Molecular Mechanisms of Oxidative Stress Relief by CAPE in ARPE-19 Cells.

Caffeic acid phenylethyl ester (CAPE) is an antioxidative agent originally derived from propolis. Oxidative stress is a significant pathogenic factor in most retinal diseases. Our previous study revealed that CAPE suppresses mitochondrial ROS production in ARPE-19 cells by regulating UCP2. The present study explores the ability of CAPE to provide longer-term protection to RPE cells and the underlying signal pathways involved. ARPE-19 cells were given CAPE pretreatment followed by t-BHP stimulation. We used in situ live cell staining with CellROX and MitoSOX to measure ROS accumulation; Annexin V-FITC/PI assay to evaluate cell apoptosis; ZO-1 immunostaining to observe tight junction integrity in the cells; RNA-seq to analyze changes in gene expression; q-PCR to validate the RNA-seq data; and Western Blot to examine MAPK signal pathway activation. CAPE significantly reduced both cellular and mitochondria ROS overproduction, restored the loss of ZO-1 expression, and inhibited apoptosis induced by t-BHP stimulation. We also demonstrated that CAPE reverses the overexpression of immediate early genes (IEGs) and activation of the p38-MAPK/CREB signal pathway. Either genetic or chemical deletion of UCP2 largely abolished the protective effects of CAPE. CAPE restrained ROS generation and preserved the tight junction structure of ARPE-19 cells against oxidative stress-induced apoptosis. These effects were mediated via UCP2 regulation of p38/MAPK-CREB-IEGs pathway.

Degradation and Deactivation of Bacterial Antibiotic Resistance Genes during Exposure to Free Chlorine, Monochloramine, Chlorine Dioxide, Ozone, Ultraviolet Light, and Hydroxyl Radical.

This work investigated degradation (measured by qPCR) and biological deactivation (measured by culture-based natural transformation) of extra- and intracellular antibiotic resistance genes (eARGs and iARGs) by free available chlorine (FAC), NH2Cl, O3, ClO2, and UV light (254 nm), and of eARGs by •OH, using a chromosomal ARG ( blt) of multidrug-resistant Bacillus subtilis 1A189. Rate constants for degradation of four 266-1017 bp amplicons adjacent to or encompassing the acfA mutation enabling blt overexpression increased in proportion to #AT+GC bps/amplicon, or in proportion to #5'-GG-3' or 5'-TT-3' doublets/amplicon, with respective values ranging from 0.59 to 2.3 (×1011 M-1 s-1) for •OH, 1.8-6.9 (×104 M-1 s-1) for O3, 3.9-9.2 (×103 M-1 s-1) for FAC, 0.35-1.2(×101 M-1 s-1) for ClO2, and 2.0-8.8 (×10-2 cm2/mJ) for UV at pH 7, and from 1.7-4.4 M-1 s-1 for NH2Cl at pH 8. For FAC, NH2Cl, O3, ClO2, and UV, ARG deactivation paralleled degradation of amplicons approximating a ∼800-1000 bp acfA-flanking sequence required for natural transformation in B. subtilis, whereas deactivation outpaced degradation for •OH. At practical disinfectant exposures, eARGs and iARGs were ≥90% degraded/deactivated by FAC, O3, and UV, but recalcitrant to NH2Cl and ClO2. iARG degradation/ deactivation always lagged cell inactivation. These findings provide a quantitative framework for evaluating ARG fate during disinfection/oxidation, and support using qPCR as a proxy for tracking ARG deactivation under carefully selected circumstances.

Examining the reliability of brain age algorithms under varying degrees of participant motion.

