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Article ; Online: Age-associated and tissue-specific decline in autophagic activity in the nematode C. elegans.

Chang, Jessica T / Hansen, Malene

2018 Volume 14, Issue 7, Page(s) 1276–1277

Abstract: Macroautophagy/autophagy is a cellular recycling process that is required for the extended life span observed in many longevity paradigms, including in the nematode C. elegans. However, little is known regarding the spatiotemporal changes in autophagic ... ...

Abstract	Macroautophagy/autophagy is a cellular recycling process that is required for the extended life span observed in many longevity paradigms, including in the nematode C. elegans. However, little is known regarding the spatiotemporal changes in autophagic activity in such long-lived mutants as well as in wild-type animals during normal aging. In a recent study, we report that autophagic activity decreases with age in several major tissues of wild-type C. elegans, including the intestine, body-wall muscle, pharynx, and nerve-ring neurons. Moreover, long-lived daf-2/insulin-signaling mutants and glp-1/Notch receptor mutants display increased autophagic activity, yet with different time- and tissue-specific differences. Notably, the intestine appears to be a critical tissue in which autophagy contributes to longevity in glp-1, but not in daf-2 mutants. Our findings indicate that autophagic degradation is reduced with age, possibly with distinct kinetics in different tissues, and that long-lived mutants increase autophagy in a tissue-specific manner, resulting in increased life span.
MeSH term(s)	Animals ; Autophagy ; Caenorhabditis elegans ; Caenorhabditis elegans Proteins ; Longevity ; Transcription Factors
Chemical Substances	Caenorhabditis elegans Proteins ; Transcription Factors
Language	English
Publishing date	2018-05-28
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Comment
ZDB-ID	2454135-7
ISSN	1554-8635 ; 1554-8627
ISSN (online)	1554-8635
ISSN	1554-8627
DOI	10.1080/15548627.2018.1445914
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Article ; Online: Assessing Tissue-Specific Autophagy Flux in Adult Caenorhabditis elegans.

Chang, Jessica T / Hansen, Malene / Kumsta, Caroline

Methods in molecular biology (Clifton, N.J.)

2020 Volume 2144, Page(s) 187–200

Abstract: The cellular recycling process of autophagy is essential for survival, development, and homeostasis. Autophagy also plays an important role in aging and has been linked to longevity in many species, including the nematode C. elegans. Study of the ... ...

Abstract	The cellular recycling process of autophagy is essential for survival, development, and homeostasis. Autophagy also plays an important role in aging and has been linked to longevity in many species, including the nematode C. elegans. Study of the physiological roles of autophagy during C. elegans aging requires methods for the spatiotemporal analysis of autophagy. Here we describe a method for assessing autophagic flux in multiple tissues of C. elegans by quantifying the pool of autophagic vesicles using fluorescently labelled Atg8/LGG-1 reporters upon autophagy inhibition using bafilomycin A
MeSH term(s)	Aging/genetics ; Animals ; Autophagy/genetics ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans/growth & development ; Caenorhabditis elegans Proteins/genetics ; Longevity/genetics ; Microtubule-Associated Proteins/genetics ; Molecular Biology/methods
Chemical Substances	Caenorhabditis elegans Proteins ; Microtubule-Associated Proteins
Language	English
Publishing date	2020-05-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-0592-9_17
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Article ; Online: Correction to: Assessing Tissue-Specific Autophagy Flux in Adult Caenorhabditis elegans.

Chang, Jessica T / Hansen, Malene / Kumsta, Caroline

Methods in molecular biology (Clifton, N.J.)

2020 Volume 2144, Page(s) C1

Abstract: Correction to: Chapter 17 in: Sean P. Curran (ed.), Aging: Methods and Protocols, Methods in Molecular Biology, vol. 2144. ...

Abstract	Correction to: Chapter 17 in: Sean P. Curran (ed.), Aging: Methods and Protocols, Methods in Molecular Biology, vol. 2144.
Language	English
Publishing date	2020-08-04
Publishing country	United States
Document type	Journal Article ; Published Erratum
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-0592-9_24
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Article ; Online: Zebrafish cerebrospinal fluid mediates cell survival through a retinoid signaling pathway.

