CM1, a Chrysin Derivative, Protects from Endotoxin-Induced Lethal Shock by Regulating the Excessive Activation of Inflammatory Responses.

Sepsis, a leading cause of death worldwide, is a harmful inflammatory condition that is primarily caused by an endotoxin released by Gram-negative bacteria. Effective targeted therapeutic strategies for sepsis are lacking. In this study, using an in vitro and in vivo mouse model, we demonstrated that CM1, a derivative of the natural polyphenol chrysin, exerts an anti-inflammatory effect by inducing the expression of the ubiquitin-editing protein TNFAIP3 and the NAD-dependent deacetylase sirtuin 1 (SIRT1). Interestingly, CM1 attenuated the Toll-like receptor 4 (TLR4)-induced production of inflammatory cytokines by inhibiting the extracellular-signal-regulated kinase (ERK)/MAPK and nuclear factor kappa B (NF-κB) signalling pathways. In addition, CM1 induced the expression of TNFAIP3 and SIRT1 on TLR4-stimulated primary macrophages; however, the anti-inflammatory effect of CM1 was abolished by the siRNA-mediated silencing of TNFAPI3 or by the genetic or pharmacologic inhibition of SIRT1. Importantly, intravenous administration of CM1 resulted in decreased susceptibility to endotoxin-induced sepsis, thereby attenuating the production of pro-inflammatory cytokines and neutrophil infiltration into the lung compared to control mice. Collectively, these findings demonstrate that CM1 has therapeutic potential for diverse inflammatory diseases, including sepsis.

SIRT1 Promotes Host Protective Immunity against Toxoplasma gondii by Controlling the FoxO-Autophagy Axis via the AMPK and PI3K/AKT Signalling Pathways.

Sirtuin 1 (SIRT1) regulates cellular processes by deacetylating non-histone targets, including transcription factors and intracellular signalling mediators; thus, its abnormal activation is closely linked to the pathophysiology of several diseases. However, its function in Toxoplasma gondii infection is unclear. We found that SIRT1 contributes to autophagy activation via the AMP-activated protein kinase (AMPK) and PI3K/AKT signalling pathways, promoting anti-Toxoplasma responses. Myeloid-specific Sirt1-/- mice exhibited an increased cyst burden in brain tissue compared to wild-type mice following infection with the avirulent ME49 strain. Consistently, the intracellular survival of T. gondii was markedly increased in Sirt1-deficient bone-marrow-derived macrophages (BMDMs). In contrast, the activation of SIRT1 by resveratrol resulted in not only the induction of autophagy but also a significantly increased anti-Toxoplasma effect. Notably, SIRT1 regulates the FoxO-autophagy axis in several human diseases. Importantly, the T. gondii-induced phosphorylation, acetylation, and cytosolic translocation of FoxO1 was enhanced in Sirt1-deficient BMDMs and the pharmacological inhibition of PI3K/AKT signalling reduced the cytosolic translocation of FoxO1 in BMDMs infected with T. gondii. Further, the CaMKK2-dependent AMPK signalling pathway is responsible for the effect of SIRT1 on the FoxO3a-autophagy axis and for its anti-Toxoplasma activity. Collectively, our findings reveal a previously unappreciated role for SIRT1 in Toxoplasma infection.

Frontalis Sling Using a Silicone Rod for Ptosis in Third Nerve Palsy: Cosmesis versus Safety.

PURPOSE: To evaluate the results of the frontalis sling operation using a silicone rod for the correction of ptosis in patients with third nerve palsy with a focus on corneal safety. METHODS: Patients with third nerve palsy who underwent the frontalis sling operation using a silicone rod between 2008 and 2019 were included in this study. The medical records of all patients were reviewed, and their clinical characteristics and postoperative outcomes were analyzed. In this retrospective, interventional case series, the main outcome measures were eyelid contour, eyelid height by margin reflex distance, and corneal status. RESULTS: Twenty-four eyes of 18 patients (12 male and six female patients) were included. The mean age at the time of surgery was 35.1 years (range, 5-64 years). Twelve patients underwent a unilateral ptosis operation, and six patients received a bilateral ptosis operation. The mean follow-up period was 32.1 months (range, 2-87 months). Most patients (21 of 24 eyes, 88%) showed poor Bell's phenomenon on preoperative examination. Satisfactory eyelid height and eyelid contour were achieved in almost all patients (mean postoperative margin reflex distance, +1.2 mm) postoperatively. Although corneal erosions were detected for several months in eight of 24 eyes after surgery, these findings were well controlled medically with artificial tear eye drops and ointments. CONCLUSIONS: Frontalis sling surgery using a silicone rod can safely and effectively correct ptosis without severe corneal complications in patients with third nerve palsy. Our study outlines a new method to define the postoperative safety outcome by specifically focusing on categorized corneal status.

