How neutrophils resist shear stress at blood vessel walls: molecular mechanisms, subcellular structures, and cell–cell interactions Daniela Begandt Walter Brendel Centre of Experimental Medicine, Department of Cardiovascular Physiology and Pathophysiology, Biomedical Center, Ludwig‐Maximilians‐Universität München, Planegg‐Martinsried, Germany. Neutrophils tend to migrate at the site of the basement membrane containing low expression of lamin 511 and collagen IV.78-81 Expression of VLA-3 (integrin α3β1, also known as CD49c/CD29)82 and VLA-6 (integrin α6β1, also known as CD49f/CD29),83 2 laminin-binding integrins on neutrophils, as well as neutrophil elastase83 are all required for migration. Correspondence: Marie-Dominique Filippi, Division of Experimental Hematology and Cancer Biology, S7.605, Cincinnati Children’s Research Foundation, 3333 Burnet Ave, Cincinnati, OH 45229; e-mail: marie-dominique.filippi@cchmc.org. PMN, polymorphonuclear neutrophil. In this study, parallel ridges/grooves were micropatterned on glass surfaces using photosensitive polyimide to create transparent substrates. The inflammatory milieu can also promote a “reverse migration” behavior. Adherence to vascular endothelium is the first step in the process of, The New England journal of medicine, 1991, Seldin and Giebisch's The Kidney (Fourth Edition), PDGF, platelet factor 4 (PF4), TGF-β, IL-4, hypoxia, fragments of fibronectin and types I and III collagen, bFGF, TNF-α, IL-8, hypoxia, heparin, fibronectin, Monocyte chemoattractant protein-1 (MCP-1), CCL2, platelet-derived endothelial cell growth factor (PD-ECGF), thrombin, TGF-β, fibronectin, and elastin, Fibrinopeptides, fibrin lysis products, C5a, formyl methionyl peptides (from bacterial proteins), platelet activating factor (PAF), TNF-α, PDGF, PF4. Increased vascular permeability caused by inflammation and the release of prostaglandin, together with a concentration gradient of chemotactic substances such as complement factors, interleukin (IL)-1, tumor necrosis factor (TNF)-α, TGF-β, platelet factor-4, and bacterial products, stimulate neutrophil migration. Phagocytes include neutrophils, eosinophils, and basophils. Fluorescence images were taken 3 h postinjection. These methods have been used to detect severe neutrophil-associated lung inflammation and infection (Xiao et al., 2010; Zhang et al., 2010), as well as the degree of neutrophil recruitment to medical-device-associated FBRs and infection (Figure 14.4) (Zhou et al., 2012b). It is then said to activate or inflame the vascular endothelium. We need to understand how leukocyte and EC interactions modulate leukocyte effector cell functions and investigate the mechanism of neutrophil migration in a tissue-specific manner in order to design targeted therapy for inflammatory conditions, both normal and pathologic. MKL1-deficient neutrophils displayed almost complete abrogation of migration in vitro due to an inability of the cells to properly retract the uropod. The authors conclude that PSGL-1–bearing microdomains scan for activated platelets present in the bloodstream.45 These dynamic interactions depend on Cdc42 activity and are necessary for efficient neutrophil transmigration in vivo.45 Importantly, during pathogenic inflammation, the uropod becomes the predominant domain for platelet interactions, such as during transfusion-related acute lung injury, suggesting that neutrophil polarization and interactions with accessory cells participate in modulating inflammation.45 This is very intriguing and merits further investigation. It will discuss how neutrophil–EC interactions and the subsequent mode of diapedesis, junctional or nonjunctional, can be context dependent and how this plasticity may be exploited clinically. Recombinant plasma gelsolin entered several clinical trials for critical care indications and community-acquired pneumonia. The final remodeling phase is characterized by synthesis and degradation of collagen and depends on a balance between factors that endorse synthesis of extracellular matrix components and enzymes that degrade these components, such as matrix metalloproteases (MMPs). During this step, the formation of the ICAM-1 adhesome and F-actin–rich filopodia emerging from ECs are critical prior to diapedesis. (2000) found that the kinetics of penetration of neutrophils through TNF-α–treated HUVECs was more rapid in a flow assay where neutrophils were perfused over the endothelium than in a static assay in which they were simply allowed to settle onto it. Consistent with our studies, the Rap1b effector Tiam1, which is also a Rac activator, has been involved in transcellular migration.76 Tiam1 deficiency in lymphocytes increased the frequency of transcellular migration across ECs in vitro and increased transcellular pore formation away from endothelial junctions.76 