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103 results on '"Seidita, G."'

1. Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exercise

Author

Robino, C., Ralf, A., Pasino, S., De Marchi, M.R., Ballantyne, K.N., Barbaro, A., Bini, C., Carnevali, E., Casarino, L., Di Gaetano, C., Fabbri, M., Ferri, G., Giardina, E., Gonzalez, A., Matullo, G., Nutini, A.L., Onofri, V., Piccinini, A., Piglionica, M., Ponzano, E., Previderè, C., Resta, N., Scarnicci, F., Seidita, G., Sorçaburu-Cigliero, S., Turrina, S., Verzeletti, A., and Kayser, M.

Published
2015
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5. WITHDRAWN: Corrigendum to ‘Development of an Italian RM Y-STR haplotype database: results of the 2013 GEFI collaborative exercise’ [Forensic. Sci. Int. Genet. 15 (2015) 56-63]

Author

Robino, C, Ralf, A, Pasino, S, De Marchi, MR, Ballantyne, KN, Barbaro, A, Bini, C, Carnevali, E, Casarino, L, Di Gaetano, C, Fabbri, M, Ferri, G, Giardina, E, Gonzalez, A, Matullo, G, Nutini, AL, Onofri, V, Piccinini, A, Piglionica, M, Ponzano, E, Previderè, C, Resta, N, Scarnicci, F, Seidita, G, Sorçaburu-Cigliero, S, Turrina, S, Verzeletti, A, and Kayser, M

Published
2018
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6. Corrigendum to 'Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exercise' [Forensic. Sci. Int. Genet. 15 (2015) 56–63] (S1872497314002245) (10.1016/j.fsigen.2014.10.008))

Author

Robino, C., Ralf, A., Pasino, S., De Marchi, M. R., Ballantyne, K. N., Barbaro, A., Bini, C., Carnevali, E., Casarino, L., Di Gaetano, C., Fabbri, M., Ferri, G., Giardina, E., Gonzalez, A., Matullo, G., Nutini, A. L., Onofri, V., Piccinini, A., Piglionica, M., Ponzano, E., Previderè, C., Resta, N., Scarnicci, F., Seidita, G., Sorçaburu-Cigliero, S., Turrina, S., Verzeletti, A., and Kayser, M.

Subjects
Socio-culturale, LS2_6
Published
2018

7. Corrigendum to 'Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exercise' [Forensic. Sci. Int. Genet. 15 (2015) 56–63]

Author

Robino, C., Ralf, A., Pasino, S., De Marchi, M. R., Ballantyne, K. N., Barbaro, A., Bini, C., Carnevali, E., Casarino, L., Di Gaetano, C., I, Fabbri, M., Ferri, G., Giardina, E., Gonzalez, A., Matullo, G., Nutini, A. L., Onofri, V., Piccinini, A., Piglionica, M., Ponzano, E., Previderè, C., Resta, N., Scarnicci, F., Seidita, G., Sorçaburu-Cigliero, S., Turrina, S., Verzeletti, A., and Kayser, M.

Subjects
RM Y-STR, Y-chromosome
Published
2018

8. Corrigendum to “Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exercise” [Forensic. Sci. Int. Genet. 15 (2015) 56–63]

Author

Robino, C., primary, Ralf, A., additional, Pasino, S., additional, De Marchi, M.R., additional, Ballantyne, K.N., additional, Barbaro, A., additional, Bini, C., additional, Carnevali, E., additional, Casarino, L., additional, Di Gaetano, C., additional, Fabbri, M., additional, Ferri, G., additional, Giardina, E., additional, Gonzalez, A., additional, Matullo, G., additional, Nutini, A.L., additional, Onofri, V., additional, Piccinini, A., additional, Piglionica, M., additional, Ponzano, E., additional, Previderè, C., additional, Resta, N., additional, Scarnicci, F., additional, Seidita, G., additional, Sorçaburu-Cigliero, S., additional, Turrina, S., additional, Verzeletti, A., additional, and Kayser, M., additional

Published
2018
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9. A new method to valued efficiency of enzyme blends for pancreas tissue digestion

Author

Salamone, M., Seidita, G., Cutitta, A., Rigogliuso, S., La Venuta, G., Mazzola, S., Bertuzzi, F., Ghersi, G., Salamone, M, Seidita, G, Cutitta, A, Rigogliuso, S, La Venuta, G, Mazzola, S, Bertuzzi, F, and Ghersi, G

Subjects
enzyme blends, pancreas tissue digestion
Abstract

One of the best successful examples of cell therapy is represented by islet transplantation since 1990. However islet isolation methods are not completely standardized yet. More than half of isolation procedures failed to isolate adequate islets for transplantation, due to variable pancreas conditions and to unpredictable enzymatic blend efficiency. Enzymes used for pancreas digestion include collagenases and neutral proteases: their composition and activity are largely variable between different batches. We set up a new in vitro method to better in vitro characterize enzymatic blend before its use in human pancreas. In our experimental approach human immortalized cells (ECV-304) or human islets were coltured within a 3-D type-I collagen gel in 96 wells plate. After one culture day, cells and/or islets were treated with different commercial enzymes (Liberase, Serva NB1 premium grade, Collagenase type P, Thermolysin, Neutral proteases) from different batches at different concentrations and for different times. Digestion of 3-D type-I collagen fibril gels were monitored by optical dense absorption to a fix l; morphology of released cells and/or islets were valued by confocal microscopy analyses. Cells were immunostained about expression of some adhesion molecules, like: integrins, cadherins and associated molecules, catenins, to appreciate cells morphology and islets aggregation modifications. Cell viability was assessed by SYTO 13 and ethidium bromide. We found that Neutral proteases is less pure and more toxic than Thermolysin: it has collagenase activity and it significantly decreases cell viability. Even viable cells after neutral proteases showed an alterated morphology with an impairment of cell to cell communications. We, also, observed an higher efficiency in extraction by Serva NB1 compared to Liberase and Collagenase type P, used in the same experimental conditions. By this in vitro method we were able to compare the minimal active enzyme concentration of different enzyme blends: we found that Liberase has its best value as number of extracted cell/alive cells in 130 mg/ml; while, in actual protocols for human pancreas is used to 1.3 mg/ml. Preliminary results showed a correlation between data achieved in cell lines with those of human islets, thus confirming the potential predictive role of this method for the selection of enzymes for human pancreas digestion purpose.

