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Epithelial tumor metastasis is preceded by an accumulation of collagen cross-links that heighten stromal stiffness and stimulate the invasive properties of tumor cells. However, the biochemical nature of collagen cross-links in cancer is still unclear. Here, we postulated that epithelial tumorigenesis is accompanied by changes in the biochemical type of collagen cross-links. Utilizing resected human lung cancer tissues and a p21CIP1/WAF1-deficient, K-rasG12D-expressing murine metastatic lung cancer model, we showed that, relative to normal lung tissues, tumor stroma contains higher levels of hydroxylysine aldehyde-derived collagen cross-links (HLCCs) and lower levels of lysine aldehyde-derived cross-links (LCCs), which are the predominant types of collagen cross-links in skeletal tissues and soft tissues, respectively. Gain- and loss-of-function studies in tumor cells showed that lysyl hydroxylase 2 (LH2), which hydroxylates telopeptidyl lysine residues on collagen, shifted the tumor stroma toward a high-HLCC, low-LCC state, increased tumor stiffness, and enhanced tumor cell invasion and metastasis. Together, our data indicate that LH2 enhances the metastatic properties of tumor cells and functions as a regulatory switch that controls the relative abundance of biochemically distinct types of collagen cross-links in the tumor stroma.

Introduction: Oncogenic RET tyrosine kinase gene fusions and activating mutations have recently been identified in lung cancers, prompting initiation of targeted therapy trials in this disease. Although RET point mutation has been identified as a driver of tumorigenesis in medullary thyroid carcinoma (MTC), no fusions have been described to date., Objective: We evaluated the role of RET fusion as an oncogenic driver in MTC., Methods: We describe a patient who died from aggressive sporadic MTC < 10 months after diagnosis. Her tumor was evaluated by means of next-generation sequencing, including an intronic capture strategy., Results: A reciprocal translocation involving RET intron 12 was identified. The fusion was validated using a targeted break apart fluorescence in situ hybridization probe, and RNA sequencing confirmed the existence of an in-frame fusion transcript joining MYH13 exon 35 with RET exon 12. Ectopic expression of fusion product in a murine Ba/F3 cell reporter model established strong oncogenicity. Three tyrosine kinase inhibitors currently used to treat MTC in clinical practice blocked tumorigenic cell growth., Conclusion: This finding represents the report of a novel RET fusion, the first of its kind described in MTC. The finding of this potential novel oncogenic mechanism has clear implications for sporadic MTC, which in the majority of cases has no driver mutation identified. The presence of a RET fusion also provides a plausible target for RET tyrosine kinase inhibitor therapies.

