Intact Blood Vessels Research Articles

Myocardin related transcription factor and galectin-3 drive lipid accumulation in human blood vessels

ObjectiveDiabetes and hypertension are important risk factors for vascular disease, including atherosclerosis. A driving factor in this process is lipid accumulation in smooth muscle cells of the vascular wall. The glucose- and mechano-sensitive transcriptional coactivator, myocardin-related transcription factor A (MRTF-A/MKL1) can promote lipid accumulation in cultured human smooth muscle cells and contribute to the formation of smooth muscle-derived foam cells. The purpose of this study was to determine if intact human blood vessels ex vivo can be used to evaluate lipid accumulation in the vascular wall, and if this process is dependent on MRTF and/or galectin-3/LGALS3. Galectin-3 is an early marker of smooth muscle transdifferentiation and a potential mediator for foam cell formation and atherosclerosis. Approach and resultsHuman mammary arteries and saphenous veins were exposed to altered cholesterol and glucose levels in an organ culture model. Accumulation of lipids, quantified by Oil Red O, was increased by cholesterol loading and elevated glucose concentrations. Pharmacological inhibition of MRTF with CCG-203971 decreased lipid accumulation, whereas adenoviral-mediated overexpression of MRTF-A had the opposite effect. Cholesterol-induced expression of galectin-3 was decreased after inhibition of MRTF. Importantly, pharmacological inhibition of galectin-3 with GB1107 reduced lipid accumulation in the vascular wall after cholesterol loading. ConclusionEx vivo organ culture of human arteries and veins can be used to evaluate lipid accumulation in the intact vascular wall, as well as adenoviral transduction and pharmacological inhibition. Although MRTF and galectin-3 may have beneficial, anti-inflammatory effects under certain circumstances, our results, which demonstrate a significant decrease in lipid accumulation, support further evaluation of MRTF- and galectin-3-inhibitors for therapeutic intervention against atherosclerotic vascular disease.

Signalling switches maintain intercellular communication in the vascular endothelium.

The single layer of cells lining all blood vessels, the endothelium, is a sophisticated signal co-ordination centre that controls a wide range of vascular functions including the regulation of blood pressure and blood flow. To co-ordinate activities, communication among cells is required for tissue level responses to emerge. While a significant form of communication occurs by the propagation of signals between cells, the mechanism of propagation in the intact endothelium is unresolved. Precision signal generation and targeted cellular manipulation was used in conjunction with high spatiotemporal mesoscale Ca2+ imaging in the endothelium of intact blood vessels. Multiple mechanisms maintain communication so that Ca2+ wave propagation occurs irrespective of the status of connectivity among cells. Between adjoining cells, regenerative IP3-induced IP3 production transmits Ca2+ signals and explains the propagated vasodilation that underlies the increased blood flow accompanying tissue activity. The inositide is itself sufficient to evoke regenerative phospholipase C-dependent Ca2+ waves across coupled cells. None of gap junctions, Ca2+ diffusion or the release of extracellular messengers is required to support this type of intercellular Ca2+ signalling. In contrast, when discontinuities exist between cells, ATP released as a diffusible extracellular messenger transmits Ca2+ signals across the discontinuity and drives propagated vasodilation. These results show that signalling switches underlie endothelial cell-to-cell signal transmission and reveal how communication is maintained in the face of endothelial damage. The findings provide a new framework for understanding wave propagation and cell signalling in the endothelium.

