The Experts below are selected from a list of 264 Experts worldwide ranked by ideXlab platform
Tadashi Yamamoto - One of the best experts on this subject based on the ideXlab platform.
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expression of the chemokine fractalkine fkn cx3cl1 by podocytes in normal and proteinuric rat Kidney Glomerulus
Nephron Experimental Nephrology, 2009Co-Authors: Koichi Katsuyama, Masaaki Nameta, Hidehiko Fujinaka, Eishin Yaoita, Yutaka Yoshida, Keiko Yamamoto, Shuichi Tomizawa, Makoto Uchiyama, Tadashi YamamotoAbstract:Background/Aims: A chemokine fractalkine (FKN/CX3CL1) is induced primarily by endothelial cells and accumulates inflammatory cells via its receptor CX3CR1. Since glomerular preferential expression of FKN/CX3CL1 gene was reported in normal human Kidney, we presumed FKN/CX3CL1 might play some roles in glomerular physiology. The purpose of this study is to examine the expression and localization of FKN/CX3CL1 in normal and proteinuric glomeruli. Methods: Normal and proteinuric rat Kidneys were studied. The gene and protein expressions of FKN/CX3CL1 and CX3CR1 were examined by real-time RT-PCR, in situ hybridization and immunohistochemistry, topic-western-blotting/">Western blotting. Results: By real-time RT-PCR, glomerular preferential expression of FKN/CX3CL1 was confirmed, whereas CX3CR1 was detected in glomeruli and cortices. The localization of FKN/CX3CL1 gene and protein were demonstrated in glomerular cells including podocytes. In nephrotic puromycin aminonucleoside (PAN) nephrosis glomeruli, increased expression of FKN/CX3CL1 in podocyte was shown by immunohistochemistry. topic-western-blotting/">Western blotting showed that in nephrotic glomeruli, the membrane-anchored form of FKN/CX3CL1 was increased while the soluble form was decreased. Conclusion: The expression of FKN/CX3CL1 in normal podocytes and the increased expression of the membrane-anchored form in nephrotic glomeruli strongly suggest that FKN/CX3CL1 may play roles in glomerular physiology such as maintaining glomerular filtration barrier.
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Expression of the chemokine fractalkine (FKN/CX3CL1) by podocytes in normal and proteinuric rat Kidney Glomerulus.
Nephron. Experimental nephrology, 2009Co-Authors: Koichi Katsuyama, Masaaki Nameta, Hidehiko Fujinaka, Eishin Yaoita, Yutaka Yoshida, Keiko Yamamoto, Shuichi Tomizawa, Makoto Uchiyama, Tadashi YamamotoAbstract:Background/Aims: A chemokine fractalkine (FKN/CX3CL1) is induced primarily by endothelial cells and accumulates inflammatory cells via its receptor CX3CR1. Since glomerular preferential expression of FKN/CX3CL1 gene was reported in normal human Kidney, we presumed FKN/CX3CL1 might play some roles in glomerular physiology. The purpose of this study is to examine the expression and localization of FKN/CX3CL1 in normal and proteinuric glomeruli. Methods: Normal and proteinuric rat Kidneys were studied. The gene and protein expressions of FKN/CX3CL1 and CX3CR1 were examined by real-time RT-PCR, in situ hybridization and immunohistochemistry, topic-western-blotting/">Western blotting. Results: By real-time RT-PCR, glomerular preferential expression of FKN/CX3CL1 was confirmed, whereas CX3CR1 was detected in glomeruli and cortices. The localization of FKN/CX3CL1 gene and protein were demonstrated in glomerular cells including podocytes. In nephrotic puromycin aminonucleoside (PAN) nephrosis glomeruli, increased expression of FKN/CX3CL1 in podocyte was shown by immunohistochemistry. topic-western-blotting/">Western blotting showed that in nephrotic glomeruli, the membrane-anchored form of FKN/CX3CL1 was increased while the soluble form was decreased. Conclusion: The expression of FKN/CX3CL1 in normal podocytes and the increased expression of the membrane-anchored form in nephrotic glomeruli strongly suggest that FKN/CX3CL1 may play roles in glomerular physiology such as maintaining glomerular filtration barrier.
