In angiogenesis, circulating mononuclear cells are recruited to vascular lesions; however, the underlying mechanisms are poorly understood.Here, we characterize the functional role of protein tyrosine kinase 7 (PTK7)-expressing CD11b(+) mononuclear cells in vitro and in vivo using a mouse model of angiogenesis. Although the frequencies of PTK7(+)CD11b(+) cells in the bone marrow remained similar after vascular endothelial growth factor-A-induced neovascularization, we observed an 11-fold increase in the cornea. Importantly, vascular endothelial growth factor-A-induced chemotaxis of PTK7(+) cells was mediated by vascular endothelial growth factor receptor 2. In a coculture with endothelial cells, PTK7(+)CD11b(+) cells stabilized the vascular network for 2 weeks by expressing high levels of angiopoietin-1. The enhanced vascular stability was abolished by knockdown of angiopoietin-1 in PTK7(+)CD11b(+) cells and could be restored by angiopoietin-1 treatment.We conclude that PTK7 expression in perivascular mononuclear cells induces vascular endothelial growth factor receptor 2 and angiopoietin-1 expression and thus contributes to vascular stabilization in angiogenesis.
We have shown previously that retinol inhibits the invasion of all-trans-retinoic acid (ATRA)-resistant human colon cancer cell lines through a retinoic acid receptor (RAR)-independent mechanism by decreasing the activity of MMP-2 and -9. The objective of the current study was to determine the mechanism by which retinol inhibits metastasis. The ATRA-resistant colon cancer cell lines HCT-116 and SW620 an were treated for 24 h with 0, 1 or 10 microM retinol. Retinol treatment decreased the activity of phosphoinositide 3-kinase (PI3K) and Akt in both cell lines. Moreover, transfection with a constitutively active Akt blocked retinol¡¯s inhibitory effect on cell invasion. To determine the effect of retinol on the PI3K/Akt signaling pathway upstream of Akt, we examined the effect of retinol on the protein levels of insulin receptor substrate 1 (IRS-1) and the PI3K subunits, p85 and p110, as well as the phosphorylation of IRS-1. Retinol did not alter the protein levels of IRS-1, p85, and p-110 or IRS-1 phosphorylation. In conclusion, retinol inhibits cell invasion by decreasing PI3K and Akt activity; however retinol does not affect PI3K or IRS-1 protein levels or phosphorylation. Supported by Research Scholar Grant # 03-233-01-CNE from the American Cancer Society.
<p>The combination treatment of JBJ-04-125-02 and osimertinib do not result in any toxicity issues associated with weight loss but osimertinib at clinically relevant dose (25 mg/kg) is too potent to exhibit additive effect when combined with JBJ-04-125-02.</p>
As FK506 binding proteins (FK506BPs) are known to play an important role in the regulation of a variety of biological processes related to cell survival, this study was designed to examined the protective effects of FK506 binding protein 12 (FK506BP) on low humidity air flow induced dry eye in a rat model using transduced PEP-1-FK506BP. After the topical application of PEP-1-FK506BP, tear volumes were markedly increased and significant prevention of cornea damage was observed compared with dry eye rats. Further, immunohistochemical analysis demonstrated that PEP-1-FK506BP markedly prevented damage to the cornea, the bulbar conjunctiva, and the palpebral conjunctiva epithelial lining compared with dry eye rats. In addition, caspase-3 and PARP expression levels were found to be decreased. These results demonstrated that topical application of PEP-1-FK506BP significantly ameliorates dry eye injury in an animal model. Thus, we suggest that PEP-1-FK506BP can be developed as a new ophthalmic drop to treat dry eye diseases.
