BUSCADOR RECOLECTA

[Organism] Homo sapiens, [Experiment type] Expression profiling by array, [Overall design] Two-condition experiment, Progressing vs regressing metastases. 3 progressing and 2 regressing metastases extracted from the same patient after M-VAX immunotherapy. One replicate per array., [Platforms (1)] GPL7088 CCDTM Hs_CCDTM36k - version 1, [Relations] BioProject PRJNA130231, 1. Sample GSM592226. Title: melanoma progressing 1. Sample type: RNA. Platform ID: GPL7088. Series: GSE24067 Progressing vs regressing melanoma metastases; GSE26383 Response to immunotherapy in melanoma metastasis., Channel 1: [Source name] melanoma, progressing [Organism] Homo sapiens [Characteristics] tissue: melanoma treatment protocol: M-VAX preservation: cryopreservation patient: 1 [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy5 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., Channel 2: [Source name] Total RNA from pooled PBMC [Organism] Homo sapiens [Characteristics] tissue: Blood preservation: cryopreservation reference: Total RNA from pooled PBMC [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy3 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., [Hybridization protocol] 6 ug of labeled, fragmented cRNA per sample was applied to microarrays in Kreatech's Hybridization buffer, and hybridized for 16 h at 42C. After hybridization, slides were washed three four times with wash solutions containg gradually decreased concentrations of SSC (1x-0x) [Scan protocol] Scanned on an Agilent DNA Microarray Scanner. Images were quantified using Agilent Feature Extraction Software (version A.9.5.3). [Description] metastasis non responding after M-VAX immunotherapy 1 [Data processing] LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Agilent software was used for feature extraction., 2. Sample GSM592227. Title: melanoma progressing 2. Sample type: RNA. Platform ID: GPL7088. Series (2): GSE24067 Progressing vs regressing melanoma metastases. GSE26383 Response to immunotherapy in melanoma metastasis., Channel 1 [Source name] melanoma, progressing [Organism] Homo sapiens [Characteristics] tissue: melanoma treatment protocol: M-VAX preservation: cryopreservation patient: 1 [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy5 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., Channel 2 [Source name] Total RNA from pooled PBMC [Organism] Homo sapiens [Characteristics] tissue: Blood preservation: cryopreservation reference: Total RNA from pooled PBMC [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy3 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., [Hybridization protocol] 6 ug of labeled, fragmented cRNA per sample was applied to microarrays in Kreatech's Hybridization buffer, and hybridized for 16 h at 42C. After hybridization, slides were washed three four times with wash solutions containg gradually decreased concentrations of SSC (1x-0x) [Scan protocol] Scanned on an Agilent DNA Microarray Scanner. Images were quantified using Agilent Feature Extraction Software (version A.9.5.3). [Description] metastasis non responding after M-VAX immunotherapy 2 [Data processing] LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Agilent software was used for feature extraction., 3. Sample GSM592228. Title: melanoma progressing 3. Sample type: RNA. Platform ID: GPL7088. Series (2): GSE24067 Progressing vs regressing melanoma metastases; GSE26383 Response to immunotherapy in melanoma metastasis, Channel 1 [Source name] melanoma, progressing [Organism] Homo sapiens [Characteristics] tissue: melanoma treatment protocol: M-VAX preservation: cryopreservation patient: 1 [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy5 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., Channel 2 [Source name] Total RNA from pooled PBMC [Organism] Homo sapiens [Characteristics] tissue: Blood preservation: cryopreservation reference: Total RNA from pooled PBMC [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy3 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., [Hybridization protocol] 6 ug of labeled, fragmented cRNA per sample was applied to microarrays in Kreatech's Hybridization buffer, and hybridized for 16 h at 42C. After hybridization, slides were washed three four times with wash solutions containg gradually decreased concentrations of SSC (1x-0x) [Scan protocol] Scanned on an Agilent DNA Microarray Scanner. Images were quantified using Agilent Feature Extraction Software (version A.9.5.3). [Description] metastasis non responding after M-VAX immunotherapy 3 [Data processing] LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Agilent software was used for feature extraction., 4. Sample GSM592229. Title: melanoma regressing 1. Sample type: RNA. Platform ID: GPL7088. Series (2): GSE24067 Progressing vs regressing melanoma