Together, these findings reveal the existence of a neutrophil signaling pathway that specifically controls the transcellular migration process. In other models, increased adhesion to ECs was also associated with increased transcellular migration.96,97 Hence, transcellular migration could be a compensatory mechanism and a highly regulated process that is favored by heightened adhesion and reduced crawling, as well as other yet-to-be-discovered factors. They have an enhanced susceptibility to infection by some bacteria (Staphylococcus aureus, Pseudomonas, and Serratia) and by some fungi (Aspergillus and Candida). High ICAM density, high integrin signaling, low Rap1b/Tiam1, and subsequent high PI3K/Akt signaling trigger neutrophil invasive protrusions and transcellular migration. The protective effect of adenosine 2A receptor activation is independent of IL-6 and TGF-β mRNA induction (59). They used 4-dimensional intravital microscopy to show that the site and order of function of PECAM and CD99 in vivo depends on the strain of mice. Understanding neutrophil migration plasticity will offer opportunities for specificity in targeting inflammation. In contrast to neutrophils, the presence and activation of both macrophages and lymphocytes in the wound is critical to the progression of the normal healing process (Fig. This suggests that early events occur mainly through release of reactive oxygen species, proteases, elastases, and other enzymes that damage tissues. The phagocytic system plays a very important role in killing pathogens early in an infection. The extravasation cascade has been well studied. Contribution: M.-D.F. However, in recent unpublished observations on static EC cultures, transmigration was not complete for about 20 min. The critical process of adherence of leukocytes to the vascular endothelium in producing injury in ischemic tissues begins early in the course of events, but the actual accumulation of neutrophils, which have been studied extensively, does not occur until 24 to 48 hours after injury. Depending on the tissue and intensity of inflammation, subsets of neutrophils can exhibit distinct life spans, synthesize various amount of cytokines,85 or undergo metabolic changes.86 Further, the neutrophil recruitment process is organ and insult specific.87,88 Neutrophil recruitment to the lung is selectin dependent in response to LPS from Escherichia coli, but not in response to LPS from Streptococcus pneumoniae or Salmonella enteritidis.89,90 Likewise, the dependency on integrins of neutrophil migration to the lungs varies with the inflammatory insult.91-94 Neutrophils also use noncanonical extravasation cascades in the lung, liver, and kidney. Okusa et al. 20 The most numerous leukocyte is the __________. Increased expression of actin depolymerizer cofilin-1 is shown to render drug-resistance in adenocarcinoma cells (Becker et al., 2014). Other recent studies have agreed that presence of flow has little effect on the proportion of neutrophils transmigrating though TNF-α–treated HUVECs (Cinamon et al., 2004). Circulating levels of gelsolin are reduced dramatically in critically ill patients, correlating strongly with the development of ARDS, decreased odds of weaning from mechanical ventilation, and death (Holm et al., 2019; Lee et al., 2006; Mounzer et al., 1999). I-R induced a reduction in expression of JAM-C at EC junctions, which increased the frequency of rTEM events.8 Interestingly, leukotriene B4 released during I-R caused neutrophil-dependent elastase activity to cleave EC JAM-C at sites of neutrophil transmigration and promote rTEM.99 Interestingly, neutrophils that had undergone rTEM exhibit enhanced ICAM-1 expression and can return to the blood circulation and infiltrate distant organs, favoring secondary organ inflammation.8 Hence, the local inflammatory milieu contributes to neutrophil migration plasticity and modulates inflammatory responses. Signals from the infection cause the endothelial cells that line the blood vessels to make a protein called selectin, which neutrophils stick to when they pass by. Hence, Rap1b limits neutrophil migration by suppressing the transcellular migration process.75 Interestingly, enhanced Akt activity specifically mediates Rap1b−/− neutrophil transcellular migration but has no effect on paracellular migration. Perspectives and conclusions: can we exploit neutrophil transmigration plasticity for clinical purposes? Neutrophils have been shown to play a fundamental role in its development and progression. Transcellular migration is favored when the EC content in stress fibers is low but the EC junctions are tight. It is also established that the process of transmigration, during which neutrophils encounter a number of interactions with ECs, causes functional changes in neutrophil effector cell functions. Note