Novel characteristics of soluble fibrin: hypercoagulability and acceleration of blood sedimentation rate mediated by its generation of erythrocyte-linked fibers.

Soluble fibrin (SF) in blood consists of monomers lacking both fibrinopeptides A with a minor population in multimeric clusters. It is a substantial component of isolated fibrinogen (fg), which spontaneously self-assembles into protofibrils progressing to fibers at sub-physiologic temperatures, a process enhanced by adsorption to hydrophobic and some metal surfaces. Comparisons of SF-rich (FR) and SF-depleted (FD) fg isolates disclosed distinct molecular imprints of each via an adsorption/desorption procedure using gold surfaced silica microplates. Accelerated plasminogen activator-induced lysis and decreased stiffness (G') of thrombin-induced FR fg clots were revealed by thomboelastography. Erythrocyte sedimentation (ESR) in afibrinogenemic plasma (Hematocrit 25-33%) was accelerated by FR fg nearly threefold that of FD fg. Stained smears disclosed frequent rouleaux formations and fibers linking stacked erythrocytes in contrast to no rouleaux by FD fg. Rouleaux formations were more pronounced at 4 °C than at ambient temperatures and at fiber-membrane contacts displayed irregular, knobby membrane contours. One of several FR fg isolates also displayed incomplete fiber networks in cell-free areas. What is more, pre-mixing FR fg with each of three monoclonal IgG anti-fg antibodies at 1.5 mol/mol fg, that inhibited fibrin polymerization, prevented rouleaux formation save occasional 2-4 erythrocyte aggregates. We conclude that spontaneously generated SF fibers bound to erythrocytes forming intercellular links culminating in rouleaux formation and ensuing ESR acceleration which in clinical settings reflects hypercoagulability. Also, the results can explain the reported fg binding to erythrocytes via ligands such as CD47, stable in vivo RBC aggregates in capillaries, and red areas of pathologic thrombi.

Molecular Basis for Surface-Initiated Non-Thrombin-Generated Clot Formation Following Viral Infection.

In this paper, we propose a model that connects two standard inflammatory responses to viral infection, namely, elevation of fibrinogen and the lipid drop shower, to the initiation of non-thrombin-generated clot formation. In order to understand the molecular basis for the formation of non-thrombin-generated clots following viral infection, human epithelial and Madin-Darby Canine Kidney (MDCK, epithelial) cells were infected with H1N1, OC43, and adenovirus, and conditioned media was collected, which was later used to treat human umbilical vein endothelial cells and human lung microvascular endothelial cells. After direct infection or after exposure to conditioned media from infected cells, tissue surfaces of both epithelial and endothelial cells, exposed to 8 mg/mL fibrinogen, were observed to initiate fibrillogenesis in the absence of thrombin. No fibers were observed after direct viral exposure of the endothelium or when the epithelium cells were exposed to SARS-CoV-2 isolated spike proteins. Heating the conditioned media to 60 °C had no effect on fibrillogenesis, indicating that the effect was not enzymatic but rather associated with relatively thermally stable inflammatory factors released soon after viral infection. Spontaneous fibrillogenesis had previously been reported and interpreted as being due to the release of the alpha C domains due to strong interactions of the interior of the fibrinogen molecule in contact with hydrophobic material surfaces rather than cleavage of the fibrinopeptides. Contact angle goniometry and immunohistochemistry were used to demonstrate that the lipids produced within the epithelium and released in the conditioned media, probably after the death of infected epithelial cells, formed a hydrophobic residue responsible for fibrillogenesis. Hence, the standard inflammatory response constitutes the ideal conditions for surface-initiated clot formation.

Enhancement of acellular biomineralization, dental pulp stem cell migration, and differentiation by hybrid fibrin gelatin scaffolds.

OBJECTIVE: The current in vitro study aims to evaluate cross-linked hydrogels with and without the addition of fibrin that could potentially be used in endodontic regeneration as a scaffold material. METHODS: Synthesis of gelatin/fibrin scaffold, and performing nanoscale characterization using cryo-electron microscopy, dynamic rheology, and XRF for structure property relations; plating dental pulp stem cells and determining mineralization, migration, and differentiation using rt-PCR, XRF, and Raman spectroscopy. RESULTS: Cryo electron imaging shows gelatin and fibrin, when gelled separately to form classical rectangular cross-linked networks, where the modulus scales inversely with the cube root of the mesh size. When gelled together, a network with a fundamentally different structure is formed, which has higher ductility and when placed as a scaffold in osteogenic media, produces twice the mineral content. Immunofluorescence, RT-PCR and Rahman Spectroscopy indicate that the hybrid gel enhances cell migration, induces odontogenic differentiation of dental pulp stem cells, and promotes formation of dentin. SIGNIFICANCE: The mechanical properties of the hybrid gel scaffold enhance in-migration of stem cells and subsequent differentiation, which are critical for regenerative procedures. Under acellular conditions, placement of the hybrid gel enhances biomineralization, which would strengthen the root if used as a scaffold for endodontic regeneration. Our in vitro findings are consistent with previous in vivo studies which show improved mineralization when bleeding is induced into the canal, given that fibrin is a primary component in blood clotting. Therefore, insertion of the hybrid gelatin-fibrin scaffold could enable more reproducible and consistent outcomes if used for regenerative endodontic treatment (RET).