Brain age algorithms using data science and machine learning techniques show promise as biomarkers for neurodegenerative disorders and aging. However, head motion during MRI scanning may compromise image quality and influence brain age estimates. We examined the effects of motion on brain age predictions in adult participants with low, high, and no motion MRI scans (Original N = 148; Analytic N = 138). Five popular algorithms were tested: brainageR, DeepBrainNet, XGBoost, ENIGMA, and pyment. Evaluation metrics, intraclass correlations (ICCs), and Bland-Altman analyses assessed reliability across motion conditions. Linear mixed models quantified motion effects. Results demonstrated motion significantly impacted brain age estimates for some algorithms, with ICCs dropping as low as 0.609 and errors increasing up to 11.5 years for high motion scans. DeepBrainNet and pyment showed greatest robustness and reliability (ICCs = 0.956-0.965). XGBoost and brainageR had the largest errors (up to 13.5 RMSE) and bias with motion. Findings indicate motion artifacts influence brain age estimates in significant ways. Furthermore, our results suggest certain algorithms like DeepBrainNet and pyment may be preferable for deployment in populations where motion during MRI acquisition is likely. Further optimization and validation of brain age algorithms is critical to use brain age as a biomarker relevant for clinical outcomes.

Enhanced inactivation of Bacillus subtilis spores during solar photolysis of free available chlorine.

Aqueous free available chlorine (FAC) can be photolyzed by sunlight and/or artificial UV light to generate various reactive oxygen species, including HO(•) and O((3)P). The influence of this chemistry on inactivation of chlorine-resistant microorganisms was investigated using Bacillus subtilis endospores as model microbial agents and simulated and natural solar radiation as light sources. Irradiation of FAC solutions markedly enhanced inactivation of B. subtilis spores in 10 mM phosphate buffer; increasing inactivation rate constants by as much as 600%, shortening inactivation curve lag phase by up to 73% and lowering CTs required for 2 log10 inactivation by as much as 71% at pH 8.0 and 10 °C. Similar results were observed at pH 7.4 and 10 °C in two drinking water samples with respective DOC concentrations and alkalinities of 0.6 and 1.2 mg C/L and 81.8 and 17.1 mg/L as CaCO3. Solar radiation alone did not inactivate B. subtilis spores under the conditions investigated. A variety of experimental data indicate that the observed enhancements in spore inactivation can be attributed to the concomitant attack of spores by HO(•) and O3, the latter of which was found to accumulate to micromolar concentrations during simulated solar irradiation of 10 mM phosphate buffer (pH 8, 10 °C) containing [FAC]0 = 8 mg/L as Cl2.

Orbital Fractures.

Orbital fractures are common in facial trauma and can be a challenge to treat. Understanding anatomy of the orbit, the clinical evaluation, indications for surgery, surgical approaches, complications, and postoperative are essential in providing appropriate treatment for patients who have sustained orbital fractures. In this article, the authors review the diagnostic evaluation, acute management, treatment options, and common complications of orbital fractures, as well as recent advancements in orbital fracture repairs.

Osteogenesis imperfecta type V: Genetic and clinical findings in eleven Chinese patients.

INTRODUCTION: Osteogenesis imperfecta (OI) type V is a rare inherited disease characterized by multiple fractures, intraosseous membrane calcification, and hypercallus formation. We investigate the causative gene, phenotype and also observe the effects of zoledronic acid in Chinese OI type V patients. METHODS: The clinical phenotype and causative gene mutation was investigated in eleven patients with type V OI. Patients were given a dose of zoledronic acid 5mg intravenously. Fracture incidence and Z-score of bone mineral density (BMD) were evaluated. Serum levels of biomarkers such as cross linked C-telopeptide of type I collagen (ß-CTX) and safety parameters were assessed. RESULTS: The c.-14C>T mutation in the 5' untranslated region of IFITM5 was detected in all patients. The phenotype was largely variable, and no significant correlation of genotype and phenotype was found. After one dose of zoledronic acid infusion, fracture incidence significantly dropped from 2fractures/year before treatment to 0fracture/year after treatment (P=0.01). Z score of lumbar spine BMD elevated from -2.6 to -1.3 (P<0.001). Serum ß-CTX level decreased by 50% (P<0.05). No serious adverse event was found. CONCLUSION: No obvious correlation was found between the genotype and phenotype. Zoledronic acid had significantly skeletal protective effects in OI of type V.