Published
2009

10. The molecular characterisation of depurinated trial DNA sample can be a model to understand the reliability of the results in forensic genetics

Author

Fattorini, P., Previdere`, C., Sorçaburu Cigliero, S., Marrubini, G., Alù, M., Barbaro, A., Carnevali, E., Carracedo, A., Casarino, L., Consoloni, L., Corato, S., Domenici, R., Fabbri, M., Giardina, E., Grignani, P., Lonero Baldassarra, S., Moratti, M., Pelotti, S., Piccinini, A., Pitacco, P., Plizza, L., Resta, N., Ricci, U., Robino, C., Salvaderi, L., Scarnicci, F., Schneider, P. M., Seidita, G., Trizzino, L., Turchi, C., Turrina, Stefania, Vatta, P., Vecchiotti, C., Verzeletti, A., and De Stefano, F.

Subjects
DNA depurination, Forensic genetics, PCR fidelity, STR typing
Published
2014

11. Biologia e Genetica

Author

De Leo, G., Fasano, S., Ginelli, E., Alessandro, R., Altruda, F., Amato, A., Antognelli, Cinzia, Barisani, D., Brancolini, C., Defilippi, P., Delrio, G., Di Bella, M. A., Dolcemascolo, G., Fontana, S., Gianguzza, M., Hirsch, E., Meneveri, R., Mezzasoma, Letizia, Minucci, S., Mirisola, M., Modesti, A., Pierantoni, R., Purrello, M., Seidita, G., Sidoti, A., Talesa, Vincenzo Nicola, Tarone, G., Tognon, M., and Tolosano, E.

Published
2013

12. The population genetic structure of Y chromosomes in 1338 italians

Author

Onofri, V., Buscemi, L., Caenazzo, L., Capelli, C., Carnevali, E., Cerri, N., Di Gaetano, C., Domenici, R., Ferri, G., Nutini, L., Pelotti, S., Previderè, C., Robino, C., Scarnicci, F., Fabbri, Matteo, Seidita, G., Roewer, L., Tagliabracci, A., and the GeFi Group

Published
2010

15. RAS residues that are distant from the GDP binding site play a critical role in dissociation factor-stimulated release of GDP

Author

VERROTTI A. C., CRECHET J. B., DI BLASI F., SEIDITA G., MIRISOLA M. G., KAVOUNIS C., NASTOPOULOS V., BURDERI E., PARMEGGIANI A., FASANO O., DE VENDITTIS, EMMANUELE, VERROTTI DI PIANELLA, Arturo, Crechet, J. B., DI BLASI, F., Seidita, G., Mirisola, M. G., Kavounis, C., Nastopoulos, V., Burderi, E., DE VENDITTIS, Emmanuele, Parmeggiani, A., Fasano, O., and Verrotti, A. C.

Published
1992

21. Analysis of the gastrinreleasing peptide receptor gene in Italian patients with autism spectrum disordersPlease cite this article as follows: Seidita G, Mirisola M, DAnna RP, Gallo A, Jensen RT, Mantey SA, Gonzalez N, Falco M, Zingale M, Elia M, Cucina L, Chiavetta V, Romano V, Cali F. 2008. Analysis of the GastrinReleasing Peptide Receptor Gene in Italian Patients With Autism Spectrum Disorders. Am J Med Genet Part B 147B:807–813.

Author

Seidita, G., Mirisola, M., DAnna, R.P., Gallo, A., Jensen, R.T., Mantey, S.A., Gonzalez, N., Falco, M., Zingale, M., Elia, M., Cucina, L., Chiavetta, V., Romano, V., and Cali, F.

Abstract

The gastrinreleasing peptide receptor GRPR was implicated for the first time in the pathogenesis of Autism spectrum disorders ASD by IshikawaBrush et al. IshikawaBrush et al. 1997: Hum Mol Genet 6: 1241–1250. Since this original observation, only one association study Marui et al. 2004: Brain Dev 26: 5–7 has further investigated, though unsuccessfully, the involvement of the GRPR gene in ASD. With the aim of contributing further information to this topic we have sequenced the entire coding region and the intronexon junctions of the GRPR gene in 149 Italian autistic patients. The results of this study led to the identification of four novel point mutations, two of which, that is, C6S and L181F, involve amino acid changes identified in two patients with ASD and Rett syndrome, respectively. Both the leucine at position 181 and the cysteine at position 6 are strongly conserved in vertebrates. C6S and L181F mutant proteins were expressed in COS7 and BALB3T3 cells, but they did not affect either GRPs binding affinity or its potency for stimulating phospholipase Cmediated production of inositol 1,4,5trisphosphate. In summary, our results do not provide support for a major role of the GRPR gene in ASD in the population of patients we have studied. However, there is a potential role of C6S and L181F mutations on GRPR function, and possibly in the pathogenesis of the autistic disorders in the two patients. © 2008 WileyLiss, Inc.

Published
2008
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22. Absence of mutations R451C and D396ter (1186 insT) in the neuroligins nos. 3 and 4, genes, respectively, in 140 Italian patients with autism spectrum disorders

Author

Romano, V., Cali, F., Mirisola, M., Gallo, A., Angileri, L., D Anna, R., Gambino, G., Ayala, Gf, Seidita, G., Alda Ragalmuto, Elia, M., ROMANO, V, CALI', F, MIRISOLA, M, GALLO, A, ANGILERI, L, D'ANNA, RP, GAMBINO, G, AYALA, GF, SEIDITA, G, RAGALMUTO, A, ELIA, M, ROMANO V, CALI' F, MIRISOLA M, GALLO A, ANGILERI L, D'ANNA RP, GAMBINO G, AYALA GF, SEIDITA G, RAGALMUTO A, and ELIA M

Abstract

ABSENCE OF MUTATIONS R451C AND D396TER (1186 INST) IN THE NEUROLIGINS NOS. 3 AND 4,GENES, RESPECTIVELY, IN 140 ITALIAN PATIENTS WITH AUTISM SPECTRUM DISORDERS..

