Screening of immune-enhancing Lactobacillus in mice by using a cell-line.

In this study, we found that it is possible to screen Lactobacillus strains that enhance the immune function of mice using HCT-8 cells. Lactobacillus were co-incubated with intestinal epithelial HCT-8 cells to detect and screen the strains that induced more interleukin-6 (IL-6) in the culture supernatant. Simultaneously, a mouse model of low immunity was established to administer the screened lactobacilli by gavage. After 4 weeks of continuous gavage, related cytokines in blood and immune cell indexes in organs were detected to comprehensively evaluate the feasibility of in vitro cell culture model for screening immune-enhancing strains. The content of IL-6 in the culture supernatant of HCT-8 cells induced by the three tested strains increased approximately 5, 8 and 15 fold compared with that of the control group. IL-6 content in serum of mice was significantly higher than that of the control group provided with cyclophosphamide (CTX). Lactobacillus paracasei ZLPC01 presented a higher ability to protect against the immune damage of CTX by decreasing the serum IgG level, increasing the transformation of mouse splenocytes, and the activity of NK cells. Furthermore, L. paracasei ZLPC01 increased cytokine content in serum (IL-6, IL-2, TNF-α and IFN-γ) and colon (IL-6 and TNF-α) in CTX-treated mice. Screening strains that enhance immunity via an in vitro cell-line is simple in operation, and the results are well correlated with those of animal experiments, which is feasible and effective in practice. In addition, L. paracasei ZLPC01 could have the potential to enhance the immunity of mice effectively through inducing intestinal cells to produce IL-6, TNF-α and other cytokines.

USP15 antagonizes CRL4CRBN-mediated ubiquitylation of glutamine synthetase and neosubstrates.

Targeted protein degradation by the ubiquitin-proteasome system represents a new strategy to destroy pathogenic proteins in human diseases, including cancer and neurodegenerative diseases. The immunomodulatory drugs (IMiDs) thalidomide, lenalidomide, and pomalidomide have revolutionized the treatment of patients with multiple myeloma (MM) and other hematologic malignancies, but almost all patients eventually develop resistance to IMiDs. CRBN, a substrate receptor of CUL4-RBX1-DDB1-CRBN (CRL4CRBN) E3 ubiquitin ligase, is a direct target for thalidomide teratogenicity and antitumor activity of IMiDs (now known as Cereblon E3 ligase modulators: CELMoDs). Despite recent advances in developing potent CELMoDs and CRBN-based proteolysis-targeting chimeras (PROTACs), many questions apart from clinical efficacy remain unanswered. CRBN is required for the action of IMiDs, but its protein expression levels do not correlate with intrinsic resistance to IMiDs in MM cells, suggesting other factors involved in regulating resistance to IMiDs. Our recent work revealed that the CRL4CRBN-p97 pathway is required for degradation of natural substrate glutamine synthetase (GS) and neosubstrates. Here, I show that USP15 is a key regulator of the CRL4CRBN-p97 pathway to control stability of GS and neosubstrates IKZF1, IKZF3, CK1-α, RNF166, GSPT1, and BRD4, all of which are crucial drug targets in different types of cancer. USP15 antagonizes ubiquitylation of CRL4CRBN target proteins, thereby preventing their degradation. Notably, USP15 is highly expressed in IMiD-resistant cells, and depletion of USP15 sensitizes these cells to lenalidomide. Inhibition of USP15 represents a valuable therapeutic opportunity to potentiate CELMoD and CRBN-based PROTAC therapies for the treatment of cancer.

Lipoic acid. Kinetics and pluripotent biological properties and derivatives.

Lipoic acid (LA) is globally known and its supplements are widely used. Despite its importance for the organism it is not considered a vitamin any more. The multiple metabolic forms and the differences in kinetics (absorption, distribution and excretion), as well as the actions of its enantiomers are analysed in the present article together with its biosynthetic path. The proteins involved in the transfer, biotransformation and activity of LA are mentioned. Furthermore, the safety and the toxicological profile of the compound are commented, together with its stability issues. Mechanisms of lipoic acid intervention in the human body are analysed considering the antioxidant and non-antioxidant characteristics of the compound. The chelating properties, the regenerative ability of other antioxidants, the co-enzyme activity and the signal transduction by the implication in various pathways will be discussed in order to be elucidated the pleiotropic effects of LA. Finally, lipoic acid integrating analogues are mentioned under the scope of the multiple pharmacological actions they acquire towards degenerative conditions.