the initial arrival of platelets to achieve hemostasis as well as neutrophils and macrophages to the healing-wound bed. EC stiffness, which promotes crawling efficiency, also favors paracellular migration.95 Conversely, high ICAM-1 expression and adhesion of neutrophils onto ECs tend to limit crawling but favor transcellular migration.96,97 Finally, CD11b-null neutrophils have defective intravascular crawling and largely fail to transmigrate, but the few cells that transmigrate do so transcellularly.11 Other models, such as Tiam1-deficient or Rap1b-deficient neutrophils, also exhibit defective crawling associated with enhanced transcellular migration.75,76 It is thus possible that transcellular migration is a rescue mechanism to enable cells that fail to reach the endothelial junction to cross the blood vessel. https://quizlet.com/38789151/connect-questions-for-ch-21-flash-cards Several imaging methods have therefore been investigated to monitor, Babich et al., 1997; van der Laken et al., 1997, Polymer Science: A Comprehensive Reference, GAGs have been widely employed as therapeutic agents due to their significant bioactivity and biological roles in tumor metastasis, tissue remodeling, neuronal development, blood clotting, and allergic response. Initially there is slow neutrophil migration mediated by tethering interactions between selectins and their endothelial cell ligands. On the surface of neutrophils and on endothelial cells in injured tissue. Figure 1.3. Asif A. Sharfuddin, Bruce A. Molitoris, in Seldin and Giebisch's The Kidney (Fourth Edition), 2008. Neutrophil activity, as an integral part of the host response, is commonly monitored by enzyme measurements and histological evaluation. Rolling leukocytes can be activated by chemoattractants such as complement C5a and platelet activating factor. There is also some suggestion that such responses translate into an effect on efficiency of migration through the endothelial monolayer. After this process, the neutrophils will release factors known as chemotactic agents to attract various cell types to the site (Table 4.3). Neutrophil migration to injured and pathogen-infected tissues is a fundamental component of innate immunity. Biopsies of human AKI have demonstrated lymphocytes in ATN and the modulatory role of T cells in the mediation of ischemic injury has been established. These cells cleanse the wound and clear it of debris. A variety of immune cells, including T cells and neutrophils, travel … Macrophages, lymphocytes, neutrophils, and platelets have to kill bacteria and clear the wound bed. The work was supported by the National Institutes of Health, National Institute of General Medical Sciences (grant GM112792) (M.-D.F.). The leading edge is made of lipids that are susceptible to extraction by cold Triton X-100 (detergent-sensitive domains) and contains CD45 and HLA. The homeostasis of the skin is regulated by an exquisitely finely tuned exchange of signaling factors among the dermal and epidermal cells. Ratner, in Polymer Science: A Comprehensive Reference, 2012, GAGs have been widely employed as therapeutic agents due to their significant bioactivity and biological roles in tumor metastasis, tissue remodeling, neuronal development, blood clotting, and allergic response. diapedesis. (a) Each layer of the skin contributes to the wound healing process and associated extracellular matrix production. On the other hand, persistence of the inflammatory phase is a hallmark of wound failure and/or excessive fibrosis. Electron microscopy and immunofluorescence images revealed that Rap1b−/− neutrophils extended long protrusions that penetrated deeper into endothelial surfaces than those formed by WT cells. During transcellular migration, EC junctions remain intact. At this ‘simple’ stage, the pleural fluid is exudative but will have a normal glucose and pH > 7.2 as bacterial invasion of the pleural space has not occurred, hence this phase is not true pleural infection, but prompt antibiotic therapy is required to prevent progress and is usually sufficient for resolution of the pleural effusion and complete recovery. In inflammation the endothelium is activated and can bind leukocytes, as a prelude to their exit from the blood vessels. (C) Immunohistochemical staining of neutrophils (400×) reveals that significantly higher numbers of neutrophils accumulated around the infected PU catheters compared to sterile catheters. TGF-β facilitates polymorphonuclear neutrophil migration from the surrounding blood vessels, from which they extrude themselves. Skin egress of neutrophils occurs via lymphatic vessels and is dependent on CD11b and CXCR4 but not CCR7. Gelsolin-deficient mice exhibit prolonged bleeding times, likely due to platelet morphological abnormalities, and impaired neutrophil migration (Witke et