Extracellular SOD modulates canonical TNFα signaling and α5β1 integrin transactivation in vascular smooth muscle cells

TNFα activates NADPH oxidase 1 (Nox1) in vascular smooth muscle cells (VSMCs). The extracellular superoxide anion (O2•-) produced is essential for the pro-inflammatory effects of the cytokine but the specific contributions of O2•- to signal transduction remain obscure. Extracellular superoxide dismutase (ecSOD, SOD3 gene) is a secreted protein that binds to cell surface heparin sulfate proteoglycans or to Fibulin-5 (Fib-5, FBLN5 gene), an extracellular matrix protein that also associates with elastin and integrins. ecSOD converts O2•- to hydrogen peroxide (H2O2) which prevents NO• inactivation, limits generation of hydroxyl radical (OH•), and creates high local concentrations of H2O2. We hypothesized that ecSOD modifies TNFα signaling in VSMCs. Knockdown of ecSOD (siSOD3) suppressed downstream TNFα signals including MAPK (JNK and ERK phosphorylation) and NF-κB activation (luciferase reporter and IκB phosphorylation), interleukin-6 (IL-6) secretion, iNOS and VCAM expression, and proliferation (Sulforhodamine B assay, PCNA western blot). These effects were associated with significant reductions in the expression of both Type1 and 2 TNFα receptors. Reduced Fib-5 expression (siFBLN5) similarly impaired NF-κB activation by TNFα, but potentiated FAK phosphorylation at Y925. siSOD3 also increased both resting and TNFα-induced phosphorylation of FAK and of glycogen synthase kinase-3β (GSK3β), a downstream target of integrin linked kinase (ILK). These effects were dependent upon α5β1 integrins and siSOD3 increased resting sulfenylation (oxidation) of both integrin subunits, while preventing TNFα-induced increases in sulfenylation. To determine how ecSOD modified TNFα-induced inflammation in intact blood vessels, mesenteric arteries from VSMC-specific ecSOD knockout (KO) mice were exposed to TNFα (10 ng/ml) in culture for 48 h. Relaxation to acetylcholine and sodium nitroprusside was impaired in WT but not ecSOD KO vessels. Thus, ecSOD association with Fib-5 supports pro-inflammatory TNFα signaling while tonically inhibiting α5β1 integrin activation.

Zero dimensional Graphene Quantum Dots self-assembled collagen 3D bio-matrices for soft tissue regeneration

The current study focuses on self-assembly of collagen fibrils with Graphene Quantum Dots (GQD) and its tissue regenerative applications. GQDs were synthesised by the liquid exfoliation method using ethyl acetoacetate (EAA) as the solvent. The photoluminescence property of the GQD was established by the UV-Vis absorption spectrum. Characteristic increase in thermal stability, enhanced tensile strength was found after the self-assembly of collagen with GQD. The GQDs self-assembled collagen bio-matrices showed 3D surface topology mimicking the extracellular matrix. The GQDs self-assembled collagen bio-matrices did not show toxicity in the structural cells namely fibroblast cells and exhibited increased biocompatibility as indicated by enhanced cell adhesion, proliferation and cell migration of fibroblast cells. Angio-proliferative effect as indicated by an enhanced blood vessel formation and intact unruptured blood vessels was observed in the GQD self-assembled collagen. The study opens a way forward for the use of zero-dimensional graphene based 3-dimensional self-assembled collagen biomatrices for soft tissue regenerative applications.

Vitamin D deficiency enhances vascular oxidative stress, inflammation, and angiotensin II levels in the microcirculation of diabetic patients

Low vitamin D (vitD) levels have been reported to be a risk factor for diabetes-related cardiovascular complications. This study examined the effects of vitD deficiency on oxidative stress (OS), inflammation, and levels of the vasoconstrictor angiotensin II (Ang II) in the microvascular tissue of type 2 diabetic patients. Patients were categorized into (i) vitD non-deficient diabetics (DNP, n = 10) and (ii) vitD-deficient diabetics (DDP, n = 10), based on their serum 25(OH)D levels. Subcutaneous fat tissues with intact blood vessels were collected during lower limb surgical procedures. The blood vessel were isolated; measurements of the antioxidant enzyme superoxide dismutase (SOD) activity, OS marker malondialdehyde (MDA), Ang II, and the inflammatory marker, TNF-α of the microvascular tissues were determined. Elevated MDA levels and reduced SOD activity, with higher levels of TNF-α and Ang II were observed in the microvascular tissues of DDP compared to DNP. VitD deficiency did not associate with glycemic parameters (fasting blood glucose and glycated hemoglobin) levels. In conclusion, vitD deficiency was correlated with higher microvascular tissue OS, inflammation, and Ang II levels in type 2 diabetic patients. This may contribute to early vasculopathy that occurs in diabetic patients, thus, may contribute to the planning of therapeutic strategies to delay or prevent cardiovascular complications.

