Renal proximal tubular epithelial cells are significantly damaged during acute kidney injury. Renal proximal tubular cell-specific autophagy-deficient mice show increased sensitivity against renal injury, while showing few pathological defects under normal fed conditions. Considering that autophagy protects the proximal tubular cells from acute renal injury, it is reasonable to assume that autophagy contributes to the maintenance of renal tubular cells under normal fed conditions. To clarify this possibility, we generated a knock out mouse model which lacks Atg7, a key autophagosome forming enzyme, in renal proximal tubular cells (Atg7^flox/flox;KAP-Cre⁺). Analysis of renal tissue from two months old Atg7^flox/flox;KAP-Cre⁺ mouse revealed an accumulation of LC3, binding protein p62/sequestosome 1 (a selective substrate for autophagy), and more interestingly, Kim-1, a biomarker for early kidney injury, in the renal proximal tubular cells under normal fed conditions. TUNEL (TdT-mediated dUTP Nick End Labeling)-positive cells were also detected in the autophagy-deficient renal tubular cells. Analysis of renal tissue from Atg7^flox/flox;KAP-Cre⁺ mice at different age points showed that tubular cells positive for p62 and Kim-1 continually increase in number in an age-dependent manner. Ultrastructural analysis of tubular cells from Atg7^flox/flox;KAP-Cre⁺ revealed the presence of intracellular inclusions and abnormal structures. These results indicated that autophagy-deficiency in the renal proximal epithelial tubular cells leads to an increase in injured cells in the kidney even under normal fed conditions.

Cathepsin D is one of the major lysosomal aspartic proteases that is essential for the normal functioning of the autophagy-lysosomal system. In the kidney, cathepsin D is enriched in renal proximal tubular epithelial cells, and its levels increase during acute kidney injury. To investigate how cathepsin D-deficiency impacts renal proximal tubular cells, we employed a conditional knockout CtsD^flox/−; Spink3^Cre mouse. Immunohistochemical analyses using anti-cathepsin D antibody revealed that cathepsin D was significantly decreased in tubular epithelial cells of the cortico-medullary region, mainly in renal proximal tubular cells of this mouse. Cathepsin D-deficient renal proximal tubular cells showed an increase of microtubule-associated protein light chain 3 (LC3; a marker for autophagosome/autolysosome)-signals and an accumulation of abnormal autophagic structures. Renal ischemia/reperfusion injury resulted in an increase of early kidney injury marker, Kidney injury molecule 1 (Kim-1), in the cathepsin D-deficient renal tubular epithelial cells of the CtsD^flox/−; Spink3^Cre mouse. Inflammation marker was also increased in the cortico-medullary region of the CtsD^flox/−; Spink3^Cre mouse. Our results indicated that lack of cathepsin D in the renal tubular epithelial cells led to an increase of sensitivity against ischemia/reperfusion injury.

Python is one of the most popular programming languages to learn and use. It is ubiquitous in fields like data analysis, artificial intelligence, image processing, robotics, and web development. Its versatility and reliability make it ideal for major companies, including Tesla and Netflix. NASA even uses it to calibrate the James Webb Space Telescope. With the advent of artificial intelligence (AI), writing a Python program can be as simple as prompting an AI assistant for a solution. For research scientists, this lowered barrier could spark a generational change, transforming every research stage from idea conception to publishing. My interest in AI-assisted programming inspired my lecture on how Python can assist PhD students in their work. This lecture begins with basic terminology and presents three mini projects as first steps for learning Python. These projects cover text extraction and analysis, statistical data analysis, and virtually staining histological samples into digital image stacks. Small variations of these projects could lead to code that extracts reagents from manuscripts, creates stunning statistical graphs, and advances histopathological methods.