Utilization of a direct sandwich ELISA kit for rapid diagnosis of adenovirus hemorrhagic cystitis in a patient with adult T-cell leukemia

We report on a case of adenovirus hemorrhagi cystitis that developed in a 49-year-old woman during intensive chemotherapy for adult T-cell leukemia. Although rapid etiologic diagnosis is essential for the effective management of viral hemorrhagic cystitis, the isolation of adenovirus from urine is often too time-cousuming. We detected the adenovirus antigen in the patient’s urine using an Adenoclone direct sandwich ELISA kit (Cambridge Bio Science), which is commonly employed for the diagnosis of adenovirus conjunctivitis. Treatment consisted of intravenous vidarabine and bladder irrigation, which resulted in prompt clinical improvement. The Adenoclone kit was useful for the rapid etiologic diagnosis of adenovirus hemmorrhagic cystitis.

Determination of okadaic acid content of dinoflagellate cells: a comparison of the HPLC-fluorescent method and two monoclonal antibody ELISA test kits

Total okadaic acid (okadaic acid plus methylokadaic acid) in acclimated clones of the dinoflagellates Prorocentrum hoffmannianum and P. lima was determined using the HPLC-fluorescent method, UBE ELISA test kit, and Rougier ELISA test kit. The nonokadaic acid-producing species. Amphidinium klebsii, Prorocentrum mexicanum, P. micans, P. cassubicum, and Gambierdiscus toxicus were examined using the same methods of analysis. All three methods yielded consistent results for P. hoffmannianum which produces only okadaic acid.
However, results of the three methods were not consistent for P. lima which produces both okadaic acid and methylokadaic acid. The UBE ELISA demonstrated little or no cross-reactivity with methylokadaic acid; whereas, the Rougier ELISA demonstrated varying degrees of cross-reactivity with that analog. Analyses of nonokadaic acid producing-species yielded negative results, with one exception. The Rougier ELISA demonstrated reactivity with extracts of G. toxicus. Since outbreaks of DSP may be caused by okadaic acid, methylokadaic acid, or a combination of these toxins, both ELISA kits may underestimate total toxin present in toxic shellfish.

Silk-based matrices and c-Kit positive cardiac progenitor cells for a cellularized silk fibroin scaffold: study of an in vivo model

The production of a cellularized silk fibroin scaffold is very difficult because it is actually impossible to differentiate cells into a well-organized cardiac tissue. Without vascularization, not only do cell masses fail to grow, but they may also exhibit an area of necrosis, indicating a lack of oxygen and nutrients. In the present study, we used the so-called tyrosine protein kinase kit (c-kit)-positive cardiac progenitor cells (CPCs) to generate cardiac cellularized silk fibroin scaffolds, multipotent cells isolated from the adult heart to date that can show some degree of differentiation toward the cardiac phenotype. To test their ability to differentiate into the cardiac phenotype in vivo as well, CPC and collagen organoid-like masses were implanted into nude mice and their behavior observed. Since the three-dimensional structure of cardiac tissue can be preserved by scaffolds, we prepared in parallel different silk fibroin scaffolds with three different geometries and tested their behavior in three different models of immunosuppressed animals.
Unfortunately, CPC cellularized silk fibroin scaffolds cannot be used in vivo. CPCs implanted alone or in collagen type I gel were destroyed by CD3+ lymphocytes’ aggregates, whereas the porous and partially oriented scaffolds elicited a consistent foreign body response characterized by giant cells. Only the electrospun meshes were resistant to the foreign body reaction. In conclusion, c-kit-positive CPCs, although expressing a good level of cardiac differentiation markers in vitro with or without fibroin meshes, are not suitable for an in vivo model of cardiac organoids because they are degraded by a T-cell-mediated immune response. Even scaffolds, which may preserve the survival of these cells in vivo, also induced a host response. However, among the tested scaffolds, the electrospun meshes (F-scaffold) induced a lower response compared to all the other tested structures.

Autocrine/Paracrine Loop Between SCF +/c-Kit + Mast Cells Promotes Cutaneous Melanoma Progression

c-Kit, or mast/stem cell growth factor receptor Kit, is a tyrosine kinase receptor structurally analogous to the colony-stimulating factor-1 (CSF-1) and platelet-derived growth factor (PDGF) CSF-1/PDGF receptor Tyr-subfamily. It binds the cytokine KITLG/SCF to regulate cell survival and proliferation, hematopoiesis, stem cell maintenance, gametogenesis, mast cell development, migration and function, and it plays an essential role in melanogenesis. SCF and c-Kit are biologically active as membrane-bound and soluble forms. They can be expressed by tumor cells and cells of the microenvironment playing a crucial role in tumor development, progression, and relapses.
To date, few investigations have concerned the role of SCF+/c-Kit+ mast cells in normal, premalignant, and malignant skin lesions that resemble steps of malignant melanoma progression. In this study, by immunolabeling reactions, we demonstrated that in melanoma lesions, SCF and c-Kit were expressed in mast cells and released by themselves, suggesting an autocrine/paracrine loop might be implicated in regulatory mechanisms of neoangiogenesis and tumor progression in human melanoma.

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Christopher Miller