Synergistic anti-proliferative activity of JQ1 and GSK2801 in triple-negative breast cancer.

BACKGROUND: Triple-negative breast cancer (TNBC) constitutes 10-20% of breast cancers and is challenging to treat due to a lack of effective targeted therapies. Previous studies in TNBC cell lines showed in vitro growth inhibition when JQ1 or GSK2801 were administered alone, and enhanced activity when co-administered. Given their respective mechanisms of actions, we hypothesized the combinatorial effect could be due to the target genes affected. Hence the target genes were characterized for their expression in the TNBC cell lines to prove the combinatorial effect of JQ1 and GSK2801. METHODS: RNASeq data sets of TNBC cell lines (MDA-MB-231, HCC-1806 and SUM-159) were analyzed to identify the differentially expressed genes in single and combined treatments. The topmost downregulated genes were characterized for their downregulated expression in the TNBC cell lines treated with JQ1 and GSK2801 under different dose concentrations and combinations. The optimal lethal doses were determined by cytotoxicity assays. The inhibitory activity of the drugs was further characterized by molecular modelling studies. RESULTS: Global expression profiling of TNBC cell lines using RNASeq revealed different expression patterns when JQ1 and GSK2801 were co-administered. Functional enrichment analyses identified several metabolic pathways (i.e., systemic lupus erythematosus, PI3K-Akt, TNF, JAK-STAT, IL-17, MAPK, Rap1 and signaling pathways) enriched with upregulated and downregulated genes when combined JQ1 and GSK2801 treatment was administered. RNASeq identified downregulation of PTPRC, MUC19, RNA5-8S5, KCNB1, RMRP, KISS1 and TAGLN (validated by RT-qPCR) and upregulation of GPR146, SCARA5, HIST2H4A, CDRT4, AQP3, MSH5-SAPCD1, SENP3-EIF4A1, CTAGE4 and RNASEK-C17orf49 when cells received both drugs. In addition to differential gene regulation, molecular modelling predicted binding of JQ1 and GSK2801 with PTPRC, MUC19, KCNB1, TAGLN and KISS1 proteins, adding another mechanism by which JQ1 and GSK2801 could elicit changes in metabolism and proliferation. CONCLUSION: JQ1-GSK2801 synergistically inhibits proliferation and results in selective gene regulation. Besides suggesting that combinatorial use could be useful therapeutics for the treatment of TNBC, the findings provide a glimpse into potential mechanisms of action for this combination therapy approach.

Impact of Gastric Acid Induced Surface Changes on Mechanical Behavior and Optical Characteristics of Dental Ceramics.

PURPOSE: To test the impact of exposure to artificial gastric acid combined with toothbrush abrasion on the properties of dental ceramics. Earlier research has indicated that immersion in artificial gastric acid has caused increased surface roughness of dental ceramics; however, the combined effects of acid immersion and toothbrush abrasion and the impact of increased surface roughness on mechanical strength and optical properties have not been studied. MATERIALS AND METHODS: Three commercially available ceramics were chosen for this study: feldspathic porcelain, lithium disilicate glass-ceramic, and monolithic zirconium oxide. The specimens (10 × 1 mm discs) were cut, thermally treated as required, and polished. Each material was divided into four groups (n = 8 per group): control (no exposure), acid only, brush only, acid + brush. The specimens were immersed in artificial gastric acid (50 ml of 0.2% [w/v] sodium chloride in 0.7% [v/v] hydrochloric acid mixed with 0.16 g of pepsin powder, pH = 2) for 2 minutes and rinsed with deionized water for 2 minutes. The procedure was repeated 6 times/day × 9 days, and specimens were stored in deionized water at 37°C. Toothbrush abrasion was performed using an ISO/ADA design brushing machine for 100 cycles/day × 9 days. The acid + brush group received both treatments. Specimens were examined under SEM and an optical microscope for morphological changes. Color and translucency were measured using spectrophotometer CIELAB coordinates (L*, a*, b*). Surface gloss was measured using a gloss meter. Surface roughness was measured using a stylus profilometer. Biaxial flexural strength was measured using a mechanical testing machine. The data were analyzed by one-way ANOVA followed by Tukey's HSD post hoc test (p < 0.05). RESULTS: Statistically significant changes were found for color, gloss, and surface roughness for porcelain and e.max specimens. No statistically significant changes were found for any properties of zirconia specimens. CONCLUSIONS: The acid treatment affected the surface roughness, color, and gloss of porcelain and e.max ceramics. The changes in translucency and mechanical strength for all materials were not statistically significant. Zirconia ceramic showed resistance to all treatments.

