Apoptosis mapping in space and time of 3D tumor ecosystems reveals transmissibility of cytotoxic cancer death

The emerging tumor-on-chip (ToC) approaches allow to address biomedical questions out of reach with classical cell culture techniques: in biomimetic 3D hydrogels they partially reconstitute ex vivo the complexity of the tumor microenvironment and the cellular dynamics involving multiple cell types (cancer cells, immune cells, fibroblasts, etc.). However, a clear bottleneck is the extraction and interpretation of the rich biological information contained, sometime hidden, in the cell co-culture videos. In this work, we develop and apply novel video analysis algorithms to automatically measure the cytotoxic effects on human cancer cells (lung and breast) induced either by doxorubicin chemotherapy drug or by autologous tumor-infiltrating cytotoxic T lymphocytes (CTL). A live fluorescent dye (red) is used to selectively pre-stain the cancer cells before co-cultures and a live fluorescent reporter for caspase activity (green) is used to monitor apoptotic cell death. The here described open-source computational method, named STAMP (spatiotemporal apoptosis mapper), extracts the temporal kinetics and the spatial maps of cancer death, by localizing and tracking cancer cells in the red channel, and by counting the red to green transition signals, over 2–3 days. The robustness and versatility of the method is demonstrated by its application to different cell models and co-culture combinations. Noteworthy, this approach reveals the strong contribution of primary cancer-associated fibroblasts (CAFs) to breast cancer chemo-resistance, proving to be a powerful strategy to investigate intercellular cross-talks and drug resistance mechanisms. Moreover, we defined a new parameter, the ‘potential of death induction’, which is computed in time and in space to quantify the impact of dying cells on neighbor cells. We found that, contrary to natural death, cancer death induced by chemotherapy or by CTL is transmissible, in that it promotes the death of nearby cancer cells, suggesting the release of diffusible factors which amplify the initial cytotoxic stimulus.     

The freshwater planarian Dugesia sicula Lepori, 1948 (Platyhelminthes, Tricladida) in Tunisia: ecology, karyology, and morphology

A study was carried out on freshwater planarians of the genus Dugesia from three localities in northern Tunisia: Joumine, Chiba, and Lebna. The three populations are fissiparous under field conditions and do not possess reproductive organs, except for some cases of hyperplastic ovaries in the Joumine population. After five months of breeding under laboratory conditions, with water temparature between 18 and 25?°C and under short photoperiod, 8.33 to 26% of specimens from the Joumine stream became sexualized, together with considerable increase of body length and width. For the other two populations the sexualization rate was very small. Extended period of darkness apparently played a positive role in this sexualization. The phenomenon of sexualization is discussed in the context of reproductive strategies. Histological and karyological studies of the sexualized animals allowed identification of these usually fissiparous planarians as representatives of Dugesia sicula Lepori, 1948. The species is here reported for the first time from Tunisia. The populations are restricted to permanent water basins resulting from the construction of dams in large rivers.     

Effectiveness of low-power laser therapy in improvement of the peripheral neuropathy induced by xenobiotics in rats

BackgroundPeripheral neuropathy (PN) is the damage and dysfunction of neurons of the peripheral nervous system. The present study was conducted to estimate the effectiveness of low-power laser therapy (LPLT) in the management of PN in a rats’ model.MethodsPN was induced by giving dichloroacetate (DCA) (250 mg/kg/day) for up to 12 weeks. Four groups of rats were used: control group, PN group, PN group treated with gabapentin and PN group treated with LPLT. The study was conducted for 8 weeks. The management of PN was estimated by behavioral tests which included hot plate and Morris water maze tests. Blood biochemical analysis were carried out.ResultsUsing of hot plate test indicated thermal hypoalgesia and using Morris water maze test showed cognitive decline in PN rats. Treatment with LPLT or gabapentin improved both the pain sensations and deteriorated memory that occurred in the PN rats. Biochemical analysis showed that LPLT significantly decreased the elevated beta-endorphin level in PN rats, while gabapentin could not reduce it. Treatment PN rats with LPLT or gabapentin shifted the high levels of TNF-α, IL-1β and IL-10 cytokines back to their normal values. Serum nitric oxide and MDA significantly increased in the PN group together with significant reduction in the rGSH level, these values were significantly improved by LPLT application while this was not the case with gabapentin treatment. Furthermore, treatment with gabapentin or LPLT significantly reduced serum ALAT and ASAT activities which are otherwise increased in the PN group. S100B, PGE2, total cholesterol, triglycerides, LDL-cholesterol, HDL-cholesterol, urea and creatinine showed insignificant changes among all groups.ConclusionsOur results showed that treatment with LPLT is more efficient than gabapentin in ameliorating the peripheral neuropathy induced by xenobiotics.