Pyroelectric Janus nanomotors for synergistic electrodynamic-photothermal-antibiotic therapies of bacterial infections.

Antibacterial electrotherapy is currently activated by external electric field or self-powered generators, but usually needs complicated power management circuits. Herein, near-infrared illumination (NIR) of pyroelectric nanoparticles (NPs) produces a built-in electric field to address the effectiveness and safety concerns in the antibacterial treatment. Janus tBT@PDA NPs were obtained by capping polydopamine (PDA) on tetragonal BaTiO3 (tBT) NPs through defining the polymerization time, followed by ciprofloxacin (CIP) loading on the PDA caps to fabricate Janus tBT@PDA-Cip NPs. NIR illumination of PDA caps creates temperature variations on tBT NPs to generate photothermal and pyroelectric effects. Finite element simulation reveals a pyroelectric potential of over 1 V and sufficient reactive oxygen species (ROS) are produced to exhibit pyroelectric dynamic therapy (PEDT). The elevated temperature on one side of the Janus NPs produces thermophoretic force to drive NP motion, which enhances interactions with bacteria and overcomes limitations in the short action distance and lifespan of ROS. The pyroelectric field accelerates CIP release through weakening the π-π stacking and electrostatic interaction with PDA and also interrupts membrane potentials of bacteria to enhance CIP invasion into bacteria. The synergistic antibacterial effect of pyroelectric tBT@PDA-Cip NPs causes the fully recovery of S. aureus-infected skin wounds and regeneration of intact epidermis, blood vessels and hair follicles, while no obvious pathological change or inflammatory lesion is detected in the major organs. Thus, the pyroelectric Janus nanomotors demonstrate synergistic PEDT/photothermal/antibiotic effects to enhance antibacterial efficacy while avoiding the necessity of excessive heat, ROS and antibiotic doses. STATEMENT OF SIGNIFICANCE: : Antibacterial treatment is challenged by antibiotics-derived side effects and the evolution of resistant strains. Phototherapy is commonly associated with excessive heat and oxidative stress, and their combinations with other agents are especially encouraged to strengthen antibacterial efficacy while alleviating the associated side effects. Electric field is another activator to generate antibacterial abilities, but usually requires complicated power management and bulk electrodes, making it inconvenient in a biological setup. To address these challenges, we propose a strategy to generate microelectric field on nanoparticles themselves and achieve synergistic electrodynamic-photothermal-antibiotic therapies. The pyroelectric effect weakens interactions between nanoparticles and antibiotics to accelerate drug release, and the built-in pyroelectric field increases membrane fluidity to enhance bacterial uptake of antibiotics.

TRPM8 channels do not regulate endothelium-dependent and -independent vascular reactivity in mice

