Automated tracking of tumor-stroma morphology in microtissues identifies functional targets within the tumor microenvironment for therapeutic intervention

Cancer-associated fibroblasts (CAFs) constitute an important part of the tumor microenvironment and promote invasion via paracrine functions and physical impact on the tumor. Although the importance of including CAFs into three-dimensional (3D) cell cultures has been acknowledged, computational support for quantitative live-cell measurements of complex cell cultures has been lacking. Here, we have developed a novel automated pipeline to model tumor-stroma interplay, track motility and quantify morphological changes of 3D co-cultures, in real-time live-cell settings. The platform consists of microtissues from prostate cancer cells, combined with CAFs in extracellular matrix that allows biochemical perturbation. Tracking of fibroblast dynamics revealed that CAFs guided the way for tumor cells to invade and increased the growth and invasiveness of tumor organoids. We utilized the platform to determine the efficacy of inhibitors in prostate cancer and the associated tumor microenvironment as a functional unit. Interestingly, certain inhibitors selectively disrupted tumor-CAF interactions, e.g. focal adhesion kinase (FAK) inhibitors specifically blocked tumor growth and invasion concurrently with fibroblast spreading and motility. This complex phenotype was not detected in other standard in vitro models. These results highlight the advantage of our approach, which recapitulates tumor histology and can significantly improve cancer target validation in vitro.     

Posterior parietal cortex activity reflects the significance of others' actions during natural viewing

The posterior parietal cortex (PPC) has been associated with multiple stimulus‐driven (e.g., processing stimulus movements, providing visual signals for the motor system), goal‐directed (e.g., directing visual attention to a target, processing behavioral priority of intentions), and action‐related functions in previous studies with non‐naturalistic paradigms. Here, we examined how these functions reflect PPC activity during natural viewing. Fourteen healthy volunteers watched a re‐edited movie during functional magnetic resonance imaging (fMRI). Participants separately annotated behavioral priority (accounting for percepts, thoughts, and emotions) they had experienced during movie episodes. Movements in the movie were quantified with computer vision and eye movements were recorded from a separate group of subjects. Our results show that while overlapping dorsomedial PPC areas respond to episodes with multiple types of stimulus content, ventrolateral PPC areas exhibit enhanced activity when viewing goal‐directed human hand actions. Furthermore, PPC activity related to viewing goal‐directed human hand actions was more accurately explained by behavioral priority than by movements of the stimulus or eye movements. Taken together, our results suggest that PPC participates in perception of goal‐directed human hand actions, supporting the view that PPC has a special role in providing visual signals for the motor system (“how”), in addition to processing visual spatial movements (“where”). Hum Brain Mapp 35:4767–4776, 2014. © 2014 Wiley Periodicals, Inc .     

Comparison of postoperative skeletal stability of maxillary segments after Le Fort I osteotomy,using patient-specific implant versus mini-plate fixation

BackgroundThree-dimensionally (3D) designed osteotomies and customised osteosynthesis are rapidly becoming standard in maxillofacial reconstructive and deformity surgery. Patient-specific implants (PSIs) have been in use for a few years in orthognathic surgery as well. In Le Fort I osteotomy, wafer-free fixation of the maxillary segment can be performed by individually manufactured cutting and drill guides together with PSIs.AimThis retrospective study was performed to compare the postoperative skeletal stability of the maxillary segment fixed by patient-specific implants versus mini-plates after Le Fort I osteotomy.PatientsFifty-one patients were divided into subgroups according to the fixation method and the advancement of the sub-spinal point. The postoperative skeletal stability of the maxillary segment was evaluated from lateral cephalometric radiographs one year postoperatively.ResultsNo statistically significant differences were found between the postoperative skeletal stability of the PSI and mini-plate fixed maxillae. Prospective studies, possibly with 3D fusion analysis, are warranted to confirm the results.ConclusionThe choice between the two fixation methods does not seem to affect the postoperative skeletal stability of the maxillary segments.     