Hypoxia promotes tau hyperphosphorylation with associated neuropathology in vascular dysfunction.

BACKGROUND: Hypertension-induced microvascular brain injury is a major vascular contributor to cognitive impairment and dementia. We hypothesized that chronic hypoxia promotes the hyperphosphorylation of tau and cell death in an accelerated spontaneously hypertensive stroke prone rat model of vascular cognitive impairment. METHODS: Hypertensive male rats (n = 13) were fed a high salt, low protein Japanese permissive diet and were compared to Wistar Kyoto control rats (n = 5). RESULTS: Using electron paramagnetic resonance oximetry to measure in vivo tissue oxygen levels and magnetic resonance imaging to assess structural brain damage, we found compromised gray (dorsolateral cortex: p = .018) and white matter (corpus callosum: p = .016; external capsule: p = .049) structural integrity, reduced cerebral blood flow (dorsolateral cortex: p = .005; hippocampus: p < .001; corpus callosum: p = .001; external capsule: p < .001) and a significant drop in cortical oxygen levels (p < .05). Consistently, we found reduced oxygen carrying neuronal neuroglobin (p = .008), suggestive of chronic cerebral hypoperfusion in high salt-fed rats. We also observed a corresponding increase in free radicals (NADPH oxidase: p = .013), p-Tau (pThr231) in dorsolateral cortex (p = .011) and hippocampus (p = .003), active interleukin-1ß (p < .001) and neurodegeneration (dorsolateral cortex: p = .043, hippocampus: p = .044). Human patients with subcortical ischemic vascular disease, a type of vascular dementia (n = 38; mean age = 68; male/female ratio = 23/15) showed reduced hippocampal volumes and cortical shrinking (p < .05) consistent with the neuronal cell death observed in our hypertensive rat model as compared to healthy controls (n = 47; mean age = 63; male/female ratio = 18/29). CONCLUSIONS: Our data support an association between hypertension-induced vascular dysfunction and the sporadic occurrence of phosphorylated tau and cell death in the rat model, correlating with patient brain atrophy, which is relevant to vascular disease.

Protocol to measure apoptosis-associated speck-like protein containing a CARD specks in human cerebrospinal fluid via imaging flow cytometry.

Apoptosis-associated speck-like protein containing a c-terminal caspase activation and recruitment domain (ASC) specks are elevated in the cerebrospinal fluid (CSF) of Alzheimer's disease and related dementias (AD/ADRDs) patients. Here, we present a flow cytometry protocol to quantify ASC specks. We describe steps for fluorescently labeling ASC specks using antibody technology, visualizing with imaging flow cytometry, and gating based on physical characteristics. CSF ASC specks levels positively correlate with phosphorylated tau (Thr181) and negatively correlate with amyloid ß ratio (42/40), thus serving as a neuroinflammatory biomarker for diagnosing AD/ADRDs. For complete details on the use and execution of this protocol, please refer to Jiang et al.1.

Evolutionary Analysis of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Reveals Genomic Divergence with Implications for Universal Vaccine Efficacy.

Coronavirus disease (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is one of the pressing contemporary public health challenges. Investigations into the genomic structure of SARS-CoV-2 may inform ongoing vaccine development efforts and/or provide insights into vaccine efficacy to fight against COVID-19. Evolutionary analysis of 540 genomes spanning 20 different countries/territories was conducted and revealed an increase in the genomic divergence across successive generations. The ancestor of the phylogeny was found to be the isolate from the 2019/2020 Wuhan outbreak. Its transmission was outlined across 20 countries/territories as per genomic similarity. Our results demonstrate faster evolving variations in the genomic structure of SARS-CoV-2 when compared to the isolates from early stages of the pandemic. Genomic alterations were predominantly located and mapped onto the reported vaccine candidates of structural genes, which are the main targets for vaccine candidates. S protein showed 34, N protein 25, E protein 2, and M protein 3 amino acid variations in 246 genomes among 540. Among identified mutations, 23 in S protein, 1 in E, 2 from M, and 7 from N protein were mapped with the reported vaccine candidates explaining the possible implications on universal vaccines. Hence, potential target regions for vaccines would be ideally chosen from the structural regions of the genome that lack high variation. The increasing variations in the genome of SARS-CoV-2 together with our observations in structural genes have important implications for the efficacy of a successful universal vaccine against SARS-CoV-2.

