Proteomic Analysis Reveals Trilaciclib-Induced Senescence.

Trilaciclib, a cyclin-dependent kinase 4/6 inhibitor, was approved as a myeloprotective agent for protecting bone marrow from chemotherapy-induced damage in extensive-stage small cell lung cancer. This is achieved through the induction of a temporary halt in the cell cycle of bone marrow cells. While it has been studied in various cancer types, its potential in hematological cancers remains unexplored. This research aimed to investigate the efficacy of trilaciclib in hematological cancers. Utilizing mass spectrometry-based proteomics, we examined the alterations induced by trilaciclib in the chronic myeloid leukemia cell line, K562. Interestingly, trilaciclib promoted senescence in these cells rather than cell death, as observed in acute myeloid leukemia, acute lymphoblastic leukemia, and myeloma cells. In K562 cells, trilaciclib hindered cell cycle progression and proliferation by stabilizing cyclin-dependent kinase 4/6 and downregulating cell cycle-related proteins, along with the concomitant activation of autophagy pathways. Additionally, trilaciclib-induced senescence was also observed in the nonsmall cell lung carcinoma cell line, A549. These findings highlight trilaciclib's potential as a therapeutic option for hematological cancers and underscore the need to carefully balance senescence induction and autophagy modulation in chronic myeloid leukemia treatment, as well as in nonsmall cell lung carcinoma cell line.

Social Sustainability of a Firm: Orientation, Practices, and Performances.

This paper investigates how firms' social sustainability practices can influence their social performance and, ultimately, financial performance. We include two corporate social sustainability practices: employee-oriented (employee well-being and equity) and socially driven (corporate social involvement) practices. Three leading social theories (social identity theory, social exchange theory, and resource-based view) are applied in explaining how firms' social practices influence intermediate and bottom-line performance outcomes. Empirical results of 212 US manufacturing firms reveal that (1) the social orientation of the firm promotes firms' social performances (employee-oriented and community-oriented outcomes) directly; (2) social orientation also indirectly promotes employee-oriented outcomes via employee well-being and equity practices, and so does community-oriented outcome via corporate social involvement practices; and (3) the firms' social performances can enhance financial performance. The theoretical and managerial implications derived from these empirical results are discussed as well.

Near Infrared and Frequency Modulated Spectroscopy as Non-Invasive Methods for Moisture Assessment of Freeze-Dried Biologics.

Near-infrared (NIR) and frequency modulated spectroscopy (FMS) were employed, for non-invasive moisture determination of a lyophilized biologic drug product (DP). Development of NIR and FMS provides a rapid non-invasive means of residual moisture measurement, and would be beneficial compared with traditional time consuming, product destructive methods such as Karl Fischer (KF). A model therapeutic enzyme in a sucrose-based formulation was employed for proof of concept studies, and NIR and FMS methods were compared side by side for residual moisture analysis. Moisture models were created using lyophilized vials and comparisons were made between the methods using different moisture preparation approaches:1) direct water droplet addition to the vial headspace, 2) use of elevated temperature (80°C), and 3) using various levels of moisture in stoppers generated during the washing and drying procedures, then lyophilizing using the stoppers and placing the sealed vials on stability. The results for direct water addition gave an average percent error for residual moisture of 5.7% for NIR and 9.4% for FMS when compared to KF. The elevated temperature method resulted in an average percent error for residual moisture of 54% for NIR and 43% for FMS compared to KF. The stopper moisture stability study, for FMS, provided an average percent error for residual moisture of 31% compared to KF. The error was greater for the elevated temperature and stopper methods, due to the low moisture values, which resulted in greater error. At this lower range of moisture (<1%) both NIR and FMS were less accurate, but from 1 to 5% their accuracy increased, based on the models used in this study. NIR and FMS methods can be used to complement KF at these lower moisture levels and models could be further improved with additional data points. NIR and FMS methods have advantages and disadvantages for residual moisture analysis when compared to each other, but both provided an accurate measurement of drug product moisture (depending on the method used for moisture increase), they can be used as process analytical technology (PAT), and both can be used for fast non-invasive moisture determination.

Personal Transformation Process of Mental Health Relief Workers in Sichuan Earthquake.

