Coagulation factor XIII: a multifunctional transglutaminase with clinical potential in a range of conditions

Coagulation Coagulation factor XIII (FXIII), a plasma transglutaminase, is best known as the final enzyme in the coagulation cascade, where it is responsible for cross-linking of fibrin. However, a growing body of evidence has demonstrated that FXIII targets a wide range of additional substrates that have important roles in health and disease. These include antifibrinolytic proteins, with cross-linking of alpha(2)-antiplasmin to fibrin, and potentially fibrinogen, being the principal mechanism(s) whereby plasmin-mediated clot degradation is minimised. FXIII also acts on endothelial cell VEGFR-2 and alpha(v)beta(3) integrin, which ultimately leads to downregulation of the antiangiogenic protein thrombospondin-1, promoting angiogenesis and neovascularisation. Under infectious disease conditions, FXIII cross-links bacterial surface proteins to. fibrinogen, resulting in immobilisation and killing, while during wound healing, FXIII induces-cross-linking of the provisional matrix. The latter process has been shown to influence the interaction of leukocytes with the provisional extracellular matrix and promote wound healing. Through these actions, there are good rationales for evaluating the therapeutic potential of FXIII in diseases in which tissue repair is dys-regulated or perturbed, including systemic sclerosis (scleroderma), invasive bacterial infections, and tissue repair, for instance healing of venous leg ulcers or myocardial injuries. Adequate levels of FXIII are also required in patients undergoing surgery to prevent or treat perioperative bleeding, and its augmentation in patients with/at risk for perioperative bleeding may also have potential clinical benefit. While there are preclinical and/or clinical data to support the use of FXIII in a range of settings, further clinical evaluation in these underexplored applications is warranted

The Effect of Secretory Factors of Adipose-Derived Stem Cells on Human Keratinocytes

The beneficial effects of adipose-derived stem cell conditioned medium (ADSC-CM) on skin regeneration have been reported. Although the mechanism of how ADSC-CM promotes skin regeneration is unclear, ADSC-CM contained various growth factors and it is an excellent raw material for skin treatment. ADSC-CM produced in a hypoxia condition of ADSC—in other words, Advanced Adipose-Derived Stem cell Protein Extract (AAPE)—has great merits for skin regeneration. In this study, human primary keratinocytes (HKs), which play fundamental roles in skin tissue, was used to examine how AAPE affects HK. HK proliferation was significantly higher in the experimental group (1.22 μg/mL) than in the control group. DNA gene chip demonstrated that AAPE in keratinocytes (p < 0.05) notably affected expression of 290 identified transcripts, which were associated with cell proliferation, cycle and migration. More keratinocyte wound healing and migration was shown in the experimental group (1.22 μg/mL). AAPE treatment significantly stimulated stress fiber formation, which was linked to the RhoA-ROCK pathway. We identified 48 protein spots in 2-D gel analysis and selected proteins were divided into 64% collagen components and 30% non-collagen components as shown by the MALDI-TOF analysis. Antibody array results contained growth factor/cytokine such as HGF, FGF-1, G-CSF, GM-CSF, IL-6, VEGF, and TGF-β3 differing from that shown by 2-D analysis. Conclusion: AAPE activates HK proliferation and migration. These results highlight the potential of the topical application of AAPE in the treatment of skin regeneration

Encephalopathy associated with autoimmune thyroid disease in patients with Graves' disease: clinical manifestations, follow-up, and outcomes

<p>Abstract</p> <p>Background</p> <p>The encephalopathy associated with autoimmune thyroid disease (EAATD) is characterized by neurological/psychiatric symptoms, high levels of anti-thyroid antibodies, increased cerebrospinal fluid protein concentration, non-specific electroencephalogram abnormalities, and responsiveness to the corticosteroid treatment in patients with an autoimmune thyroid disease. Almost all EAATD patients are affected by Hashimoto's thyroiditis (HT), although fourteen EAATD patients with Graves' disease (GD) have been also reported.</p> <p>Methods</p> <p>We have recorded and analyzed the clinical, biological, radiological, and electrophysiological findings and the data on the therapeutic management of all GD patients with EAATD reported so far as well as the clinical outcomes in those followed-up in the long term.</p> <p>Results</p> <p>Twelve of the fourteen patients with EAATD and GD were women. The majority of GD patients with EAATD presented with mild hyperthyroidism at EAATD onset or shortly before it. Active anti-thyroid autoimmunity was detected in all cases. Most of the patients dramatically responded to corticosteroids. The long term clinical outcome was benign but EAATD can relapse, especially at the time of corticosteroid dose tapering or withdrawal. GD and HT patients with EAATD present with a similar clinical, biological, radiological, and electrophysiological picture and require an unaffected EAATD management.</p> <p>Conclusions</p> <p>GD and HT equally represent the possible background condition for the development of EAATD, which should be considered in the differential diagnosis of all patients with encephalopathy of unknown origin and an autoimmune thyroid disease, regardless of the nature of the underlying autoimmune thyroid disease.</p