Fracture resistance of yttria-stabilized zirconia dental implant abutments.

PURPOSE: An in vitro study was performed to assess the effect of different degrees of clinical reduction of zirconia abutments on the failure load of clinical assemblies. MATERIALS AND METHODS: Zirconia abutments (Y-TZP Ceramic Abutment, Astra Tech) were prepared with 0, 0.5, or 1 mm of external axial reduction starting 1 mm above the height-of-contour. Abutments (n = 10) were attached to implant analogs (25 Ncm torque) embedded in a stainless steel cylinder using Field's metal. Fracture loads (N) were determined when assemblies were loaded at 60 degrees off-axis until failure (Instron, CHS = 0.1 mm/min). Groups were statistically compared using ANOVA (p < 0.05). RESULTS: Fracture loads for all assemblies displayed large variations within groups. There were no statistically significant differences (p > 0.05) among different abutment groups with a mean fracture load of 429 N (+/-140) for the control group, 576 N (+/-120) for 0.5-mm margins, and 547 (+/-139) for 1.0-mm margins. All fractures occurred at the interface where the abutment was connected to the analog. CONCLUSION: In this in vitro study of simulated ultimate assembly strength, the preparation of zirconia abutments did not significantly impair the fracture resistance of simulated implant assemblies. All implant abutments fractured at rates higher than the maximum incisal forces (90-370 N) estimated to occur in the anterior region of the mouth.

Clinical predictors of the medical interventions provided to patients with low back pain in the emergency department.

BACKGROUND: Low back pain is a common complaint in emergency departments (ED), where deviations from standard of care have been noted. OBJECTIVE: To relate the ordering of advanced imaging and opioid prescriptions with the presentation of low back pain in ED. METHODS: Six hundred adults with low back pain from three centers were prospectively analyzed for history, examination, diagnosis, and the ordering of tests and treatments. RESULTS: Of 559 cases the onset of pain was less than one week in 79.2%; however, most had prior low back pain, 63.5% having warning signs of a potential serious condition, and 83.9% had psychosocial risk factors. Computer tomography (CT) or magnetic resonance imaging (MRI) were ordered in 16.6%, opioids were prescribed in 52.6%, and hospital admission in 4.5%. A one-year follow-up of 158 patients found 40.8% received subsequent spine care and 5.1% had a medically serious condition. Caucasian race, age 50 years or older, warning signs, and radicular findings were associated with advanced imaging. Severe pain and psychosocial factors were associated with opioid prescribing. CONCLUSIONS: Most patients present to the ED with acute exacerbations of chronic low back pain. Risk factors for a serious condition are common, but rarely do they develop. Racial disparities and psychosocial factors had concerning relationships with clinical decision-making.

An in vitro assessment of prefabricated fiber post systems.

BACKGROUND: The authors conducted a study to characterize the in vitro retention, fracture and light transmission behavior of four different fiber-reinforced resin-based composite root canal posts. METHODS: The authors divided 44 teeth into four groups according to the type of post they would receive: parallel fiberglass posts, double-tapered fiber quartz posts, tapered fiberglass posts and two different types of parallel fiberglass posts. They prepared teeth and cemented posts with dual-cured resin cement. After the specimens aged, the authors conducted pull-out tests. For fracture testing, they loaded posts at 45 degrees in a universal testing machine. They determined load-to-fracture values and relative stiffness. They measured percentage of light transmission by means of a light microscope coupled with a spectrometer. RESULTS: Both tapered posts showed lower retention than did the parallel posts. Scanning electron microscropic analysis indicated that more cement adhered to the surfaces of the parallel fiberglass post than to those of the tapered fiberglass post. All posts demonstrated some plastic behavior, with the double-tapered fiber quartz post being stiffer than the others. CONCLUSION: Parallel fiber-reinforced composite posts showed better retention than did tapered posts when a dual-cured resin-based cement was used. Translucent prefabricated posts have limited light transmission. CLINICAL IMPLICATIONS: Parallel prefabricated fiber posts luted with dual-cured resin-based cement seem to be suitable for clinical application in endodontically treated teeth that require post-and-core restoration, showing good retention and acceptable fracture resistance.