Objective: We sought to characterize the relationship between serum 25-hydroxy vitamin D (25-OH D) levels and implantation and clinical pregnancy rates in women who undergo a euploid blastocyst embryo transfer., Study Design: This retrospective cohort study, conducted in an academic setting, included 529 cycles in which comprehensive chromosome screening was performed as part of routine infertility care with an autologous transfer of 1 or 2 euploid blastocysts. After excluding repeat cycles there were 517 unique cycles representing 517 women for evaluation. Vitamin D levels from serum samples obtained on the day of ovulation trigger in the fresh in vitro fertilization cycle were analyzed. The primary outcome was ongoing pregnancy rate as defined by sonographic presence of fetal heart rate at >8 weeks' gestation., Results: For the population as a whole, serum vitamin D ranges and pregnancy outcomes did not correlate. Furthermore, pregnancy rates did not differ when comparing women in different strata of vitamin D levels (<20, 20-29.9, and ≥30 ng/mL). No meaningful breakpoint for vitamin D levels and ongoing pregnancy rate was identified using receiver operating characteristic analysis with the resultant line possessing an area under the curve of 0.502. Multivariate logistic regression controlling for age, transfer order, race, season, and body mass index did not yield a different result. The study was powered to detect an 18% difference in ongoing pregnancy rates between patients grouped by the 3 vitamin D ranges., Conclusion: In women undergoing euploid embryo transfer, vitamin D status was unrelated to pregnancy outcomes. Measuring serum 25-OH vitamin D levels does not predict the likelihood that euploid blastocysts will implant. These results may not apply to women who do not undergo extended embryo culture, blastocyst biopsy for comprehensive chromosome screening, and euploid embryo transfer., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Currently, there is no in vitro or ex vivo model that can isolate circulating tumor cells (CTCs). Recently, we developed a four-dimensional (4D) lung cancer model that allows for the isolation of CTCs. We postulated that these cells have different properties than parental (2D) cells., Materials and Methods: We obtained CTCs by growing A549, H1299, 393P, and 344SQ cell lines on the 4D lung model. The CTCs were functionally characterized in vitro and gene expression of the cell adhesion molecules was compared with respective 2D cells. Integrin beta 4 (ITGB4) was further investigated by stably transfecting the A549 and H1299 cells., Results: We found that all cell lines produced CTCs, and that CTCs from the 4D model were less adherent to the plastic and have a slower growth rate than respective 2D cells (P < 0.01). Most of the cell adhesion molecules were downregulated (P < 0.05) in CTCs, and ITGB4 was the common molecule, significantly more underexpressed in CTCs from all cell lines than their respective 2D cells. The modulation of ITGB4 led to a differential function of 2D cells., Conclusions: CTCs from the 4D model have different transcriptional, translational, and in vitro characteristics than the same cells grown on a petri dish, and these CTCs from the 4D model have the properties of CTCs that are responsible for metastasis., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The in vivo validation of cancer mutations and genes identified in cancer genomics is resource-intensive because of the low throughput of animal experiments. We describe a mouse model that allows multiple cancer mutations to be validated in each animal line. Animal lines are generated with multiple candidate cancer mutations using transposons. The candidate cancer genes are tagged and randomly expressed in somatic cells, allowing easy identification of the cancer genes involved in the generated tumours. This system presents a useful, generalised and efficient means for animal validation of cancer genes.

In this article, we examine the relationships between supervisor-level factors and abusive supervision. Drawing from social learning theory (Bandura, 1973), we argue that supervisors' history of family aggression indirectly impacts abusive supervision via both hostile cognitions and hostile affect, with angry rumination functioning as a first-stage moderator. Using multisource data, we tested the proposed relationships in a series of 4 studies, each providing evidence of constructive replication. In Study 1, we found positive relationships between supervisors' history of family aggression, hostile affect, explicit hostile cognitions, and abusive supervision. We obtained the same pattern of results in Studies 2, 3, and 4 using an implicit measure of hostile cognitions and controlling for previously established antecedents of abusive supervision. Angry rumination moderated the indirect relationship between supervisors' history of family aggression and abusive supervision via hostile affect only. Overall, the results highlight the important role of supervisor-level factors in the abusive supervision dynamics., (PsycINFO Database Record (c) 2014 APA, all rights reserved.)

Aberrant AKT activation is prevalent across multiple human cancer lineages providing an important new target for therapy. Twenty-two independent phosphorylation sites have been identified on specific AKT isoforms likely contributing to differential isoform regulation. However, the mechanisms regulating phosphorylation of individual AKT isoform molecules have not been elucidated because of the lack of robust approaches able to assess phosphorylation of multiple sites on a single AKT molecule. Using a nanofluidic proteomic immunoassay (NIA), consisting of isoelectric focusing followed by sensitive chemiluminescence detection, we demonstrate that under basal and ligand-induced conditions that the pattern of phosphorylation events is markedly different between AKT1 and AKT2. Indeed, there are at least 12 AKT1 peaks and at least 5 AKT2 peaks consistent with complex combinations of phosphorylation of different sites on individual AKT molecules. Following insulin stimulation, AKT1 was phosphorylated at Thr308 in the T-loop and Ser473 in the hydrophobic domain. In contrast, AKT2 was only phosphorylated at the equivalent sites (Thr309 and Ser474) at low levels. Further, Thr308 and Ser473 phosphorylation occurred predominantly on the same AKT1 molecules, whereas Thr309 and Ser474 were phosphorylated primarily on different AKT2 molecules. Although basal AKT2 phosphorylation was sensitive to inhibition of phosphatidylinositol 3-kinase (PI3K), basal AKT1 phosphorylation was essentially resistant. PI3K inhibition decreased pThr451 on AKT2 but not pThr450 on AKT1. Thus, NIA technology provides an ability to characterize coordinate phosphorylation of individual AKT molecules providing important information about AKT isoform-specific phosphorylation, which is required for optimal development and implementation of drugs targeting aberrant AKT activation.