al., 1995). These cells had reduced expression of the myosin light chain 9 component of the myosin II complex and overexpression of CD11b integrin. Overall design of the microfluidic devices for neutrophils separation from whole blood and chemotaxis assay. REVERSE MIGRATION OF NEUTROPHILS. Knockdown of caveolin-1 in ECs specifically reduced transcellular migration.49 Consistently, another group reported that high levels of caveolin-1 in ECs favored the transcellular path, whereas its downregulation promoted the paracellular route.96. Interestingly, the few cells that did migrate exhibited enhanced directional persistence.17 Another important regulator of F-actin dynamics is the nonmuscle myosin class II protein Myh9.18 Using stimulated emission depletion nanoscopy, the authors show that Myh9 is localized in lamellipodia and the uropod. Chemotaxis of cells into the wound milieu is followed by cellular activation, which signifies the phenotypic alteration of cellular, biochemical, and functional properties induced by local mediators (Fig. The height (5 or 3 μm) and the repeat spacing (6–14 μm) of the ridges were systematically changed to investigate the effect of microgeometry on neutrophil migration. They consist of fibers made of DNA, proteins, and other material from neutrophil granules. Several imaging methods have therefore been investigated to monitor neutrophil migration. This binding is due to molecules called cell adhesion molecules (CAM) The first step in the migration of neutrophils from blood vessels into the tissues? The healing process can be broadly classified into four phases: hemostasis, inflammation, proliferation, and remodeling. S2A), because the counts of migrating neutrophil toward tumor supernatants were lower in wells where rmCXCL5 was added. Mice deficient for gelsolin exhibit impaired bronchodilation; airway smooth muscle from these mice exhibit impaired relaxation and impaired actin depolymerization (Mikami et al., 2017). The results of this study suggest that optimization of both surface chemistry and topography may be important when designing biomaterials for tissue engineering. Wound healing in skin is characterized by an orderly sequence of events beginning immediately after injury. Neutrophils likely play a modest role as an effector cell in the initiation and extension phases, while T cells, B cells, and macrophages probably have modulatory roles in the extension and repair phases. Additionally, chemotactic peptide receptor agonists were found useful for imaging infection and associated inflammatory cell accumulation in vivo (Babich et al., 1997; van der Laken et al., 1997). Increased vascular permeability caused by inflammation and the release of prostaglandin, together with a concentration gradient of chemotactic substances such as complement factors, interleukin (IL)-1, tumor necrosis factor (TNF)-α, TGF-β, platelet factor-4, and bacterial products, stimulate, : results from defects in molecules found on the surface of neutrophils (integrins) that allow neutrophils to bind (adhere) to the cells lining the blood vessels (vascular endothelium). There is also significant protection from ischemic injury and mortality by blockade of the shared ligand to all three selectins (E-, P-, and L-selectin), which seem to be dependent on the presence of a key fucosyl sugar on the selectin ligand (43, 196). TGF-β facilitates polymorphonuclear, Illustration by Katherine R. Parrish, ACNP, CNS, h after injury. have also shown that although macrophages are required for the full extent of the ischemic renal injury, activation of their adenosine 2A receptors reduces neutrophil accumulation and provides protection against injury, as seen in experiments with macrophage depleted and adenosine 2A receptors deficient mice. Conversely, neutrophils are the main producers of CXCL2, and this chemokine is critical for correct breaching of EC junctions.68, Despite a number of studies providing convincing evidence that neutrophils migrate transcellularly in vivo, the physiological purpose of this process has remained controversial, perhaps because of the difficulty to investigate this phenomenon.69 Studies using transmission electron microscopy of tissue sections clearly showed that in skin tissues, neutrophils take the transcellular route in response to the bacterial chemoattractant formyl-Met-Leu-Phe (fMLP) in vivo.70 The relative contribution of each route of migration depends on the tissue and the vascular bed in vivo, as well as the EC model used for in vitro studies. Stem cell transplantation offers a potential cure (Cole et al., 2013), is possible when the patient has a suitable related or unrelated donor, and is recommended in patients with absent or very low oxidative burst (Kuhns et al., 2010). Then the neutrophils begin to squeeze through the wall of