Abstract Retinol inhibits the growth of all-trans-retinoic acid (ATRA)-resistant human colon cancer cell lines through a retinoic acid receptor (RAR)-independent mechanism. The objectives of the current study were to determine if retinol inhibited the invasion of ATRA-resistant colon cancer cells independent of RAR and the effects of retinol on matrix metalloproteinases (MMPs). Retinol inhibited the migration and invasion of two ATRA-resistant colon cancer cell lines, HCT-116 and SW620, in a dose-dependent manner. To determine if transcription, particularly RAR-mediated transcription, or translation of new genes was required for retinol to inhibit cell invasion, cells were treated with retinol and cycloheximide, actinomycin D, or an RAR pan-antagonist. Treatment of cells with retinol and cycloheximide, actinomycin D, or an RAR pan-antagonist did not block the ability of retinol to inhibit cell invasion. In addition, retinol decreased MMP-1 mRNA levels in both cell lines, MMP-2 mRNA levels in the SW620 cell line, and MMP-7 and -9 mRNA levels in the HCT-116 cell line. Retinol also decreased the activity of MMP-2 and -9 and MMP-9 protein levels while increasing tissue inhibitor of MMP-1 media levels. In conclusion, retinol reduces the metastatic potential of ATRA-resistant colon cancer cells via a novel RAR-independent mechanism that may involve decreased MMP mRNA levels and activity.
C. elegans exhibits a directional migration toward a remembered temperature setpoint (Ts) by activating thermo-sensorimotor neurons. While cryophilic thermotaxis is well reproduced, thermophilic thermotaxis requires very stringent temperature regulations - otherwise, worms exhibit random migration in colder side of Ts. Here, we introduce a thermal stimulus device developed to control worms with different thermotactic behaviors on both colder and warmer sides of the Ts. On a linear gradient, the worm population displayed a Gaussian distribution near Ts but in a skewed shape with a peak shifted to the colder side due to their atactic motion in colder temperature than Ts. By repetitive application of thermal gradient-reversals, we found that their population density became higher near Ts because the speed at which the worms accumulate toward Ts was much faster than that of the dispersion by diffusion to the cold side, resulting in forced aggregation of worms at the desired temperature.
Traumatic brain injury (TBI) by an external physical impact results in compromised brain function via undesired neuronal death. Following the injury, resident and peripheral immune cells, astrocytes, and neural stem cells (NSCs) cooperatively contribute to the recovery of the neuronal function after TBI. However, excessive pro-inflammatory responses of immune cells, and the disappearance of endogenous NSCs at the injury site during the acute phase of TBI, can exacerbate TBI progression leading to incomplete healing. Therefore, positive outcomes may depend on early interventions to control the injury-associated cellular milieu in the early phase of injury. Here, we explore electrical stimulation (ES) of the injury site in a rodent model (male Sprague-Dawley rats) to investigate its overall effect on the constituent brain cell phenotype and composition during the acute phase of TBI. Our data showed that a brief ES for 1 hr on day 2 of TBI promoted anti-inflammatory phenotypes of microglia as assessed by CD206 expression and increased the population of NSCs and Nestin+ astrocytes at 7 days post-TBI. Also, ES effectively increased the number of viable neurons when compared to the unstimulated control group. Given the salience of microglia and neural stem cells for healing after TBI, our results strongly support the potential benefit of the therapeutic use of ES during the acute phase of TBI to regulate neuroinflammation and to enhance neuroregeneration.
<p>This file includes Supplementary Figures S1-S9. Figure S1. PARP1 is a partner of the RAP80-BRCA1 complex and promotes BRCA1 PARsylation. Figure S2. In vitro PARP1-driven PARsylation of BRCA1 fragments. Figure S3. Identification of a BRCA1 sequence required for optimal mono- and poly- ADP-ribosylation of the BRCA1 F3.7 fragment. Figure S4. The BRCA1 D5 sequence is not required for IRIF localization of BRCA1 in S/G2 phase nuclei. Figure S5. Olaparib treatment leads to increased BRCA1 chromatin association at sites near an I-SceI induced DSB. Figure S6. (a) RAP80 depletion and/or Olaparib treatment does not interfere with the interaction between BARD1 and BACH1, CtIP or RAD51. (b) The BRCA1-D5 sequence is required for a stable interaction between BRCA1 and RAP80 after IR, but not for the interaction between BRCA1 and BACH1 or CtIP. (c) RAP80 is not required for BRCA1 PARsylation in cells. (d) The RAP80-BRCA1 complex contains both PARsylated and unmodified BRCA1 molecules. Figure S7. PID is required for efficient RAP80 binding to PAR and PARsylated BRCA1, but not required for RAP80 IRIF localization. Figure S8. PARP1 activity is required for normal HRR tuning. Figure S9. Effects of RAD51 or EXO1/DNA2L depletion on STGC of cells expressing endogenous BRCA1, BRCA1-WT or BRCA1-D5, or depleted of BRCA1.</p>
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