25. The molecular characterization of a depurinated trial DNA sample can be a model to understand the reliability of the results in forensic genetics

Author

Gregorio Seidita, Peter M. Schneider, Paola Pitacco, Silvia Corato, Eugenia Carnevali, Carla Vecchiotti, Pierangela Grignani, Solange Sorçaburu-Cigliero, Milena Alù, Anna Barbaro, Stefania Turrina, Andrea Verzeletti, Francesco De Stefano, Francesca Scarnicci, Laura Plizza, Stefania Lonero Baldassarra, Matteo Fabbri, Angel Carracedo, Carlo Previderè, Ranieri Domenici, Nicoletta Resta, Paolo Vatta, L. Casarino, Chiara Turchi, Lara Consoloni, Lucia Trizzino, Carlo Robino, Ugo Ricci, Vanessa Nicolin, Paolo Fattorini, Marco Moratti, Giorgio Marrubini, Luca Salvaderi, Emiliano Giardina, Susi Pelotti, Andrea Piccinini, Fattorini, Paolo, Previderè, Carlo, Sorçaburu-Cigliero, Solange, Marrubini, Giorgio, Alù, Milena, Barbaro, Anna M., Carnevali, Eugenia, Carracedo, Angel, Casarino, Lucia, Consoloni, Lara, Corato, Silvia, Domenici, Ranieri, Fabbri, Matteo, Giardina, Emiliano, Grignani, Pierangela, Baldassarra, Stefania Lonero, Moratti, Marco, Nicolin, Vanessa, Pelotti, Susi, Piccinini, Andrea, Pitacco, Paola, Plizza, Laura, Resta, Nicoletta, Ricci, Ugo, Robino, Carlo, Salvaderi, Luca, Scarnicci, Francesca, Schneider, Peter M., Seidita, Gregorio, Trizzino, Lucia, Turchi, Chiara, Turrina, Stefania, Vatta, Paolo, Vecchiotti, Carla, Verzeletti, Andrea, De Stefano, Francesco, Previderè, C, Sorçaburu Cigliero, S, Marrubini, G, Alù, M, Barbaro, Am, Carnevali, E, Carracedo, A, Casarino, L, Consoloni, L, Corato, S, Domenici, R, Fabbri, M, Giardina, E, Grignani, P, Baldassarra, Sl, Moratti, M, Pelotti, S, Piccinini, A, Pitacco, P, Plizza, L, Resta, N, Ricci, U, Robino, C, Salvaderi, L, Scarnicci, F, Schneider, Pm, Seidita, G, Trizzino, L, Turchi, C, Turrina, S, Vatta, P, Vecchiotti, C, Verzeletti, A, De Stefano, F., Fattorini, P., Previderè, C., Sorçaburu-Cigliero, S., Marrubini, G., Alù, M., Barbaro, A., Carnevali, E., Carracedo, A., Casarino, L., Consoloni, L., Corato, S., Domenici, R., Fabbri, M., Giardina, E., Grignani, P., Baldassarra, S., Moratti, M., Nicolin, V., Pelotti, S., Piccinini, A., Pitacco, P., Plizza, L., Resta, N., Ricci, U., Robino, C., Salvaderi, L., Scarnicci, F., Schneider, P., Seidita, G., Trizzino, L., Turchi, C., Turrina, S., Vatta, P., Vecchiotti, C., and Verzeletti, A.

Subjects
DNA depurination, Forensic genetics, PCR fidelity, STR typing, Biochemistry, Clinical Biochemistry, Genotyping Techniques, DNA damage, Sample (material), Reproducibility of Result, Biology, Polymerase Chain Reaction, NO, Analytical Chemistry, law.invention, forensic genetics, Settore MED/43 - Medicina Legale, law, Settore BIO/13 - Biologia Applicata, Genotype, Humans, Polymerase chain reaction, Protocol (science), Genetics, Medicine (all), Reproducibility of Results, Forensic genetic, DNA, Amplicon, DNA Fingerprinting, Settore BIO/18 - Genetica, DNA depurination, Forensic genetics, PCR fidelity, STR typing, DNA profiling, Settore MED/03 - Genetica Medica, Microsatellite Repeat, Genotyping Technique, Microsatellite Repeats, Human
Abstract

The role of DNA damage in PCR processivity/fidelity is a relevant topic in molecular investigation of aged/forensic samples. In order to reproduce one of the most common lesions occurring in postmortem tissues, a new protocol based on aqueous hydrolysis of the DNA was developed in vitro. Twenty-five forensic laboratories were then provided with 3.0 μg of a trial sample (TS) exhibiting, in mean, the loss of 1 base of 20, and a molecular weight below 300 bp. Each participating laboratory could freely choose any combination of methods, leading to the quantification and to the definition of the STR profile of the TS, through the documentation of each step of the analytical approaches selected. The results of the TS quantification by qPCR showed significant differences in the amount of DNA recorded by the participating laboratories using different commercial kits. These data show that only DNA quantification "relative" to the used kit (probe) is possible, being the "absolute" amount of DNA inversely related to the length of the target region (r(2) = 0.891). In addition, our results indicate that the absence of a shared stable and certified reference quantitative standard is also likely involved. STR profiling was carried out selecting five different commercial kits and amplifying the TS for a total number of 212 multiplex PCRs, thus representing an interesting overview of the different analytical protocols used by the participating laboratories. Nine laboratories decided to characterize the TS using a single kit, with a number of amplifications varying from 2 to 12, obtaining only partial STR profiles. Most of the participants determined partial or full profiles using a combination of two or more kits, and a number of amplifications varying from 2 to 27. The performance of each laboratory was described in terms of number of correctly characterized loci, dropped-out markers, unreliable genotypes, and incorrect results. The incidence of unreliable and incorrect genotypes was found to be higher for participants carrying out a limited number of amplifications, insufficient to define the correct genotypes from damaged DNA samples such as the TS. Finally, from a dataset containing about 4500 amplicons, the frequency of PCR artifacts (allele dropout, allele drop-in, and allelic imbalance) was calculated for each kit showing that the new chemistry of the kits is not able to overcome the concern of template-related factors. The results of this collaborative exercise emphasize the advantages of using a standardized degraded DNA sample in the definition of which analytical parameters are critical for the outcome of the STR profiles.

