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Singh A.V.,University of Milan | Rahman A.,Instituto Of Moda Burgo | Sudhir Kumar N.V.G.,Instituto Of Moda Burgo | Aditi A.S.,University of Pune | And 5 more authors.
Materials and Design | Year: 2012

The nature's intrigue plans to create structures from nano-micro to mesoscale has inspired researchers to design artificial object with novel and extra ordinary properties. Recently, the convergence of biomaterials and polymer advances from nano- to micro-scale with new experimental and computational tools has provided the opportunity to constitute increasingly complex fabrics for the textiles industry. In this regard, learning lessons from efficient natural processes to design smart fabrics, mimicking natural phenomena could revolutionize the textile industry for the design of interactive apparel. Here, we review 10 bio-inspired strategies to imply textile industry, to change the face of fashion and fabrics, based upon current advances in science to enrich diverse areas of textile industry. In each case, we present examples demonstrating nature's design and subsequent parallel advances in biomimetic materials and polymer sciences, combining interdisciplinary engineering principles to mimic nature inspired designs into fabrics. We predict as advances in science of biomimesis continue to unfold and uncover finer details, bioinspired emulations may increasingly give way to benefit in designing smart fabrics. © 2011 Elsevier Ltd.


Sala E.,COGENTECH | De Marco A.,COGENTECH
Protein Expression and Purification | Year: 2010

Testing several different growth and purification conditions by means of a small-scale screening is a valuable approach for identifying optimal protocols for large preparative protein productions. The availability of new mini columns for size exclusion chromatography (SEC) enables the coupling of parallel affinity purification using magnetic beads with the aggregation analysis performed by gel-filtration. Only 35 μL of bead eluate are sufficient to run both experiments and each SEC cycle needs no more than 30 min, washing and equilibration inclusive. We have shown that SEC ideally completes the SDS-PAGE analysis since it allows estimating the amount of soluble aggregates recovered in the elution fraction after affinity purification, although larger aggregates seem to be lost in the pre-column filters. SEC is also useful to identify low molecular mass contaminants that can be underestimated as they migrate together with the front of the acrylamide gel.© 2010 Published by Elsevier Inc.


Chiarella P.,Biomedical University of Rome | Edelmann B.,University of Kiel | Fazio V.M.,Biomedical University of Rome | Sawyer A.M.,Monoclonal Antibody Core Facility | de Marco A.,Cogentech
Biotechnology Letters | Year: 2010

An aluminium hydroxide adjuvant induced a more elevated and rapid immune responses against short peptides conjugated to the Keyhole Lympet Hemocyanin carrier than immuneasy adjuvant. Furthermore, since carrier proteins may compete with the fused or chemically linked polypeptides in eliciting antigen-specific immune response, we classified the immunogenicity of the most common carrier proteins used in molecular biology for antigen expression and mouse immunisation. The disulfide isomerase protein A gave a carrier with the lowest immunogenicity whilst disulfide isomerase protein C gave the highest immunogenicity and therefore should be avoided as a fusion partner. Using this protein as a model, we identified and located the immunodominant epitopes along its sequence. These results now enable the combination of carrier and immunisation conditions to be optimized. © 2010 Springer Science+Business Media B.V.


Dolgikh V.V.,All Russian Institute for Plant Protection | Senderskiy I.V.,All Russian Institute for Plant Protection | Pavlova O.A.,All Russian Institute for Plant Protection | Naumov A.M.,All Russian Institute for Plant Protection | Beznoussenko G.V.,Cogentech
Eukaryotic Cell | Year: 2011

Microsporidia are a group of fungus-related intracellular parasites with severely reduced metabolic machinery. They lack canonical mitochondria, a Krebs cycle, and a respiratory chain but possess genes encoding glycolysis enzymes, a glycerol phosphate shuttle, and ATP/ADP carriers to import host ATP. The recent finding of alternative oxidase genes in two clades suggests that microsporidial mitosomes may retain an alternative respiratory pathway. We expressed the fragments of mitochondrial chaperone Hsp70 (mitHsp70), mitochondrial glycerol-3-phosphate dehydrogenase (mitG3PDH), and alternative oxidase (AOX) from the microspo-ridium Antonospora (Paranosema) locustae in Escherichia coli. Immunoblotting with antibodies against recombinant polypeptides demonstrated specific accumulation of both metabolic enzymes inA. locustae spores. At the same time comparable amounts of mitochondrial Hsp70 were found in spores and in stages of intracellular development as well. Immunoelectron microscopy of ultrathin cryosections of spores confirmed mitosomal localization of the studied proteins. Small amounts of enzymes of an alternative respiratory chain in mero-gonial and early sporogonial stages, alongside their accumulation in mature spores, suggest conspicuous changes in components and functions of mitosomes during the life cycle of microsporidia and the important role of these organelles in parasite energy metabolism, at least at the final stages of sporogenesis.© 2011, American Society for Microbiology. All Rights Reserved.