Melanoma is a highly lethal malignancy notorious for its aggressive clinical course and eventual resistance to existing therapies. Currently, we possess a limited understanding of the genetic events driving melanoma progression, and much effort is focused on identifying pro-metastatic aberrations or perturbed signaling networks that constitute new therapeutic targets. In this study, we validate and assess the mechanism by which homeobox transcription factor A1 (HOXA1), a pro-invasion oncogene previously identified in a metastasis screen by our group, contributes to melanoma progression. Transcriptome and pathway profiling analyses of cells expressing HOXA1 reveals upregulation of factors involved in diverse cytokine pathways that include the transforming growth factor beta (TGFβ) signaling axis, which we further demonstrate to be required for HOXA1-mediated cell invasion in melanoma cells. Transcriptome profiling also shows HOXA1's ability to potently downregulate expression of microphthalmia-associated transcription factor (MITF) and other genes required for melanocyte differentiation, suggesting a mechanism by which HOXA1 expression de-differentiates cells into a pro-invasive cell state concomitant with TGFβ activation. Our analysis of publicly available data sets indicate that the HOXA1-induced gene signature successfully categorizes melanoma specimens based on their metastatic potential and, importantly, is capable of stratifying melanoma patient risk for metastasis based on expression in primary tumors. Together, these validation data and mechanistic insights suggest that patients whose primary tumors express HOXA1 are among a high-risk metastasis subgroup that should be considered for anti-TGFβ therapy in adjuvant settings. Moreover, further analysis of HOXA1 target genes in melanoma may reveal new pathways or targets amenable to therapeutic intervention.