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Identification and localization of novel genes preferentially expressed in human Kidney Glomerulus
Nephrology (Carlton Vic.), 2008Co-Authors: Lino Munoz Cuellar, Masahito Miyamoto, Masayuki Tasaki, Hidehiko Fujinaka, Eishin Yaoita, Yutaka Yoshida, Keiko Yamamoto, Linning Zhao, Ismail Tamer, Tadashi YamamotoAbstract:SUMMARY Aim: To find novel genes abundantly and preferentially expressed in human Glomerulus, we constructed a glomerular cDNA library and verified the reliability of our database by comparison with the Stanford Microarray Database (SMD), followed by reverse transcription polymerase chain reaction (RT-PCR) and in situ hybridization (ISH). Methods: RNA was extracted from normal human glomeruli, and the cDNA library was constructed by plasmid cloning. Out of 5 × 103 clones from the library, 91 UniGene clusters of more than three clones were identified as ‘glomerular-abundant genes’. All these genes were referred to the SMD, and 18 genes were defined as ‘glomerular preferential genes’. Four unknown genes –IFI27, CRHBP, FLJ10154 and SEMA5B– were selected for RT-PCR to compare expression in the Glomerulus with that in the cortex and medulla, and for ISH to examine glomerular localization. Also, three unknown genes that were glomerular abundant but not listed in the SMD –DDX5, HSPC138, and MGC10940– were selected for RT-PCR and ISH. Finally, a Kidney biopsy topic-specimen/">specimen of crescentic glomerulonephritis was used for ISH to examine glomerular expression for CRHBP mRNA. Results: Among the selected seven glomerular-abundant genes, six were confirmed as ‘glomerular preferential genes’ by RT-PCR. By ISH, all these genes were demonstrated in podocytes. The expression of CRHBP mRNA in a single living podocyte was not changed between normal and crescentic Glomerulus. Conclusion: Glomerular preferential expression and podocyte localization of these novel genes have been demonstrated for the first time. Because some of these genes were not listed in SMD, our database can be a useful tool to find novel human glomerular genes.
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Human Kidney Glomerulus proteome and biomarker discovery of Kidney diseases
Proteomics Clinical Applications, 2008Co-Authors: Yutaka Yoshida, Izumi Taguchi, Masahito Miyamoto, Hidehiko Fujinaka, Eishin Yaoita, Bo Xu, Ying Zhang, Tadashi YamamotoAbstract:The Kidney Glomerulus is the site of plasma filtration and production of primary topic-urine/">urine in the Kidney. The structure not only plays a pivotal role in ultrafiltration of plasma into topic-urine/">urine but also is the locus of Kidney diseases progressing to chronic renal failure. Patients afflicted with these glomerular diseases frequently progress to irreversible loss of renal function and inevitably require replacement therapies. The diagnosis and treatment of glomerular diseases are now based on clinical manifestations, urinary protein excretion level, and renal pathology of needle biopsy topic-specimen/">specimens. The molecular mechanisms underlying the progression of glomerular diseases are still obscure despite a great number of clinical and experimental studies. Proteomics is a particularly promising approach for the discovery of proteins relevant to physiological and pathophysiological processes, and has been recently employed in nephrology. Although until now most efforts of proteomic analysis have been conducted with topic-urine/">urine, the biological fluid that is easily collected without invasive procedures, proteomic analysis of the Glomerulus, the tissue most proximal to the disease loci, is the most straightforward approach. In this review, we attempt to outline the current status of clinical proteomics of the Glomerulus and provide a perspective of protein biomarker discovery of glomerular diseases.
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proteomic analysis of human Kidney Glomerulus with fluoresent 2d difference gel electrophoresis 2d dige using saturation labeling
Seibutsu Butsuri Kagaku, 2006Co-Authors: Yutaka Yoshdia, Tadashi YamamotoAbstract:We have attempted to separate a minimal amount of proteins of glomeluli isolated from biopsy tissues of human Kidney on large format 2-DE gels by exploiting extremely sensitive fluorescent dyes (Cy3- and Cy-5 saturation dye). Only 2.5μg glomerular proteins corresponding to 2-3 glomeruli, gave highly resolved 2-DE profiles, suggesting the applicability of 2-DE to analyze Glomerulus topic-proteome/">proteome in biopsy topic-specimen/">specimens. We propose a platform for highly quantitative analysis of differential protein expression in the Glomerulus in health and disease with the saturation dye as a detection tool.
Yutaka Yoshida - One of the best experts on this subject based on the ideXlab platform.