Foremost among the ion channels that control vascular tone is the transient receptor potential (TRP) cation channels. The TRPM subfamily of the TRP channels includes eight members, expressed in various excitable and non-excitable cells and involved in diverse functions, such as sensing membrane potential, cold, taste, redox, osmolarity, and cell survival. Previous analysis of TRPM gene expression showed the highest expression of TRPM8 in intact blood vessels compared with other isoforms (TRPM1-7). Studies have also revealed that activating TRPM8 by menthol and icilin modulates endothelium-dependent and independent vascular reactivity. However, the non-selectivity of pharmacological TRPM8 modulators constitutes a significant limitation in the prior studies aimed at delineating TRPM8-dependent vasoregulation. In this study, we demonstrate that acetylcholine (ACH)- and sodium nitroprusside (SNP)-induced reduction in arterial pressure was unaltered in mice lacking TRPM8. We generated endothelial-specific knockout (KO) of TRPM8 (ecTRPM8-/-) by breeding TRPM8 floxed (TRPM8fl/fl) with Cdh5-Cre (VE-Cadherin-Cre) mice. Unlike global TRPM8 KO, ecTRPM8-/- mice were not resistant to cold temperatures. Phenylephrine-induced contraction of renal and mesenteric arteries isolated from TRPM8fl/fl and ecTRPM8-/- mice were similar. Also, ACH- and SNP-induced relaxation of phenylephrine-precontracted renal and mesenteric arteries from TRPM8fl/fl and ecTRPM8-/- mice were comparable. These findings suggest that TRPM8 channels do not regulate endothelium-dependent and -independent vasorelaxation in mice. Disclosure of Funding: NHLBI: R01 HL151735-01 and R01HL151735-S This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Endothelial PAR2 activation evokes resistance artery relaxation.

Protease-activated receptor-1 & -2 (PAR1 and PAR2) are expressed widely in cardiovascular tissues including endothelial and smooth muscle cells. PAR1 and PAR2 may regulate blood pressure via changes in vascular contraction or relaxation mediated by endothelial Ca2+ signaling, but the mechanisms are incompletely understood. By using single-cell Ca2+ imaging across hundreds of endothelial cells in intact blood vessels, we explored PAR-mediated regulation of blood vessel function using PAR1 and PAR2 activators. We show that PAR2 activation evoked multicellular Ca2+ waves that propagated across the endothelium. The PAR2-evoked Ca2+ waves were temporally distinct from those generated by muscarinic receptor activation. PAR2 activated distinct clusters of endothelial cells, and these cells were different from those activated by muscarinic receptor stimulation. These results indicate that distinct cell clusters facilitate spatial segregation of endothelial signal processing. We also demonstrate that PAR2 is a phospholipase C-coupled receptor that evokes Ca2+ release from the IP3 -sensitive store in endothelial cells. A physiological consequence of this PAR2 signaling system is endothelium-dependent relaxation. Conversely, PAR1 activation did not trigger endothelial cell Ca2+ signalingnor relax or contract mesenteric arteries. Neither did PAR1 activators alter the response to PAR2 or muscarinic receptor activation. Collectively, these results suggest that endothelial PAR2 but not PAR1 evokes mesenteric artery relaxation by evoking IP3 -mediated Ca2+ release from the internal store. Sensing mediated by PAR2 receptors is distributed to spatially separated clusters of endothelial cells.

Upper Ureteral Reconstruction with a Tapered Descending Colon after Failed Pyeloplasties in a 13-Year-Old Boy

An 11-year-old boy was referred for further management of a 6-cm-long grossly stenosed ureter following two failed left ureteropelvic junction (UPJ) obstruction repairs elsewhere. A tapered segment of the descending colon (TDC) was used successfully for ureteral reconstruction. The UPJ was exposed through a left flank incision. The stenosed segment was excised; both ends appeared severely inflamed and thickened. Tissue interposition was required and ureteroplasty with a TDC was performed by incising the peritoneum adjacent to the excised ureter to mobilize the descending colon to the retroperitoneal space. To prepare the TDC, an 8-cm segment of the colon with intact blood vessels was isolated, tapered, and sutured into a funnel shape using a 14-Fr catheter as a temporary stent. After colocolostomy, the colon was returned to the abdominal cavity, the peritoneum was closed carefully to prevent vascular compromise, and the TDC was anastomosed to the ureter and renal calyx with interrupted absorbable sutures. A double J stent (DJS) and percutaneous nephrostomy tube were placed. Postoperative recovery was uneventful. The DJS was removed on day 50 after confirming smooth urine flow through both the ureter-TDC and calyx-TDC anastomoses. Diuretic renography performed 68 days postoperatively was unobstructed. The patient is currently well after 12 months follow-up. This would appear to be the first report of a TDC being used to create a funnel-shaped segment to reconstruct a long, grossly stenosed ureter. The TDC is simpler than the re-tubularizing colon but requires monitoring for postoperative mucus-related complications and malignant transformation.