After Sichuan earthquake, a few dedicated teams of psychological volunteers have been committed to helping the survivors of the disaster for a long period. Their personal transformation experiences were absent in the literatures. The purpose of this qualitative research was to adopt a qualitative research to examine individuals' lived experiences of personal transformation after long-term disaster mental health services. The study interviewed 10 psychological counselors, 3 psychiatric nurses, 4 psychiatrist, and 6 social workers. We built an explorative model to understand disaster relief workers personal transformation after long-term periods in disaster settings. The model proposes that existential learning is a key mediator factor from negative transformation to positive transformation. Cultivating the existential learning though training, caring and support involves a number of processes that includes acceptance, being authentic, and being mindful of here and now, which can transform their suffering to growth.

Hydrogen Peroxide Induced Protein Oxidation During Storage and Lyophilization Process.

Although the impact of hydrogen peroxide (HP) on proteins in liquid solutions has been studied extensively, the impact during lyophilization has been largely overlooked. The purpose of this work was to investigate the effect of HP on lyophilized proteins and HP removal by lyophilization. A protein formulation at 5 mg/mL and its placebo were spiked with HP up to 5.0 ppm and then lyophilized. HP concentration, protein oxidation, and aggregation were monitored before and after lyophilization, as well as during storage at 25°C. The lyophilization process removed on average 94.1% of HP from protein formulation, but only 72.5% from the placebo. There were also significant increases in protein oxidization and aggregation. The oxidation increment correlated with the decrease of HP concentration in both the protein formulation and placebo at all temperatures. Protein oxidation at different freezing temperatures was also studied in follow-up studies. Data from these studies suggest that (1) HP has a significant impact on oxidation and aggregation of protein during lyophilization; (2) significant oxidation can occur even when the protein formulation is frozen; (3) the oxidized protein is more prone to aggregation during lyophilization process.

A Microscopic and Biomarker Evaluation of Embolic Filter Debris Collected During Carotid Artery Stenting.

PURPOSE: To evaluate and characterize debris retrieved from the cerebral embolic protection devices (EPDs) used during carotid artery stenting (CAS) and compare debris size, volume, tissue types, cellular composition, and protein biomarker expression in symptomatic and asymptomatic patients. METHODS: Distal protection filters were retrieved from 22 consecutive patients (mean age 71.6 years, range 52-85; 16 men) undergoing elective CAS between July 2012 and February 2014 for >70% internal carotid artery stenosis (mean 85.4% ± 10.3%). Six patients were symptomatic. The debris within each EPD was visually characterized using stereomicroscopy and then processed for histology and immunohistochemistry. Biomarkers were immunohistochemically measured to evaluate plaque stability [matrix metalloproteinase-9 (MMP-9)], inflammation [glycoprotein CD68 and interleukin-6 (IL-6)], or phenotype [smooth muscle (SM)-actin and type IV collagen]. The immunohistochemical results were measured using semiquantitative grading criteria based on both staining intensity and distribution in the samples. RESULTS: Macroscopic debris was visible in 5/22 EPDs; 3 of the 5 filters came from symptomatic patients. Microscopic debris was detected in all filters and ranged in size from 0.01 to 8.57 mm(2). Debris consisted of calcified, fibrous, and necrotic tissue, as well as fibrin and foam cells with no significant difference between the symptomatic and asymptomatic groups. There was no association between the degree or type of embolic material and stenosis severity, carotid tortuosity, calcium grade, soft plaque, or arch type. Symptomatic patients had a larger volume of debris (8.24 vs 0.58 mm(3), p<0.01), mean particle size (1.30 vs 0.32 mm(2), p<0.001), and expression of biomarkers IL-6 (2.17 vs 0.81, p<0.05), CD68 (2.00 vs 0.38, p<0.01), SM-actin (1.00 vs 0.25, p=0.055), type IV collagen (1.17 vs 0.25,p=0.082), and MMP-9 (1.00 vs 0.06, p<0.05). CONCLUSION: Histological analysis revealed particulate embolization in all EPDs used during CAS. Symptomatic patients had a larger volume of embolic debris, mean particle size, and the biomarkers associated with inflammation, necrotic core, and diminished fibrous cap.

Overcoming resistance of targeted EGFR monotherapy by inhibition of STAT3 escape pathway in soft tissue sarcoma.