Published
2014

26. GENETICA DEL CANCRO

Author

ALESSANDRO, Riccardo, DE LEO, Giacomo, DE LEO,G, FASANO,S, GINELLI, E, ALESSANDRO,R, ALTRUDA,F, AMATO,A, ANTOGNELLI,C, BARISANI,D, BRANCOLINI,C, DE FILIPPI,P, DELRIO,G, DOLCEMASCOLO,G, FONTANA,S, GIANGUZZA,M, HIRSCH,E, MENEVERI,R, MEZZASOMA,L, MINUCCI,S, MODESTI,A, PIERANTONI,R, PURRELLO,M, SEIDITA,G, SIDOTI,A, TALESA,VN, TARONE,G, TOGNON,M, TOLOSANO,E, GINELLI,E, ALESSANDRO R., ALTRUDA F., AMATO A., BRANCOLINI C., DE LEO G., DEFILIPPI P., DELRIO G., DI BELLA MA., DOLCEMASCOLO G., FASANO S., FONTANA S., GIANGUZZA M., GINELLI E., HIRSCH E., MENEVERI R., MINUCCI S., MIRISOLA M., MODESTI A., PIERANTONI R., PURRELLO M., SEIDITA G., SIDOTI A., TARONE G., TOGNON M., TOLOSANO E., ALESSANDRO R, and DE LEO G

Subjects
BIOLOGIA CANCRO, GENETICA MOLECOLARE, Settore BIO/13 - Biologia Applicata, cancerogenesi, NEOPLASIA, mutazioni, ONCOGENI, Cancro, ONCOSOPPRESSORI
Published
2013

27. RIPRODUZIONE E CICLO CELLULARE

Author

BRANCOLINI,C, FASANO,S, DE LEO, Giacomo, DE LEO,g, GINELLI,E, FASANO,S, BARNCOLINI,C, DE LEO,G, ALESSANDRO R., ALTRUDA F., AMATO A., BRANCOLINI C., DE LEO G., DEFILIPPI P., DELRIO G., DI BELLA MA., DOLCEMASCOLO G., FASANO S., FONTANA S., GIANGUZZA M., GINELLI E., HIRSCH E., MENEVERI R., MINUCCI S., MIRISOLA M., MODESTI A., PIERANTONI R., PURRELLO M., SEIDITA G., SIDOTI A., TARONE G., TOGNON M., TOLOSANO E., BRANCOLINI C, FASANO S, DE LEO G, GINELLI, E, ALESSANDRO,R, ALTRUDA,F, AMATO,A, ANTOGNELLI,C, BARISANI,D, BRANCOLINI,C, DE FILIPPI,P, DELRIO,G, DOLCEMASCOLO,G, FONTANA,S, GIANGUZZA,M, HIRSCH,E, MENEVERI,R, MEZZASOMA,L, MINUCCI,S, MODESTI,A, PIERANTONI,R, PURRELLO,M, SEIDITA,G, SIDOTI,A, TALESA,VN, TARONE,G, TOGNON,M, and TOLOSANO,E

Subjects
BIOLOGIA DELLO SVILUPPO, Settore BIO/13 - Biologia Applicata, eucarioti, ciclo cellulare, BIOLOGIA CELLULARE, Riproduzione
Published
2009

28. Funzione cellulare e traffico intracellulare

Author

TARONE,G, DE FILIPPI,P, FONTANA, Simona, ALESSANDRO, Riccardo, DE LEO, Giacomo, DE LEO, G, GINELLI, E, FASANO, S, TARONE, G, DE LEO,G, ALESSANDRO, R, FONTANA, S, DEFILIPPI, P, ALESSANDRO R., ALTRUDA F., AMATO A., BRANCOLINI C., DE LEO G., DEFILIPPI P., DELRIO G., DI BELLA MA., DOLCEMASCOLO G., FASANO S., FONTANA S., GIANGUZZA M., GINELLI E., HIRSCH E., MENEVERI R., MINUCCI S., MIRISOLA M., MODESTI A., PIERANTONI R., PURRELLO M., SEIDITA G., SIDOTI A., TARONE G., TOGNON M., TOLOSANO E., TARONE G, DE FILIPPI P, FONTANA S, ALESSANDRO R, DE LEO G, FASANO,S, ALESSANDRO,R, ALTRUDA,F, AMATO,A, ANTOGNELLI,C, BARISANI,D, BRANCOLINI,C, DE FILIPPI,P, DELRIO,G, DOLCEMASCOLO,G, FONTANA,S, GIANGUZZA,M, HIRSCH,E, MENEVERI,R, MEZZASOMA,L, MINUCCI,S, MODESTI,A, PIERANTONI,R, PURRELLO,M, SEIDITA,G, SIDOTI,A, TALESA,VN, TARONE,G, TOGNON,M, and TOLOSANO,E

Subjects
MEMBRANE BIOLOGICHE, TRASPORTO DI MEMBRANA, Settore BIO/13 - Biologia Applicata, BIOLOGIA CELLULARE, ADESIONE CELLULARE
Published
2008

29. Mutazioni: tipi, origini, conseguenze

Author

DE LEO, Giacomo, DI BELLA, Maria Antonietta, SEIDITA, Gregorio, FASANO, S, TOGNON, M, ALESSANDRO R., ALTRUDA F., AMATO A., BRANCOLINI C., DE LEO G., DEFILIPPI P., DELRIO G., DI BELLA MA., DOLCEMASCOLO G., FASANO S., FONTANA S., GIANGUZZA M., GINELLI E., HIRSCH E., MENEVERI R., MINUCCI S., MIRISOLA M., MODESTI A., PIERANTONI R., PURRELLO M., SEIDITA G., SIDOTI A., TARONE G., TOGNON M., TOLOSANO E., DE LEO G, DI BELLA MA, FASANO S, TOGNON M, SEIDITA G, DE LEO, G, GINELLI, E, FASANO, S, DI BELLA, MA, TOGNON, M, and SEIDITA, G