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profiling and annotation of human Kidney Glomerulus proteome
Proteome Science, 2013Co-Authors: Yutaka Yoshida, Masaaki Nameta, Tomoo Makiguchi, Toshikazu Ikoma, Sameh Magdeldin, Hidehiko Fujinaka, Bo Xu, Ying Zhang, Eishin YaoitaAbstract:Background The comprehensive analysis of human Kidney Glomerulus we previously performed using highly purified glomeruli, provided a dataset of 6,686 unique proteins representing 2,966 distinct genes. This dataset, however, contained considerable topic-redundancy/">redundancy resulting from identification criteria under which all the proteins matched with the same set of peptides and its subset were reported as identified proteins. In this study we reanalyzed the raw data using the Mascot search engine and highly stringent criteria in order to select proteins with the highest scores matching peptides with scores exceeding the “Identity Threshold” and one or more unique peptides. This enabled us to exclude proteins with lower scores which only matched the same set of peptides or its subset. This approach provided a high-confidence, non-redundant dataset of identified proteins for extensive profiling, annotation, and comparison with other topic-proteome/">proteome datasets that can provide biologically relevant knowledge of Glomerulus topic-proteome/">proteome.
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expression of the chemokine fractalkine fkn cx3cl1 by podocytes in normal and proteinuric rat Kidney Glomerulus
Nephron Experimental Nephrology, 2009Co-Authors: Koichi Katsuyama, Masaaki Nameta, Hidehiko Fujinaka, Eishin Yaoita, Yutaka Yoshida, Keiko Yamamoto, Shuichi Tomizawa, Makoto Uchiyama, Tadashi YamamotoAbstract:Background/Aims: A chemokine fractalkine (FKN/CX3CL1) is induced primarily by endothelial cells and accumulates inflammatory cells via its receptor CX3CR1. Since glomerular preferential expression of FKN/CX3CL1 gene was reported in normal human Kidney, we presumed FKN/CX3CL1 might play some roles in glomerular physiology. The purpose of this study is to examine the expression and localization of FKN/CX3CL1 in normal and proteinuric glomeruli. Methods: Normal and proteinuric rat Kidneys were studied. The gene and protein expressions of FKN/CX3CL1 and CX3CR1 were examined by real-time RT-PCR, in situ hybridization and immunohistochemistry, topic-western-blotting/">Western blotting. Results: By real-time RT-PCR, glomerular preferential expression of FKN/CX3CL1 was confirmed, whereas CX3CR1 was detected in glomeruli and cortices. The localization of FKN/CX3CL1 gene and protein were demonstrated in glomerular cells including podocytes. In nephrotic puromycin aminonucleoside (PAN) nephrosis glomeruli, increased expression of FKN/CX3CL1 in podocyte was shown by immunohistochemistry. topic-western-blotting/">Western blotting showed that in nephrotic glomeruli, the membrane-anchored form of FKN/CX3CL1 was increased while the soluble form was decreased. Conclusion: The expression of FKN/CX3CL1 in normal podocytes and the increased expression of the membrane-anchored form in nephrotic glomeruli strongly suggest that FKN/CX3CL1 may play roles in glomerular physiology such as maintaining glomerular filtration barrier.
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Expression of the chemokine fractalkine (FKN/CX3CL1) by podocytes in normal and proteinuric rat Kidney Glomerulus.
Nephron. Experimental nephrology, 2009Co-Authors: Koichi Katsuyama, Masaaki Nameta, Hidehiko Fujinaka, Eishin Yaoita, Yutaka Yoshida, Keiko Yamamoto, Shuichi Tomizawa, Makoto Uchiyama, Tadashi YamamotoAbstract:Background/Aims: A chemokine fractalkine (FKN/CX3CL1) is induced primarily by endothelial cells and accumulates inflammatory cells via its receptor CX3CR1. Since glomerular preferential expression of FKN/CX3CL1 gene was reported in normal human Kidney, we presumed FKN/CX3CL1 might play some roles in glomerular physiology. The purpose of this study is to examine the expression and localization of FKN/CX3CL1 in normal and proteinuric glomeruli. Methods: Normal and proteinuric rat Kidneys were studied. The gene and protein expressions of FKN/CX3CL1 and CX3CR1 were examined by real-time RT-PCR, in situ hybridization and immunohistochemistry, topic-western-blotting/">Western blotting. Results: By real-time RT-PCR, glomerular preferential expression of FKN/CX3CL1 was confirmed, whereas CX3CR1 was detected in glomeruli and cortices. The localization of FKN/CX3CL1 gene and protein were demonstrated in glomerular cells including podocytes. In nephrotic puromycin aminonucleoside (PAN) nephrosis glomeruli, increased expression of FKN/CX3CL1 in podocyte was shown by immunohistochemistry. topic-western-blotting/">Western blotting showed that in nephrotic glomeruli, the membrane-anchored form of FKN/CX3CL1 was increased while the soluble form was decreased. Conclusion: The expression of FKN/CX3CL1 in normal podocytes and the increased expression of the membrane-anchored form in nephrotic glomeruli strongly suggest that FKN/CX3CL1 may play roles in glomerular physiology such as maintaining glomerular filtration barrier.
