Microbiota as Drivers and as Therapeutic Targets in Ocular and Tissue Specific Autoimmunity

Autoimmune uveitis is a major cause of blindness in humans. Activation of retina-specific autoreactive T cells by commensal microbiota has been shown to trigger uveitis in mice. Although a culprit microbe and/or its immunogenic antigen remains to be identified, studies from inducible and spontaneous mouse models suggest the potential of microbiota-modulating therapies for treating ocular autoimmune disease. In this review, we summarize recent findings on the contribution of microbiota to T cell-driven, tissue-specific autoimmunity, with an emphasis on autoimmune uveitis, and analyze microbiota-altering interventions, including antibiotics, probiotics, and microbiota-derived metabolites (e.g., short-chain fatty acids), which have been shown to be effective in other autoimmune diseases. We also discuss the need to explore more translational animal models as well as to integrate various datasets (microbiomic, transcriptomic, proteomic, metabolomic, and other cellular measurements) to gain a better understanding of how microbiota can directly or indirectly modulate the immune system and contribute to the onset of disease. It is hoped that deeper understanding of these interactions may lead to more effective treatment interventions.

Rapid Differentiation of Epithelial Cell Types in Forensic Samples Using Intrinsic Fluorescence and Morphological Signatures

Establishing the tissue source of epithelial cells within a biological sample is an important capability for forensic laboratories. In this study we used Imaging Flow Cytometry (IFC) to analyze individual cells recovered from buccal, contact epithelial, and vaginal samples that had been dried between 24 hours and more than eight weeks. Measurements capturing the size, shape, and fluorescent properties of cells were collected in an automated manner and then used to build a multivariate statistical framework for differentiating cells based on tissue type. Results showed that cell populations from the three tissue types could be differentiated using a Discriminant Function plot of IFC measurements. Epidermal cells were distinguished from vaginal and buccal cells with an average classification accuracy of ~94%. Ultimately, cellular measurements such as these, which can be obtained non-destructively, may provide probative information for many types of biological samples and complement results from standard genetic profiling techniques.

High-Speed Broadband Real-Time Monitoring of Cell Viscoelasticity Reveals Oscillatory Myosin Activity

This article presents the investigation of the dynamic behavior of the cytoskeleton of live human cells, enabled by a recently-developed control-based approach on scanning probe microscope (SPM). Mechanical behaviors of live cells play an important role in various cell physiological and pathological activities, and have been studied via various techniques and approaches. Studies of evolutions of mechanical properties of live cell, however, are still rather limited and scarce, due to the limitations of current instruments including SPM for single cellular measurements. Particularly, currently nanomechanical measurements using SPM is too slow to excite the mechanical behavior and then measure the corresponding response of life biological species over a large frequency range (broadband). Moreover, large uncertainty is induced in the in-liquid nanomechanical measurement using SPM, as in the indentation quantification, the effects of the acceleration force from the cantilever motion and the hydrodynamic force are not accounted for. The main contribution of this article is the use of a control-based nanomechanical protocol to interrogate the viscoelasticity oscillation of live human prostate cancer cell (PC-3 cells) and its dependence on myosin activities. The experiment results show that as the oscillation of static elastic modulus reported earlier in the literature, the oscillation of dynamic viscoelastic modulus measured is also periodic with a 200-second period. Moreover, as the elastic modulus oscillation, both the amplitude and the period of the viscoelasticity oscillation also strongly depend on the myosin activities, and closely regulated by the calcium density of the cytoskeleton.

Tanshinone II A Relieves Adriamycin-induced Myocardial Injury in Rat Model

<p>As an effective antineoplastic agent, adriamycin (ADR) remains a use for the treatment of cancer. However, it is limited by the serous cardiotoxicity. Tanshinone II A is the main effective component of Salvia miltiorrhiza Bunge which has been used for treatment of cardiovascular diseases. The purpose of this study is to evaluate the protective effect of Tanshinone II A on adriamycin-induced myocardial injury in rat and explore the mechanism of this effect. Male Wistar rats (200 ± 20 g) were divided into three groups, control (CON) group, adriamycin (ADR) group and ADR + Tanshinone II A (TRA) group. At the end of the 4 week treatment period, cardiac function was evaluated by transthoracic echocardiography. Molecular and cellular measurements were performed in atrial muscle to examine histopathological changes, the formation of fibrosis, Inflammation and apoptosis. Cardiac dysfunction was induced by adriamycin, as indicated by significant decreases in ventricular fractional shortening and ejection fraction. This adriamycin-induced cardiac dysfunction was prevented by the treatment of Tanshinone II A. Adriamycin induced pathological changes and fibrosis, activated apoptosis (increased TUNEL index, apoptotic DNA fragmentation,and caspase-3 activity and decreased Bcl-2/Bax ratio), inflammation and suppressed phosphorylation status (eIF2α and PERK) in atrial. All these molecular and cellular alterations induced by ADR were not found in the rats treated with Tanshinone II A. These findings demonstrate clearly that Tanshinone II A protects the cardiomyocytes against the ADR-induced cardiomyopathy by preventing the activation of cardiac fibrosis and apoptosis, and the effects are probably mediated through ERS pathway.</p>