Subjects
Settore BIO/13 - Biologia Applicata, genetica
Published
2007

30. Le basi dell'organizzazione biologica

Author

DI BELLA, Maria Antonietta, FONTANA, Simona, ALESSANDRO, Riccardo, SEIDITA, Gregorio, DE LEO, Giacomo, ALESSANDRO R., ALTRUDA F., AMATO A., BRANCOLINI C., DE LEO G., DEFILIPPI P., DELRIO G., DI BELLA MA., DOLCEMASCOLO G., FASANO S., FONTANA S., GIANGUZZA M., GINELLI E., HIRSCH E., MENEVERI R., MINUCCI S., MIRISOLA M., MODESTI A., PIERANTONI R., PURRELLO M., SEIDITA G., SIDOTI A., TARONE G., TOGNON M., TOLOSANO E., DI BELLA MA, FONTANA S, ALESSANDRO R, SEIDITA G, DE LEO G, DE LEO,G, FASANO,S, GINELLI, E, ALESSANDRO,R, ALTRUDA,F, AMATO,A, ANTOGNELLI,C, BARISANI,D, BRANCOLINI,C, DE FILIPPI,P, DELRIO,G, DI BELLA, MA, DOLCEMASCOLO,G, FONTANA,S, GIANGUZZA,M, HIRSCH,E, MENEVERI,R, MEZZASOMA,L, MINUCCI,S, MIRISOLA,MG, MODESTI,A, PIERANTONI,R, PURRELLO,M, SEIDITA,G, SIDOTI,A, TALESA,VN, TARONE,G, TOGNON,M, TOLOSANO,E, DI BELLA,MA, SEIDITA, G, DE LEO, G, GINELLI,E, FONTANA, S, and ALESSANDRO, R

Subjects
Settore BIO/13 - Biologia Applicata, BIOLOGIA, BIOLOGIA GENERALE, CELLULA, EVOLUZIONE
Published
2007

31. Analysis of the gastrin-releasing peptide receptor gene in Italian patients with autism spectrum disorders

Author

Alessia Gallo, Robert T. Jensen, Mario G. Mirisola, Filippo Cali, M. Falco, Gregorio Seidita, Samuel A. Mantey, L. Cucina, Nieves González, Marinella Zingale, Rosalba D'Anna, Maurizio Elia, V. Chiavetta, Valentino Romano, SEIDITA, G, MIRISOLA, MG, D'ANNA, R, GALLO, A, JENSEN, RT, MANTEY, SA, GONZALEZ, N, FALCO, M, ZINGALE, M, ELIA, M, CUCINA, L, CHIAVETTA, V, ROMANO, V, and CALI, F

Subjects
Adult, Male, medicine.medical_specialty, BALB 3T3 Cells, Adolescent, DNA Mutational Analysis, Population, Rett syndrome, Biology, Mice, Cellular and Molecular Neuroscience, Exon, Settore BIO/13 - Biologia Applicata, Internal medicine, Gastrin-releasing peptide, Chlorocebus aethiops, medicine, Gastrin-releasing peptide receptor, Animals, Humans, Point Mutation, Autistic Disorder, Child, autism, gastrin-releasing peptide receptor, signal transduction,G-protein-coupled receptor, association study, education, Gene, Genetics (clinical), Aged, Genetics, education.field_of_study, Point mutation, Middle Aged, medicine.disease, Pedigree, Receptors, Bombesin, Developmental disorder, Psychiatry and Mental health, Endocrinology, Italy, Case-Control Studies, COS Cells, Female
Abstract

The gastrin-releasing peptide receptor (GRPR) was implicated for the first time in the pathogenesis of Autism spectrum disorders (ASD) by Ishikawa-Brush et al. [Ishikawa-Brush et al. (1997): Hum Mol Genet 6: 1241-1250]. Since this original observation, only one association study [Marui et al. (2004): Brain Dev 26: 5-7] has further investigated, though unsuccessfully, the involvement of the GRPR gene in ASD. With the aim of contributing further information to this topic we have sequenced the entire coding region and the intron/exon junctions of the GRPR gene in 149 Italian autistic patients. The results of this study led to the identification of four novel point mutations, two of which, that is, C6S and L181F, involve amino acid changes identified in two patients with ASD and Rett syndrome, respectively. Both the leucine at position 181 and the cysteine at position 6 are strongly conserved in vertebrates. C6S and L181F mutant proteins were expressed in COS-7 and BALB/3T3 cells, but they did not affect either GRP's binding affinity or its potency for stimulating phospholipase C-mediated production of inositol 1,4,5-trisphosphate. In summary, our results do not provide support for a major role of the GRPR gene in ASD in the population of patients we have studied. However, there is a potential role of C6S and L181F mutations on GRPR function, and possibly in the pathogenesis of the autistic disorders in the two patients.

Published
2008

32. Development of an Italian RM Y-STR haplotype database: Results of the 2013 GEFI collaborative exercise

Author

Andrea Piccinini, Solange Sorçaburu-Cigliero, Gregorio Seidita, A. Gonzalez, Andrea Verzeletti, Emiliano Giardina, E. Ponzano, Marilidia Piglionica, Carlo Robino, Giuseppe Matullo, Arwin Ralf, Manfred Kayser, Francesca Scarnicci, Carla Bini, A.L. Nutini, Valerio Onofri, C. Di Gaetano, Gianmarco Ferri, Stefania Turrina, Nicoletta Resta, M. De Marchi, Matteo Fabbri, Kaye N. Ballantyne, L. Casarino, A. Barbaro, Eugenia Carnevali, S Pasino, Carlo Previderè, Robino, C., Ralf, A., Pasino, S., De Marchi, M., Ballantyne, K., Barbaro, A., Bini, C., Carnevali, E., Casarino, L., Di Gaetano, C., Fabbri, M., Ferri, G., Giardina, E., Gonzalez, A., Matullo, G., Nutini, A., Onofri, V., Piccinini, A., Piglionica, M., Ponzano, E., Previderè, C., Resta, N., Scarnicci, F., Seidita, G., Sorçaburu-Cigliero, S., Turrina, S., Verzeletti, A., Kayser, M., and Genetic Identification

Subjects
Quality Control, Mutation rate, Regional Italian, Lineage differentiation, DNA Primer, Y-chromosome, Rapidly mutating Y-STRs (RM Y-STRs), Haplotype, Relative differentiation, Italy, Biology, computer.software_genre, Pathology and Forensic Medicine, Genetic, Databases, Genetic, Genetics, Haplotype, Italy, Lineage differentiation, Rapidly mutating Y-STRs (RM Y-STRs), Relative differentiation, Y-chromosome, Humans, Y-STR, Cooperative Behavior, DNA Primers, Chromosomes, Human, Y, Database, Base Sequence, Medicine (all), humanities, Forensic science, Haplotypes, Microsatellite, Haplotype estimation, computer, Human
Abstract