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Identification and localization of novel genes preferentially expressed in human Kidney Glomerulus
Nephrology (Carlton Vic.), 2008Co-Authors: Lino Munoz Cuellar, Masahito Miyamoto, Masayuki Tasaki, Hidehiko Fujinaka, Eishin Yaoita, Yutaka Yoshida, Keiko Yamamoto, Linning Zhao, Ismail Tamer, Tadashi YamamotoAbstract:SUMMARY Aim: To find novel genes abundantly and preferentially expressed in human Glomerulus, we constructed a glomerular cDNA library and verified the reliability of our database by comparison with the Stanford Microarray Database (SMD), followed by reverse transcription polymerase chain reaction (RT-PCR) and in situ hybridization (ISH). Methods: RNA was extracted from normal human glomeruli, and the cDNA library was constructed by plasmid cloning. Out of 5 × 103 clones from the library, 91 UniGene clusters of more than three clones were identified as ‘glomerular-abundant genes’. All these genes were referred to the SMD, and 18 genes were defined as ‘glomerular preferential genes’. Four unknown genes –IFI27, CRHBP, FLJ10154 and SEMA5B– were selected for RT-PCR to compare expression in the Glomerulus with that in the cortex and medulla, and for ISH to examine glomerular localization. Also, three unknown genes that were glomerular abundant but not listed in the SMD –DDX5, HSPC138, and MGC10940– were selected for RT-PCR and ISH. Finally, a Kidney biopsy topic-specimen/">specimen of crescentic glomerulonephritis was used for ISH to examine glomerular expression for CRHBP mRNA. Results: Among the selected seven glomerular-abundant genes, six were confirmed as ‘glomerular preferential genes’ by RT-PCR. By ISH, all these genes were demonstrated in podocytes. The expression of CRHBP mRNA in a single living podocyte was not changed between normal and crescentic Glomerulus. Conclusion: Glomerular preferential expression and podocyte localization of these novel genes have been demonstrated for the first time. Because some of these genes were not listed in SMD, our database can be a useful tool to find novel human glomerular genes.
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Human Kidney Glomerulus proteome and biomarker discovery of Kidney diseases
Proteomics Clinical Applications, 2008Co-Authors: Yutaka Yoshida, Izumi Taguchi, Masahito Miyamoto, Hidehiko Fujinaka, Eishin Yaoita, Bo Xu, Ying Zhang, Tadashi YamamotoAbstract:The Kidney Glomerulus is the site of plasma filtration and production of primary topic-urine/">urine in the Kidney. The structure not only plays a pivotal role in ultrafiltration of plasma into topic-urine/">urine but also is the locus of Kidney diseases progressing to chronic renal failure. Patients afflicted with these glomerular diseases frequently progress to irreversible loss of renal function and inevitably require replacement therapies. The diagnosis and treatment of glomerular diseases are now based on clinical manifestations, urinary protein excretion level, and renal pathology of needle biopsy topic-specimen/">specimens. The molecular mechanisms underlying the progression of glomerular diseases are still obscure despite a great number of clinical and experimental studies. Proteomics is a particularly promising approach for the discovery of proteins relevant to physiological and pathophysiological processes, and has been recently employed in nephrology. Although until now most efforts of proteomic analysis have been conducted with topic-urine/">urine, the biological fluid that is easily collected without invasive procedures, proteomic analysis of the Glomerulus, the tissue most proximal to the disease loci, is the most straightforward approach. In this review, we attempt to outline the current status of clinical proteomics of the Glomerulus and provide a perspective of protein biomarker discovery of glomerular diseases.
Masaaki Nameta - One of the best experts on this subject based on the ideXlab platform.
