The Experts below are selected from a list of 162 Experts worldwide ranked by ideXlab platform
David J. Griffiths - One of the best experts on this subject based on the ideXlab platform.
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Jaagsiekte sheep retrovirus infection of Lung Slice cultures
Retrovirology, 2015Co-Authors: Chris Cousens, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Jeanie Finlayson, Mark P Dagleish, David J. GriffithsAbstract:BackgroundJaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible neoplastic disease of sheep. OPA is an economically important veterinary disease and is also a valuable naturally occurring animal model of human Lung cancer, with which it shares a similar histological appearance and the activation of common cell signaling pathways. Interestingly, the JSRV Env protein is directly oncogenic and capable of driving cellular transformation in vivo and in vitro . Previous studies of JSRV infection in cell culture have been hindered by the lack of a permissive cell line for the virus. Here, we investigated the ability of JSRV to infect Slices of ovine Lung tissue cultured ex vivo .ResultsWe describe the use of precision cut Lung Slices from healthy sheep to study JSRV infection and transformation ex vivo . Following optimization of the culture system we characterized JSRV infection of Lung Slices and compared the phenotype of infected cells to natural field cases and to experimentally-induced OPA tumors from sheep. JSRV was able to infect cells within Lung Slices, to produce new infectious virions and induce cell proliferation. Immunohistochemical labeling revealed that infected Lung Slice cells express markers of type II pneumocytes and phosphorylated Akt and ERK1/2. These features closely resemble the phenotype of natural and experimentally-derived OPA in sheep, indicating that Lung Slice culture provides an authentic ex vivo model of OPA.ConclusionsWe conclude that we have established an ex vivo model of JSRV infection. This model will be valuable for future studies of JSRV replication and early events in oncogenesis and provides a novel platform for studies of JSRV-induced Lung cancer.
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jaagsiekte sheep retrovirus infection of Lung Slice cultures
Retrovirology, 2015Co-Authors: Chris Cousens, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Jeanie Finlayson, Mark P Dagleish, David J. GriffithsAbstract:Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible neoplastic disease of sheep. OPA is an economically important veterinary disease and is also a valuable naturally occurring animal model of human Lung cancer, with which it shares a similar histological appearance and the activation of common cell signaling pathways. Interestingly, the JSRV Env protein is directly oncogenic and capable of driving cellular transformation in vivo and in vitro. Previous studies of JSRV infection in cell culture have been hindered by the lack of a permissive cell line for the virus. Here, we investigated the ability of JSRV to infect Slices of ovine Lung tissue cultured ex vivo. We describe the use of precision cut Lung Slices from healthy sheep to study JSRV infection and transformation ex vivo. Following optimization of the culture system we characterized JSRV infection of Lung Slices and compared the phenotype of infected cells to natural field cases and to experimentally-induced OPA tumors from sheep. JSRV was able to infect cells within Lung Slices, to produce new infectious virions and induce cell proliferation. Immunohistochemical labeling revealed that infected Lung Slice cells express markers of type II pneumocytes and phosphorylated Akt and ERK1/2. These features closely resemble the phenotype of natural and experimentally-derived OPA in sheep, indicating that Lung Slice culture provides an authentic ex vivo model of OPA. We conclude that we have established an ex vivo model of JSRV infection. This model will be valuable for future studies of JSRV replication and early events in oncogenesis and provides a novel platform for studies of JSRV-induced Lung cancer.
Markus Friden - One of the best experts on this subject based on the ideXlab platform.