School professionals have a unique vantage point for identifying child maltreatment and they are a frequent source of referral to child protective services. Disturbingly, past studies have found that maltreatment concerns reported by educators go unsubstantiated by child protective services at much higher rates than suspected maltreatment reported by other professionals. This study explores whether there are systematic differences in the characteristics of cases reported by educators as compared to other professionals and examines whether such variation might account for differences in investigation outcome. Analyses were based on 7,725 cases of suspected maltreatment referred by professionals to child protective services from the Canadian Incidence Study of Reported Child Abuse and Neglect - 2003 a national database on the characteristics of children and families investigated by child protective services. School professionals were responsible for 35.8% of professional referrals. Reports by educators were much more likely to be unsubstantiated (45.3%) than those by other professionals (28.4%) in subsequent child protective investigation. Cases reported by educators were found to contain significantly more child risk factors (e.g., child emotional and behavioural problems) and fewer caregiver and family risk factors (e.g., caregiver mental health problem, single parent family) than cases reported by other professionals. Even controlling for these differences, educator-reported concerns were still 1.84, 95% CI [1.41, 2.40] times as likely to be unsubstantiated as reports from other professionals. Contrary to the notion that educators are mostly reporting non-severe cases, suspected/substantiated cases reported by school professionals were more likely to be judged as chronic and more likely to involve families with a previous child protection history. Results are concerning for the capacity of the education and child protection systems to work together to meet their shared goal of promoting healthy child development. Additional research is needed on the way in which child risks and problems influence child protective service, particularly in the context of chronic abuse and neglect and lack of availability of child and family mental health interventions. Potential problems with credibility of school professionals as reporters of child maltreatment concerns also warrant further investigation., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Nuclear genetic disorders causing mitochondrial DNA (mtDNA) depletion are clinically and genetically heterogeneous, and the molecular etiology remains undiagnosed in the majority of cases. Through whole-exome sequencing, we identified recessive nonsense and splicing mutations in FBXL4 segregating in three unrelated consanguineous kindreds in which affected children present with a fatal encephalopathy, lactic acidosis, and severe mtDNA depletion in muscle. We show that FBXL4 is an F-box protein that colocalizes with mitochondria and that loss-of-function and splice mutations in this protein result in a severe respiratory chain deficiency, loss of mitochondrial membrane potential, and a disturbance of the dynamic mitochondrial network and nucleoid distribution in fibroblasts from affected individuals. Expression of the wild-type FBXL4 transcript in cell lines from two subjects fully rescued the levels of mtDNA copy number, leading to a correction of the mitochondrial biochemical deficit. Together our data demonstrate that mutations in FBXL4 are disease causing and establish FBXL4 as a mitochondrial protein with a possible role in maintaining mtDNA integrity and stability., (Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: To determine whether blastocyst biopsy and rapid quantitative real-time polymerase chain reaction (qPCR)-based comprehensive chromosome screening (CCS) improves in vitro fertilization (IVF) implantation and delivery rates., Design: Randomized controlled trial., Setting: Academic reproductive medicine center., Patient(s): Infertile couples in whom the female partner (or oocyte donor) is between the ages of 21 and 42 years who are attempting conception through IVF., Intervention(s): Embryonic aneuploidy screening., Main Outcome Measure(s): Sustained implantation and delivery rates., Result(s): We transferred 134 blastocysts to 72 patients in the study (CCS) group and 163 blastocysts to 83 patients in the routine care (control) group. Sustained implantation rates (probability that an embryo will implant and progress to delivery) were statistically significantly higher in the CCS group (89 of 134; 66.4%) compared with those from the control group (78 of 163; 47.9%). Delivery rates per cycle were also statistically significantly higher in the CCS group. Sixty one of 72 treatment cycles using CCS led to delivery (84.7%), and 56 of 83 (67.5%) control cycles ultimately delivered. Outcomes were excellent in both groups, but use of CCS clearly improved patient outcomes., Conclusion(s): Blastocyst biopsy with rapid qPCR-based comprehensive chromosomal screening results in statistically significantly improved IVF outcomes, as evidenced by meaningful increases in sustained implantation and delivery rates., Clinical Trial Registration Number: NCT01219283., (Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

A consistent requirement for all preimplantation genetic testing is the need to obtain DNA from the oocyte or embryo. Currently this sample is attained through biopsy of one or both polar bodies, blastomere biopsy at the cleavage stage, or trophectoderm biopsy after blastulation. Selecting the optimal time for biopsy requires careful consideration. Polar body biopsy is less invasive and provides more time for analysis but fails to capture as many as one in three embryonic aneuploidies. Additionally, the inability to readily distinguish nondysjunction from premature separation of sister chromatids greatly limits the predictive value of the technique and may lead to an overdiagnosis of aneuploidy in as many as 45% of cases with first polar-body errors. Cleavage-stage biopsy provides adequate samples but is detrimental to the embryo. The adverse effect of blastomere biopsy may result in approximately two of every five reproductively competent embryos losing their ability to implant and sustain development. Trophectoderm biopsy does not adversely impact the embryos. However, for the majority of clinical programs without a genetics laboratory, vitrification would be necessary to allow time for the genetic analysis. Although this extends the time required for treatment, clinical outcomes are equivalent after transfer of euploid blasts during fresh IVF and cryopreserved embryo transfer cycles, so that excellent outcomes are maintained. At present the blastocyst stage is the optimal time to perform biopsies for preimplantation genetic testing., (Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