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profiling and annotation of human Kidney Glomerulus proteome
Proteome Science, 2013Co-Authors: Yutaka Yoshida, Masaaki Nameta, Tomoo Makiguchi, Toshikazu Ikoma, Sameh Magdeldin, Hidehiko Fujinaka, Bo Xu, Ying Zhang, Eishin YaoitaAbstract:Background The comprehensive analysis of human Kidney Glomerulus we previously performed using highly purified glomeruli, provided a dataset of 6,686 unique proteins representing 2,966 distinct genes. This dataset, however, contained considerable topic-redundancy/">redundancy resulting from identification criteria under which all the proteins matched with the same set of peptides and its subset were reported as identified proteins. In this study we reanalyzed the raw data using the Mascot search engine and highly stringent criteria in order to select proteins with the highest scores matching peptides with scores exceeding the “Identity Threshold” and one or more unique peptides. This enabled us to exclude proteins with lower scores which only matched the same set of peptides or its subset. This approach provided a high-confidence, non-redundant dataset of identified proteins for extensive profiling, annotation, and comparison with other topic-proteome/">proteome datasets that can provide biologically relevant knowledge of Glomerulus topic-proteome/">proteome.
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expression of the chemokine fractalkine fkn cx3cl1 by podocytes in normal and proteinuric rat Kidney Glomerulus
Nephron Experimental Nephrology, 2009Co-Authors: Koichi Katsuyama, Masaaki Nameta, Hidehiko Fujinaka, Eishin Yaoita, Yutaka Yoshida, Keiko Yamamoto, Shuichi Tomizawa, Makoto Uchiyama, Tadashi YamamotoAbstract:Background/Aims: A chemokine fractalkine (FKN/CX3CL1) is induced primarily by endothelial cells and accumulates inflammatory cells via its receptor CX3CR1. Since glomerular preferential expression of FKN/CX3CL1 gene was reported in normal human Kidney, we presumed FKN/CX3CL1 might play some roles in glomerular physiology. The purpose of this study is to examine the expression and localization of FKN/CX3CL1 in normal and proteinuric glomeruli. Methods: Normal and proteinuric rat Kidneys were studied. The gene and protein expressions of FKN/CX3CL1 and CX3CR1 were examined by real-time RT-PCR, in situ hybridization and immunohistochemistry, topic-western-blotting/">Western blotting. Results: By real-time RT-PCR, glomerular preferential expression of FKN/CX3CL1 was confirmed, whereas CX3CR1 was detected in glomeruli and cortices. The localization of FKN/CX3CL1 gene and protein were demonstrated in glomerular cells including podocytes. In nephrotic puromycin aminonucleoside (PAN) nephrosis glomeruli, increased expression of FKN/CX3CL1 in podocyte was shown by immunohistochemistry. topic-western-blotting/">Western blotting showed that in nephrotic glomeruli, the membrane-anchored form of FKN/CX3CL1 was increased while the soluble form was decreased. Conclusion: The expression of FKN/CX3CL1 in normal podocytes and the increased expression of the membrane-anchored form in nephrotic glomeruli strongly suggest that FKN/CX3CL1 may play roles in glomerular physiology such as maintaining glomerular filtration barrier.
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Expression of the chemokine fractalkine (FKN/CX3CL1) by podocytes in normal and proteinuric rat Kidney Glomerulus.
Nephron. Experimental nephrology, 2009Co-Authors: Koichi Katsuyama, Masaaki Nameta, Hidehiko Fujinaka, Eishin Yaoita, Yutaka Yoshida, Keiko Yamamoto, Shuichi Tomizawa, Makoto Uchiyama, Tadashi YamamotoAbstract:Background/Aims: A chemokine fractalkine (FKN/CX3CL1) is induced primarily by endothelial cells and accumulates inflammatory cells via its receptor CX3CR1. Since glomerular preferential expression of FKN/CX3CL1 gene was reported in normal human Kidney, we presumed FKN/CX3CL1 might play some roles in glomerular physiology. The purpose of this study is to examine the expression and localization of FKN/CX3CL1 in normal and proteinuric glomeruli. Methods: Normal and proteinuric rat Kidneys were studied. The gene and protein expressions of FKN/CX3CL1 and CX3CR1 were examined by real-time RT-PCR, in situ hybridization and immunohistochemistry, topic-western-blotting/">Western blotting. Results: By real-time RT-PCR, glomerular preferential expression of FKN/CX3CL1 was confirmed, whereas CX3CR1 was detected in glomeruli and cortices. The localization of FKN/CX3CL1 gene and protein were demonstrated in glomerular cells including podocytes. In nephrotic puromycin aminonucleoside (PAN) nephrosis glomeruli, increased expression of FKN/CX3CL1 in podocyte was shown by immunohistochemistry. topic-western-blotting/">Western blotting showed that in nephrotic glomeruli, the membrane-anchored form of FKN/CX3CL1 was increased while the soluble form was decreased. Conclusion: The expression of FKN/CX3CL1 in normal podocytes and the increased expression of the membrane-anchored form in nephrotic glomeruli strongly suggest that FKN/CX3CL1 may play roles in glomerular physiology such as maintaining glomerular filtration barrier.