Although epidermal growth factor receptor (EGFR) is often over-expressed in soft tissue sarcoma (STS), a phase II trial using an EGFR inhibitor gefitinib showed a low response rate. This study identified a new secondary resistance mechanism of gefitinib in STS, and developed new strategies to improve the effectiveness of EGFR inhibition particularly by blocking the STAT3 pathway.We demonstrated that seven STS cell lines of diverse histological origin showed resistance to gefitinib despite blockade of phosphorylated EGFR (pEGFR) and downstream signal transducers (pAKT and pERK) in PI3K/AKT and RAS/ERK pathways. Gefitinib exposure was not associated with decrease in the ratio of pSTAT3/pSTAT1. The relative STAT3 abundance and activation may be responsible for the drug resistance. We therefore hypothesized that the addition of a STAT3 inhibitor could overcome the STAT3 escape pathway.We found that the addition of STAT3 inhibitor S3I-201 to gefitinib achieved synergistic anti-proliferative and pro-apoptotic effects in all three STS cell lines examined. This was confirmed in a fibrosarcoma xenografted mouse model, where the tumours from the combination group (418mm3) were significantly smaller than those from untreated (1032mm3) or single drug (912 and 798mm3) groups.Our findings may have clinical implications for optimising EGFR-targeted therapy in STS.

Randomized, controlled clinical trial of bilayer ceramic and metal-ceramic crown performance.

PURPOSE: Analyzing the clinical performance of restorative materials is important, as there is an expectation that these materials and procedures will restore teeth and do no harm. The objective of this research study was to characterize the clinical performance of metal-ceramic crowns, core ceramic crowns, and core ceramic/veneer ceramic crowns based on 11 clinical criteria. MATERIALS AND METHODS: An IRB-approved, randomized, controlled clinical trial was conducted as a single-blind pilot study. The following three types of full crowns were fabricated: (1) metal-ceramic crown (MC) made from a Pd-Au-Ag-Sn-In alloy (Argedent 62) and a glass-ceramic veneer (IPS d.SIGN veneer); (2) non-veneered (glazed) lithium disilicate glass-ceramic crown (LDC) (IPS e.max Press core and e.max Ceram Glaze); and (3) veneered lithia disilicate glass-ceramic crown (LDC/V) with glass-ceramic veneer (IPS Empress 2 core and IPS Eris). Single-unit crowns were randomly assigned. Patients were recalled for each of 3 years and were evaluated by two calibrated clinicians. Thirty-six crowns were placed in 31 patients. A total of 12 crowns of each of the three crown types were studied. Eleven criteria were evaluated: tissue health, marginal integrity, secondary caries, proximal contact, anatomic contour, occlusion, surface texture, cracks/chips (fractures), color match, tooth sensitivity, and wear (of crowns and opposing enamel). Numerical rankings ranged from 1 to 4, with 4 being excellent, and 1 indicating a need for immediate replacement. Statistical analysis of the numerical rankings was performed using a Fisher's exact test. RESULTS: There was no statistically significant difference between performance of the core ceramic crowns and the two veneered crowns at year 1 and year 2 (p > 0.05). All crowns were rated either as excellent or good for each of the clinical criteria; however, between years 2 and 3, gradual roughening of the occlusal surface occurred in some of the ceramic-ceramic crowns, possibly caused by dissolution and wear of the glaze. Statistically significant differences in surface texture (p= 0.0013) and crown wear (p= 0.0078) were found at year 3 between the metal-ceramic crowns and the lithium-disilicate-based crowns. CONCLUSION: Based on the 11 criteria, the clinical performance of ceramic-ceramic crowns was comparable to that of the metal-ceramic crowns after 2 years; however, gradual roughening occurred between years 2 and 3, which resulted in differences in surface texture and wear.

Three years in vivo wear: core-ceramic, veneers, and enamel antagonists.