We conducted 2 studies of coworker dyads to test a theoretical model exploring why and under what circumstances employees are the targets of workplace exclusion. Adopting a victim precipitation perspective, we integrate belongingness and social exchange theories to propose that employees who display workplace incivility are distrusted and therefore are targets of workplace exclusion. Highlighting the importance of the context of the perpetrator-target relationship, we also find support for the postulation that this mediated relationship is strengthened when the target employee is perceived to be a weak exchange partner and is attenuated when he or she is viewed as a valuable exchange partner. Theoretical and practical implications are discussed., (PsycINFO Database Record (c) 2013 APA, all rights reserved.)

Endometrial cancer is the most common gynecological malignancy, with more than 280,000 cases occurring annually worldwide. Although previous studies have identified important common somatic mutations in endometrial cancer, they have primarily focused on a small set of known cancer genes and have thus provided a limited view of the molecular basis underlying this disease. Here we have developed an integrated systems-biology approach to identifying novel cancer genes contributing to endometrial tumorigenesis. We first performed whole-exome sequencing on 13 endometrial cancers and matched normal samples, systematically identifying somatic alterations with high precision and sensitivity. We then combined bioinformatics prioritization with high-throughput screening (including both shRNA-mediated knockdown and expression of wild-type and mutant constructs) in a highly sensitive cell viability assay. Our results revealed 12 potential driver cancer genes including 10 tumor-suppressor candidates (ARID1A, INHBA, KMO, TTLL5, GRM8, IGFBP3, AKTIP, PHKA2, TRPS1, and WNT11) and two oncogene candidates (ERBB3 and RPS6KC1). The results in the "sensor" cell line were recapitulated by siRNA-mediated knockdown in endometrial cancer cell lines. Focusing on ARID1A, we integrated mutation profiles with functional proteomics in 222 endometrial cancer samples, demonstrating that ARID1A mutations frequently co-occur with mutations in the phosphatidylinositol 3-kinase (PI3K) pathway and are associated with PI3K pathway activation. siRNA knockdown in endometrial cancer cell lines increased AKT phosphorylation supporting ARID1A as a novel regulator of PI3K pathway activity. Our study presents the first unbiased view of somatic coding mutations in endometrial cancer and provides functional evidence for diverse driver genes and mutations in this disease.

Objective: Fathers are seldom the focus of efforts to address child maltreatment and little is currently known about the effectiveness of intervention for this population. To address this gap, we examined the efficacy of a community-based group treatment program for fathers who had abused or neglected their children or exposed their children to domestic violence., Methods: Using a sample of 98 group participants, we examined the magnitude and clinical significance of pre- to post-intervention changes in parenting, co-parenting and generalized aggression., Results: Intervention led to considerable changes in fathers' over-reactivity to children's misbehavior and respect for their partner's commitment and judgment, with results being statistically significant, medium in size, moving mean scores into the normative range and with 36-43% of men who initially scored in the clinical range recovering by the end of intervention. Changes in other domains were also evident though of lesser magnitude., Conclusions: Although this study is limited in length of follow-up and the lack of a control group, results are promising for continued development of fathering interventions for this population of high-risk men., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Clinical and genomic evidence suggests that the metastatic potential of a primary tumor may be dictated by prometastatic events that have additional oncogenic capability. To test this "deterministic" hypothesis, we adopted a comparative oncogenomics-guided function-based strategy involving: (1) comparison of global transcriptomes of two genetically engineered mouse models with contrasting metastatic potential, (2) genomic and transcriptomic profiles of human melanoma, (3) functional genetic screen for enhancers of cell invasion, and (4) evidence of expression selection in human melanoma tissues. This integrated effort identified six genes that are potently proinvasive and oncogenic. Furthermore, we show that one such gene, ACP5, confers spontaneous metastasis in vivo, engages a key pathway governing metastasis, and is prognostic in human primary melanomas., (Copyright © 2011 Elsevier Inc. All rights reserved.)

