Constitutive renal Rel/nuclear factor-κB expression in Lewis polycystic kidney disease rats

AIM： To determine the temporal expression and pattern of Rel/nuclear factor （NF）-κB proteins in renal tissue in polycystic kidney disease （PKD）. METHODS： The renal expression of Rel/NF-κB proteins was determined by immunohistochemistry, immunofuorescence and immunoblot analysis in Lewis polycystic kidney rats （LPK, a genetic ortholog of human nephronopthsis-9） from postnatal weeks 3 to 20. At each timepoint, renal disease progression and the mRNA expression of NF-κB-dependent genes （TNFa and CCL2） were determined. NF-κB was also histologically assessed in human PKD tissue.RESULTS： Progressive kidney enlargement in LPK rats was accompanied by increased renal cell proliferation and interstitial monocyte accumulation （peaking at weeks 3 and 10 respectively）, and progressive interstitial fibrosis （with a smooth muscle actin and Sirius Red deposition significantly increased compared to Lewis kidneys from weeks 3 to 6 onwards）. Rel/NF-κB proteins （phosphorylated-p105, p65, p50, c-Rel and RelB） were expressed in cystic epithelial cells （CECs） of LPK kidneys as early as postnatal week 3 and sustained until late-stage disease at week 20. From weeks 10 to 20, nuclear p65, p50, RelB and cytoplasmic IκBa protein levels, and TNFa and CCL2 expression, were upregulated in LPK compared to Lewis kidneys. NF-κB proteins were consistently expressed in CECs of human PKD. The DNA damage marker γ-H2AX was also identifed in the CECs of LPK and human polycystic kidneys. CONCLUSION： Several NF-κB proteins are consistently expressed in CECs in human and experimental PKD. These data suggest that the upregulation of both the canonical and non-canonical pathways of NF-κB signaling may be a constitutive and early pathological feature of cystic renal diseases.

Critical size limit of biodegradable nanopartides for enhanced lymph node trafficking and paracortex penetration

Lymph node (LN) targeti ng through interstitial drain age of nan oparticles (NPs) is an attractive strategy to stimulate a pote nt immune respo nse, as LNs are the primary site for lymphocyte priming by antigen presenting cells (APCs) and triggering of an adaptive immune response. NP size has been shown to influence the efficiency of LN-targeting and retention after subcutaneous injection. For clinical translation, biodegradable NPs are preferred as carrier for vaccine delivery. However, the selective "size gateM for effective LN-drainage, particularly the kinetics of LN trafficking, is less well defined. This is partly due to the challenge in generating size-controlled NPs from biodegradable polymers in the sub-100-nm range. Here, we report the preparation of three sets of poly(lactic-co-glycolic)-b-poly(ethylene-glycol)(PLGA-b-PEG) NPs with number average diameters of 20-, 40-, and 100-nm and narrow size distributions using flash nanoprecipitation. Using NPs labeled with a near-infrared dye, we showed that 20-nm NPs drain rapidly across proximal and distal LNs following subcutaneous inoculation in mice and are retai ned in LNs more effectively than NPs with a nu mber average diameter of 40-nm. The drain age of 100-nm NPs was n egligible. Furthermore, the 20-nm NPs showed the highest degree of penetration around the paracortex region and had enhanced access to dendritic cells in the LNs. Together, these data confirmed that small, size-controlled PLGA-b-PEG NPs at the lower threshold of about 30-nm are most effective for LN trafficking, retention, and APC uptake after s.c. administration. This report could inform the design of LN-targeted NP carrier for the delivery of therapeutic or prophylactic vaccines.

Mitochondrial-triggered immune responses mechanistically connect drug-induced steatohepatitis and cardiomyopathy associated with nonalcoholic steatohepatitis

In our prior publication[1],we elucidated IL-33-dependent mechanisms that regulate drug-induced liver injury.In this commentary,we mechanistically connected IL-33-dependent drug-induced steatohepatitis with nonalcoholic steatohepatitis.Drug-induced steatohepatitis(DISH)is a rare form of drug-induced liver injury caused by drugs that are capable of inducing metabolic injury,steatosis or steatohepatitis[2].

Deep learning-based analysis of macaque corneal sub-basal nerve fibers in confocal microscopy images

Background:To develop and validate a deep learning-based approach to the fully-automated analysis of macaque corneal sub-basal nerves using in vivo confocal microscopy(IVCM).Methods:IVCM was used to collect 108 images from 35 macaques.58 of the images from 22 macaques were used to evaluate different deep convolutional neural network(CNN)architectures for the automatic analysis of sub-basal nerves relative to manual tracings.The remaining images were used to independently assess correlations and interobserver performance relative to three readers.Results:Correlation scores using the coefficient of determination between readers and the best CNN averaged 0.80.For inter-observer comparison,inter-correlation coefficients(ICCs)between the three expert readers and the automated approach were 0.75,0.85 and 0.92.The ICC between all four observers was 0.84,the same as the average between the CNN and individual readers.Conclusions:Deep learning-based segmentation of sub-basal nerves in IVCM images shows high to very high correlation to manual segmentations in macaque data and is indistinguishable across readers.As quantitative measurements of corneal sub-basal nerves are important biomarkers for disease screening and management,the reported work offers utility to a variety of research and clinical studies using IVCM.