Antibody-mediated Prevention of Fusarium Mycotoxins in the Field

Fusarium mycotoxins directly accumulated in grains during the infection of wheat and other cereal crops by Fusarium head blight (FHB) pathogens are detrimental to humans and domesticated animals. Prevention of the mycotoxins via the development of FHB-resistant varieties has been a challenge due to the scarcity of natural resistance against FHB pathogens. Various antibodies specific to Fusarium fungi and mycotoxins are widely used in immunoassays and antibody-mediated resistance in planta against Fusarium pathogens has been demonstrated. Antibodies fused to antifungal proteins have been shown to confer a very significantly enhanced Fusarium resistance in transgenic plants. Thus, antibody fusions hold great promise as an effective tool for the prevention of mycotoxin contaminations in cereal grains. This review highlights the utilization of protective antibodies derived from phage display to increase endogenous resistance of wheat to FHB pathogens and consequently to reduce mycotoxins in field. The role played by Fusarium-specific antibody in the resistance is also discussed

Antikörper vermittelte Fusarien-Resistenz in transgenen Pflanzen

Pathogenic fungi lead to large yield losses in agriculture and in ornamental plant production. They attack plants as well as seeds and contaminate them with mycotoxins and other metabolites. Resistant wild type species are often not available, so the only possibility to combat fungal pathogen is the use of fungicides which include risks for environment and the consumers as well as costs. The use of recombinant antibodies in transgenic plant to generate fungal pathogen resistant plants is a promising way to solve these problems in a consumer-friendly way. The goal of this work was to generate scFv antibody fragments against surface structures of Fusarium. These scFvs were be fused to antifungal peptides and proteins (AFP) and expressed in plants and should mediate resistance against the pathogenic fungi in planta. To generate scFv antibodies chicken were immunized with different fungal antigen-preparations. The total-RNA and the mRNA was isolated from spleen cells, the cDNS was synthesized, the VH- and VL-domains were amplified and assembled into a phagemid vector via restriction sites to generate scFv libraries. The diversity of the libraries was determined and the most promising libraries were combined and panned via Phage Display against fungal antigens. Soluble scFvs were tested for antigen binding activity after the third round of panning and the best binding scFvs were expressed in bacteria. The purified scFvs CWPD2, FPCWPA5 and SGB3 were characterised by ELISA, immunoblot and immunofluorescence. The scFvs were fused to different AFP and the scFvs as well as the AFP-scFv-fusions were cloned into different vector- and expression systems and their inhibitory effect was demonstrated in vitro. Resistent tests on stable transformed F2-plants of Arabidopsis demonstrated that the AFP-scFv constructs AG-scFv CWPD2, RS-scFv CWPD2, Lacto-scFv CWPD2 and Chi-scFv CWPD2 mediated antibody-resistance against F. oxysporum f.sp. matthiolae in A. thaliana cv. Columbia plants

Antikörper vermittelte Fusarien-Resistenz in transgenen Pflanzen

Pathogenic fungi lead to large yield losses in agriculture and in ornamental plant production. They attack plants as well as seeds and contaminate them with mycotoxins and other metabolites. Resistant wild type species are often not available, so the only possibility to combat fungal pathogen is the use of fungicides which include risks for environment and the consumers as well as costs. The use of recombinant antibodies in transgenic plant to generate fungal pathogen resistant plants is a promising way to solve these problems in a consumer-friendly way. The goal of this work was to generate scFv antibody fragments against surface structures of Fusarium. These scFvs were be fused to antifungal peptides and proteins (AFP) and expressed in plants and should mediate resistance against the pathogenic fungi in planta. To generate scFv antibodies chicken were immunized with different fungal antigen-preparations. The total-RNA and the mRNA was isolated from spleen cells, the cDNS was synthesized, the VH- and VL-domains were amplified and assembled into a phagemid vector via restriction sites to generate scFv libraries. The diversity of the libraries was determined and the most promising libraries were combined and panned via Phage Display against fungal antigens. Soluble scFvs were tested for antigen binding activity after the third round of panning and the best binding scFvs were expressed in bacteria. The purified scFvs CWPD2, FPCWPA5 and SGB3 were characterised by ELISA, immunoblot and immunofluorescence. The scFvs were fused to different AFP and the scFvs as well as the AFP-scFv-fusions were cloned into different vector- and expression systems and their inhibitory effect was demonstrated in vitro. Resistent tests on stable transformed F2-plants of Arabidopsis demonstrated that the AFP-scFv constructs AG-scFv CWPD2, RS-scFv CWPD2, Lacto-scFv CWPD2 and Chi-scFv CWPD2 mediated antibody-resistance against F. oxysporum f.sp. matthiolae in A. thaliana cv. Columbia plants