In Vitro Toxicity of Bone Graft Materials to Human Mineralizing Cells.

Bone graft materials from synthetic, bovine, and human sources were analyzed and tested for in vitro cytotoxicity on dental pulp stem cells (DPSCs) and osteosarcoma cells (Saos-2). Raman spectroscopy indicated significant amounts of collagen only in human bone-derived materials, where the mineral to protein ratio was 3.55 ± 0.45, consistent with bone. X-ray fluorescence revealed tungsten (W) concentrations of 463 ± 73, 400 ± 77, and 92 ± 42 ppm in synthetic, bovine, and human bone chips, respectively. When these chips were added to DPSCs on tissue culture plastic, the doubling times after two days were the same as the controls, 16.5 ± 0.5 h. Those cultured with synthetic or bovine chips were 96.5 ± 8.1 and 25.2 ± 1.4 h, respectively. Saos-2 was more sensitive. During the first two days with allogeneic or bovine graft materials, cell numbers declined. When DPSC were cultured on collagen, allogeneic and bovine bone chips did not increase doubling times. We propose cytotoxicity was associated with tungsten, where only the concentration in human bone chips was below 184 ppm, the value reported as cytotoxic in vitro. Cells on collagen were resistant to bone chips, possibly due to tungsten adsorption by collagen.

Controlling the Anionic Ratio and Gradient in Kesterite Technology.

Accurate anionic control during the formation of chalcogenide solid solutions is fundamental for tuning the physicochemical properties of this class of materials. Compositional grading is the key aspect of band gap engineering and is especially valuable at the device interfaces for an optimum band alignment, for controlling interface defects and recombination and for optimizing the formation of carrier-selective contacts. However, a simple and reliable technique that allows standardizing anionic compositional profiles is currently missing for kesterites and the feasibility of achieving a compositional gradient remains a challenging task. This work aims at addressing these issues by a simple and innovative technique. It basically consists of first preparing a pure sulfide absorber with a specific thickness followed by the synthesis of a pure selenide part of complementary thickness on top of it. Specifically, the technique is applied to the synthesis of Cu2ZnSn(S,Se)4 and Cu2ZnGe(S,Se)4 kesterite absorbers, and a series of characterizations are performed to understand the anionic redistribution within the absorbers. For identical processing conditions, different Se incorporation dynamics is identified for Sn- and Ge-based kesterites, leading to a homogeneous or graded composition in depth. It is first demonstrated that for Sn-based kesterite the anionic composition can be perfectly controlled through the thicknesses ratio of the sulfide and selenide absorber parts. Then, it is demonstrated that for Ge-based kesterite an anionic (Se-S) gradient is obtained and that by adjusting the processing conditions the composition at the back side can be finely tuned. This technique represents an innovative approach that will help to improve the compositional reproducibility and determine a band gap grading strategy pathway for kesterites. Furthermore, due to its simplicity and reliability, the proposed methodology could be extended to other chalcogenide materials.

Fibers Generated by Plasma Des-AA Fibrin Monomers and Protofibril/Fibrinogen Clusters Bind Platelets: Clinical and Nonclinical Implications.

Objective Soluble fibrin (SF) is a substantial component of plasma fibrinogen (fg), but its composition, functions, and clinical relevance remain unclear. The study aimed to evaluate the molecular composition and procoagulant function(s) of SF. Materials and Methods Cryoprecipitable, SF-rich (FR) and cryosoluble, SF-depleted (FD) fg isolates were prepared and adsorbed on one hydrophilic and two hydrophobic surfaces and scanned by atomic force microscopy (AFM). Standard procedures were used for fibrin polymerization, crosslinking by factor XIII, electrophoresis, and platelet adhesion. Results Relative to FD fg, thrombin-induced polymerization of FR fg was accelerated and that induced by reptilase was markedly delayed, attributable to its decreased (fibrinopeptide A) FpA. FR fg adsorption to each surface yielded polymeric clusters and co-cryoprecipitable solitary monomers. Cluster components were crosslinked by factor XIII and comprised ≤21% of FR fg. In contrast to FD fg, FR fg adsorption on hydrophobic surfaces resulted in fiber generation enabled by both clusters and solitary monomers. This began with numerous short protofibrils, which following prolonged adsorption increased in number and length and culminated in surface-linked three-dimensional fiber networks that bound platelets. Conclusion The abundance of adsorbed protofibrils resulted from (1) protofibril/fg clusters whose fg was dissociated during adsorption, and (2) adsorbed des-AA monomers that attracted solution counterparts initiating protofibril assembly and elongation by their continued incorporation. The substantial presence of both components in transfused plasma and cryoprecipitate augments hemostasis by accelerating thrombin-induced fibrin polymerization and by tightly anchoring the resulting clot to the underlying wound or to other abnormal vascular surfaces.