Small-world connectivity dictates collective endothelial cell signaling

Every blood vessel is lined by a single layer of highly specialized, yet adaptable and multifunctional endothelial cells. These cells, the endothelium, control vascular contractility, hemostasis, and inflammation and regulate the exchange of oxygen, nutrients, and waste products between circulating blood and tissue. To control each function, the endothelium processes endlessly arriving requests from multiple sources using separate clusters of cells specialized to detect specific stimuli. A well-developed but poorly understood communication system operates between cells to integrate multiple lines of information and coordinate endothelial responses. Here, the nature of the communication network has been addressed using single-cell Ca2+ imaging across thousands of endothelial cells in intact blood vessels. Cell activities were cross-correlated and compared to a stochastic model to determine network connections. Highly correlated Ca2+ activities occurred in scattered cell clusters, and network communication links between them exhibited unexpectedly short path lengths. The number of connections between cells (degree distribution) followed a power-law relationship revealing a scale-free network topology. The path length and degree distribution revealed an endothelial network with a “small-world” configuration. The small-world configuration confers particularly dynamic endothelial properties including high signal-propagation speed, stability, and a high degree of synchronizability. Local activation of small clusters of cells revealed that the short path lengths and rapid signal transmission were achieved by shortcuts via connecting extensions to nonlocal cells. These findings reveal that the endothelial network design is effective for local and global efficiency in the interaction of the cells and rapid and robust communication between endothelial cells in order to efficiently control cardiovascular activity.

Blood vessel occlusion by Cryptococcus neoformans is a mechanism for haemorrhagic dissemination of infection.

Meningitis caused by infectious pathogens is associated with vessel damage and infarct formation, however the physiological cause is often unknown. Cryptococcus neoformans is a human fungal pathogen and causative agent of cryptococcal meningitis, where vascular events are observed in up to 30% of patients, predominantly in severe infection. Therefore, we aimed to investigate how infection may lead to vessel damage and associated pathogen dissemination using a zebrafish model that permitted noninvasive in vivo imaging. We find that cryptococcal cells become trapped within the vasculature (dependent on their size) and proliferate there resulting in vasodilation. Localised cryptococcal growth, originating from a small number of cryptococcal cells in the vasculature was associated with sites of dissemination and simultaneously with loss of blood vessel integrity. Using a cell-cell junction tension reporter we identified dissemination from intact blood vessels and where vessel rupture occurred. Finally, we manipulated blood vessel tension via cell junctions and found increased tension resulted in increased dissemination. Our data suggest that global vascular vasodilation occurs following infection, resulting in increased vessel tension which subsequently increases dissemination events, representing a positive feedback loop. Thus, we identify a mechanism for blood vessel damage during cryptococcal infection that may represent a cause of vascular damage and cortical infarction during cryptococcal meningitis.

Caveolar disruption with methyl-β-cyclodextrin causes endothelium-dependent contractions in Wistar rat carotid arteries.