Analysis of neonatal heart rate variability by a microprocessor-based on-line system

The effects of different rejection logic limits on the results of analysis of neonatal heart rate variability from electrocardiograms by a microprocessor-based system were studied on fourteen infants after normal labor and on ten infants after elective cesarean section. In addition to the infant's movements and crying, two further main sources of error in the calculation of variability indices were detected: premature beats and problems in the shape of QRS complexes in a neonatal electrocardiogram. No noteworthy problem was observed in the calculation of the interval index (II), which describes the long term variability of heart rate. In the calculation of the differential indices (DI), which describe the short term component of heart rate, distorted QRS complexes created very high DIs when the rejection logic was ten beats per minute (bpm). When stricter rejection limits were used (five bpm), the DI values even in these cases fell within the normal range. The DI values calculated using a rejection limit of five bpm were always lower than those calculated using a rejection limit of ten bpm, but the reference values of neonatal II and DI after normal labor were similar to those presented previously using a rejection limit of ten bpm. In conclusion, the neonatal electrocardiogram appears to be a noisy signal, comparable with the abdominal fetal electrocardiogram, and strict rejection limits are useful in processing it for indices of variability.     

Chalone-induced inhibition in DNA synthesis of human psoriatic epidermal cells cultured in diffusion chambers in mice.

Pieces of human psoriatic skin and suspensions of mouse bone marrow cells as non-epidermal control tissue were cultured for four days in diffusion chambers implanted intraperitoneally in 40 mice. The mice were divided into three groups. On the second and third day of the culture partially purified epidermal extract containing epidermal chalone, granulocyte extract containing granulocyte chalone, and Hanks' solution were injected intraperitoneally into the mice of the three groups, respectively. On the fourth day, incorporation of 3H-TdR into the diffusion chamber cultures and various epithelia and several non-epidermal tissues of the host animal was studied autoradiographically and radiochemically. The results showed that the epidermal chalone induced a marked, statistically significant, depression in the labelling index of the cultured psoriatic epidermal cells and in the number of labelled cells in the host ear epidermis (31% and 62%, respectively); the other host epithelia were not significantly inhibited. A crude granulocyte extract used in most experiments was clearly toxic; the semipurified granulocyte chalone used in one experiment had no effect on the psoriatic and cultures, but affected weakly the cultured bone marrow cells. Neither the epidermal nor the granulocyte extract had any inhibitory effect on the non-epidermal tissues studied.     

Metabolomics of Synovial Fluid and Infrapatellar Fat Pad in Patients with Osteoarthritis or Rheumatoid Arthritis

Inflammation - Osteoarthritis (OA) and autoimmune-driven rheumatoid arthritis (RA) are inflammatory joint diseases with complex and insufficiently understood pathogeneses. Our objective was to...     

pH-triggered surface charge-reversal nanoparticles alleviate experimental murine colitis via selective accumulation in inflamed colon regions

In this study, we developed pH-triggered surface charge-reversal lipid nanoparticles (LNPs), loaded with budesonide, which could precisely deliver the drug to inflamed colon segments for the treatment of ulcerative colitis. Polyethyleneimine (PEI) was used to render LNPs cationic (PEI-LNPs), and Eudragit® S100 (ES) was coated on PEI-LNPs to obtain pH-triggered charge-reversal LNPs (ES-PEI-LNPs). ES coating avoided a burst drug release under acidic conditions mimicking the stomach and early small intestine environments and showed a sustained release in the colon. The surface charge of ES-PEI-LNPs switched from negative to positive under colonic conditions owing to pH-triggered removal of the ES coating. Bioimaging of the mouse gastrointestinal tract and confocal analysis of colon tissues revealed that ES-PEI-LNPs selectively accumulated in an inflamed colon. Furthermore, ES-PEI-LNPs mitigated experimental colitis in mice. These results suggest that the pH-triggered charge-reversal LNPs could be a promising drug carrier for ulcerative colitis therapy and other colon-targeted treatments.     

Cryptococcosis during systemic glucocorticosteroid treatment

Cryptococcosis is an opportunistic infection caused by a fungus, Cryptococcus neoformans. It is usually seen in immunocompromised patients with AIDS, leukaemia, lymphoma, sarcoidosis or immunosuppressive treatments. We describe a patient who was treated with systemic glucocorticosteroids for 4 years because of lung sarcoidosis. During the last year of treatment, a papular eruption developed which later became ulcerative. In a histopathological examination of a skin biopsy, there was granulomatous inflammation, and the disease was treated as sarcoidosis without success. After 1 year's unsuccessful treatment, another skin biopsy and skin fungal culture revealed C. neoformans. Cryptococcal antigen was found in blood and cerebrospinal fluid, too. The patient was successfully treated first with an amphotericin-B-flucytosine combination and later with fluconazole.