Cavitation and fatal hemoptysis after immunotherapy for advanced lung adenocarcinoma: A case report.

Immune checkpoint inhibitor (ICI)-related massive hemoptysis with cavitation has rarely been identified. Here, we report a case of advanced lung adenocarcinoma with lethal bleeding after eight cycles of pembrolizumab. A 55-year-old male was diagnosed with stage IV non-small cell lung cancer (NSCLC). Following confirmation of high programmed death-ligand 1 (PD-L1) expression of 60% cancer cells, he subsequently received pembrolizumab monotherapy. His symptoms and chest images significantly improved after four cycles of therapy. However, after eight cycles of immunotherapy, he presented with recurrence of bloody sputum and shortness of breath. Pembrolizumab was discontinued and a diagnosis of checkpoint inhibitor-associated pneumonitis (CIP) was made. When the CIP was absorbed after glucocorticoid therapy, the patient died of sudden massive hemoptysis with cavitation in the lesion. KEY POINTS: Although checkpoint inhibitor associated pneumonitis was the leading cause of ICI-related death, clinicians should be alerted to the finding that more attention should be given to hemoptysis attributed to ICI therapy in advanced lung cancer.

1,5-Bis(4-bromo-phen-yl)-3-phenyl-pentane-1,5-dione.

The asymmetric unit of the title compound, C(23)H(18)Br(2)O(2), contains two independent mol-ecules with slightly different conformations. In the absence of classical inter-molecular inter-actions, the crystal packing is stabilized by van der Waals forces.

Ultrasound-Guided Percutaneous Core Needle Biopsy for the Diagnosis of Pancreatic Disease.

Few studies have evaluated the performance of percutaneous core needle biopsies of the pancreas. This article is an overview of the advantages, disadvantages, predictive power and complications associated with percutaneous ultrasound-guided core needle biopsies of pancreatic lesions. A comprehensive literature search of Medline (using PubMed as the search engine) and EMBASE was done to identify suitable studies up to March 2017. A study of quantitative pre-operative pancreatic biopsy data was reported. Lesion location, mean or median number of passes, inadequate tissue or technical failures and complications were assessed for all cases by reviewing clinical notes and post-procedural imaging. The analysis included 13 studies, mostly of a retrospective nature. The sensitivity (mean: 94.42%, range: 90%-100%) and specificity (mean: 97.94%, range: 94.7%-100%) of the procedure were high, and the mean accuracy of diagnosis was 95.76 (range: 91-100). Furthermore, the procedure had a high negative predictive value of approximately 76.26%. Of the 13 reported studies, 7.3% were inadequate or technical failure cases. The mean rate of complications was 2.08%, which seemed similar to the lower limit of this rate for endoscopic ultrasound-guided fine-needle aspirations. The risk of tumor seeding with ultrasound-guided core needle biopsies was not reported in the included articles. With the development of technology, ultrasound-guided percutaneous core needle biopsy for pancreatic lesions is increasingly available and has optimal diagnostic power in pancreatic neoplasms.

Correction: Manipulation of cell adhesion and dynamics using RGD functionalized polymers.

Correction for 'Manipulation of cell adhesion and dynamics using RGD functionalized polymers' by Juyi Li et al., J. Mater. Chem. B, 2017, DOI: .

Manipulation of cell adhesion and dynamics using RGD functionalized polymers.

We have successfully synthesized an ABA tri-block co-polymer of poly(methacrylic acid)-block-poly(2-hydroxyethyl methacrylate)-block-poly(methacrylic acid), having Mw = 100k and 272k where we were able to insert RDG or RGD peptide sequences using thiol-acrylate Michael addition. A soft silicone stamp was then used to imprint a 0.4-micron wide grating of the copolymer with a period of 10 microns. The samples were then examined with atomic force microscopy after application of an external electric field and the pattern was observed to stretch by a factor of five. Cells plated onto these substrates showed clear preference for the striped patterns formed under the influence of the external field, and no preferential attachment to the patterns formed in the absence of the field. Cell migration experiments, using the agarose droplet method, performed on spun cast copolymer films showed minimal migration and adhesion on the substrates without peptides or those with only with the RDG peptide, while good adhesion and significant outward migration was observed for cells plated on the copolymers with the RGD sequence. Taken together these results confirmed our hypothesis that a smart biomimetic polymer substrate could be constructed where functional domains could be revealed selectively allowing us to mimic the natural design of engineered tissue constructs.