Proactive personality and job performance: a social capital perspective.

This study of 126 employee-supervisor dyads examined a mediated model of the relationship between proactive personality and job performance. The model, informed by the social capital perspective, suggests that proactive employees reap performance benefits by means of developing social networks that provide them the resources and latitude to pursue high-level initiatives. Structural equation modeling suggested that the relationship between proactive personality and job performance is mediated by network building and initiative taking on the part of the employee.

The Partnership Access Line: evaluating a child psychiatry consult program in Washington State.

OBJECTIVE: To evaluate a telephone-based child mental health consult service for primary care providers (PCPs). DESIGN: Record review, provider surveys, and Medicaid database analysis. SETTING: Washington State Partnership Access Line (PAL) program. PARTICIPANTS: A total of 2285 PAL consultations by 592 PCPs between April 1, 2008, and April 30, 2011. INTERVENTIONS: Primary care provider-initiated consultations with PAL service. MAIN OUTCOME MEASURES: The PAL call characteristics, PCP feedback surveys, and Medicaid claims between April 2007 and December 2009 for fee-for-service Medicaid children before and after a PAL call. RESULTS: Sixty-nine percent of calls were about children with serious emotional disturbances, and 66% of calls were about children taking psychiatric medications. Primary care providers nearly always received new psychosocial treatment advice (87% of calls) and were more likely to receive advice to start rather than stop a medication (46% vs 24% of calls). Primary care provider feedback surveys reported uniformly positive satisfaction with the program. Among Medicaid children, there was significant increases in attention-deficit/hyperactivity disorder and antidepressant medication use after the PAL call but no significant change in reimbursements for mental health medications (P < .05). Children with a history of foster care experienced a 132% increase in outpatient mental health visits after the PAL call (P < .05). CONCLUSIONS: Primary care providers used PAL for psychosocial and medication treatment assistance for particularly high-needs children and were satisfied with the service. Furthermore, PAL was associated with increased use of outpatient mental health care for some children.

Second opinions improve ADHD prescribing in a medicaid-insured community population.

OBJECTIVE: The appropriate use of psychotropic medications in youths is an important public health concern. In this article, we describe a review process developed to monitor the use of stimulants and atomoxetine for attention-deficit/hyperactivity syndrome (ADHD) in youths receiving fee-for-service Medicaid services. METHOD: Washington State Medicaid developed threshold safety parameters for ADHD medications through a process involving the community. A second opinion was mandated when safety thresholds based on dose, combination therapies, or age was exceeded. Use and cost were compared 2 years before and after the program began. RESULTS: From May 2006 to April 2008, 5.35% of ADHD prescriptions exceeded safety thresholds, resulting in 1,046 second-opinion reviews. Of those, 538 (51.4%) resulted in a prescription adjustment. Adjustments were made to primary care physician (52%), psychiatrist (50%), nurse practitioner (54%), and physician assistant-written (51%) prescriptions. When the preperiod and postperiod were compared, second opinions reduced ADHD medication at high doses (53%), in combinations (44%), and for patients 5 years of age and younger (23%). The review process resulted in a savings of $1.2 million, with 538 fewer patients exceeding safety thresholds. This was a 10:1 return over administrative costs; however, the overall Medicaid expenditures for ADHD medication still increased because of higher unit costs and the preferential use by clinicians of newer brands entering the market. CONCLUSIONS: A statewide second-opinion process reduced outlier ADHD medication prescription practices and was cost-effective. Suggestions for process and quality improvements in prescribing to children diagnosed with ADHD are discussed.

Nonfluorinated volatile copper(I) 1,3-diketiminates as precursors for Cu metal deposition via atomic layer deposition.

Novel nonfluorinated Cu(diketiminate)L complexes with L = neutral olefinic ligand have been prepared as stable, volatile Cu(I) precursors for the deposition of copper films by an atomic layer deposition (ALD) process. Among them, the complexes of 4-a and 5-a are the most volatile and stable at low temperature (55 degrees C). A clean, conformal copper film was deposited at 120 degrees C in an ALD process. These Cu(I) complexes are the first examples of nonfluorinated copper(I) diketiminates that can be readily applied to an industrial microelectronic fabrication process.