The effects of using spacer grafts on lower-eyelid retraction surgery in patients with facial nerve palsy.

PURPOSE: To evaluate the effects of the concomitant use of spacer grafts in lateral tarsal strip surgery in patients with facial nerve palsy-related lower-eyelid retraction. METHODS: Patients who underwent lateral tarsal strip surgery to correct facial nerve palsy-related lower-eyelid retraction were retrospectively reviewed. Postoperative decreases in marginal reflex distance-2 values at 1, 2 and 6 months were measured along with the effects of spacer grafts. RESULTS: Forty-five patients (28 males) were included (mean age: 59.56 years). Mean preoperative marginal reflex distance-2 was 6.87 ± 1.34 mm. Twenty patients underwent lateral tarsal strip surgery only (lateral tarsal strip-only); 25 patients underwent lateral tarsal strip surgery using spacer grafts (lateral tarsal strip + graft). Median (interquartile range) follow-up duration was 12.0 (6.0-23.0) months. Retraction was significantly improved and maintained at 1, 2 and 6 months postoperatively in all patients (mean marginal reflex distance-2: 3.78 ± 1.06 mm, 4.30 ± 1.23 mm and 4.72 ± 1.11 mm, respectively). Surgical outcomes were significantly better in the lateral tarsal strip + graft than in the lateral tarsal strip-only group (Δmarginal reflex distance-2: 3.92 vs. 2.05 mm at 1 month, p < 0.001; 3.38 vs. 1.61 mm at 2 months, p = 0.001; 2.88 vs. 1.69 at 6 months, p = 0.042). Subgroup analyses by spacer graft type revealed no significant differences. CONCLUSION: The concomitant use of spacer material in lateral tarsal strip surgery yielded better surgical outcomes than lateral tarsal strip surgery alone. The use of spacer grafts should be considered for correcting severe facial nerve palsy-related lower-eyelid retraction.

The effect of lacrimal drainage abnormality on the surgical outcomes of congenital lacrimal fistula and vice versa.

PURPOSE: To evaluate the various surgical modalities of congenital lacrimal fistula and assess the mutual effect of lacrimal fistula and lacrimal drainage abnormality on the individual surgical outcomes. METHODS: In this retrospective cohort and case-control study, 74 eyes from 63 patients with lacrimal fistula who underwent surgical management between 2000 and 2015 at three medical centers were enrolled. The data collected included sex, age, preoperative symptoms, presence of concurrent lacrimal drainage abnormality, surgical methods, and surgical outcomes. The main outcome measures were treatment outcomes based on lacrimal drainage patency and symptom improvement, surgical outcome of fistulectomy according to the presence of lacrimal drainage abnormality, and surgical outcome of lacrimal drainage abnormality according to the presence of fistula. RESULTS: The mean age at the time of surgery was 9.2 (SD, ±8.8) years and the mean follow-up duration was 14.4 (SD, ±19.5) months. All eyes (37/37) with fistula without lacrimal drainage abnormality demonstrated surgical success after simple fistulectomy. Patients with concurrent lacrimal drainage abnormalities showed more frequent surgical failure than those with fistula alone (p = 0.009). However, the presence of fistula did not affect the outcomes of surgery for lacrimal drainage abnormality (p = 0.179). CONCLUSION: Simple fistulectomy is sufficient for sole asymptomatic or pauci-symptomatic lacrimal fistula. Symptomatic fistula as well as those accompanied with lacrimal drainage abnormality underwent fistulectomy and lacrimal drainage system surgery. Patients with accompanying lacrimal drainage system abnormalities showed less favorable outcomes. Meticulous preoperative examination of the lacrimal drainage system is critical for surgical planning and prognosis prediction.