We developed a model of the relationships among aggressive norms, abusive supervision, psychological distress, family undermining, and supervisor-directed deviance. We tested the model in 2 studies using multisource data: a 3-wave investigation of 184 full-time employees (Study 1) and a 2-wave investigation of 188 restaurant workers (Study 2). Results revealed that (a) abusive supervision mediated the relationship between aggressive norms and psychological distress, (b) psychological distress mediated the effects of abusive supervision on spouse undermining, (c) abusive supervision had a direct positive relationship with supervisor-directed deviance, (d) the positive relationship between psychological distress and spouse undermining was stronger for men as opposed to women, and (e) employees engaged in relationship-oriented occupations reported greater levels of abusive supervision and psychological distress. Implications for theory and practice are discussed.

Measurement of glutathione (GSH) and glutathione disulfide (GSSG) is a crucial tool to assess cellular redox state. Herein we report a direct approach to determine intracellular GSH based on a rapid chromatographic separation coupled with acidic potassium permanganate chemiluminescence detection, which was extended to GSSG by incorporating thiol blocking and disulfide bond reduction. Importantly, this simple procedure avoids derivatisation of GSH (thus minimising auto-oxidation) and overcomes problems encountered when deriving the concentration of GSSG from 'total GSH'. The linear range and limit of detection for both analytes were 7.5 × 10(-7) to 1 × 10(-5) M, and 5 × 10(-7) M, respectively. GSH and GSSG were determined in cultured muscle cells treated for 24 h with glucose oxidase (0, 15, 30, 100, 250 and 500 mU mL(-1)), which exposed them to a continuous source of reactive oxygen species (ROS). Both analyte concentrations were greater in myotubes treated with 100 or 250 mU mL(-1) glucose oxidase (compared to untreated controls), but were significantly lower in myotubes treated with 500 mU mL(-1) (p < 0.05), which was rationalised by considering measurements of H(2)O(2) and cell viability. However, the GSH/GSSG ratio in myotubes treated with 100, 250 and 500 mU mL(-1) glucose oxidase exhibited a dose-dependent decrease that reflected the increase in intracellular ROS.

The authors develop and test theoretical extensions of the relationships of task conflict, relationship conflict, and 2 dimensions of team effectiveness (performance and team-member satisfaction) among 2 samples of work teams in Taiwan and Indonesia. Findings show that relationship conflict moderates the task conflict-team performance relationship. Specifically, the relationship is curvilinear in the shape of an inverted U when relationship conflict is low, but the relationship is linear and negative when relationship conflict is high. The results for team-member satisfaction are more equivocal, but the findings provide some evidence that relationship conflict exacerbates the negative relationship between task conflict and team-member satisfaction., (PsycINFO Database Record (c) 2011 APA, all rights reserved.)

Effective clinical management of prostate cancer (PCA) has been challenged by significant intratumoural heterogeneity on the genomic and pathological levels and limited understanding of the genetic elements governing disease progression. Here, we exploited the experimental merits of the mouse to test the hypothesis that pathways constraining progression might be activated in indolent Pten-null mouse prostate tumours and that inactivation of such progression barriers in mice would engender a metastasis-prone condition. Comparative transcriptomic and canonical pathway analyses, followed by biochemical confirmation, of normal prostate epithelium versus poorly progressive Pten-null prostate cancers revealed robust activation of the TGFβ/BMP-SMAD4 signalling axis. The functional relevance of SMAD4 was further supported by emergence of invasive, metastatic and lethal prostate cancers with 100% penetrance upon genetic deletion of Smad4 in the Pten-null mouse prostate. Pathological and molecular analysis as well as transcriptomic knowledge-based pathway profiling of emerging tumours identified cell proliferation and invasion as two cardinal tumour biological features in the metastatic Smad4/Pten-null PCA model. Follow-on pathological and functional assessment confirmed cyclin D1 and SPP1 as key mediators of these biological processes, which together with PTEN and SMAD4, form a four-gene signature that is prognostic of prostate-specific antigen (PSA) biochemical recurrence and lethal metastasis in human PCA. This model-informed progression analysis, together with genetic, functional and translational studies, establishes SMAD4 as a key regulator of PCA progression in mice and humans.