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development of a novel Lung Slice methodology for profiling of inhaled compounds
Journal of Pharmaceutical Sciences, 2016Co-Authors: Erica Backstrom, Anders Lundqvist, Elin Boger, Petter Svanberg, Par Ewing, Margareta Hammarlundudenaes, Markus FridenAbstract:The challenge of defining the concentration of unbound drug at the Lung target site after inhalation limits the possibility to optimize target exposure by compound design. In this study, a novel rat Lung Slice methodology has been developed and applied to study drug uptake in Lung tissue, and the mechanisms by which this occurs. Freshly prepared Lung Slices (500 μm) from drug-naive rats were incubated with drugs followed by determination of the unbound drug volume of distribution in Lung (Vu,Lung), as the total concentration of drug in Slices divided by the buffer (unbound) concentration. Vu,Lung determined for a set of inhaled drug compounds ranged from 2.21 mL/g for salbutamol to 2970 mL/g for dibasic compound A. Co-incubation with monensin, a modulator of lysosomal pH, resulted in inhibition of tissue uptake of basic propranolol to 13%, indicating extensive lysosomal trapping. Partitioning into cells was particularly high for the cation MPP+ and the dibasic compound A, likely because of the carrier-mediated transport and lysosomal trapping. The results show that different factors are important for tissue uptake and the presented method can be used for profiling of inhaled compounds, leading to a greater understanding of distribution and exposure of drug in the Lung.
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development of a Lung Slice methodology to study the mechanisms of Lung uptake and lysosomal trapping
European Respiratory Journal, 2014Co-Authors: Erica Backstrom, Anders Lundqvist, Elin Boger, Petter Svanberg, Par Ewing, Margareta Hammarlundudenaes, Markus FridenAbstract:Introduction: Inhalation drug projects face the challenge of identifying the Lung target site unbound drug concentration to better understand the pharmacokinetic/pharmacodynamic (PK/PD) relationships. This study describes the development and application of a new Lung Slice methodology to study drug uptake in the rat Lung and the mechanisms by which it occurs. Methods: Slices (500 µm) from agarose-inflated rat Lungs were incubated for 6 hours with prototypic basic, neutral, and acidic drugs (propranolol, diazepam, and indomethacin, respectively). Drug concentrations were measured in buffer and tissue using LC-MS/MS. The unbound drug volume of distribution in the Lung (Vu,Lung) was calculated by dividing the amount, i.e. total concentration of drug in Slices by the buffer concentration, taking into account the volume of the airway spaces estimated with 14C-inulin. Co-incubation with monensin, a known modulator of lysosomal pH, was made to assess the impact of lysosomal trapping. Results: Vu,Lung for propranolol, diazepam and indomethacin was 297, 19.3, and 11.2 ml/g, respectively. The uptake of propranolol was concentration-dependently inhibited by monensin and reached 98% inhibition at the highest concentration of monensin. The uptake of diazepam and indomethacin was unaffected by monensin. Conclusions: The uptake of propranolol was related to remarkably extensive lysosomal trapping in Lung tissue which did not occur for diazepam and indomethacin. The presented methodology to study drug distribution in the Lung can facilitate and improve the knowledge of PK/PD relationships of Lung targeted drugs.
Chris Cousens - One of the best experts on this subject based on the ideXlab platform.
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Jaagsiekte sheep retrovirus infection of Lung Slice cultures
Retrovirology, 2015Co-Authors: Chris Cousens, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Jeanie Finlayson, Mark P Dagleish, David J. GriffithsAbstract:BackgroundJaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible neoplastic disease of sheep. OPA is an economically important veterinary disease and is also a valuable naturally occurring animal model of human Lung cancer, with which it shares a similar histological appearance and the activation of common cell signaling pathways. Interestingly, the JSRV Env protein is directly oncogenic and capable of driving cellular transformation in vivo and in vitro . Previous studies of JSRV infection in cell culture have been hindered by the lack of a permissive cell line for the virus. Here, we investigated the ability of JSRV to infect Slices of ovine Lung tissue cultured ex vivo .ResultsWe describe the use of precision cut Lung Slices from healthy sheep to study JSRV infection and transformation ex vivo . Following optimization of the culture system we characterized JSRV infection of Lung Slices and compared the phenotype of infected cells to natural field cases and to experimentally-induced OPA tumors from sheep. JSRV was able to infect cells within Lung Slices, to produce new infectious virions and induce cell proliferation. Immunohistochemical labeling revealed that infected Lung Slice cells express markers of type II pneumocytes and phosphorylated Akt and ERK1/2. These features closely resemble the phenotype of natural and experimentally-derived OPA in sheep, indicating that Lung Slice culture provides an authentic ex vivo model of OPA.ConclusionsWe conclude that we have established an ex vivo model of JSRV infection. This model will be valuable for future studies of JSRV replication and early events in oncogenesis and provides a novel platform for studies of JSRV-induced Lung cancer.