Recently introduced rapidly mutating Y-chromosomal short tandem repeat (RM Y-STR) loci, displaying a multiple-fold higher mutation rate relative to any other Y-STRs, including those conventionally used in forensic casework, have been demonstrated to improve the resolution of male lineage differentiation and to allow male relative separation usually impossible with standard Y-STRs. However, large and geographically-detailed frequency haplotype databases are required to estimate the statistical weight of RM Y-STR haplotype matches if observed in forensic casework. With this in mind, the Italian Working Group (GEFI) of the International Society for Forensic Genetics launched a collaborative exercise aimed at generating an Italian quality controlled forensic RM Y-STR haplotype database. Overall 1509 male individuals from 13 regional populations covering northern, central and southern areas of the Italian peninsula plus Sicily were collected, including both "rural'' and "urban'' samples classified according to population density in the sampling area. A subset of individuals was additionally genotyped for Y-STR loci included in the Yfiler and PowerPlex Y23 (PPY23) systems (75% and 62%, respectively), allowing the comparison of RM and conventional Y-STRs. Considering the whole set of 13 RM Y-STRs, 1501 unique haplotypes were observed among the 1509 sampled Italian men with a haplotype diversity of 0.999996, largely superior to Yfiler and PPY23 with 0.999914 and 0.999950, respectively. AMOVA indicated that 99.996% of the haplotype variation was within populations, confirming that genetic-geographic structure is almost undetected by RM Y-STRs. Haplotype sharing among regional Italian populations was not observed at all with the complete set of 13 RM Y-STRs. Haplotype sharing within Italian populations was very rare (0.27% non-unique haplotypes), and lower in urban (0.22%) than rural (0.29%) areas. Additionally, 422 father-son pairs were investigated, and 20.1% of them could be discriminated by the whole set of 13 RM Y-STRs, which was very close to the theoretically expected estimate of 19.5% given the mutation rates of the markers used. Results obtained from a high-coverage Italian haplotype dataset confirm on the regional scale the exceptional ability of RM Y-STRs to resolve male lineages previously observed globally, and attest the unsurpassed value of RM Y-STRs for male-relative differentiation purposes. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

Published
2014

33. MUTAZIONI: TIPI, ORIGINI, CONSEGUENZE

Author

DE LEO, Giacomo, DI BELLA, Maria Antonietta, BARISANI,D, TOGNON,M, DE LEO,G, FASANO,S, GINELLI, E, ALESSANDRO,R, ALTRUDA,F, AMATO,A, ANTOGNELLI,C, BARISANI,D, BRANCOLINI,C, DE FILIPPI,P, DELRIO,G, DI BELLA, MA, DOLCEMASCOLO,G, FONTANA,S, GIANGUZZA,M, HIRSCH,E, MENEVERI,R, MEZZASOMA,L, MINUCCI,S, MIRISOLA,MG, MODESTI,A, PIERANTONI,R, PURRELLO,M, SEIDITA,G, SIDOTI,A, TALESA,VN, TARONE,G, TOGNON,M, and TOLOSANO,E

Subjects
RICOMBINAZIONE, RIPARAZIONE DNA, GENETICA MOLECOLARE, TECNOLOGIE GENETICHE, CARIOTIPO
Published
2013

34. GENETICA UMANA

Author

DE LEO, Giacomo, DI BELLA, Maria Antonietta, SEIDITA, Gregorio, BARISANI,D, TOGNON,M, DE LEO,G, FASANO,S, GINELLI, E, ALESSANDRO,R, ALTRUDA,F, AMATO,A, ANTOGNELLI,C, BARISANI,D, BRANCOLINI,C, DE FILIPPI,P, DELRIO,G, DI BELLA, MA, DOLCEMASCOLO,G, FONTANA,S, GIANGUZZA,M, HIRSCH,E, MENEVERI,R, MEZZASOMA,L, MINUCCI,S, MIRISOLA,MG, MODESTI,A, PIERANTONI,R, PURRELLO,M, SEIDITA,G, SIDOTI,A, TALESA,VN, TARONE,G, TOGNON,M, and TOLOSANO,E

Subjects
EREDITA', GENETICA IMMUNOGLOBULINE
Published
2013

35. LA GENETICA GENERALE

Author

DE LEO, Giacomo, DI BELLA, Maria Antonietta, MIRISOLA, Mario Giuseppe, DE LEO,G, FASANO,S, GINELLI, E, ALESSANDRO,R, ALTRUDA,F, AMATO,A, ANTOGNELLI,C, BARISANI,D, BRANCOLINI,C, DE FILIPPI,P, DELRIO,G, DOLCEMASCOLO,G, FONTANA,S, GIANGUZZA,M, HIRSCH,E, MENEVERI,R, MEZZASOMA,L, MINUCCI,S, MIRISOLA, MG, MODESTI,A, PIERANTONI,R, PURRELLO,M, SEIDITA,G, SIDOTI,A, TALESA,VN, TARONE,G, TOGNON,M, TOLOSANO,E, DI BELLA, MA, and MIRISOLA,MG

Subjects
Settore BIO/13 - Biologia Applicata, GENETICA, GENETICA AMBIENTALE
Published
2013

36. Mutagenic alteration of the distal switch II region of RAS blocks CDC25-dependent signaling functions

Author

F Di Blasi, Mario G. Mirisola, Arturo C. Verrotti, Ottavio Fasano, Gregorio Seidita, Mirisola, M. G., Seidita, G., VERROTTI DI PIANELLA, Arturo, DI BLASI, F., and Fasano, O.