The analysis of DKK1 polymorphisms in relation to skeletal phenotypes and bone response to alendronate treatment in Chinese postmenopausal women.

AIM: To investigate the correlation between DKK1 polymorphisms with bone phenotypes and response to alendronate treatment. MATERIALS & METHODS: Five tag single nucleotide polymorphisms of DKK1 were analyzed in 639 Chinese postmenopausal women with osteoporosis or osteopenia. Bone mineral density (BMD), ß-CTX and ALP were measured before and after alendronate treatment. RESULTS: Genotypes at rs1896367, rs1528877 and rs2241529 correlated to baseline BMD (p < 0.05). rs1528877 and rs2241529 polymorphisms correlated to baseline ß-CTX levels (p < 0.05). rs2241529 polymorphisms of DKK1 had a small influence on the skeletal response to alendronate treatment (p < 0.05). CONCLUSION: DKK1 polymorphisms may correlate to baseline BMD and serum ß-CTX levels, but present a weak effect on the response to alendronate.

Gorham-Stout disease: radiological, histological, and clinical features of 12 cases and review of literature.

Gorham-Stout disease (GSD) is an exceedingly rare disease characterized by progressive osteolysis and angiomatosis. We investigate the features of this disease and evaluate the effects of bisphosphonates (BPs) on it. The clinical, radiological, and pathological characteristics of 12 patients diagnosed with GSD were summarized. Immunohistochemical staining with specific lymphatic endothelial markers (D2-40), vascular markers (CD 31, CD 34), and vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor 3 (VEGFR-3) was performed in specimens of bone biopsy. Patients were treated with either BPs or conjunction therapy of radiation and BPs. The effects of BPs were evaluated by the change of radiological progression, bone mineral density (BMD) and bone turnover biomarkers. To further evaluate the prognosis, a literature review was done. Idiopathic massive osteolysis was found in all patients, including 11 polyostotic and one mono-ostotic osteolysis. Soft tissue lymphangioma was presented in four patents. Four patients were complicated with chylothorax. Endothelial cells lining the proliferative vessels were positive for CD31 and CD34 and D2-40. Mild expression of VEGF and VEGFR-3 was also revealed. Stabilization in osteolysis and improvement in BMD were observed after single therapy with BPs or combined with radiotherapy. High mortality rate was found in patients with chylothorax. Spontaneous, progressive osteolysis is the most typical sign of GSD. BPs and radiotherapy can contribute to the clinical stabilization in bone lesion of GSD. The complicated chylothorax possibly indicates poor prognosis.

Enzymatically Degradable Polyester-Based Adhesives.

A designed 3,4-dihydroxyphenylalanine (DOPA) mimetic enzymatic degradable synthetic adhesive with good adhesion to soft tissue and metals made by a simple two-step reaction is presented in this article. This adhesive has degradable polycaprolactone-type of repeat units together with glycidyl methacrylate (GMA) and oligo(ethylene glycol) methacrylate (OEGMA) on the polymer backbone. Radical initiated copolymerization of 2-methylene-1,3-dioxepane (MDO), glycidyl methacrylate (GMA) and OEGMA followed by immobilization of catechol group on epoxy rings of GMA provided the adhesive material. Fe(acac)3 was proved to be the most effective cross-linking agent with lap shear strength of 13.13 ± 1.74 kPa and 218.4 ± 16.0 kPa on soft tissue (porcine skin) and metal (aluminum), respectively. The cross-linked adhesive showed good adhesion stability in pH 7 PBS buffer at 37 °C for at least 1 week. Because of the high adhesive strength, enzymatic degradability, and low toxicity, the material is a promising candidate for future studies as medical glue.

SOST polymorphisms and response to alendronate treatment in postmenopausal Chinese women with osteoporosis.