Bianchi F.,The FIRC Institute for Molecular Oncology Foundation | Bianchi F.,University of Milan | Nicassio F.,The FIRC Institute for Molecular Oncology Foundation | Marzi M.,The FIRC Institute for Molecular Oncology Foundation | And 9 more authors.
EMBO Molecular Medicine | Year: 2011

Lung cancer is the first cause of cancer mortality worldwide, and its early detection is currently the main available strategy to improve disease prognosis. While early diagnosis can be successfully achieved through tomography-based population screenings in high-risk individuals, simple methodologies are needed for effective cancer prevention programs. We developed a test, based on the detection of 34 microRNAs (miRNAs) from serum, that could identify patients with early stage non-small cell lung carcinomas (NSCLCs) in a population of asymptomatic high-risk individuals with 80% accuracy. The signature could assign disease probability accurately either in asymptomatic or symptomatic patients, is able to distinguish between benign and malignant lesions, and to capture the onset of the malignant disease in individual patients over time. Thus, our test displays a number of features of clinical relevance that project its utility in programs for the early detection of NSCLC. © 2011 EMBO Molecular Medicine.


Colombo M.,Fondazione IRCCS Instituto Nazionale dei Tumori | de Vecchi G.,Fondazione IRCCS Instituto Nazionale dei Tumori | Caleca L.,Fondazione IRCCS Instituto Nazionale dei Tumori | Foglia C.,Fondazione IRCCS Instituto Nazionale dei Tumori | And 9 more authors.
PLoS ONE | Year: 2013

Several unclassified variants (UVs) have been identified in splicing regions of disease-associated genes and their characterization as pathogenic mutations or benign polymorphisms is crucial for the understanding of their role in disease development. In this study, 24 UVs located at BRCA1 and BRCA2 splice sites were characterized by transcripts analysis. These results were used to evaluate the ability of nine bioinformatics programs in predicting genetic variants causing aberrant splicing (spliceogenic variants) and the nature of aberrant transcripts. Eleven variants in BRCA1 and 8 in BRCA2, including 8 not previously characterized at transcript level, were ascertained to affect mRNA splicing. Of these, 16 led to the synthesis of aberrant transcripts containing premature termination codons (PTCs), 2 to the up-regulation of naturally occurring alternative transcripts containing PTCs, and one to an in-frame deletion within the region coding for the DNA binding domain of BRCA2, causing the loss of the ability to bind the partner protein DSS1 and ssDNA. For each computational program, we evaluated the rate of non-informative analyses, i.e. those that did not recognize the natural splice sites in the wild-type sequence, and the rate of false positive predictions, i.e., variants incorrectly classified as spliceogenic, as a measure of their specificity, under conditions setting sensitivity of predictions to 100%. The programs that performed better were Human Splicing Finder and Automated Splice Site Analyses, both exhibiting 100% informativeness and specificity. For 10 mutations the activation of cryptic splice sites was observed, but we were unable to derive simple criteria to select, among the different cryptic sites predicted by the bioinformatics analyses, those actually used. Consistent with previous reports, our study provides evidences that in silico tools can be used for selecting splice site variants for in vitro analyses. However, the latter remain mandatory for the characterization of the nature of aberrant transcripts. © 2013 Colombo et al.


Beesley J.,Queensland Institute of Medical Research | Johnatty S.E.,Queensland Institute of Medical Research | Chen X.,Queensland Institute of Medical Research | Spurdle A.B.,Queensland Institute of Medical Research | And 9 more authors.
Breast Cancer Research and Treatment | Year: 2011

ABCC11 is an ATP-binding cassette transporter responsible for the transport of a diverse range of lipophilic compounds. A single nucleotide polymorphism (SNP) encoding an amino acid change has recently been shown to determine whether cerumen (earwax) is wet or dry. We hypothesised that this ABCC11 SNP may be associated with breast cancer risk because an association has been reported between wet earwax and increased risk of breast cancer. We therefore analysed the frequency of the functional SNP in 1342 cases and 2256 controls from two breast cancer studies of Caucasian women but found no evidence for an association with breast cancer risk. © 2010 Springer Science+Business Media, LLC.