the blood vessel to reach the damaged area. Myosin filaments associate with bundles of actin and generate cortical tension, which suppresses F-actin protrusions. The trafficking of neutrophils was generally considered to be a 1‐way process until recently. In the vessel wall, activated neutrophils can degranulate and generate free radicals to cause damage and undergo NETosis. Leukocyte extravasation (also commonly known as leukocyte adhesion cascade or diapedesis – the passage of cells through the intact vessel wall) is the movement of leukocytes out of the circulatory system and towards the site of tissue damage or infection. Neutrophils can migrate away from inflammatory foci within tissues.7 In addition, transmigrating neutrophils can undergo abluminal to luminal migration across the endothelium, which is called reverse transendothelial migration (rTEM).8 rTEM is predominantly paracellular and specifically depends on JAM-C. Figure 14.4. Schematic drawing of a partial-thickness wound in the inflammatory phase. (A) The paracellular route. Finally, once the neutrophils have passed the EC barrier, they must cross a layer of pericytes77 lining the blood vessel and then breach the perivascular basement membrane. Vascular endothelial growth factor-A (VEGF-A) and basic fibroblast growth factor (FGF2 or bFGF) are the key growth factors in angiogenesis. Killing of bacteria by neutrophils involves a burst of respiratory activity to generate superoxide (Rosenzweig and Holland, 2011). The findings have now been validated in vivo, showing that Rap1b−/− neutrophils use the transcellular migration at higher frequency in response to intradermal inflammatory challenge (Chanchal SurChowdhury and M.-D.F., manuscript submitted March 2019). Floating neutrophils are first captured onto the EC surface in response to inflammatory cytokines and bacteria-derived peptides via upregulation of adhesive molecules on the endothelial luminal surface. Natalia V. Bogatcheva, Roberto F. Machado, in Reference Module in Biomedical Sciences, 2020. Once firmly attached, leukocytes flatten and polarize and then crawl onto the endothelial apical surface in search of a permissive site of extravasation.11,12, Leukocyte polarization is necessary for directional migration (Figure 2). Defects in any of the enzymes involved in the respiratory burst can lead to a defect in intracellular killing by neutrophils. One is activated at the cell front and involves high levels of PI3K-PIP3 and subsequent Rac1/2-mediated actin polymerization formation. Paracellular diapedesis is itself a multistep process, which involves engagement of several adhesion molecules, including ICAM-1/2, vascular cell adhesion molecule 1, junctional adhesion molecule A (JAM-A) and JAM-C, PECAM-1, CD99, and endothelial cell–selective adhesion molecule.5,56 This was demonstrated by studies using genetic mouse models and intravital microscopy, which identified the exact location where leukocyte transmigration is blocked. The uropod is enriched with detergent-resistant lipid domains containing CD44, CD43, l-selectin, heavily glycosylated proteins (eg, PSGL-1), and integrins.19-22 In addition, Mac-1 in neutrophils or lymphocyte function–associated antigen-1 in lymphocytes accumulates along the cell sides and at the uropod during active migration in vitro.22-27 This spatial reorganization is functionally important for leukocyte crawling. However, these methods cannot accurately depict dynamic cellular responses. Pro-inflammatory cytokines, including IL-6, IL-8 and TNF-alpha create intercellular ‘gaps’ by producing changes in the anatomical shape of pleural mesothelial cells, and this further enhances permeability and encourages movement of fluid resulting in the accumulation of an effusion. These studies are now being validated and further investigated using new genetic mouse models and advances in intravital imaging. The fact that increased gelsolin expression is noted in IPF lung (Oikonomou et al., 2009) suggests gelsolin involvement into the pathogenesis of this disease. doi: https://doi.org/10.1182/blood-2018-12-844605. ESAM, endothelial cell–selective adhesion molecule. These cells cleanse the wound and clear it of debris. In addition, parallel ridges/grooves can be used to control the direction and rate of cell migration on the surface. In blood flow, while RBCs accumulate in the center of blood vessels and develop a cell-depleted peripheral layer (CDPL) near the vessel wall, leukocytes and platelets flow in CDPL. Neutrophils are the first blood leukocytes to enter the wound site, and their numbers peak at 24–48 h after injury. This phenomenon was first observed in neutrophils by Cinamon et al,23 but it also occurs during lymphocyte crawling.25,48,49 Live-cell imaging combined with immunofluorescence demonstrated that crawling leukocytes