer solution in the presence of 5 mM [Fe(CN)6]3-/4- (the frequency range: 100 mHz to 100 kHz, potential: REST, amplitude: 10 mV). The comparisons of Nyquist plots revealed that the diameters of semicircle increased in the case of electrodes modified with the artificial receptors, when compared with an unmodified electrode. This is probably because the artificial receptors are proteins which inhibit interfacial electron transfer. When insulin was added to the solution, the diameters of semicircle further increased, while no drastic increase was observed when C-peptide was added. Comparison of the responses between electrodes modified with 3GαL and αL3G revealed that the electrode modified with 3GαL exhibited a better S/N ratio in response to insulin. In case of 3GαL, the side of αCT segment was immobilized on the gold surface and L1CR domain whose size is much larger than αCT segment kept rather free. In contrast, in case of αL3G, the side of larger domain of L1CR was already fixed and the smaller part of αCT segment kept rather free. Therefore, it was considered that the effect of structural change induced by the recognition of insulin on the electrochemical property of electrode should be more significant in the case of 3GαL than the case of αL3G. From these results, it has been concluded that insulin can be measured electrochemically with the electrode modified with the protein-based artificial receptors.

The Forms and Meaning of Ancient Judahite Sacrificial System: A Discussion Based on Leviticus 1-7

Sacrifices are performed in almost every human society of which ancient Israel is not exempted. Since sacrifices do not take place anywhere and anyhow, the article describes sacred places and objects in ancient Israelite society.This article looks at the forms of the sacrifices and their meaning as described in Leviticus 1-7. It helps one to understand the purposes of performing these sacrifices.This article also shows the roles of both the priest and the offerors in performing sacrifices. The article, in addition, seeks to explain the role of blood in the ancient Israelite sacrificial system.The article demonstrates that sacrifice was a means of restoring a broken relationship between YHWH and his people caused by their sins. Keywords: forms and meaning, ancient Israel, sacrificial system, Leviticus 1-7