Chiarella P.,Biomedical University of Rome | Leuener M.,European Molecular Biology Laboratory | Fasci C.,European Molecular Biology Laboratory | de Marco A.,Cogentech | And 3 more authors.
Biotechnology Progress | Year: 2011

We have previously demonstrated how to transform the conventional method of hybridoma production and screening into a fast, high-throughput technology. Nevertheless, there were still open questions related to automated procedures and immunization protocols that we address now by comparing the hybridoma production work-flow in automated and manually executed processes. In addition, since the animals' antibody responses to single or multiple antigen challenge affect monoclonal antibody throughput, different immunization and fusion strategies were tested. Specifically, the results obtained with multiplexing (multiple target antigens injected into a single animal) and single antigen immunization followed by splenocyte pooling immediately before fusion were compared with conventional methods. The results presented here demonstrate that the optimal protocol consists of automated somatic-cell fusion and hybridoma dilution followed by manual plating of hybridoma cells. Additionally, more specific and productive hybridoma clones were obtained with multiplexed immunization in a single animal with respect to the splenocyte pooling from single antigen immunized animals. However, in terms of overall antibody yield, the conventional method consisting of single immunization for each single animal assured ten times more specific hybridoma cell lines than the strategy based on the multiple antigen immunization followed by separate fusion step. In conclusion, the most productive approach for recovering a large number of suitable antibodies relies on single antigen immunization followed by automated fusion and dilution steps and manual plating. © 2011 American Institute of Chemical Engineers.


Fiorino A.,Fondazione IRCCS Instituto Nazionale dei Tumori | Manenti G.,Fondazione IRCCS Instituto Nazionale dei Tumori | Gamba B.,Fondazione IRCCS Instituto Nazionale dei Tumori | Bucci G.,Cogentech | And 8 more authors.
International Journal of Biochemistry and Cell Biology | Year: 2016

Retina-derived POU domain Factor 1 (RPF-1), a member of POU transcription factor family, is encoded by POU6F2 gene, addressed by interstitial deletions at chromosome 7p14 in Wilms tumor (WT). Its expression has been detected in developing kidney and nervous system, suggesting an early role for this gene in regulating development of these organs. To investigate into its functions and determine its role in transcriptional regulation, we generated an inducible stable transfectant from HEK293 cells. RPF-1 showed nuclear localization, elevated stability, and transactivation of promoters featuring POU consensus sites, and led to reduced cell proliferation and in vivo tumor growth. By addressing the whole transcriptome regulated by its induction, we could detect a gross alteration of gene expression that is consistent with promoter occupancy predicted by genome-wide Chip-chip analysis. Comparison of bound regulatory regions with differentially expressed genes allowed identification of 217 candidate targets. Enrichment of divergent octamers in predicted regulatory regions revealed promiscuous binding to bipartite POUS and POUH consensus half-sites with intervening spacers. Gel-shift competition assay confirmed the specificity of RPF-1 binding to consensus motifs, and demonstrated that the Ser-rich region upstream of the POU domain is indispensable to achieve DNA-binding. Promoter-reporter activity addressing a few target genes indicated a dependence by RPF-1 on transcriptional response. In agreement with its expression in developing kidney and nervous system, the induced transcriptome appears to indicate a function for this protein in early renal differentiation and neuronal cell fate, providing a resource for understanding its role in the processes thereby regulated. © 2016 Elsevier Ltd


PubMed | Cogentech, Fondazione IRCCS Instituto Nazionale dei Tumori and University of Milan
Type: | Journal: The international journal of biochemistry & cell biology | Year: 2016

Retina-derived POU domain Factor 1 (RPF-1), a member of POU transcription factor family, is encoded by POU6F2 gene, addressed by interstitial deletions at chromosome 7p14 in Wilms tumor (WT). Its expression has been detected in developing kidney and nervous system, suggesting an early role for this gene in regulating development of these organs. To investigate into its functions and determine its role in transcriptional regulation, we generated an inducible stable transfectant from HEK293 cells. RPF-1 showed nuclear localization, elevated stability, and transactivation of promoters featuring POU consensus sites, and led to reduced cell proliferation and in vivo tumor growth. By addressing the whole transcriptome regulated by its induction, we could detect a gross alteration of gene expression that is consistent with promoter occupancy predicted by genome-wide Chip-chip analysis. Comparison of bound regulatory regions with differentially expressed genes allowed identification of 217 candidate targets. Enrichment of divergent octamers in predicted regulatory regions revealed promiscuous binding to bipartite POUS and POUH consensus half-sites with intervening spacers. Gel-shift competition assay confirmed the specificity of RPF-1 binding to consensus motifs, and demonstrated that the Ser-rich region upstream of the POU domain is indispensable to achieve DNA-binding. Promoter-reporter activity addressing a few target genes indicated a dependence by RPF-1 on transcriptional response. In agreement with its expression in developing kidney and nervous system, the induced transcriptome appears to indicate a function for this protein in early renal differentiation and neuronal cell fate, providing a resource for understanding its role in the processes thereby regulated.

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