Protective effects of fermented rice extract on ulcerative colitis induced by dextran sodium sulfate in mice.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease (IBD), characterized by the gut mucosal ulceration. Growing evidence indicates that dysregulation of immune response to the commensal microbiota involves the pathogenesis of IBD. Previous studies have demonstrated the favorable probiotic effects of fermented rice extracts through triple fermentation with Saccharomyces cerevisiae and Weissella cibaria (FRe). Thus, the therapeutic potential of FRe for UC was examined. Dextran sodium sulfate UC mice model was orally administered distilled water as a control, sulfasalazine, or FRe at 300, 200, and 100 mg/kg, once a day for a week. The UC control exhibited body weight loss, bloody stools, and colonic shortening. However, the FRe, especially at 300 mg/kg, led to a reduction in weight loss, disease activity index scores, and colon weight, and an increase in colorectal length. The histopathological analyses revealed mild changes involved in the colonic crypt and mucosal damages in the FRe groups, along with inhibited inflammation. Indeed, the FRe reduced neutrophil infiltration and production of proinflammatory cytokines (i.e., tumor necrosis factor-α, interleukin-6/-8). This was accompanied by the down-regulation of nuclear factor-kappa B. The gene expression responsible for the intestinal barrier integrity (i.e., Zonna occludens-1/-2, Claudin-1, Occludin, Mucin-1/-2) was up-regulated in the FRe groups. In addition, the FRe reduced lipid peroxidation and enhanced antioxidant activity. Interestingly, the microbiota dysbiosis was attenuated in the FRe groups, and the number of beneficial bacteria, Lactobacilli and Bifidobacteria, was increased. It suggests that the FRe potently ameliorate UC as a functional food.

Wheat Extract Oil (WEO) Attenuates UVB-Induced Photoaging via Collagen Synthesis in Human Keratinocytes and Hairless Mice.

The efficacy of wheat extract oil (WEO), standardized to glucosylceramides, for protecting against ultraviolet B (UVB)-induced damage of skin barrier function was assessed using the SHK-1 hairless mouse model and two human skin cell lines, namely, CCD-986sk and HeCaT. The ability of repeated oral administration of 30, 60, and 120 mg of WEO/kg/day for 12 weeks to prevent skin damage of SKH-1 hairless mice induced by UVB irradiation was evaluated. The results demonstrated that UVB-induced water evaporation (transepidermal water loss, TEWL) was significantly decreased by WEO. Similarly, UVB-induced losses in moisture and skin elasticity were improved by WEO supplementation. WEO attenuated the tissue procollagen type I, hyaluronic acid (HA), and ceramide reductions induced by UVB treatment as well. Collagen concentrations in skin tissue were increased in the WEO-treated mice, while UVB-induced epidermal thickening was reduced. In vitro studies using HeCaT human keratinocytes confirmed increased HA and collagen synthesis in response to WEO treatment. This may occur via WEO suppression of matrix metallopeptidase-1 (MMP-1), since its induction by UVB treatment was diminished in treated CCD-986sk cells. Oral administration of WEO improves skin barrier function in UVB-irradiated mice by attenuating damage typically observed in photoaging. This research further clarifies the clinical benefits previously observed by dietary WEO consumption.

Activating transcription factor 3 represses inflammatory responses by binding to the p65 subunit of NF-κB.

Activating transcription factor 3 (ATF3) is induced by inflammatory responses, cell death, cytokines, and oxidative stress conditions. ATF3 is a negative regulator in the Toll-like receptor 4 signalling pathway. The principal molecule in this pathway is nuclear factor κB (NF-κB) that translocates into the nucleus to initiate the transcription of inflammatory mediators. However, scarce data are available regarding the interaction of ATF3 and p65, a part of the NF-κB dimer. Therefore, we studied the mechanism of regulation of p65 by ATF3 in RAW 264.7 cells. First, LPS-mediated NF-κB activation was confirmed, and then the direct interaction of ATF3 and p65 was observed through immunoprecipitation (IP). The presence of histone deacetylase 1 (HDAC1) was also detected in the complex. In ATF3 deficient cells, NF-κB activity was up-regulated and HDAC1 was not detected by IP. These observations suggest that p65 is attenuated by ATF3 such that ATF3 recruits HDAC1 to the ATF3/p65 complex and facilitates the deacetylation of p65. Likewise, inflammatory response genes were induced by translocated NF-κB in ATF3-deficient cells. Cumulatively, we uncovered a novel mechanism for the negative regulation of NF-κB by ATF3 via direct interaction with p65.