OBJECTIVES: Test the hypotheses that there are equivalent wear rates for enamel-versus-enamel and ceramic-versus-enamel, analyzing the in vivo wear of crown ceramics, their natural enamel antagonists, and the corresponding two contralateral teeth; and, that bite force does not correlate with the wear. METHODS: A controlled, clinical trial was conducted involving patients needing full coverage crowns opposing enamel antagonists. Bite forces were measured using a bilateral gnathodynamometer. Single-unit restorations of metal/ceramic (Argedent 62, Argen Corp/IPS d.SIGN veneer); or, core-ceramic/veneer from either, Empress2/Eris, or e.max Press core/e.max Ceram glaze (ceramics: Ivoclar Vivadent, USA) were randomly assigned, fabricated and cemented. Impressions were made of the ceramic crowns, as well as each maxillary and mandibular quadrant at one week (baseline) and one, two and three years. Resulting models were scanned (3D laser scanner). Maximum wear was calculated by superimposing baseline with annual images. RESULTS: There were a total of thirty-six crowns required for thirty-one patients. Each restoration had three associated enamel teeth: crown, (1) antagonist, (2) contralateral and (3) contralateral-antagonist. SAS PROC MIXED (α=0.05) indicated no statistical significance for mean maximum wear among crown ceramics, enamel antagonists and contralaterals. However, enamel wear was statistically significant in relation to intraoral location (p=0.04) and among years (p<0.02). Analyzed alone, the enamel contralateral-antagonist exhibited significantly greater wear (p<0.001). Considering all wear sites, there was no correlation with bite force (p=0.15). SIGNIFICANCE: The ceramics and their antagonists exhibited in vivo wear rates within the range of normal enamel. Future studies should examine the wear implications of the contralateral-antagonist enamel.

Interval estimation in two-stage, drop-the-losers clinical trials with flexible treatment selection.

In a two-stage, drop-the-losers clinical trial, researchers choose the 'best' among a number of treatments at an interim analysis after the first stage. The selected treatment continues to the second stage for confirmation of efficacy, and the remaining treatments (the 'losers') are dropped from the study. Wu et al. (Biometrika 2010; 97:405-418) showed how to construct confidence limits for the mean difference between the selected treatment and the control when the treatment is chosen after the first stage based on the highest efficacy in the primary clinical endpoint. In this article, we show how to construct a lower confidence limit for the mean difference when the treatment is chosen based on first-stage safety data, early endpoint efficacy data, a combination of safety and efficacy data or any other prespecified selection rule. The result extends the applicability of drop-the-losers designs, for in practice, the 'best' treatment often is not chosen for efficacy alone.

Development and temporal organization of repetitive behavior in an animal model.

Despite repetitive behaviors being a common feature of a number of clinical disorders and ubiquitous in normative development, little attention has been given to their ontogeny or temporal dynamics. We characterized these features in a mouse model of repetitive behavior to identify discrete trajectories of development and developmental changes in temporal dynamics. Three qualitatively distinct trajectory groups were identified which allowed for an examination of the interaction between temporal organization and developmental trajectory. Significant differences in temporal dynamics were found across development and among trajectory groups. Significant interactions of trajectory group and developmental period on temporal organization were also found. The combination of group-based trajectory modeling and a novel method for analysis and graphic depiction of temporal organization allowed for the exploration of the interplay between these two fundamental behavioral processes. Such methods may be useful tools in the assessment and treatment of repetitive behavior in clinical populations.

Open label aripiprazole in the treatment of youth with tic disorders.

BACKGROUND: Primarily safe and efficacious treatments for chronic tic disorders are needed. Also needed are such treatments that target co-morbid conditions. Aripiprazole, a dopaminergic/serotonergic agent with partial agonist properties at the D2 dopamine receptor and 5-hydrdoxytryptamine 1A (5-HT(1A)) receptor and antagonist properties at the 5-HT(2A) receptor, holds promise in both regards. OBJECTIVE: This was an open-label, flexible-dose study to evaluate the safety of aripiprazole in children and adolescents with a primary diagnosis of a chronic tic disorder with/without co-morbid disorder(s). METHOD: Sixteen children (15 males) aged 8-17 years participated in the 6-week trial. Ratings for tic, obsessive compulsive disorder (OCD), attention-deficit/hyperactivity disorder (ADHD), and side effects were administered weekly. Baseline and exit laboratory measures, electrocardiograms (ECGs), weight, and height were obtained. RESULTS: The average daily aripiprazole dose was 3.3 mg (range 1.25-7.5 mg). Significant pre-and posttreatment differences were ascertained for the Yale Global Tic Severity Scale motor (p < or = 0.0001), phonic (p < or = 0.0001), and total tic (p < or = 0.0001) scores. Results of other rating scales suggested significant improvements in co-morbid disorders as well, including OCD, ADHD, and depressive disorders. Although aripiprazole was well tolerated, increases in weight were found. CONCLUSION: In this preliminary open-label trial, aripiprazole was a well-tolerated treatment for tics and co-morbid OCD and ADHD symptoms. Improvements in co-morbid conditions may be secondary to tic reduction or to specific to aripiprazole therapy; however, further study is warranted.