When it escapes early detection, malignant melanoma becomes a highly lethal and treatment-refractory cancer. Melastatin is greatly downregulated in metastatic melanomas and is widely believed to function as a melanoma tumor suppressor. Here we report that tumor suppressive activity is not mediated by melastatin but instead by a microRNA (miR-211) hosted within an intron of melastatin. Increasing expression of miR-211 but not melastatin reduced migration and invasion of malignant and highly invasive human melanomas characterized by low levels of melastatin and miR-211. An unbiased network analysis of melanoma-expressed genes filtered for their roles in metastasis identified three central node genes: IGF2R, TGFBR2, and NFAT5. Expression of these genes was reduced by miR-211, and knockdown of each gene phenocopied the effects of increased miR-211 on melanoma invasiveness. These data implicate miR-211 as a suppressor of melanoma invasion whose expression is silenced or selected against via suppression of the entire melastatin locus during human melanoma progression., (Copyright © 2010 Elsevier Inc. All rights reserved.)

A cardinal feature of malignant melanoma is its metastatic propensity. An incomplete view of the genetic events driving metastatic progression has been a major barrier to rational development of effective therapeutics and prognostic diagnostics for melanoma patients. In this study, we conducted global genomic characterization of primary and metastatic melanomas to examine the genomic landscape associated with metastatic progression. In addition to uncovering three genomic subclasses of metastastic melanomas, we delineated 39 focal and recurrent regions of amplification and deletions, many of which encompassed resident genes that have not been implicated in cancer or metastasis. To identify progression-associated metastasis gene candidates, we applied a statistical approach, Integrative Genome Comparison (IGC), to define 32 genomic regions of interest that were significantly altered in metastatic relative to primary melanomas, encompassing 30 resident genes with statistically significant expression deregulation. Functional assays on a subset of these candidates, including MET, ASPM, AKAP9, IMP3, PRKCA, RPA3, and SCAP2, validated their pro-invasion activities in human melanoma cells. Validity of the IGC approach was further reinforced by tissue microarray analysis of Survivin showing significant increased protein expression in thick versus thin primary cutaneous melanomas, and a progression correlation with lymph node metastases. Together, these functional validation results and correlative analysis of human tissues support the thesis that integrated genomic and pathological analyses of staged melanomas provide a productive entry point for discovery of melanoma metastases genes.

Golgi phosphoprotein 3 (GOLPH3; also known as GPP34/GMx33/MIDAS) represents an exciting new class of oncoproteins involved in vesicular trafficking. Encoded by a gene residing on human chromosome 5p13, which is frequently amplified in multiple solid tumor types, GOLPH3 was initially discovered as a phosphorylated protein localized to the Golgi apparatus. Recent functional, cell biological, and biochemical analyses show that GOLPH3 can function as an oncoprotein to promote cell transformation and tumor growth by enhancing activity of the mammalian target of rapamycin, a serine/threonine protein kinase known to regulate cell growth, proliferation, and survival. Although its precise mode of action in cancer remains to be elucidated, the fact that GOLPH3 has been implicated in protein trafficking, receptor recycling, and glycosylation points to potential links of these cellular processes to tumorigenesis. Understanding how these processes may be deregulated and contribute to cancer pathogenesis and drug response will uncover new avenues for therapeutic intervention.