Mechanism of Cisplatin-Induced Cytotoxicity Is Correlated to Impaired Metabolism Due to Mitochondrial ROS Generation.

The chemotherapeutic use of cisplatin is limited by its severe side effects. In this study, by conducting different omics data analyses, we demonstrated that cisplatin induces cell death in a proximal tubular cell line by suppressing glycolysis- and tricarboxylic acid (TCA)/mitochondria-related genes. Furthermore, analysis of the urine from cisplatin-treated rats revealed the lower expression levels of enzymes involved in glycolysis, TCA cycle, and genes related to mitochondrial stability and confirmed the cisplatin-related metabolic abnormalities. Additionally, an increase in the level of p53, which directly inhibits glycolysis, has been observed. Inhibition of p53 restored glycolysis and significantly reduced the rate of cell death at 24 h and 48 h due to p53 inhibition. The foremost reason of cisplatin-related cytotoxicity has been correlated to the generation of mitochondrial reactive oxygen species (ROS) that influence multiple pathways. Abnormalities in these pathways resulted in the collapse of mitochondrial energy production, which in turn sensitized the cells to death. The quenching of ROS led to the amelioration of the affected pathways. Considering these observations, it can be concluded that there is a significant correlation between cisplatin and metabolic dysfunctions involving mROS as the major player.

Risk factors predicting the need for additional surgery in consecutive esotropia.

PURPOSE: To evaluate the risk factors predicting surgical treatment in consecutive esotropia occurring after surgery for intermittent exotropia. METHODS: The clinical records of 52 patients with consecutive esotropia who underwent exotropia surgery were retrospectively reviewed. All patients demonstrated consecutive esotropia with diplopia or suppression for more than 1 month after surgery for exotropia. Patients were divided into two groups (the surgical group and nonsurgical group) depending on whether surgery was required for consecutive esotropia. Surgery for esotropia was performed only in patients with more than 10 prism diopters (PD) esodeviation that persisted for a minimum of 6 months, those who had suppression in 1 eye or diplopia, and those who could not achieve fine stereopsis. The nonsurgical treatment up to 6 months postoperatively was part-time patching and prism therapy in both groups. Patient characteristics were evaluated in the two groups. RESULTS: The surgical group was composed of 17 patients and the nonsurgical group was composed of 35 patients. Age, gender, refractive error, best-corrected visual acuity, and postoperative overcorrection at 1 day were not significantly different in the two groups (P > .05). However, the distance strabismic angle at 1 month postoperatively was 2.5 ± 3.8 PD esodeviation (range: 14 PD esotropia to 4 PD exotropia) in the nonsurgical group and 5.4 ± 5.1 PD esodeviation (range: 20 PD esotropia to orthotropia) in the surgical group; these values were statistically significant (P < .05). CONCLUSIONS: The clinically significant risk factor affecting the surgical decision for consecutive esotropia was a large esotropic angle at 1 month postoperatively in this study.

Glial proliferation after vitrectomy for a macular hole: a spectral domain optical coherence tomography study.

PURPOSE: To characterize eyes with glial proliferation after macular hole (MH) surgery. METHODS: We retrospectively reviewed patients who underwent vitrectomy for an idiopathic MH using spectral domain optical coherence tomography (SD-OCT). The pattern of the closed MH after surgery was categorized according to the presence (group 1) or absence (group 2) of apparent glial proliferation, which was determined by glial tissue reflectivity involving the external limiting membrane (ELM) and photoreceptor layers. Photoreceptor layer defect areas were categorized as mild or severe. Best-corrected visual acuity (BCVA) and pre- and postoperative OCT parameters were compared between the two groups. RESULTS: Among 30 eyes followed-up for a median of 11 months, seven (23 %) were assigned to group 1 and 23 (77 %) to group 2. The median age was higher in group 1 (70 years) than in group 2 (63 years). The postoperative BCVA was poorer in group 1 than in group 2 at 3 months and at the final examination (P = 0.022 and P < 0.001 respectively). The median preoperative basal hole diameter in group 1 (1,219 µm) was larger than that of group 2 (590 µm) (P = 0.002). The MH index (hole height/basal hole diameter) was smaller in group 1 than in group 2 (P = 0.012). At the final examination, group 1 had larger mild and severe photoreceptor layer defect areas (medians 1,300 µm and 207 µm respectively) than group 2 (medians 110 µm and 70 µm respectively) (P < 0.001 and P < 0.001 respectively). CONCLUSIONS: Eyes with glial proliferation after surgery for MH had different preoperative characteristics than eyes with no evidence of glial proliferation. In addition to a large hole diameter, other factors such as a small MH index and advanced age could be involved in the development of glial proliferation.

