Treating Melanoma in Situ During a Pandemic with Telemedicine and a Combination of Imiquimod, 5-Fluorouracil, and Tretinoin.

The recent coronavirus disease 2019 (COVID-19) pandemic has created a quandary for the physician in terms of evaluating and treating cutaneous skin cancers, particularly melanomas. At the onset of the pandemic, many planned medical and surgical visits for skin cancers were postponed. Physicians and patients have had to balance the risk of exposure to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with that of worsening morbidity and mortality due to delays in skin cancer treatments. We present a male patient who had two melanoma-in-situs (MISs) that were treated during the COVID-19 pandemic with a combination of topical imiquimod 5% cream, 5-fluorouracil 2% solution, and tretinoin 0.1% cream. The successful treatments occurred without in-person visits and with the aid of telemedicine. Although surgery is the standard for the treatment of melanoma in situ, this case demonstrates an effective viable treatment modality for MIS during a pandemic situation.

Tensile strength, growth factor content and proliferation activities for two platelet concentrates of platelet-rich fibrin and concentrated growth factor.

BACKGROUND/PURPOSE: Platelet-rich fibrin (PRF) can be obtained by centrifuging fresh blood in the absence of anticoagulants. Varying the centrifugation speeds may produce tougher and richer concentrated growth factors (CGF). This study examines tensile strength, growth factor content, and the potential of CGF and PRF in promoting periodontal cell proliferation. MATERIALS AND METHODS: Blood (40 mL/subject) was collected from 44 healthy subjects. PRF and CGF were prepared by centrifuging at 3000 rpm and switching speeds ranging within 3000 rpm, respectively. Fibrin strip was prepared and its tensile strength was measured. Transforming growth factor beta 1 (TGF-ß1), platelet-derived growth factor BB (PDGF-BB), and epidermal growth factor (EGF) in the residual serum and fibrin clots were determined by enzyme-linked immunosorbent assay, and their effects on the proliferation of hFOB1.19 osteoblasts and human gingival fibroblasts were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. RESULTS: Compared with PRF, tensile strength of CGF was significantly higher. Concentrations and amounts of PDGF-BB and EGF in CGF were significantly higher than those in PRF. Osteoblast number was significantly higher in the cultures with fetal bovine serum (FBS, 10%) and with PRF or CGF fibrin clots (5%, 10%, and 50%) compared to that without FBS. Moreover, osteoblast number in CGF, regardless of the preparation of 10% and 50%, was significantly greater than that in PRF. Similar findings were also observed for gingival fibroblasts among the various subjects. CONCLUSION: Varying centrifugation speeds can modify the tensile strength and biological activities of platelet fibrin clots.

Crystallographic studies of the binding of ligands to the dicarboxylate site of Complex II, and the identity of the ligand in the "oxaloacetate-inhibited" state.

Mitochondrial Complex II (succinate:ubiquinone oxidoreductase) is purified in a partially inactivated state, which can be activated by removal of tightly bound oxaloacetate (E.B. Kearney, et al., Biochem. Biophys. Res. Commun. 49 1115-1121). We crystallized Complex II in the presence of oxaloacetate or with the endogenous inhibitor bound. The structure showed a ligand essentially identical to the "malate-like intermediate" found in Shewanella Flavocytochrome c crystallized with fumarate (P. Taylor, et al., Nat. Struct. Biol. 6 1108-1112) Crystallization of Complex II in the presence of excess fumarate also gave the malate-like intermediate or a mixture of that and fumarate at the active site. In order to more conveniently monitor the occupation state of the dicarboxylate site, we are developing a library of UV/Vis spectral effects induced by binding different ligands to the site. Treatment with fumarate results in rapid development of the fumarate difference spectrum and then a very slow conversion into a species spectrally similar to the OAA-liganded complex. Complex II is known to be capable of oxidizing malate to the enol form of oxaloacetate (Y.O. Belikova, et al., Biochim. Biophys. Acta 936 1-9). The observations above suggest it may also be capable of interconverting fumarate and malate. It may be useful for understanding the mechanism and regulation of the enzyme to identify the malate-like intermediate and its pathway of formation from oxaloacetate or fumarate.

3-nitropropionic acid is a suicide inhibitor of mitochondrial respiration that, upon oxidation by complex II, forms a covalent adduct with a catalytic base arginine in the active site of the enzyme.

We report three new structures of mitochondrial respiratory Complex II (succinate ubiquinone oxidoreductase, E.C. 1.3.5.1) at up to 2.1 A resolution, with various inhibitors. The structures define the conformation of the bound inhibitors and suggest the residues involved in substrate binding and catalysis at the dicarboxylate site. In particular they support the role of Arg(297) as a general base catalyst accepting a proton in the dehydrogenation of succinate. The dicarboxylate ligand in oxaloacetate-containing crystals appears to be the same as that reported for Shewanella flavocytochrome c treated with fumarate. The plant and fungal toxin 3-nitropropionic acid, an irreversible inactivator of succinate dehydrogenase, forms a covalent adduct with the side chain of Arg(297). The modification eliminates a trypsin cleavage site in the flavoprotein, and tandem mass spectroscopic analysis of the new fragment shows the mass of Arg(297) to be increased by 83 Da and to have the potential of losing 44 Da, consistent with decarboxylation, during fragmentation.

Crystallization of mitochondrial respiratory complex II from chicken heart: a membrane-protein complex diffracting to 2.0 A.

A procedure is presented for preparation of diffraction-quality crystals of a vertebrate mitochondrial respiratory complex II. The crystals have the potential to diffract to at least 2.0 A with optimization of post-crystal-growth treatment and cryoprotection. This should allow determination of the structure of this important and medically relevant membrane-protein complex at near-atomic resolution and provide great detail of the mode of binding of substrates and inhibitors at the two substrate-binding sites.

Innovative therapies in wound healing.

BACKGROUND: Apligraf is a bioengineered skin product composed of neonatal fibroblasts and keratinocytes. The FDA has approved Apligraf for the treatment of chronic venous ulcers and diabetic ulcers. OBJECTIVE: We review the development of bioengineered skin, examine the cellular activities of various growth factors that may facilitate wound healing, and discuss the results of clinical trials with a particular construct, Apligraf, as proof of principle. CONCLUSION: Bioengineered skin acts as a "smart" delivery system for growth factors and other stimulatory substances. Not only does it present a novel treatment for chronic and diabetic ulcers, but it could also be considered for application to other types of acute wounds.

Growth factors, signal transduction, and cellular responses.

The extraordinary advances in the field of growth factors and signal transduction have created new and promising therapeutic interventions. We intend to explain the difficult nomenclatures associated with growth factors and their mechanisms of action.

Treatment of Venous Ulcers Using a Bilayered Living Skin Construct.

Venous ulcers are a common cause of pain and suffering in the elderly US population. Up to 1 million individuals suffer from this disease, and the true costs in terms of medical care and quality of life are only beginning to be appreciated. Bioengineered skin constructs are being used increasingly to treat venous ulcers with some success and can be used if traditional first-line treatment methods fail.