Chang, Jessica T / Lehtinen, Maria K / Sive, Hazel

Developmental neurobiology

2016 Volume 76, Issue 1, Page(s) 75–92

Abstract: Cerebrospinal fluid (CSF) includes conserved factors whose function is largely unexplored. To assess the role of CSF during embryonic development, CSF was repeatedly drained from embryonic zebrafish brain ventricles soon after their inflation. Removal of ...

Abstract	Cerebrospinal fluid (CSF) includes conserved factors whose function is largely unexplored. To assess the role of CSF during embryonic development, CSF was repeatedly drained from embryonic zebrafish brain ventricles soon after their inflation. Removal of CSF increased cell death in the diencephalon, indicating a survival function. Factors within the CSF are required for neuroepithelial cell survival as injected mouse CSF but not artificial CSF could prevent cell death after CSF depletion. Mass spectrometry analysis of the CSF identified retinol binding protein 4 (Rbp4), which transports retinol, the precursor to retinoic acid (RA). Consistent with a role for Rbp4 in cell survival, inhibition of Rbp4 or RA synthesis increased neuroepithelial cell death. Conversely, ventricle injection of exogenous human RBP4 plus retinol, or RA alone prevented cell death after CSF depletion. Zebrafish rbp4 is highly expressed in the yolk syncytial layer, suggesting Rbp4 protein and retinol/RA precursors can be transported into the CSF from the yolk. In accord with this suggestion, injection of human RBP4 protein into the yolk prevents neuroepithelial cell death in rbp4 loss-of-function embryos. Together, these data support the model that Rbp4 and RA precursors are present within the CSF and used for synthesis of RA, which promotes embryonic neuroepithelial survival.
MeSH term(s)	Animals ; Cell Death ; Cell Survival ; Cerebral Ventricles/metabolism ; Retinoids/metabolism ; Retinol-Binding Proteins, Plasma/metabolism ; Signal Transduction/physiology ; Tretinoin/cerebrospinal fluid ; Zebrafish/metabolism ; Zebrafish Proteins/metabolism
Chemical Substances	Rbp4 protein, zebrafish ; Retinoids ; Retinol-Binding Proteins, Plasma ; Zebrafish Proteins ; Tretinoin (5688UTC01R)
Language	English
Publishing date	2016-01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2256184-5
ISSN	1932-846X ; 1097-4695 ; 1932-8451 ; 0022-3034
ISSN (online)	1932-846X ; 1097-4695
ISSN	1932-8451 ; 0022-3034
DOI	10.1002/dneu.22300
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Zs.A 853: Show issues

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Article ; Online: Manual drainage of the zebrafish embryonic brain ventricles.

Chang, Jessica T / Sive, Hazel

Journal of visualized experiments : JoVE

2012 , Issue 70, Page(s) e4243

Abstract: Cerebrospinal fluid (CSF) is a protein rich fluid contained within the brain ventricles. It is present during early vertebrate embryonic development and persists throughout life. Adult CSF is thought to cushion the brain, remove waste, and carry secreted ...