Caveolae are organizing centers for cellular signal transduction in endothelial cells (ED) and smooth muscle cells (SMCs) in the blood vessels. Myography was used to investigate the effects of a caveolar disruption using methyl-β-cyclodextrin (MBCD) on maxi-K channels in rat carotid arteries. Incubation of carotid segments with MBCD augmented contractions in response to BaK (chemical channel agonist) but not those induced by depolarizing high potassium physiological saline (KPSS). In contrast, incubation with cholesterol-saturated MBCD (Ch-MBCD) abolished the effects of MBCD. Mechanical removal of endothelial cells by MBCD triggered a small contraction in response to BaK. Incubation with nitroarginine methyl ester (L-NAME) inhibited nitric oxide (NO) release, causing increased contractions in response to BaK, and this effect was reversed by pretreatment with MBCD. These results suggest that MBCD inhibits endothelial NO release. Contrastingly, inhibition of maxi-K channels with iberiotoxin enhanced contractions in response to BaK. Likewise, L-NAME decreased the contractile effect of iberiotoxin, as in the ED-denuded arteries. Transmission electron microscopy (TEM) showed the presence and absence of caveolae in intact blood vessels before and after MBCD treatment, respectively, whereas histology confirmed ED removal after the treatment. Caveolar disruption using MBCD impairs ED-dependent relaxation by inhibiting the release of NO from the ED and altered the contractility of SMCs independent of the ED due to reduced contribution of maxi-K channels to the SMC membrane potential, causing depolarization and increasing carotid artery contraction. These findings might help to understand the physiological role of the maxi-K channels in rat carotid arteries.

Protein fucosylation is required for Notch dependent vascular integrity in zebrafish

The onset of circulation in a developing embryo requires intact blood vessels to prevent hemorrhage. The development of endothelial cells, and their subsequent recruitment of perivascular mural cells are important processes to establish and maintain vascular integrity. These processes are genetically controlled during development, and mutations that affect endothelial cell specification, pattern formation, or maturation through the addition of mural cells can result in early developmental hemorrhage. We created a strong loss of function allele of the zebrafish GDP-mannose 4,6 dehydratase (gmds) gene that is required for the de novo synthesis of GDP-fucose, and homozygous embryos display cerebral hemorrhages. Our data demonstrate that gmds mutants have early defects in vascular patterning with ectopic branches observed at time of hemorrhage. Subsequently, defects in the number of mural cells that line the vasculature are observed. Moreover, activation of Notch signaling rescued hemorrhage phenotypes in gmds mutants, highlighting a potential downstream pathway that requires protein fucosylation for vascular integrity. Finally, supplementation with fucose can rescue hemorrhage frequency in gmds mutants, demonstrating that synthesis of GDP-fucose via an alternative (salvage) pathway may provide an avenue toward therapeutic correction of phenotypes observed due to defects in de novo GDP-fucose synthesis. Together, these data are consistent with a novel role for the de novo and salvage protein fucosylation pathways in regulating vascular integrity through a Notch dependent mechanism.

Abstract 1581: Nammisomes: Preclinical validation of a platform for selective and synchronized delivery of novel immunotherapy prodrug combinations to tumors

Abstract The approval of checkpoint inhibitors, CAR-T cell therapies, and other immunotherapies has led to a significant improvement in cancer patient survival rates over the past 7 years. However, the pace of improvement has slowed lately as the field moves beyond the minority of patients with immune primed or accessible cancers. It is generally expected that rationale combinations of immunotherapies that provide mechanisms to both induce as well as maintain the immune responses in the face of tumor-derived immune checkpoint mediators, will be required to effectively treat these patients. Systemic exposure to such combinations often can give rise to severe toxicity profiles that limit their therapeutic efficacy. Nammisomes are an innovative and unique platform designed to deliver novel immune modulating prodrugs more selectively and synchronously to the tumor site. The immune active agents are conjugated to lipids to create prodrugs that self-assemble into a liposomal bilayer. By mixing multiple prodrugs at the desired ratio, co-formulations are produced with the optimum stoichiometry. These Nammisomes are iv injected and selectively enter tumors through the leaky vasculature within tumors, while remaining in circulation in normal tissues with intact blood vessels, The enriched phagocytic activity in tumor environments causes the release of the active agents within tumors relative to normal tissues limiting systemic exposure and toxicities. Preclinical efficacy and safety data of Nammisomes delivering combinations of a TLR agonist and a checkpoint inhibitor showing anti-tumor efficacy and tolerability in syngeneic mouse models will be presented. In summary, Nammisomes will be highlighted as a combination therapeutic modality to demonstrate the potential of this approach in anti-cancer therapy. Citation Format: David R. Stover, Dhruba Bharali, Tahmineh Safaie, Bruce A. Hay, Rajesh K. Sharma, Scott Ayers. Nammisomes: Preclinical validation of a platform for selective and synchronized delivery of novel immunotherapy prodrug combinations to tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1581.