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in depth proteomic profiling of the normal human Kidney Glomerulus using two dimensional protein prefractionation in combination with liquid chromatography tandem mass spectrometry
Journal of Proteome Research, 2007Co-Authors: Masahito Miyamoto, Izumi Taguchi, Yoshimi Nagasaka, Masaaki Nameta, Masayuki Tasaki, Hiroshi Sezaki, Bo Xu, Yutaka Yoshida, Ying Zhang, Lino Munoz CuellarAbstract:The Kidney Glomerulus plays a pivotal role in ultrafiltration of plasma into topic-urine/">urine and also is the locus of Kidney disease progressing to chronic renal failure. We have focused proteomic analysis on the Glomerulus that is most proximal to the disease locus. In the present study, we aimed to provide a confident, in-depth profiling of the Glomerulus topic-proteome/">proteome. The glomeruli were highly purified from the Kidney cortex from a male, 68-year-old patient who underwent nephroureterectomy due to ureter carcinoma. The patient was normal in clinical examinations including serum creatinine and urea levels and liver function, and did not receive any chemotherapy and radiotherapy. The cortical tissue was histologically normal, and no significant deposition of immunoglobulins and complement C3 was observed. We employed a novel strategy of protein separation using 1D (SDS-PAGE) and 2D (solution-phase IEF in combination with SDS-PAGE) prefractionation prior to the shotgun analysis with LC−MS/MS. The protein prefractionatio...
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In-depth proteomic profiling of the normal human Kidney Glomerulus using two-dimensional protein prefractionation in combination with liquid chromatography-tandem mass spectrometry.
Journal of proteome research, 2007Co-Authors: Masahito Miyamoto, Izumi Taguchi, Yoshimi Nagasaka, Masaaki Nameta, Masayuki Tasaki, Hiroshi Sezaki, Yutaka Yoshida, Ying Zhang, Lino Munoz CuellarAbstract:The Kidney Glomerulus plays a pivotal role in ultrafiltration of plasma into topic-urine/">urine and also is the locus of Kidney disease progressing to chronic renal failure. We have focused proteomic analysis on the Glomerulus that is most proximal to the disease locus. In the present study, we aimed to provide a confident, in-depth profiling of the Glomerulus topic-proteome/">proteome. The glomeruli were highly purified from the Kidney cortex from a male, 68-year-old patient who underwent nephroureterectomy due to ureter carcinoma. The patient was normal in clinical examinations including serum creatinine and urea levels and liver function, and did not receive any chemotherapy and radiotherapy. The cortical tissue was histologically normal, and no significant deposition of immunoglobulins and complement C3 was observed. We employed a novel strategy of protein separation using 1D (SDS-PAGE) and 2D (solution-phase IEF in combination with SDS-PAGE) prefractionation prior to the shotgun analysis with LC-MS/MS. The protein prefractionation produced 90 fractions, and eventually provided a confident set of identified proteins consisting of 6686 unique proteins (3679 proteins with two or more peptide matches and 3007 proteins with one peptide match), representing 2966 distinct genes. All the identified proteins were annotated and classified in terms of molecular function and biological process, compiled into 1D and 2D protein arrays, consisting of 15 and 75 sections, corresponding to the protein fractions which were defined by MW and pI range, and deposited on a Web-based database (http://www.hkupp.org). The most remarkable feature of the Glomerulus topic-proteome/">proteome was a high incidence of identification of cytoskeleton-related proteins, presumably reflecting the well-developed, cytoskeletal organization of glomerular cells related to their physiological functions.
Thomas Benzing - One of the best experts on this subject based on the ideXlab platform.