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jaagsiekte sheep retrovirus infection of Lung Slice cultures
Retrovirology, 2015Co-Authors: Chris Cousens, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Jeanie Finlayson, Mark P Dagleish, David J. GriffithsAbstract:Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible neoplastic disease of sheep. OPA is an economically important veterinary disease and is also a valuable naturally occurring animal model of human Lung cancer, with which it shares a similar histological appearance and the activation of common cell signaling pathways. Interestingly, the JSRV Env protein is directly oncogenic and capable of driving cellular transformation in vivo and in vitro. Previous studies of JSRV infection in cell culture have been hindered by the lack of a permissive cell line for the virus. Here, we investigated the ability of JSRV to infect Slices of ovine Lung tissue cultured ex vivo. We describe the use of precision cut Lung Slices from healthy sheep to study JSRV infection and transformation ex vivo. Following optimization of the culture system we characterized JSRV infection of Lung Slices and compared the phenotype of infected cells to natural field cases and to experimentally-induced OPA tumors from sheep. JSRV was able to infect cells within Lung Slices, to produce new infectious virions and induce cell proliferation. Immunohistochemical labeling revealed that infected Lung Slice cells express markers of type II pneumocytes and phosphorylated Akt and ERK1/2. These features closely resemble the phenotype of natural and experimentally-derived OPA in sheep, indicating that Lung Slice culture provides an authentic ex vivo model of OPA. We conclude that we have established an ex vivo model of JSRV infection. This model will be valuable for future studies of JSRV replication and early events in oncogenesis and provides a novel platform for studies of JSRV-induced Lung cancer.
Charline Alleaume - One of the best experts on this subject based on the ideXlab platform.
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Jaagsiekte sheep retrovirus infection of Lung Slice cultures
Retrovirology, 2015Co-Authors: Chris Cousens, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Jeanie Finlayson, Mark P Dagleish, David J. GriffithsAbstract:BackgroundJaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible neoplastic disease of sheep. OPA is an economically important veterinary disease and is also a valuable naturally occurring animal model of human Lung cancer, with which it shares a similar histological appearance and the activation of common cell signaling pathways. Interestingly, the JSRV Env protein is directly oncogenic and capable of driving cellular transformation in vivo and in vitro . Previous studies of JSRV infection in cell culture have been hindered by the lack of a permissive cell line for the virus. Here, we investigated the ability of JSRV to infect Slices of ovine Lung tissue cultured ex vivo .ResultsWe describe the use of precision cut Lung Slices from healthy sheep to study JSRV infection and transformation ex vivo . Following optimization of the culture system we characterized JSRV infection of Lung Slices and compared the phenotype of infected cells to natural field cases and to experimentally-induced OPA tumors from sheep. JSRV was able to infect cells within Lung Slices, to produce new infectious virions and induce cell proliferation. Immunohistochemical labeling revealed that infected Lung Slice cells express markers of type II pneumocytes and phosphorylated Akt and ERK1/2. These features closely resemble the phenotype of natural and experimentally-derived OPA in sheep, indicating that Lung Slice culture provides an authentic ex vivo model of OPA.ConclusionsWe conclude that we have established an ex vivo model of JSRV infection. This model will be valuable for future studies of JSRV replication and early events in oncogenesis and provides a novel platform for studies of JSRV-induced Lung cancer.
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jaagsiekte sheep retrovirus infection of Lung Slice cultures
Retrovirology, 2015Co-Authors: Chris Cousens, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Jeanie Finlayson, Mark P Dagleish, David J. GriffithsAbstract:Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible neoplastic disease of sheep. OPA is an economically important veterinary disease and is also a valuable naturally occurring animal model of human Lung cancer, with which it shares a similar histological appearance and the activation of common cell signaling pathways. Interestingly, the JSRV Env protein is directly oncogenic and capable of driving cellular transformation in vivo and in vitro. Previous studies of JSRV infection in cell culture have been hindered by the lack of a permissive cell line for the virus. Here, we investigated the ability of JSRV to infect Slices of ovine Lung tissue cultured ex vivo. We describe the use of precision cut Lung Slices from healthy sheep to study JSRV infection and transformation ex vivo. Following optimization of the culture system we characterized JSRV infection of Lung Slices and compared the phenotype of infected cells to natural field cases and to experimentally-induced OPA tumors from sheep. JSRV was able to infect cells within Lung Slices, to produce new infectious virions and induce cell proliferation. Immunohistochemical labeling revealed that infected Lung Slice cells express markers of type II pneumocytes and phosphorylated Akt and ERK1/2. These features closely resemble the phenotype of natural and experimentally-derived OPA in sheep, indicating that Lung Slice culture provides an authentic ex vivo model of OPA. We conclude that we have established an ex vivo model of JSRV infection. This model will be valuable for future studies of JSRV replication and early events in oncogenesis and provides a novel platform for studies of JSRV-induced Lung cancer.