Can indocyanine green perfusion imaging detect microangiopathy in mastectomy skin flaps?

Intraoperative vascular imaging using indocyanine green (ICG) angiography may better predict flap viability than clinical judgement alone. Intraoperative ICG angiography was used in a chronic corticosteroid user undergoing bilateral nipple sparing mastectomy with expander placement. Large blood vessels were visualized, however, the skin surrounding these vessels remained dark. The flap demarcated to full-thickness necrosis that matched the intraoperative SPY findings. Visualization of intact blood vessels may not be sufficient to rule out flap necrosis in some patients. In these circumstances, interpretation of perfusion with consideration of patient factors will be required to accurately predict flap viability.

Caffeic acid improves glucose utilization and maintains tissue ultrastructural morphology while modulating metabolic activities implicated in neurodegenerative disorders in isolated rat brains.

Impaired glucose utilization has been implicated in the pathophysiology of neurodegenerative diseases. The neuroprotective effect of caffeic acid (CA) was investigated in the isolated rat brain by determining its ability to promote glucose uptake, mitigate redox imbalance, modulate purinergic and cholinergic activities, elemental distribution, and maintain tissue morphology. Isolated rat brains were incubated for 2 hours with glucose, CA and glucose, and metformin and glucose. There was an increased glucose uptake, glutathione level, superoxide dismutase, and catalase activities in brain tissues incubated with CA compared to the controls. Incubation with CA also led to significantly decreased levels of malondialdehyde, nitric oxide, acetylcholinesterase, butyrylcholinesterase, and ATPase activities. Electron microscopy (scanning electron microscopy and transmission electron microscopy) analysis portrayed a maintenance of tissue ultrastructural morphology in 2CA-incubated tissues as indicated by the intact synaptic vesicles, blood vessels, dendritic and neuronal network, mitochondria, and presynaptic membrane. CA also restored altered elemental levels in brain tissues. These results indicate the stimulatory potential of CA on brain glucose utilization with simultaneous neuroprotective activities.

A versatile technique for high-resolution three-dimensional imaging of human arterial segments using microcomputed tomography

BackgroundQuantitative methods for evaluating microstructure of arterial specimens typically rely on histologic techniques that involve random sampling, which cannot account for the unique spatial distribution of features in three dimensions. MethodsTo overcome this limitation, we demonstrate a nondestructive method for three-dimensional imaging of intact human blood vessels using microcomputed tomography (microCT). Human artery segments were dehydrated and stained in an iodine solution then imaged with a standard laboratory microCT scanner. Image visualization and segmentation was performed using commercially available and open source software. ResultsStaining of cadaveric vessels with iodine enabled clear visualization of the arterial wall with microCT, preserved tissue morphology, and generated high-resolution images with a voxel size of 5.4 μm. Various components of the arterial wall were segmented using a combination of manual and automatic thresholding algorithms. ConclusionsOur approach allows for spatial mapping of human artery tissue samples that can guide targeted histologic analysis of smaller tissue segments, provide geometric data to inform finite element models, quantify degree of atherosclerosis, and help to evaluate the foreign body response to intravascular medical implants. (JVS–Vascular Science 2021;2:13-19.) Clinical RelevanceIn this article, we describe a powerful technique for whole artery analysis of pathologic human tissue specimens that provides high-resolution spatial detail regarding composition of the blood vessel wall. The protocol described here is a valuable adjunct that can be used as a research tool to inform finite element modeling of arteries, quantify pathologic response (ie, neointimal hyperplasia and vascular calcification), and evaluate the tissue/device interface of implanted medical devices.