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strangers on a train atypical glutamate receptors in the Kidney Glomerulus focus on functional nmda receptors with atypical properties are expressed in podocytes
American Journal of Physiology-cell Physiology, 2011Co-Authors: Michael Köttgen, Thomas BenzingAbstract:just more than a decade ago, a fascinating cell type of the Kidney Glomerulus was catapulted into the focus of Kidney research. Landmark genetic studies identified gene defects of Kidney podocytes as cause of topic-proteinuria/">proteinuria and end-stage renal failure ([1][1], [12][2], [14][3], [27][4]) and initiated a
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Strangers on a Train*: atypical glutamate receptors in the Kidney Glomerulus. Focus on “Functional NMDA receptors with atypical properties are expressed in podocytes”
American journal of physiology. Cell physiology, 2010Co-Authors: Michael Köttgen, Thomas BenzingAbstract:just more than a decade ago, a fascinating cell type of the Kidney Glomerulus was catapulted into the focus of Kidney research. Landmark genetic studies identified gene defects of Kidney podocytes as cause of topic-proteinuria/">proteinuria and end-stage renal failure ([1][1], [12][2], [14][3], [27][4]) and initiated a
Riccardo Bruni - One of the best experts on this subject based on the ideXlab platform.
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ultrasmall polymeric nanocarriers for drug delivery to podocytes in Kidney Glomerulus
Journal of Controlled Release, 2017Co-Authors: Riccardo Bruni, Paolo Possenti, Carlotta Bordignon, Min Li, Stefania OrdaniniAbstract:Abstract We explored the use of new drug-loaded topic-nanocarrier/">nanocarriers and their targeted delivery to the Kidney Glomerulus and in particular to podocytes, in order to overcome the failure of current therapeutic regimens in patients with proteinuric (i.e. abnormal amount of proteins in the topic-urine/">urine) diseases. Podocytes are glomerular cells which are mainly responsible for glomerular filtration and are primarily or secondarily involved in chronic Kidney diseases. Therefore, the possibility to utilise a podocyte-targeted drug delivery could represent a major breakthrough in Kidney disease research, particularly in terms of dosage reduction and elimination of systemic side effects of current therapies. Four-arm star-shaped polymers, with/without a hydrophobic poly- e -caprolactone core and a brush-like polyethylene glycol (PEG) hydrophilic shell, were synthesised by controlled/living polymerisation (ROP and ATRP) to allow the formation of stable ultrasmall colloidal nanomaterials of tuneable size (5–30 nm), which are able to cross the glomerular filtration barrier (GFB). The effects of these nanomaterials on glomerular cells were evaluated in vitro. Nanomaterial accumulation and permeability in the Kidney Glomerulus were also assessed in mice under physiological and pathological conditions. Drug (dexamethasone) encapsulation was performed in order to test loading capacity, release kinetics, and podocyte repairing effects. The marked efficacy of these drug-loaded topic-nanocarrier/">nanocarriers in repairing damaged podocytes may pave the way for developing a cell-targeted administration of new and traditional drugs, increasing efficacy and limiting side effects.
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Unimolecular polymeric nanocarriers for targeting podocytes in Kidney Glomerulus
2016Co-Authors: Riccardo BruniAbstract:L’obiettivo dello studio qui presentato e lo sviluppo di nuovi nanomateriali per il trattamento diretto delle disfunzioni dei glomeruli nei podociti. Tali disfunzioni rappresentano uno dei maggiori problemi di salute nel mondo. Lo studio esplora l’uso di nuovi topic-nanocarrier/">nanocarrier caricati farmaceuticamente e la loro capacita di raggiungere i glomeruli renali ed in particolare i podociti, in maniera tale da superare i problemi terapeutici in pazienti con disfunzioni proteinuriche. La sfida piu significativa e stato lo sviluppo di nanomateriali colloidali in grado di attraversare la barriera di filtrazione glomerulare (GFB) in maniera tale di colpire i podociti. Come sopra accennato la barriera e composta da una struttura a tre strati cosi definita: cellule endoteliali fenestrate, podociti e membrana basale nel mezzo che, in condizioni fisiologiche ha una dimensione di pori di 6-10nm, cioe una dimensione generalmente troppo piccola per la permeazione di alcune proteine e di nanoparticelle biomedicali. Lo studio si focalizza sulla progettazione di nanovettori polimerici ultra piccoli (
