Siniscalco's Group

department of experimental medicine – division of pharmacology - Second University of Naples - Italy
Group Leader
Dario Siniscalco
Catia Giordano, Livio Luongo, Vito de Novellis, Sabatino Maione, Francesco Rossi
Scientific interests
Stem cell activation. Use of human mesenchymal stem cells as therapeutic tool in neurodegeneration.
Stem cells therapy for the neuronal recovery after central and peripheral nervous system injury.
Studying of neuronal apoptosis. Role of bcl-2 family, caspases, and cell cycle regulator genes.
Apoptotic genes expression. Apoptosis and neuropathic pain. Role of reactive oxygen species in apoptosis.
Molecular regulation of neural stem cell death. Apoptosis in neural precursor cells.
Osteoclastic cell activation.
Role of metabotropic glutamate receptors in the neuropathic pain pathway. Role of ozone in neuropathic pain treatment.
Protein expression at low temperature.
Expression methods for heterologous proteins; chemical and mechanical cellular lysis; sonication; protein extraction and purification; SDS PAGE; Western blotting; dot immunobinding; methods for the determination of protein concentration; analysis of protein activity; ELISA assay.
Genetic engineering, technology of DNA recombination. Enzymatic catalysis on DNA; gene cloning; extraction and purification of plasmidic and genomic DNA; extraction and purification of RNA (from cell culture, tissue, formalin-fixed, paraffin-embedded tissue sections); PCR; RT-PCR; Real Time PCR; primers design; DNA sequencing; DNA and RNA gel electrophoresis; methods for the determination of nucleic acids concentration; methods for studying gene expression variation. Using of mice knock out.
Stem cell culture, human mesenchymal bone marrow-derived stem cell culture; cell culture (bacteria, neuronal, viral); human monocytes; human tumour cells; human osteoclast cells; growth curves; preparation of competent cells; chemical transformation; electroporation; coniugation; viral infection and amplification; chemical and physical sterilization. In vitro methods for studying apoptosis.
HPLC; GC-MS; UV-VIS spectrophotometry; solvent extraction.
Immunoistochemistry; immunocytochemistry; fixation, dehydration/hydration; slides preparation; using of criotome and microtome; optical microscopy; fluorescent microscopy; antigen retrieval; double/triple staining; signal amplification (i.e. cy3, cy3 plus, strep-cy3, FITC, DAPI, Quantum Dots, TSA); TUNEL reaction. Multi-label quantum dot immunohistochemical detection procedure.
PubMed, ChemSketch, Macplasmap, Chemfinder, Molecular Analyst (BioRad), Quantity One (BioRad), Basic Local Alignment Search Tool (BLAST), FASTA, PROW (Protein Reviews on the Web), Lasergene (DNASTAR), EditSeq, Oligo 4.05 (Primer Analysis Software – National Biosciences), Primer Express (Applied Biosystems), CLC DNA Workbench, Beacon Designer 3.01 (Premier Biosoft International), MicroPlate Manager (BioRad), KJ-Junior (Bio-Tek), Soft Max Pro (Molecular Devices), Axiovision 2.0 e 3.1 (Zeiss), IM1000 (Leica), NanoDrop 3.0.0, NanoDrop 1000 V 3.2.1.
Data analysis; preparing data for presentation in peer-reviewed journals and at professional meetings; preparing of manuscripts for submission to peer-reviewed journals. Preparing of lab meeting, progress reports, talks, grant proposals. Tutor activity for internal students. Lab safety.
relevant publications
1. Siniscalco D. Transplantation of human mesenchymal stem cells in the study of neuropathic pain. Methods Mol Biol. 2010; 617:337-45.
2. Siniscalco D, Giordano C, Galderisi U, Luongo L, Alessio N, Di Bernardo G, de Novellis V, Rossi F, Maione S. Intra-brain microinjection of human mesenchymal stem cells decreases allodynia in neuropathic mice. Cell Mol Life Sci. 2010, 67: 655–669.
3. D'Agostino B, Sullo N, Siniscalco D, De Angelis A, Rossi F. Mesenchymal stem cell therapy for the treatment of chronic obstructive pulmonary disease. Expert Opin Biol Ther. 2010; 10(5):681-7.
4. D. Siniscalco, F. Rossi, S. Maione. Stem cell therapy for neuropathic pain treatment. Journal of Stem Cells and Regenerative Medicine 2008, vol III (1).
Scientific instruments
Cell culture fully equipped lab.
Molecular biology lab: thermal cyclers, gel electrophoresis systems, Gel Doc 2000 UV System,  UV-Vis spectrophotometers, ultracentrifuges,  microplate reader, western blot analysis system.

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