AIM: To investigate the association between SOST gene polymorphisms and response to alendronate treatment. MATERIALS & METHODS: 639 Chinese postmenopausal women with osteoporosis or osteopenia received alendronate treatment. Polymorphisms of SOST were analyzed. Bone mineral density (BMD), serum ALP and ß-CTX levels were measured. The correlation of SOST polymorphisms with changes of BMD and bone biomarkers after treatment was analyzed. RESULTS: rs1234612 and rs851054 polymorphisms were correlated to baseline lumbar spine BMD (p < 0.05). After 12 months of treatment rs1234612 and rs865429 polymorphisms were correlated to BMD changes at the lumbar spine (p < 0.05) or femoral neck (p < 0.05), respectively. CONCLUSION: The polymorphisms of SOST are genetic factors affecting bone health and response to alendronate in Chinese postmenopausal women.

Novel mutations in FKBP10 and PLOD2 cause rare Bruck syndrome in Chinese patients.

Bruck syndrome (BS) is an extremely rare form of osteogenesis imperfecta characterized by congenital joint contracture, multiple fractures and short stature. We described the phenotypes of BS in two Chinese patients for the first time. The novel compound heterozygous mutations c.764_772dupACGTCCTCC (p.255_257dupHisValLeu) in exon 5 and c.1405G>T (p.Gly469X) in exon 9 of FKBP10 were identified in one proband. The novel compound heterozygous mutations c.1624delT (p.Tyr542Thrfs*18) in exon 14 and c.1880T>C (p.Val627Ala) in exon 17 of PLOD2 were identified in another probrand. Intravenous zoledronate was a potent agent for these patients, confirmed the efficacy of bisphosphonates on this disease. In conclusion, the novel causative mutations identified in the patients expand the genotypic spectrum of BS.

Association of farnesyl diphosphate synthase polymorphisms and response to alendronate treatment in Chinese postmenopausal women with osteoporosis.

BACKGROUND: Genetic factors are important in the pathogenesis of osteoporosis, but less is known about the genetic determinants of osteoporosis treatment. We aimed to explore the association between the gene polymorphisms of key enzyme farnesyl diphosphate synthase (FDPS) in mevalonate signaling pathway of osteoclast and response to alendronate therapy in osteoporotic postmenopausal women in China. METHODS: The study group comprised 639 postmenopausal women aged (62.2 ± 7.0) years with osteoporosis or osteopenia who had been randomly assigned to low dose group (70 mg/2 w) or standard dose group (70 mg/w) of alendronate in this 1-year study. We identified allelic variant of the FDPS gene using the polymerase chain reaction and restriction enzyme Faul. Before and after treatment, serum levels of calcium, phosphate, alkaline phosphatase (ALP), cross linked C-telopeptide of type I collagen (ß-CTX) were detected. Bone mineral density (BMD) at lumbar spine and proximal femur was measured. The association was analyzed between the polymorphisms of FDPS gene and the changes of BMD, bone turnover biomarkers after the treatment. RESULTS: The FDPS rs2297480 polymorphisms were associated with baseline BMD at femoral neck, and patients with CC genotype had significantly higher baseline femoral neck BMD ((733.6 ± 84.1) mg/cm(2)) than those with AC genotypes ((703.0 ± 86.9) mg/cm(2)) and AA genotypes ((649.8 ± 62.4) mg/cm(2)) (P < 0.01). No significant difference in BMD at lumbar spine was observed among different genotypes of FDPS. The percentage change of serum ALP level was significantly lower in patients with CC genotype (-22.9%) than that in those with AC genotype (-24.1%) and AA genotype (-29.8%) of FDPS after 12 months of alendronate treatment (P < 0.05). Neither percentage change of BMD nor ß-CTX level after alendronate treatment had association with FDPS genotype. CONCLUSIONS: FDPS gene was probably a candidate gene to predict femoral neck BMD at baseline. FDPS gene alleles could predict change percentage of ALP after treatment of alendronate, but possibly had no significant relationship with the responsiveness of BMD to alendronate therapy.