Spatiotemporal Quantification of Metastatic Tumour Cell Growth and Distribution in Lymph Nodes by Whole-Mount Tissue 3D Imaging

Lymph nodes (LNs) are a common site of metastasis in many solid cancers. Tumour cells can migrate to LNs for further metastatic colonization of distant organs, indicating poor prognosis and requiring different clinical interventions. The current histopathological diagnostic methods used for the detection of clinical lymph node metastasis (LNM) still have some limitations, such as incomplete observation. To obtain a complete picture of tumour-metastasized LNs at spatial and temporal scales, we used 3D imaging of solvent-cleared organs (uDISCO) and 3D rapid immunostaining. MC38 cells tagged with EGFP were injected into the left footpad of C57BL/6 mice. Draining lymph nodes (DLNs) obtained from these mice were cleared using uDISCO. Metastatic colorectal cancer (CRC) cells in various regions of DLNs from mice at different time points were quantified using whole-mount tissue 3D imaging. The results revealed several stages of tumour cell growth and distribution in LNs: 1) invasion of lymphatic vessels (LVs) and blood vessels (BVs); 2) dispersion outside LVs and BVs for proliferation and expansion; and 3) re-entry into BVs and efferent lymphatic vessels (ELVs) for further distant metastasis. Moreover, these data demonstrated that mouse fibroblast cells (MFCs) could not only promote the LNM of tumour cells but could also metastasize to LNs together with tumour cells, thus providing a “soil” for tumour cell colonization. In conclusion, whole-mount tissue 3D imaging and spatiotemporal analysis of LNM may together constitute an auxiliary method to improve the accuracy of clinical LNM detection in the future.

Study on Phytoconstituents and Antimicrobial Potential of Sapindus mukorossi Fruit Extract

Sapindus mukorossiis well known for its folk medicinal values. The fruit is valued for the saponins (10.1%) present in the pericarp which constitutes up to 56.5% of the drupe known for inhibiting tumour cell growth. Recently many of the pharmacological actions of this plant have been explored which include the antimicrobial molluscicidal and insecticidal actions.Thefruitsareofconsiderableimportancefor their medicinal value for treatingseveraldiseaseslikeexcessivesalivation,pimples,epilepsy,chlorosis,migranes, eczemaand psoriasis etc.The powdered seeds are employed in the treatment ofdentalcaries, arthritis,commoncold, constipation and nausea. Keeping in view the medicinal importance of the fruit, it was thought worthwhile to carry out systematic study on phytochemical investigation and antimicrobial potential of fruit extract of Sapindus mukorossi.Fruit were dried, powdered and extracted with differentsolventsystemswithincreasingpolarity. The extracts were screened for the antibacterialandantifungalactivity.Predominance of antibacterial activity was observed at dose level of 50μg/ml of petroleum ether extract. Maximum zone of inhibition (19mm)&(18mm)wasobservedagainstSalmonellatyphiiand Shigelladysenteriarespectively as compared to standard drug Ampicillin20μg/ml.Sinceantibacterialactivity was observed in 50μg/ml concentration of petroleum ether extract, it was chosen forexploringitsantifungalpotentialalso.Maximum zone of inhibition was observed against Aspergillus sulfurous as compared to standard drug ketoconazole 20μg/ml. From the above study, it could be concluded that Sapindus mukorossipetroleum ether fruit extracts(50μg/ml) have good antimicrobial potential and can be explored further to isolate active principlesfrom the same.

The Synthesis of 3H-Labelled 8-Azido-N6-Benzyladenine and Related Compounds for Photoaffinity Labelling of Cytokinin-Binding Proteins

The biology of the group of plant hormones termed cytokinins is reviewed to reveal areas where further studies of cytokinin-binding proteins could be significant. Such areas include: inhibition of human tumour cell growth by cytokinin ribosides, the role of cytokinins in the development of diverse micro-organisms including the cyanobacteria and Mycobacterium tuberculosis, the very rapid responses of plant cells to exogenous cytokinins, and other aspects of cytokinin plant biology. Photoaffinity labelling (PAL) coupled to the recent advances in HPLC of proteins and mass spectral analysis and sequencing of proteins, may have relevance to these areas. To facilitate PAL, we present experimental details for two methods for synthesis of 8-azido-N6-benzyladenine, which has the azido affinity group in the preferred position of the purine ring. Synthesis from [2-3H]adenosine yielded the above-mentioned PAL reagent with 3H in the purine ring and also gave labelled 9-riboside and 8-azido-N6,9-dibenzyladenine. 8-Azido-N6-benzyladenine was also prepared from 6,8-dichloropurine by a facile synthesis, which would allow a label to be sited in the benzyl group where substituents can also be introduced to vary cytokinin activity. The use of inactive cytokinin analogues in assessing the significance of PAL is discussed.

The role of the interleukin (IL)-6/IL-6 receptor axis in cancer

Interleukin-6 (IL-6) is a pleiotropic cytokine that activates a classic signalling pathway upon binding to its membrane-bound receptor (IL-6R). Alternatively, IL-6 may ‘trans-signal’ in a manner that is facilitated by its binding to a soluble derivative of the IL-6 receptor (sIL-6R). Resultant signal transduction is, respectively, driven by the association of IL-6/IL-6R or IL-6/sIL-6R complex with the membrane-associated signal transducer, gp130 (Glycoprotein 130). Distinct JAK (Janus tyrosine kinase)/STAT (signal transducers and activators of transcription) and other signalling pathways are activated as a consequence. Of translational relevance, overexpression of IL-6 has been documented in several neoplastic disorders, including but not limited to colorectal, ovarian and breast cancer and several haematological malignancies. This review attempts to summarise our current understanding of the role of IL-6 in cancer development. In short, these studies have shown important roles for IL-6 signalling in tumour cell growth and survival, angiogenesis, immunomodulation of the tumour microenvironment, stromal cell activation, and ultimate disease progression. Given this background, we also consider the potential for therapeutic targeting of this system in cancer.