Abstract	Cerebrospinal fluid (CSF) is a protein rich fluid contained within the brain ventricles. It is present during early vertebrate embryonic development and persists throughout life. Adult CSF is thought to cushion the brain, remove waste, and carry secreted molecules(1,2). In the adult and older embryo, the majority of CSF is made by the choroid plexus, a series of highly vascularized secretory regions located adjacent to the brain ventricles(3-5). In zebrafish, the choroid plexus is fully formed at 144 hours post fertilization (hpf)(6). Prior to this, in both zebrafish and other vertebrate embryos including mouse, a significant amount of embryonic CSF (eCSF) is present . These data and studies in chick suggest that the neuroepithelium is secretory early in development and may be the major source of eCSF prior to choroid plexus development(7). eCSF contains about three times more protein than adult CSF, suggesting that it may have an important role during development(8,9). Studies in chick and mouse demonstrate that secreted factors in the eCSF, fluid pressure, or a combination of these, are important for neurogenesis, gene expression, cell proliferation, and cell survival in the neuroepithelium(10-20). Proteomic analyses of human, rat, mouse, and chick eCSF have identified many proteins that may be necessary for CSF function. These include extracellular matrix components, apolipoproteins, osmotic pressure regulating proteins, and proteins involved in cell death and proliferation(21-24). However, the complex functions of the eCSF are largely unknown. We have developed a method for removing eCSF from zebrafish brain ventricles, thus allowing for identification of eCSF components and for analysis of the eCSF requirement during development. Although more eCSF can be collected from other vertebrate systems with larger embryos, eCSF can be collected from the earliest stages of zebrafish development, and under genetic or environmental conditions that lead to abnormal brain ventricle volume or morphology. Removal and collection of eCSF allows for mass spectrometric analysis, investigation of eCSF function, and reintroduction of select factors into the ventricles to assay their function. Thus the accessibility of the early zebrafish embryo allows for detailed analysis of eCSF function during development.
MeSH term(s)	Animals ; Cerebral Ventricles/chemistry ; Cerebral Ventricles/embryology ; Cerebral Ventricles/surgery ; Cerebrospinal Fluid/chemistry ; Drainage/methods ; Zebrafish/embryology
Language	English
Publishing date	2012-12-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/4243
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Article ; Online: An assay for permeability of the zebrafish embryonic neuroepithelium.

Chang, Jessica T / Sive, Hazel

Journal of visualized experiments : JoVE

2012 , Issue 68, Page(s) e4242

Abstract: The brain ventricular system is conserved among vertebrates and is composed of a series of interconnected cavities called brain ventricles, which form during the earliest stages of brain development and are maintained throughout the animal's life. The ... ...

Abstract	The brain ventricular system is conserved among vertebrates and is composed of a series of interconnected cavities called brain ventricles, which form during the earliest stages of brain development and are maintained throughout the animal's life. The brain ventricular system is found in vertebrates, and the ventricles develop after neural tube formation, when the central lumen fills with cerebrospinal fluid (CSF) (1,2). CSF is a protein rich fluid that is essential for normal brain development and function(3-6). In zebrafish, brain ventricle inflation begins at approximately 18 hr post fertilization (hpf), after the neural tube is closed. Multiple processes are associated with brain ventricle formation, including formation of a neuroepithelium, tight junction formation that regulates permeability and CSF production. We showed that the Na,K-ATPase is required for brain ventricle inflation, impacting all these processes (7,8), while claudin 5a is necessary for tight junction formation (9). Additionally, we showed that "relaxation" of the embryonic neuroepithelium, via inhibition of myosin, is associated with brain ventricle inflation. To investigate the regulation of permeability during zebrafish brain ventricle inflation, we developed a ventricular dye retention assay. This method uses brain ventricle injection in a living zebrafish embryo, a technique previously developed in our lab(10), to fluorescently label the cerebrospinal fluid. Embryos are then imaged over time as the fluorescent dye moves through the brain ventricles and neuroepithelium. The distance the dye front moves away from the basal (non-luminal) side of the neuroepithelium over time is quantified and is a measure of neuroepithelial permeability (Figure 1). We observe that dyes 70 kDa and smaller will move through the neuroepithelium and can be detected outside the embryonic zebrafish brain at 24 hpf (Figure 2). This dye retention assay can be used to analyze neuroepithelial permeability in a variety of different genetic backgrounds, at different times during development, and after environmental perturbations. It may also be useful in examining pathological accumulation of CSF. Overall, this technique allows investigators to analyze the role and regulation of permeability during development and disease.
MeSH term(s)	Animals ; Cerebral Ventricles/embryology ; Cerebral Ventricles/metabolism ; Epithelium/embryology ; Epithelium/metabolism ; Fluorescent Dyes/chemistry ; Fluorescent Dyes/pharmacokinetics ; Microinjections ; Zebrafish/embryology
Chemical Substances	Fluorescent Dyes
Language	English
Publishing date	2012-10-24
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/4242
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Article ; Online: Hormetic heat stress and HSF-1 induce autophagy to improve survival and proteostasis in C. elegans.

Kumsta, Caroline / Chang, Jessica T / Schmalz, Jessica / Hansen, Malene

Nature communications

2017 Volume 8, Page(s) 14337

Abstract: Stress-response pathways have evolved to maintain cellular homeostasis and to ensure the survival of organisms under changing environmental conditions. Whereas severe stress is detrimental, mild stress can be beneficial for health and survival, known as ... ...