Translation of Collagen Ultrastructure to Biomaterial Fabrication for Material-Independent but Highly Efficient Topographic Immunomodulation.

Supplement-free induction of cellular differentiation and polarization solely through the topography of materials is an auspicious strategy but has so far significantly lagged behind the efficiency and intensity of media-supplementation-based protocols. Consistent with the idea that 3D structural motifs in the extracellular matrix possess immunomodulatory capacity as part of the natural healing process, it is found in this study that human-monocyte-derived macrophages show a strong M2a-like prohealing polarization when cultured on type I rat-tail collagen fibers but not on collagen I films. Therefore, it is hypothesized that highly aligned nanofibrils also of synthetic polymers, if packed into larger bundles in 3D topographical biomimetic similarity to native collagen I, would induce a localized macrophage polarization. For the automated fabrication of such bundles in a 3D printing manner, the strategy of "melt electrofibrillation" is pioneered by the integration of flow-directed polymer phase separation into melt electrowriting and subsequent selective dissolution of the matrix polymer postprocessing. This process yields nanofiber bundles with a remarkable structural similarity to native collagen I fibers, particularly for medical-grade poly(ε-caprolactone). These biomimetic fibrillar structures indeed induce a pronounced elongation of human-monocyte-derived macrophages and unprecedentedly trigger their M2-like polarization similar in efficacy as interleukin-4 treatment.

Immunomodulatory effects of polysaccharides enzymatic hydrolysis from Hericium erinaceus on the MODE-K/DCs co-culture model.

The aim of this study was to explore the indirect immunomodulatory activities and its mechanism of enzymatic hydrolysis of Hericium erinaceus polysaccharides (EHEP) in the MODE-K/DCs co-culture model. According to the TEER value, transmission of phenol red and AKP activity of MODE-K cells, single model was established in order to evaluate the eligibility of MODE-K cells monolayer. Then the MODE-K/DCs co-culture model was set up and HEP and EHEP were added into the apical chamber, DCs were obtained for the expression of key surface markers, the ability of phagocytosis, the morphology, the secretion of cytokines and the production of target proteins. We found that after 21 d of culture, the MODE-K cells monolayer became intact and dense, which can be used for the MODE-K/DCs co-culture model. Under the treatment of HEP and EHEP, immature DCs become into mature DCs with the high expression of CD86 and MHCII, the low antigens up-taking, the typical morphology, the more content of IL-12 and TNF-α and the high level of TLR4, MyD88 and NF-κB proteins. However, compared with HEP, EHEP showed the better immunomodulatory activities. These findings indicated that EHEP could indirectly affect the immune function of DCs in the MODE-K/DCs co-culture model.

The regenerative mechanisms of platelet-rich plasma: A review.

Orthobiologics continue to gain popularity in many areas of medical science, especially in the field of regenerative medicine. Platelet-rich plasma derivatives are orthobiologic tools of particular interest. These biologic products can be obtained via centrifugation of a patient's whole blood and the components can then be subsequently isolated, concentrated and ultimately administered into injured tissues, particularly in areas where standard healing is disrupted. The elevated concentration of platelets above the basal value enables accelerated growth of various tissues with minimal side effects. The application of autologous orthobiologics is a relatively new biotechnology undergoing expansion which continues to reveal optimistic results in the stimulation and enhanced healing of various sorts of tissue injuries. The local release of growth factors and cytokines contained in platelet alpha granules accelerates and ameliorates tissue repair processes, mimicking and supporting standard wound healing. This effect is greatly enhanced upon combination with the fibrinolytic system, which are essential for complete regeneration. Fibrinolytic reactions can dictate proper cellular recruitment of certain cell populations such as mesenchymal stem cells and other immunomodulatory agents. Additionally, these reactions also control proteolytic activity in areas of wound healing and regenerative processes of mesodermal tissues including bone, cartilage, and muscle, which makes it particularly valuable for musculoskeletal health, for instance. Although many investigations have demonstrated significant results with platelet-rich plasma derivatives, further studies are still warranted.