Ermore, perfusates exclude the influence of other organs. It should be noted that, a biomarker will only be fully relevant once it can be validated in the whole organism. The candidate 10781694 biomarkers identified in organ perfusates should be validated in bodily fluids relevant to the disease condition of the specific organ. To maintain physiological kidney function, an oncotic agent was included in the perfusates to create “physiological” colloid osmotic pressure. In previous studies, BSA was the most commonly used oncotic agent. However, it was not appropriate for this study because high amounts of BSA would be filtered into the urinary tract and interfere with the detection of kidney origin proteins. Dextran was used as an oncotic agent in this study. As a result, many protein preparation methods could not be used because dextran is readily precipitated by ethanol, acetonitrile, and acetone and would be retained by the reversed-phase chromatography column. However, dextran does not affect SDS-PAGE, which makes in-gel digestion of the proteins for LC-MS/MS analysis possible.Supporting InformationTable S1 The identified proteins in the isolated rat kidney perfusion-driven urine. (XLS) Table S2 The total human kidney origin proteins in urine.(XLS)Table S3 The human kidney origin proteins in urine wereclassified in terms of molecular function and biological process. (XLS)DiscussionThis study aimed to identify human kidney origin proteins in urine. We present an approach to profile the isolated rat kidney perfusion-driven urine proteome, to match the identified ratAuthor ContributionsConceived and designed the experiments: YHG. Performed the experiments: LLJ. Analyzed the data: LLJ. Contributed reagents/materials/ analysis tools: XDL CS LLW MLL ZGG. Wrote the paper: YHG LLJ. Gave suggestions for modification of the manuscript: ZHL.
Esophageal adenocarcinomas (AC) of the gastroesophageal junction are believed to be mainly induced by gastroesophageal reflux, while squamous cell carcinomas (SCC) are mainly attributed to alcohol and tobacco Title Loaded From File consumption [1,2]. In the last decades, the incidence of AC has raised dramatically in western countries [3]. Despite multimodal therapeutic strategies, overall outcome for patients with gastroesophageal cancer remains poor. So novel therapeutic concepts are urgently needed, and insights into the pathophysiology of disease are a prerequisite for their development. Like many other carcinomas, esophageal cancers mainly spread through the lymphatic system, and lymphovascular invasion of tumor cells is associated with diminished prognosis of patients [4]. Today striking evidence exists that tumors may establish not only their own blood vessels supply, but might also induce the formation of new lymphatic vessels (lymphangiogenesis) and to migrate actively into this newly formed vessels to promote their spread [5].So possible inhibition of this process might be of benefit for patients, especially as recent data suggest that the process of lymphangiogenesis and lymphatic vessel invasion (LVI) is not only limited to Title Loaded From File primary tumors, but is also evident in lymph node metastases, resulting in further spread of tumor cells [6]. The formation of tumor associated lymphatic vessels is a complex process and currently only partially understood. During embryonic angiogenesis, platelets seem to play a key role in separating blood and lymphatic vasculature [7]. But there is increasing evidence that platelets might interact with th.Ermore, perfusates exclude the influence of other organs. It should be noted that, a biomarker will only be fully relevant once it can be validated in the whole organism. The candidate 10781694 biomarkers identified in organ perfusates should be validated in bodily fluids relevant to the disease condition of the specific organ. To maintain physiological kidney function, an oncotic agent was included in the perfusates to create “physiological” colloid osmotic pressure. In previous studies, BSA was the most commonly used oncotic agent. However, it was not appropriate for this study because high amounts of BSA would be filtered into the urinary tract and interfere with the detection of kidney origin proteins. Dextran was used as an oncotic agent in this study. As a result, many protein preparation methods could not be used because dextran is readily precipitated by ethanol, acetonitrile, and acetone and would be retained by the reversed-phase chromatography column. However, dextran does not affect SDS-PAGE, which makes in-gel digestion of the proteins for LC-MS/MS analysis possible.Supporting InformationTable S1 The identified proteins in the isolated rat kidney perfusion-driven urine. (XLS) Table S2 The total human kidney origin proteins in urine.(XLS)Table S3 The human kidney origin proteins in urine wereclassified in terms of molecular function and biological process. (XLS)DiscussionThis study aimed to identify human kidney origin proteins in urine. We present an approach to profile the isolated rat kidney perfusion-driven urine proteome, to match the identified ratAuthor ContributionsConceived and designed the experiments: YHG. Performed the experiments: LLJ. Analyzed the data: LLJ. Contributed reagents/materials/ analysis tools: XDL CS LLW MLL ZGG. Wrote the paper: YHG LLJ. Gave suggestions for modification of the manuscript: ZHL.
Esophageal adenocarcinomas (AC) of the gastroesophageal junction are believed to be mainly induced by gastroesophageal reflux, while squamous cell carcinomas (SCC) are mainly attributed to alcohol and tobacco consumption [1,2]. In the last decades, the incidence of AC has raised dramatically in western countries [3]. Despite multimodal therapeutic strategies, overall outcome for patients with gastroesophageal cancer remains poor. So novel therapeutic concepts are urgently needed, and insights into the pathophysiology of disease are a prerequisite for their development. Like many other carcinomas, esophageal cancers mainly spread through the lymphatic system, and lymphovascular invasion of tumor cells is associated with diminished prognosis of patients [4]. Today striking evidence exists that tumors may establish not only their own blood vessels supply, but might also induce the formation of new lymphatic vessels (lymphangiogenesis) and to migrate actively into this newly formed vessels to promote their spread [5].So possible inhibition of this process might be of benefit for patients, especially as recent data suggest that the process of lymphangiogenesis and lymphatic vessel invasion (LVI) is not only limited to primary tumors, but is also evident in lymph node metastases, resulting in further spread of tumor cells [6]. The formation of tumor associated lymphatic vessels is a complex process and currently only partially understood. During embryonic angiogenesis, platelets seem to play a key role in separating blood and lymphatic vasculature [7]. But there is increasing evidence that platelets might interact with th.