Abstract	Stress-response pathways have evolved to maintain cellular homeostasis and to ensure the survival of organisms under changing environmental conditions. Whereas severe stress is detrimental, mild stress can be beneficial for health and survival, known as hormesis. Although the universally conserved heat-shock response regulated by transcription factor HSF-1 has been implicated as an effector mechanism, the role and possible interplay with other cellular processes, such as autophagy, remains poorly understood. Here we show that autophagy is induced in multiple tissues of Caenorhabditis elegans following hormetic heat stress or HSF-1 overexpression. Autophagy-related genes are required for the thermoresistance and longevity of animals exposed to hormetic heat shock or HSF-1 overexpression. Hormetic heat shock also reduces the progressive accumulation of PolyQ aggregates in an autophagy-dependent manner. These findings demonstrate that autophagy contributes to stress resistance and hormesis, and reveal a requirement for autophagy in HSF-1-regulated functions in the heat-shock response, proteostasis and ageing.
MeSH term(s)	Animals ; Autophagy ; Caenorhabditis elegans/genetics ; Caenorhabditis elegans/metabolism ; Caenorhabditis elegans Proteins/metabolism ; Gene Expression Regulation ; Green Fluorescent Proteins/metabolism ; Heat-Shock Response ; Hormesis ; Peptides/metabolism ; Protein Aggregates ; Proteostasis ; Survival Analysis ; Transcription Factors/metabolism
Chemical Substances	Caenorhabditis elegans Proteins ; Peptides ; Protein Aggregates ; Transcription Factors ; heat shock factor-1, C elegans ; Green Fluorescent Proteins (147336-22-9) ; polyglutamine (26700-71-0)
Language	English
Publishing date	2017-02-15
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	2041-1723
ISSN (online)	2041-1723
DOI	10.1038/ncomms14337
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Article: Manual drainage of the zebrafish embryonic brain ventricles

Chang, Jessica T / Sive, Hazel

Journal of visualized experiments. 2012 Dec. 16, , no. 70

2012

Abstract	Cerebrospinal fluid (CSF) is a protein rich fluid contained within the brain ventricles. It is present during early vertebrate embryonic development and persists throughout life. Adult CSF is thought to cushion the brain, remove waste, and carry secreted molecules1,2. In the adult and older embryo, the majority of CSF is made by the choroid plexus, a series of highly vascularized secretory regions located adjacent to the brain ventricles3-5. In zebrafish, the choroid plexus is fully formed at 144 hours post fertilization (hpf)6. Prior to this, in both zebrafish and other vertebrate embryos including mouse, a significant amount of embryonic CSF (eCSF) is present . These data and studies in chick suggest that the neuroepithelium is secretory early in development and may be the major source of eCSF prior to choroid plexus development7. eCSF contains about three times more protein than adult CSF, suggesting that it may have an important role during development8,9. Studies in chick and mouse demonstrate that secreted factors in the eCSF, fluid pressure, or a combination of these, are important for neurogenesis, gene expression, cell proliferation, and cell survival in the neuroepithelium10-20. Proteomic analyses of human, rat, mouse, and chick eCSF have identified many proteins that may be necessary for CSF function. These include extracellular matrix components, apolipoproteins, osmotic pressure regulating proteins, and proteins involved in cell death and proliferation21-24. However, the complex functions of the eCSF are largely unknown. We have developed a method for removing eCSF from zebrafish brain ventricles, thus allowing for identification of eCSF components and for analysis of the eCSF requirement during development. Although more eCSF can be collected from other vertebrate systems with larger embryos, eCSF can be collected from the earliest stages of zebrafish development, and under genetic or environmental conditions that lead to abnormal brain ventricle volume or morphology. Removal and collection of eCSF allows for mass spectrometric analysis, investigation of eCSF function, and reintroduction of select factors into the ventricles to assay their function. Thus the accessibility of the early zebrafish embryo allows for detailed analysis of eCSF function during development.
Keywords	Danio rerio ; adults ; apolipoproteins ; brain ; cell death ; cell proliferation ; cell viability ; cerebrospinal fluid ; chicks ; choroid plexus ; drainage ; embryo (animal) ; embryogenesis ; environmental factors ; extracellular matrix ; gene expression ; mass spectrometry ; mice ; neurogenesis ; osmotic pressure ; rats ; wastes
Language	English
Dates of publication	2012-1216
Size	p. e4243.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/4243
Database	NAL-Catalogue (AGRICOLA)