Henny M Martineau - One of the best experts on this subject based on the ideXlab platform.
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Jaagsiekte sheep retrovirus infection of Lung Slice cultures
Retrovirology, 2015Co-Authors: Chris Cousens, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Jeanie Finlayson, Mark P Dagleish, David J. GriffithsAbstract:BackgroundJaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible neoplastic disease of sheep. OPA is an economically important veterinary disease and is also a valuable naturally occurring animal model of human Lung cancer, with which it shares a similar histological appearance and the activation of common cell signaling pathways. Interestingly, the JSRV Env protein is directly oncogenic and capable of driving cellular transformation in vivo and in vitro . Previous studies of JSRV infection in cell culture have been hindered by the lack of a permissive cell line for the virus. Here, we investigated the ability of JSRV to infect Slices of ovine Lung tissue cultured ex vivo .ResultsWe describe the use of precision cut Lung Slices from healthy sheep to study JSRV infection and transformation ex vivo . Following optimization of the culture system we characterized JSRV infection of Lung Slices and compared the phenotype of infected cells to natural field cases and to experimentally-induced OPA tumors from sheep. JSRV was able to infect cells within Lung Slices, to produce new infectious virions and induce cell proliferation. Immunohistochemical labeling revealed that infected Lung Slice cells express markers of type II pneumocytes and phosphorylated Akt and ERK1/2. These features closely resemble the phenotype of natural and experimentally-derived OPA in sheep, indicating that Lung Slice culture provides an authentic ex vivo model of OPA.ConclusionsWe conclude that we have established an ex vivo model of JSRV infection. This model will be valuable for future studies of JSRV replication and early events in oncogenesis and provides a novel platform for studies of JSRV-induced Lung cancer.
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jaagsiekte sheep retrovirus infection of Lung Slice cultures
Retrovirology, 2015Co-Authors: Chris Cousens, Charline Alleaume, Esther Bijsmans, Henny M Martineau, Jeanie Finlayson, Mark P Dagleish, David J. GriffithsAbstract:Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible neoplastic disease of sheep. OPA is an economically important veterinary disease and is also a valuable naturally occurring animal model of human Lung cancer, with which it shares a similar histological appearance and the activation of common cell signaling pathways. Interestingly, the JSRV Env protein is directly oncogenic and capable of driving cellular transformation in vivo and in vitro. Previous studies of JSRV infection in cell culture have been hindered by the lack of a permissive cell line for the virus. Here, we investigated the ability of JSRV to infect Slices of ovine Lung tissue cultured ex vivo. We describe the use of precision cut Lung Slices from healthy sheep to study JSRV infection and transformation ex vivo. Following optimization of the culture system we characterized JSRV infection of Lung Slices and compared the phenotype of infected cells to natural field cases and to experimentally-induced OPA tumors from sheep. JSRV was able to infect cells within Lung Slices, to produce new infectious virions and induce cell proliferation. Immunohistochemical labeling revealed that infected Lung Slice cells express markers of type II pneumocytes and phosphorylated Akt and ERK1/2. These features closely resemble the phenotype of natural and experimentally-derived OPA in sheep, indicating that Lung Slice culture provides an authentic ex vivo model of OPA. We conclude that we have established an ex vivo model of JSRV infection. This model will be valuable for future studies of JSRV replication and early events in oncogenesis and provides a novel platform for studies of JSRV-induced Lung cancer.