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unimolecular polymeric nanocarriers for targeting podocytes in Kidney Glomerulus
2016Co-Authors: Riccardo BruniAbstract:L’obiettivo dello studio qui presentato e lo sviluppo di nuovi nanomateriali per il trattamento diretto delle disfunzioni dei glomeruli nei podociti. Tali disfunzioni rappresentano uno dei maggiori problemi di salute nel mondo. Lo studio esplora l’uso di nuovi topic-nanocarrier/">nanocarrier caricati farmaceuticamente e la loro capacita di raggiungere i glomeruli renali ed in particolare i podociti, in maniera tale da superare i problemi terapeutici in pazienti con disfunzioni proteinuriche. La sfida piu significativa e stato lo sviluppo di nanomateriali colloidali in grado di attraversare la barriera di filtrazione glomerulare (GFB) in maniera tale di colpire i podociti. Come sopra accennato la barriera e composta da una struttura a tre strati cosi definita: cellule endoteliali fenestrate, podociti e membrana basale nel mezzo che, in condizioni fisiologiche ha una dimensione di pori di 6-10nm, cioe una dimensione generalmente troppo piccola per la permeazione di alcune proteine e di nanoparticelle biomedicali. Lo studio si focalizza sulla progettazione di nanovettori polimerici ultra piccoli (<30nm) per valutare se, in tal modo, i trasportatori sono capaci di permeare attraverso la barriera glomerulare, la quale si ipotizza che aumenti la propria capacita di filtrazione sotto condizione patologica. I nanomateriali colloidali ultra piccoli sono sintetizzati con successo da multi braccia idrofobicche bio degradabili composte da poli estere ( ad esempio poli epsilon caprolattone) copolimerizzato con poly(ethylene glicol) (PEG) a configurazione a pettine, struttura che serve come densa corona idrofilica. Il principale risultato di questo lavoro puo essere riassunto come segue: a) formazione di una libreria di polimeri anfifilici multi braccia che sono stati sintetizzati da polimerizzazioni Atom Transfer Radical (ATRP) di PEG metacrilato, partendo da iniziatori ATRP commerciali a 4 braccia e anche con macro iniziatori auto prodotti multi braccia costituiti da poli (epsilon caprolattone). Qualora fosse necessario un monomero fluorescente ( rodamina metacrilata) questo e stato copolimerizzato per ottenere materiale tracciabile per test biologici. Le cinetiche ATRP e distribuzione dei pesi molecolari dei polimeri sintetizzati sono state studiate in maniera tale da identificare le condizioni di reazione ottimali (solvente, catalizzatore, legante, concentrazione e temperatura) per controllare la polimerizzazione e anche in maniera tale da ottenere polimerizzazione quasi monodisperse. b) queste polimerizzazioni sono caratterizzate da analisi NMR, IR e GPC al fine di identificare la composizione, peso molecolare e polidispersione. Dispersione in acqua e tamponi fisiologici di tali polimeri sono stati analizzati attraverso l’uso di dynamic light scattering (DLS) per valutare dimensione media e distribuzione della dimensione. Un sottoinsieme di polimeri i quali presentano proprieta ottimali chimico fisiche sono stati identificati e ulteriormente usati per test biologici. In particolare sono stati selezionati nanovettori uni molecolari di dimensione fissata < 30nm ( compatibili con la filtrazione renale). c) test in vitro sui podociti sono stati portati a termine per caratterizzare la citotossicita dei polimeri, il danneggiamento del citoscheletro cellulare con conseguente cambiamento morfologico ed internalizzazione. In seguito sono stati eseguiti test in vivo su topi sani e malati da topic-proteinuria/">proteinuria indotta per analizzare la biodistribuzione del materiale, eseguendo somministrazione intra venosa. I polimeri selezionati hanno mostrano un profilo biocompatibile e una capacita di attraversare la barriera glomerulare renale dipendente dall’estensione del danneggiamento della stessa. d) un farmaco idrofobico, desametasone, e stato incapsulato con successo nel core delle nanoparticelle e i profili di rilascio sono stati analizzati in condizioni sink. I podociti danneggiati sono stati riparati con successo attraverso il trasporto controllato del desametasone, avendo come obiettivo lo sviluppo di nuovi nanoterapici rene-specifici. L’efficacia di questi topic-nanocarrier/">nanocarrier, caricati farmaceuticamente nella riparazione di podociti danneggiati, e grazie alle loro capacita di attraversare le GFB possono aprire la strada per lo sviluppo di nuovi farmaci e il miglioramento di quelli tradizionali per la somministrazione a target cellulare, nei casi di disfunzione cronica renale, incrementando l’efficacia e limitando gli effetti indesiderati rispetto ai farmaci attualmente in uso.