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Article: An assay for permeability of the zebrafish embryonic neuroepithelium

Chang, Jessica T / Sive, Hazel

Journal of visualized experiments. 2012 Oct. 24, , no. 68

2012

Abstract	The brain ventricular system is conserved among vertebrates and is composed of a series of interconnected cavities called brain ventricles, which form during the earliest stages of brain development and are maintained throughout the animal's life. The brain ventricular system is found in vertebrates, and the ventricles develop after neural tube formation, when the central lumen fills with cerebrospinal fluid (CSF) 1,2. CSF is a protein rich fluid that is essential for normal brain development and function3-6. In zebrafish, brain ventricle inflation begins at approximately 18 hr post fertilization (hpf), after the neural tube is closed. Multiple processes are associated with brain ventricle formation, including formation of a neuroepithelium, tight junction formation that regulates permeability and CSF production. We showed that the Na,K-ATPase is required for brain ventricle inflation, impacting all these processes 7,8, while claudin 5a is necessary for tight junction formation 9. Additionally, we showed that "relaxation" of the embryonic neuroepithelium, via inhibition of myosin, is associated with brain ventricle inflation. To investigate the regulation of permeability during zebrafish brain ventricle inflation, we developed a ventricular dye retention assay. This method uses brain ventricle injection in a living zebrafish embryo, a technique previously developed in our lab10, to fluorescently label the cerebrospinal fluid. Embryos are then imaged over time as the fluorescent dye moves through the brain ventricles and neuroepithelium. The distance the dye front moves away from the basal (non-luminal) side of the neuroepithelium over time is quantified and is a measure of neuroepithelial permeability (Figure 1). We observe that dyes 70 kDa and smaller will move through the neuroepithelium and can be detected outside the embryonic zebrafish brain at 24 hpf (Figure 2). This dye retention assay can be used to analyze neuroepithelial permeability in a variety of different genetic backgrounds, at different times during development, and after environmental perturbations. It may also be useful in examining pathological accumulation of CSF. Overall, this technique allows investigators to analyze the role and regulation of permeability during development and disease.
Keywords	Danio rerio ; brain ; cerebrospinal fluid ; embryo (animal) ; fluorescent dyes ; genetic background ; myosin ; permeability ; sodium-potassium-exchanging ATPase ; tight junctions ; vertebrates
Language	English
Dates of publication	2012-1024
Size	p. e4242.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/4242
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Spatiotemporal regulation of autophagy during

Chang, Jessica T / Kumsta, Caroline / Hellman, Andrew B / Adams, Linnea M / Hansen, Malene

eLife

2017 Volume 6

Abstract: Autophagy has been linked to longevity in many species, but the underlying mechanisms are unclear. Using a GFP-tagged and a new tandem-tagged Atg8/LGG-1 reporter, we quantified autophagic vesicles and performed autophagic flux assays in multiple tissues ... ...

Abstract	Autophagy has been linked to longevity in many species, but the underlying mechanisms are unclear. Using a GFP-tagged and a new tandem-tagged Atg8/LGG-1 reporter, we quantified autophagic vesicles and performed autophagic flux assays in multiple tissues of wild-type
MeSH term(s)	Aging ; Animal Structures/chemistry ; Animals ; Autophagosomes/metabolism ; Autophagy ; Caenorhabditis elegans/cytology ; Caenorhabditis elegans/physiology ; Caenorhabditis elegans Proteins/analysis ; Microtubule-Associated Proteins/analysis ; Models, Animal ; Spatio-Temporal Analysis
Chemical Substances	Caenorhabditis elegans Proteins ; LGG-1 protein, C elegans ; Microtubule-Associated Proteins
Language	English
Publishing date	2017-07-04
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.18459
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