Subjects
Adenosine monophosphate, biology, Cdc25, Saccharomyces cerevisiae, Mutagenesis (molecular biology technique), Cell Biology, biology.organism_classification, Biochemistry, Molecular biology, Nucleotide exchange factor, Adenylyl cyclase, chemistry.chemical_compound, Ras-GRF1, chemistry, biology.protein, Ras2, Molecular Biology
Abstract

We have explored the role of the distal switch II region of the yeast RAS2 protein in determining the response to the nucleotide exchange factor CDC25. We first constructed yeast tester strains in which the deletion of the chromosomal CDC25, RAS1, and RAS2 genes, in combination with the chromosomal suppressor CRI4, resulted in detectable phenotypes in vivo and in vitro. Phenotypes included impaired growth at 37 degrees C, defective glucose-induced cyclic AMP signaling, and low adenylyl cyclase activity of membrane preparations. Tester strains were subsequently used for the reintroduction of various combinations of wild-type and mutated RAS2 and CDC25 genes by genetic techniques, as well as for in vitro reconstitution assays with the corresponding proteins. CDC25 restored both growth and glucose-induced cyclic AMP signaling in the presence, but not in the absence of wild-type RAS2. A gene encoding a RAS2 protein with a mutationally altered switch II region was expressed but was ineffective in reintegrating exchange factor-dependent responses in vivo. Wild-type, but not mutagenically altered, RAS2 proteins were stimulated by exchange factors in vitro. We conclude that the conserved distal switch II region is required for CDC25-dependent activation of RAS.

Published
1994

37. La persona nel diritto civile

Author

ZERBO, Stefania, ARGO, Antonina, De Luca, L, Zagra, M., Zagra, M, Argo, A, Madea, B, Procaccianti, P, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Milone, L, Pascali, V, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, and Zerbo, S

Subjects
Settore MED/43 - Medicina Legale, capacità di agire, capacità giuridica
Published
2011

38. Accertamento e trattamento sanitario obbligatorio

Author

Zagra, M, ARGO, Antonina, Zagra, M, Argo, A, Madea, B, Procaccianti, P, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Milone, L, Pascali, V, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, and Zerbo, S

Subjects
TSO, Settore MED/43 - Medicina Legale
Published
2011

39. Attività clinica : colpa medica e responsabilità professionale

Author

Zagra, M, De Luca, L, Triolo, V., ZERBO, Stefania, ARGO, Antonina, FLERES, Pierangela, Zagra, M. Argo, A, Madea, B, Procaccianti, P, Argo, A, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Madea, B, Milone, L, Pascali, V, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, Zagra, M, and Zerbo, S

Subjects
Settore MED/43 - Medicina Legale, colpa medica
Published
2011

40. TIPIZZAZIONE DEL DNA DI DITTERI CAMPIONATI SU UN CORPO RINVENUTO IN SICILIA

Author

Piscionieri,D, Triolo,V, SEIDITA, Gregorio, NICOSIA, Aldo, PROCACCIANTI, Salvatore, CARNEVALI, DOBOSZ,E, LANCIA,M, Piscionieri,D, Seidita,G, Nicosia,A, Triolo,V, and Procaccianti,S

Subjects
Tanatologia,entomologia forense
Abstract

L’entomologia forense, come ormai ampiamente noto, viene utilizzata routinariamente nelle indagini forensi, in particolare in quei casi nei quali i classici dati tanatocronologici (ipostasi, rigidità, temperatura corporea, ecc.) non possono essere d’ausilio per l’identificazione dell’epoca della morte. Gli insetti campionati su resti cadaverici, per potere essere utilizzati a fini forensi, necessitano di una precisa identificazione di specie, dato che ciascuna specie si caratterizza per una differente durata del ciclo vitale. Gli insetti che colonizzano il cadavere, soprattutto nelle prime fasi di colonizzazione, hanno una distribuzione abbastanza uniforme in tutto il mondo, tuttavia è possibile a volte rinvenire delle differenze correlate alla fauna locale. In questo lavoro viene presentato un case report in cui dimostriamo l’utile integrazione tra i dati di entomologia forense e quelli di tassonomia molecolare.

Published
2011

41. Conoscenze fondamentali della patologia forense per il sanitario non specialista in medicina legale

Author

PROCACCIANTI, Paolo, MILONE, Livio, ARGO, Antonina, FLERES, Pierangela, Zagra, M, Madea, B., Zagra, M, Argo, A, Madea, B, Procaccianti, P, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Milone, L, Pascali, V, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, and Zerbo, S

Subjects
lesività medico legale, Settore MED/43 - Medicina Legale
Published
2011

42. Medicina e nursing delle evidenze

Author

Zagra, M, ARGO, Antonina, Zagra, M, Argo, A, Madea, B, Procaccianti, P, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Milone, L, Pascali, V, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, and Zerbo, S

Subjects
Settore MED/43 - Medicina Legale, nursing delle evidenze
Published
2011

43. Informatica, web e telemedicina

Author

Zagra, M, ZERBO, Stefania, ARGO, Antonina, Zagra, M, Argo, A, Madea, B, Procaccianti, P, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Madea, B, Milone, L, Pascali, V, Procaccianti, P, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, and Zerbo, S

Subjects
telemedicina, teleconsulenza, Settore MED/43 - Medicina Legale
Published
2011

44. Referto/rapporto e denunce sanitarie obbligatorie

Author

Zagra, M, ARGO, Antonina, SORTINO, Cettina, Zagra, M, Argo, A, Madea, B, Procaccianti, P, Argo, A, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Madea, B, Milone, L, Pascali, V, Procaccianti, P, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, Zagra, M, and Zerbo, S

Subjects
Settore MED/43 - Medicina Legale, referto, denuncia di reato
Published
2011

45. Genetica e laboratorio di medicina legale

Author

PROCACCIANTI, Salvatore, SEIDITA, Gregorio, PROCACCIANTI, Paolo, Zagra,M, Argo, A, Madea, Procaccianti, P, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Madea, B, Milone, L, Pascali, V, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, Zagra, M, and Zerbo, S

Subjects
Settore MED/43 - Medicina Legale, DNA, genetica forense
Published
2011

46. Consenso informato

Author

Zagra, M, Triolo, V, ARGO, Antonina, ZERBO, Stefania, Zagra, M, Argo, A, Madea, B, Procaccianti, P, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Milone, L, Pascali, V, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, and Zerbo, S

Subjects
consenso informato, atto medico, Settore MED/43 - Medicina Legale
Published
2011