metastases; GSE26383 Response to immunotherapy in melanoma metastasis., Channel 1 [Source name] melanoma, regressing [Organism] Homo sapiens [Characteristics] tissue: melanoma treatment protocol: M-VAX preservation: cryopreservation patient: 1 [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy5 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., Channel 2 [Source name] Total RNA from pooled PBMC [Organism] Homo sapiens [Characteristics] tissue: Blood preservation: cryopreservation reference: Total RNA from pooled PBMC [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy3 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., Hybridization protocol 6 ug of labeled, fragmented cRNA per sample was applied to microarrays in Kreatech's Hybridization buffer, and hybridized for 16 h at 42C. After hybridization, slides were washed three four times with wash solutions containg gradually decreased concentrations of SSC (1x-0x) [Scan protocol] Scanned on an Agilent DNA Microarray Scanner. Images were quantified using Agilent Feature Extraction Software (version A.9.5.3). [Description] metastasis responding after M-VAX immunotherapy 1 [Data processing] LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Agilent software was used for feature extraction., 5. Sample GSM592230. Title: melanoma regressing 2. Sample type: RNA. Platform ID: GPL7088. Series (2): GSE24067 Progressing vs regressing melanoma metastases; GSE26383 Response to immunotherapy in melanoma metastasis., Channel 1 [Source name] melanoma, regressing [Organism] Homo sapiens [Characteristics] tissue: melanoma treatment protocol: M-VAX preservation: cryopreservation patient: 1 [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy5 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., Channel 2 [Source name] Total RNA from pooled PBMC [Organism] Homo sapiens [Characteristics] tissue: Blood preservation: cryopreservation reference: Total RNA from pooled PBMC [Extracted molecule] total RNA [Extraction protocol] Total RNA extracted using Qiagen's RNEasy Mini kit following manufacturer's instructions [Label] Cy3 [Label protocol] 0.5 µg of total RNA was primed with 0.25 ug oligo dT-T7 primer (5’ AAA CGA CGG CCA GTG AAT TGT AAT ACG ACT CAC TAT AGG CGC T(15) 3’) at 70°C for 3 min, then reverse transcribed at 42°C for 90 min in the presence of 200 U SuperScript II RTase (Invitrogen), and 1 mM dNTP. Second strand synthesis was done with the 5’ primer switch method using 0.25 ug Ts/SMARTII primers (5’ AAG CAG TGG TAA CAA CGC AGA GTA CGC GGG 3’) for priming at 65°C for 1 min, and subsequent primer extension at 75°C for 30 min using the Advantage cDNA Polymerase Mix (Clontech) and 0.2mM dNTP. Double stranded cDNA subjected to in vitro transcription at 42°C for 6h using Megascript T7 kit (Ambion). After two rounds of linear amplification, cRNA was chemically labeled using ULS Cy3/Cy5 Fluorescent Labeling Kit (Kreatech), and fragmented by Fragmentation Reagents kit (Ambion)., [Hybridization protocol] 6 ug of labeled, fragmented cRNA per sample was applied to microarrays in Kreatech's Hybridization buffer, and hybridized for 16 h at 42C. After hybridization, slides were washed three four times with wash solutions containg gradually decreased concentrations of SSC (1x-0x) [Scan protocol] Scanned on an Agilent DNA Microarray Scanner. Images were quantified using Agilent Feature Extraction Software (version A.9.5.3). [Description] metastasis responding after M-VAX immunotherapy 2 [Data processing] LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Agilent software was used for feature extraction., We documented the transcriptional pattern of 3 progressing and 2 regressing synchronous melanoma metastases from the same patient following M-VAX treatment.

Plano general de la sala de lectura de la biblioteca del Instituto Cervantes de Hamburgo y sus usuarios

Plano general de la sala de lectura de la Biblioteca Ana María Matute del Instituto Cervantes de Casablanca, que recibe su actual nombre en 2011

Plano general: un usuario leyendo en la hemeroteca del Instituto Cervantes de Berlín

Plano general de la biblioteca Gerardo Diego del Instituto Cervantes de Bruselas

Plano general de la sala de proyecciones de la biblioteca Gerardo Diego del Instituto Cervantes de Bruselas

Plano general de algunas estanterías de la biblioteca Gonzalo Rojas del Instituto Cervantes de Bremen

Plano medio de las estanterías de la sala de lectura de la biblioteca del Instituto Cervantes de Moscú

Plano medio del mostrador de la sala de lectura de la biblioteca del Instituto Cervantes de Moscú. En la pared, un cuadro con una fotografía de Miguel Delibes, escritor español que da nombre a la biblioteca

Plano medio del mostrador de la sala de lectura de la biblioteca del Instituto Cervantes de Moscú. Al fondo, las estanterías de libros