Identifying temporally differentially expressed genes through functional principal components analysis.

Time course gene microarray is an important tool to identify genes with differential expressions over time. Traditional analysis of variance (ANOVA) type of longitudinal investigation may not be applicable because of irregular time intervals and possible missingness due to contamination in microarray experiments. Functional principal components analysis is proposed to test hypotheses in the change of the mean curves. A permutation test under a mild assumption is used to make the method more robust. The proposed method outperforms the recently developed extraction of differential gene expression and a 2-way mixed effects ANOVA under reasonable gene expression models in simulation. Real data on transcriptional profiles of blood cells microarray from treated and untreated individuals were used to illustrate this method.

An optimal DNA pooling strategy for progressive fine mapping.

We present a cost-effective DNA pooling strategy for fine mapping of a single Mendelian gene in controlled crosses. The theoretical argument suggests that it is potentially possible for a single-stage pooling approach to reduce the overall experimental expense considerably by balancing costs for genotyping and sample collection. Further, the genotyping burden can be reduced through multi-stage pooling. Numerical results are provided for practical guidelines. For example, the genotyping effort can be reduced to only a small fraction of that needed for individual genotyping at a small loss of estimation accuracy or at a cost of increasing sample sizes slightly when recombination rates are 0.5% or less. An optimal two-stage pooling scheme can reduce the amount of genotyping to 19.5%, 14.5% and 6.4% of individual genotyping efforts for identifying a gene within 1, 0.5, and 0.1 cM, respectively. Finally, we use a genetic data set for mapping the rice xl(t) gene to demonstrate the feasibility and efficiency of the DNA pooling strategy. Taken together, the results demonstrate that this DNA pooling strategy can greatly reduce the genotyping burden and the overall cost in fine mapping experiments.

Four-year clinical performance of a lithia disilicate-based core ceramic for posterior fixed partial dentures.

PURPOSE: The objective of this research was to test the hypothesis that 3-unit fixed partial dentures (FPDs) made from a moderately high-strength core ceramic will adequately resist fracture in posterior regions if fabricated with a minimal connector size of 4 mm. MATERIALS AND METHODS: Thirty ceramic FPD core frameworks were prepared using a hot-pressing technique and a lithia disilicate-based core ceramic. The maximum occlusal force was measured for each patient prior to tooth preparation. Connector heights and widths were measured for each FPD. Patients were recalled annually after cementation for 4 years and evaluated using 11 clinical criteria. All FPDs were examined by 2 independent clinicians, and rankings for each criterion were made from 1 to 4 (4 = excellent; 1 = unacceptable). RESULTS: The fracture rate was approximately 3% per year, and the proportion of good overall ratings in the nonfractured FPDs was reduced by more than 6% per year, where a good overall rating was defined to be a rank of 3 or 4 in all 11 criteria. There was little evidence that the use of either resin-reinforced glass-ionomer cement (Protec CEM) or dual-cure resin cement (Variolink II) made any difference in terms of fracture rate or overall rating (P= .30, .63, .97, and .71 for the 4 years, respectively). From a fracture resistance perspective, 4 of the 30 ceramic FPDs fractured within the 4-year evaluation period, representing an 86.7% success rate. Another FPD was replaced because of a caries lesion on 1 abutment tooth away from the margin. One FPD fracture was associated with the subject having the greatest occlusal force (1,031 N). The other 2 fractures were associated with FPDs that exhibited connector heights of less than 3 mm. All criteria were ranked good to excellent during the 4-year period for the remaining FPDs. CONCLUSION: Fractured FPDs were associated with a connector height of less than 4 mm; thus, the hypothesis was accepted.

Procedural learning and cognitive flexibility in a mouse model of restricted, repetitive behaviour.