generate numerous finger-like protrusions that extend underneath the cells and at the cell periphery. Loss of CD2AP stimulates the dynamics of ICAM-1 clustering, which facilitates the formation of ICAM-1 complexes on the endothelial surface. Neutrophils are the first line of cellular defense against invading microorganisms by crossing the endothelial cell (EC) barrier to reach inflamed tissues.1,2 Neutrophil tissue infiltration is critical for pathogen elimination and tissue repair and must be tightly regulated, as aberrant neutrophil accumulation into tissues causes tissue damage, as seen during acute lung injury, multiple organ dysfunction syndrome, vascular inflammation, and arthritis.3. Because of these biological roles, GAGs have become promising materials for constructing tissue scaffolds. Singbartl et al. Controlling the TGF-β signaling is a strategy being used to prevent excessive wound healing, which leads to hypertrophic scars. Macrophages, lymphocytes, neutrophils, and platelets have to kill bacteria and clear the wound bed. Caveolin-1 is more enriched around lymphocytes that take the transcellular route than around those migrating at the junction. Paracellular migration is favored when the EC content in stress fibers is high, which helps open EC junctions. 8 rTEM is predominantly … It also increases EC stress fibers, which in turn can destabilize EC junctions, thus favoring paracellular migration.95 Conversely, soft substrate promotes significantly more transcellular diapedesis by reducing stress fibers in ECs and thus their stiffness. These interactions with the endothelium are mediated through endothelial adhesion molecules that are upregulated during ischemic conditions (241). (241) found that platelet P-selectin—not endothelial P-selectin—was the main determinant in neutrophil-mediated ischemic renal injury. Within 6 h of injury, circulating immune cells migrate into the wound. The random motility coefficient μ, 9.8 × 10− 9 cm2/s, was the greatest at a ridge height of 5 μm and spacing of 10 μm, about 10 times faster than on smooth glass surface. This type of migration is most prevalent during sterile inflammation caused by ischemia-reperfusion injury (I-R), suggesting that rTEM is a pathological process. CD2AP in ECs destabilizes ICAM clusters and limits transcellular migration. It is now clear that neutrophil transmigration depends on the organ and the inflammatory insult. While normally found in the blood stream, neutrophils migrate rapidly to sites of infection in tissue. Marie-Dominique Filippi; Neutrophil transendothelial migration: updates and new perspectives. Unligated molecules are recruited to the EC border via certain types of vesicles called the endothelial lateral border recycling compartment (LBRC).65 Finally, during this process, neutrophils generate nuclear lobes that insert into the lamellipodia at the front of the cells, which bend the endothelial actin network and squeeze through the EC barrier.66 Contractile stresses exerted by neutrophils and ECs are also necessary to perturb endothelial junctional tensions in order to open gaps for transmigration and for neutrophils to push themselves across the gap.67 Hence, diapedesis can be mechanically regulated by transmigrating leukocytes and proinflammatory signals that increase EC contractility. Defects in neutrophil numbers are mostly secondary; however, congenital neutropenia does occur, leaving affected individuals susceptible to recurrent infections. The locally formed fibrin clot serves as a scaffold for migrating cells such as neutrophils, monocytes, fibroblasts, and endothelial cells (Fig. In a series of experiments, Rabb and colleagues showed that T-cell–deficient nu/nu mice are protected against ischemic injury, along with attenuation of ICAM-1 expression (42). Complex Migration of Leukocytes. I refer the readers to excellent reviews on this topic.87,88 The exact molecular mechanisms of these events are still unclear. In addition, the effect of surface chemistry on neutrophil migration was studied by deposition of a thin layer of “inert”, biocompatible metal such as Au–Pd alloy and titanium on patterned substrates. Neutrophils also known as neutrophilic granulocytes or polymorphonuclear leukocytes are a type of white blood cells (leukocytes) that normally make up the largest number of circulating white blood cells 1).Neutrophils are considered to be the first line of defense during inflammation and infections. Conflict-of-interest disclosure: The author declares no competing financial interests. The subsequent stimulation of neutrophils by chemokines that are presented by the endothelial luminal surface triggers the activation of leukocyte integrins, enabling firm adhesion and arrest of neutrophils on the endothelial surface.
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