Comparison of clinical features between two different types of exotropia before 12 months of age based on stereopsis outcome.

PURPOSE: To compare the clinical outcomes and characteristics of 2 different types of early-onset exotropia on the basis of stereopsis outcome. DESIGN: Retrospective case series. PARTICIPANTS: A total of 24 patients with newly diagnosed exotropia before 12 months of age and at least 1 year of follow-up after surgery. METHODS: The clinical records of all patients were reviewed. Patients were divided into 2 groups according to stereopsis. The presumable early-onset intermittent exotropia (EIE) group was composed of 6 patients (25%) who showed excellent stereopsis of ≥60 arc seconds. The primary infantile exotropia (PIE) group was composed of 18 patients (75%) who showed stereopsis of ≤100 arc seconds. We compared the preoperative and postoperative clinical features of the 2 groups. MAIN OUTCOME MEASURES: The age at onset and visit, age at surgery, constancy before surgery, preoperative and postoperative angles of deviation, distant suppression, reoperation rate, and presence of dissociative vertical deviation (DVD) and inferior oblique overaction (IOOA). RESULTS: The age at the stereopsis test was 6.8 years in the EIE group and 6.4 years in the PIE group (P=0.41). There was no statistical difference in the mean preoperative exodeviation (32.8 prism diopters [PD] in the EIE group vs. 34.7 PD in the PIE group, P=0.58) and postoperative deviation at the stereopsis test (1.0 PD in the EIE group vs. 2.0 PD in the PIE group, P=0.97). The reoperation rate was 33% in the EIE group and 27% in the PIE group (P=1.00). There was no statistical difference in constancy between the EIE and PIE groups (33% vs. 56%, respectively, P=0.64). However, DVD (61%) and IOOA (56%) were noted only in the PIE group (P=0.016, P=0.024, respectively), and distant suppression was noted only in the PIE group (61%, P=0.016). CONCLUSIONS: The results indicate that excellent sensory outcome was observed in only 25% of patients with exotropia before 12 months of age, but motor outcome and reoperation rate were not different between the 2 types of exotropia. We observed DVD, IOOA, and distant suppression only in the PIE group. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Doxorubicin induces cytotoxicity through upregulation of pERK-dependent ATF3.

Although doxorubicin is commonly used in the treatment of many cancer types, its use in chemotherapy has been limited, largely because of its severe side effects, including cardiotoxicity and nephrotoxicity. In this study, we aimed to identify the mechanism of doxorubicin-induced cytotoxicity by using the human kidney proximal tubule cell line HK-2. Furthermore, we investigated the role of activating transcription factor 3 (ATF3) as a mediator of doxorubicin-induced cytotoxicity by using wild-type mouse embryonic fibroblasts (MEF) cells and ATF3 knockout (KO) cells. In HK-2 cells, doxorubicin decreased cell viability in a dose-dependent manner and induced an increase in cells in the sub G1 and G2/M phases at all doses. Doxorubicin treatment showed the following dose-dependent effects: increase in the secretion of tumor necrosis factor alpha; decrease in the expression of phosphorylated protein kinase A and Bcl-2; and increase in the expression of phosphorylated signal transducer and activator of transcription 3, phosphorylated extracellular signal-regulated kinase (ERK), and ATF3. Based on these results, we suggest that doxorubicin induces cytotoxicity through an ERK-dependent pathway, and ATF3 plays a pivotal role as a transcriptional regulator in this process.