Restricted, repetitive behaviours (e.g., stereotypies, compulsions, rituals) in neurodevelopmental disorders have been linked to alterations in cortico-basal ganglia circuitry. Cognitive processes mediated by this circuitry (e.g., procedural learning, executive function) are likely to be impaired in individuals exhibiting high rates of repetitive behaviour. To test this hypothesis, we assessed both procedural learning and cognitive flexibility (reversal learning) using a T-maze task in deer mice (Peromyscus maniculatus) exhibiting various rates of repetitive behaviour (vertical jumping and backward somersaulting). These mice exhibited high rates of stereotypy when reared in standard rodent cages, and such behaviour was significantly attenuated by housing them in larger more complex environments. Mice reared in complex environments exhibited significantly better procedural and reversal learning than standard caged mice. Thus, early experience associated with the prevention and attenuation of stereotypy was associated with better striatally mediated learning and cognitive flexibility. Stereotypy score was significantly correlated with the number of errors made in reversal learning, and interacted with housing condition to affect overall cognitive performance. Our findings support the applicability of the deer mouse model of spontaneous stereotypy to a wider range of restricted, repetitive behaviour (e.g., insistence on sameness) typical of neurodevelopmental disorders.

D-cycloserine does not enhance exposure-response prevention therapy in obsessive-compulsive disorder.

Obsessive-compulsive disorder is a common, chronic, and oftentimes disabling disorder. The only established first-line treatments for obsessive-compulsive disorder are exposure and response prevention therapy and the serotonin reuptake inhibitors. Many patients do not experience complete symptom resolution with either modality and require augmentation approaches. Recent animal and clinical data suggest that D-cycloserine, a partial agonist that acts at the strychnine-insensitive glycine-recognition site of the N-methyl-D-aspartate receptor complex, may enhance extinction learning that occurs in exposure-based psychotherapies. Given this, this study examined if D-cycloserine (250 mg) enhances the overall efficacy and rate of change of exposure and response prevention therapy for adult obsessive-compulsive disorder. Participants were 24 adults meeting Diagnostic and Statistical Manual of Mental Disorders-IV criteria for obsessive-compulsive disorder. The study design was a randomized, double-blinded, placebo-controlled augmentation trial examining exposure and response prevention therapy+D-cycloserine versus exposure and response prevention therapy+placebo. All patients received 12 weekly sessions of exposure and response prevention treatment. The first session involved building a ritual hierarchy and providing psychoeducation about obsessive-compulsive disorder. The second session involved a practice exposure. Sessions 3-12 involved exposure and response prevention exercises. D-cycloserine or placebo (250 mg) was taken 4 h before every session. No significant group differences were found across outcome variables. The rate of improvement did not differ between groups. The present results fail to support the use of D-cycloserine with exposure and response prevention therapy for adult obsessive-compulsive disorder. As this study is the first to explore this question and a number of methodological issues must be considered when interpreting the findings, the conclusions that may be drawn from our results are limited.

Ethnic differences in the reliability and validity of a Panic Disorder Screen.

PURPOSE: The purpose of this study was to compare the reliability and validity of the Brief Panic Disorder Screen (BPDS) in samples of African American and Caucasian American primary care patients. METHODS: The BPDS was administered to 295 patients scheduled to visit a primary care clinic for medical reasons. The presence or absence of a panic disorder diagnosis was established during a clinical interview with a psychiatrist. Measures of reliability (internal consistency) and validity (criterion validity) were compared between the two ethnic subgroups. FINDINGS: The BPDS demonstrated greater reliability and validity for Caucasians than African Americans. This effect was maintained even after controlling for group differences in key demographic variables. Differences between ethnic groups were apparent in both those with and those without panic disorder (PD). BPDS responses of African Americans with PD demonstrated very low internal consistency whereas a high rate of false positive PD diagnoses was related to higher than expected BPDS scores among African Americans without PD, particularly on the level of fear felt when experiencing shortness of breath or heart palpitations. DISCUSSION: These findings support the notion that cultural differences in the language and meaning associated with anxiety disorders contribute to the difficulty of accurately diagnosing PD in primary care populations. Additional research is needed to provide a better understanding of the cultural aspects of the anxiety experience. Such research would facilitate the development of better screening tools for panic and other anxiety disorders for ethnic minority primary care populations.