47. Cartella clinica

Author

Zagra, M, ARGO, Antonina, ZERBO, Stefania, Zagra, M, Argo, A, Madea, B, Procaccianti, P, Bertol, E, Bonifacio, A, Cecchi, R, Dall’Acqua, P, De Luca, L, Di Vella, G, Fleres, P, Guarino, R, Madea, B, Milone, L, Pascali, V, Procaccianti, P, Procaccianti, S, Scorretti, C, Seidita, G, Sortino, C, Trignano, C, Triolo, V, Vella, M, and Zerbo, S

Subjects
Settore MED/43 - Medicina Legale, Cartella clinica, falso materiale e ideologico
Published
2011

48. A new method to value efficiency of enzyme blends for pancreatic tissue digestion

Author

Salvatrice Rigogliuso, Gregorio Seidita, Monica Salamone, Giulio Ghersi, Federico Bertuzzi, Angela Cuttitta, Salvatore Mazzola, Salamone, M, Seidita, G, Cuttitta, A, Rigogliuso, S, Mazzola, S, Bertuzzi, F, and Ghersi, G

Subjects
endocrine system, medicine.medical_specialty, Proteases, Islets transplantation, medicine.medical_treatment, Collagenase, Islets of Langerhans Transplantation, Thermolysin, Cell Separation, Cell Line, Islets of Langerhans, Clostridium histolyticum, Settore BIO/10 - Biochimica, Internal medicine, medicine, Humans, Collagenases, Pancreas, Transplantation, Islet cell transplantation, geography, Evaluation alive cell, geography.geographical_feature_category, biology, Pancreatic islets, REcombinant protein, Proteolytic enzymes, Endothelial Cells, proteolytic enzymes, biology.organism_classification, Islet, medicine.anatomical_structure, Endocrinology, Biochemistry, Gelatinases, Surgery, Collagen, Gels, Peptide Hydrolases
Abstract

Islet transplantation, since the 90’s, has been resulting to be one of the best successful example of human cell therapy. Nevertheless, islet isolation procedure is not completely standardized; in fact, more than fifty percent of islets procedures don’t arrive to their transplantation. This is due both to the variability of donor’s pancreas and to an unpredictable enzymatic blend efficiency. Enzymes used in pancreas digestion are extracted from Clostridium histolyticum bacteria and digest several substrates. In particular they have strong collagenolytic activity compared to vertebrate collagenases. However, several impediments persist in human islet isolation success probably due to the variability in composition and concentration of collagenases used during the digestion phase‏. In islets isolation’s process neutral protease is also used and plays an important role. However, it should be considered as double-edged sword: it contributes to accelerate the tissue dissociation but, sometimes, its action could result decreased in islet yield, through their fragmentation, breakdown, inactivation‏. Proteases’ activities cannot be preciously adjusted in a narrow range since it doesn’t exist any approach showing the best possible dosage and composition of enzymes that can be used in a successful process of human islets pancreas extraction. At the moment, the available data on commercial enzymatic activity are not sufficient to predict their efficiency in the process of pancreas digestion and consequently it is very difficult to select which enzyme batches can be successfully used. For these reasons we apply to generate an innovative evaluation assay to select enzymes useful in islet pancreas isolation procedure.

Published
2010

49. A new method to value efficiency of enzyme blends for pancreas tissue digestion

Author

SALAMONE, Monica, SEIDITA, Gregorio, Cuttitta, A, RIGOGLIUSO, Salvatrice, Mazzola, S, Bertuzzi, F, GHERSI, Giulio, Salamone, M, Seidita, G, Cuttitta, A, Rigogliuso, S, Mazzola, S, Bertuzzi, F, and Ghersi, G

Subjects
Settore BIO/10 - Biochimica, Pancreatic enzyme, cell therapy, collagenases
Abstract

In pancreatic islets isolation for cell therapy the major enzymes used are obtained from Clostridium hystoliticum: class I and class II collagenases. They are used in a defined tissue dissociation enzyme mixture together with neutral protease (Dispase) or thermolysin (Thermostable Neutral Protease). However, just to now, people working in islets production found variable outcomes in isolation procedures mainly due to large variability in enzymatic blend composition and efficacy. Using electrophoresis and gelatin zymography approaches together with densitometry evaluation assays we compared the composition, stability and auto-digestion processes of C. hystoliticum collagenases, Neutral protease and Thermolysin from Roches and Serva. Moreover, we have assessed: –the stability of enzymes when they are in solution at different temperatures (-20 C; 0 C and room temperature); –their digestive activity when applied at different working temperatures (25 C; 30 C; 37 C and 42 C). Our results shown a heterogeneous composition of the different enzymatic blend enzymes analyzed. Furthermore, heterogeneity is observed among different batch enzymes; we found several more proteins and/or fragments compare to HPLC profiles published by vendors. In gelatin zymographyes several digestive bands were catalytic, showing very high complex degradative patterns, in part active even in condition of calcium deprivation. Additionally, in neutral protease from Serva (and not in Thermolysin) contaminants with gelatinolytic activities were detected. An auto-digestive/inactivation processes of enzymes occurred at different working temperatures, reduced by lowering temperature up to 25 C. These data taken together strongly imply a not controlled digestive processes due to several contaminants in enzyme blends and to autocatalytic processes. Moreover, the presence of low molecular weight gelatine/ degradative activities obstacles the possibility to control islet digestion due to aspecific catalytic activities. Generation of recombinant collagenases probably could be of help to overcome the variability in the extractive processes.

Published
2010

50. C. HYSTOLITICUM RECOMBINANT COLLAGENASES AND METHOD FOR THE MANUFACTURE THEREOF

Author

Bertuzzi, F, Cuttitta, A, Mazzola, S, Salamone, M, GHERSI, Giulio, SEIDITA, Gregorio, Bertuzzi, F, Cuttitta, A, Ghersi, G, Mazzola, S, Salamone, M, and Seidita, G

Subjects
Settore BIO/13 - Biologia Applicata, COLLAGENASES, Settore BIO/10 - Biochimica, CELL TERAPY, DIABET 1, RECOMBINANT PROTEIN
Abstract

The present invention relates to the production of recombinant collagenases, and in particular describes a method for the production of recombinant clostridium histolyticum collagenases Col characterized by a yield higher than approximately 140 mg/l of culture of said collagenases in soluble, biologically active form, collagenases produced by this method, compositions comprising these collagenases and the use thereof.

Published
2010

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