Connexins: Key Players in the Control of Vascular Plasticity and Function.

Of the 21 members of the connexin family, 4 (Cx37, Cx40, Cx43, and Cx45) are expressed in the endothelium and/or smooth muscle of intact blood vessels to a variable and dynamically regulated degree. Full-length connexins oligomerize and form channel structures connecting the cytosol of adjacent cells (gap junctions) or the cytosol with the extracellular space (hemichannels). The different connexins vary mainly with regard to length and sequence of their cytosolic COOH-terminal tails. These COOH-terminal parts, which in the case of Cx43 are also translated as independent short isoforms, are involved in various cellular signaling cascades and regulate cell functions. This review focuses on channel-dependent and -independent effects of connexins in vascular cells. Channels play an essential role in coordinating and synchronizing endothelial and smooth muscle activity and in their interplay, in the control of vasomotor actions of blood vessels including endothelial cell reactivity to agonist stimulation, nitric oxide-dependent dilation, and endothelial-derived hyperpolarizing factor-type responses. Further channel-dependent and -independent roles of connexins in blood vessel function range from basic processes of vascular remodeling and angiogenesis to vascular permeability and interactions with leukocytes with the vessel wall. Together, these connexin functions constitute an often underestimated basis for the enormous plasticity of vascular morphology and function enabling the required dynamic adaptation of the vascular system to varying tissue demands.

Huge Lipoma in the Floor of the Mouth: A Case Report

Lipomas are adipose mesenchymal neoplasms. The oral cavity is not commonly affected. representing about0.5% to 5% of all benign oral tumors. The clinical presentation is typically as an asymptomatic yellowish mass.The overlying epithelium is intact and superficial blood vessels are usually evident over the tumour. Although benign in nature their progressive growth may cause interference with speech and mastication due to tumour’s dimension. The present report shows the case of a 52-year old male who presented with a large intraoral lipoma with extension to the right submandibular region. Bangladesh J Otorhinolaryngol; April 2016; 22(1): 58-61

An automated method for segmentation and quantification of blood vessels in histology images

Alzheimer's disease (AD) is a chronic neuro-degenerative disease that adversely affect many people on a global scale. Despite different diagnostic and therapeutic treatment, there is no cure for AD. The brain is one of the most complex organ and researchers are still trying to understand so as to find a cure. Objective: To complement the efforts of clinical researchers engaged in research in alzheimer's disease, accurate segmentation and quantification of blood vessels in brain images is required. Method: For robust segmentation of blood vessels even in the presence of colour variation, we introduce a fully automated morphological tool that can extract and quantify vessels from haematoxylin and diaminobenzidine stained histology brain image. The method, exploits saturation channel of stained image slides, ISODATA threshold method is applied to obtain a binary image. This helps in eliminating background and remaining with only blood vessels. A one-stage procedure that includes eliminating small artefacts is performed on the binary mask. The intensity of the image is transformed. Joining is performed to deal with fragmentation of intact blood vessels on the images, and artefactual appearance of the blood vessel structures. The artefactual fragments based on measured incoherence with neighbouring tissue are removed. The vessels are then labelled to facilitate quantification. Morphometric measurements are used during the vessel quantification assess both vessels with lumen and vessels without lumen. We have quantified the diameter of blood vessels. Results: The image processing technique is developed in close collaboration with neuroscientist experts to help clinician. We have evaluated our proposed approach qualitatively. The method was validated against their manual quantification results. Qualitative results show that the method can indeed segment the blood vessels in the presence of colour variations and artefacts. The quantitative method produces fairly better results.