Genome-wide copy number analyses of human cancers identified a frequent 5p13 amplification in several solid tumour types, including lung (56%), ovarian (38%), breast (32%), prostate (37%) and melanoma (32%). Here, using integrative analysis of a genomic profile of the region, we identify a Golgi protein, GOLPH3, as a candidate targeted for amplification. Gain- and loss-of-function studies in vitro and in vivo validated GOLPH3 as a potent oncogene. Physically, GOLPH3 localizes to the trans-Golgi network and interacts with components of the retromer complex, which in yeast has been linked to target of rapamycin (TOR) signalling. Mechanistically, GOLPH3 regulates cell size, enhances growth-factor-induced mTOR (also known as FRAP1) signalling in human cancer cells, and alters the response to an mTOR inhibitor in vivo. Thus, genomic and genetic, biological, functional and biochemical data in yeast and humans establishes GOLPH3 as a new oncogene that is commonly targeted for amplification in human cancer, and is capable of modulating the response to rapamycin, a cancer drug in clinical use.

The Tuberous Sclerosis Complex component, TSC1, functions as a tumor suppressor via its regulation of diverse cellular processes, particularly cell growth. TSC1 exists in a complex with TSC2 and functions primarily as a key negative regulator of mammalian target of rapamycin complex 1 (mTORC1) signaling and protein synthesis, although the TSC1/TSC2 complex also shows mTORC1-independent outputs to other pathways. Here, we explored the role of TSC1 in various aspects of stem cell biology and dissected the extent to which TSC1 functions are executed via mTORC1-dependent versus mTORC1-independent pathways. Using hematopoietic stem cells (HSCs) as a model system, we demonstrate that somatic deletion of TSC1 produces striking stem cell and derivative effector cell phenotypes characterized by increased HSC cell cycling, mobilization, marked progressive depletion, defective long-term repopulating potential, and hematopoietic lineage developmental aberrations. On the mechanistic level, we further establish that TSC1 regulation of HSC quiescence and long-term repopulating potential and hematopoietic lineage development is mediated through mTORC1 signaling. In contrast, TSC1 regulation of HSC mobilization is effected in an mTORC1-independent manner, and gene profiling and functional analyses reveals the actin-bundling protein FSCN1 as a key TSC1/TSC2 target in the regulation of HSC mobilization. Thus, TSC1 is a critical regulator of HSC self-renewal, mobilization, and multilineage development and executes these actions via both mTORC1-dependent and -independent pathways.

Background: Recent developments in anaesthesia and patient demographics have potentially changed the practice of obstetric general anaesthesia. There are few contemporary data on Australasian practice of general anaesthesia for caesarean section, especially relating to airway management, anaesthetic techniques and complications., Methods: Using a standardised case record form, a prospective observational study was conducted during 2005-06 in 13 maternity hospitals dealing with approximately 49 500 deliveries per annum. Patient demographics, airway management, anaesthetic techniques and major complications were evaluated in those given general anaesthesia., Results: Data were obtained from 1095 women receiving general anaesthesia for caesarean section, 47% of which were classified as category 1 and 18% as category 4. Tracheal intubation was planned in all cases with rapid-sequence induction used in 97%. A grade 3 or 4 laryngoscopic view was obtained in 3.6 and 0.6% of cases respectively, with 3.3% considered a difficult intubation. There were four failed intubations (0.4%, 95% CI 0.01-0.9%), of which three were subsequently managed using a laryngeal mask airway. Antacid prophylaxis was used in 94% of elective cases and 64% of emergencies. Regurgitation of gastric contents was noted in eight cases (0.7%, 95% CI 0.2-1.2%), with one confirmed case of aspiration (0.1%, 95% CI 0.002-0.5%). There were no cases of serious airway-related morbidity., Conclusions: General anaesthesia is most commonly used in emergency situations. Tracheal intubation after rapid-sequence induction remains the predominant approach to airway management in Australasia. The incidence of failed intubation is consistent with previous studies. Aspiration prophylaxis is not routinely used for emergency surgery.

Background: The obstetric population is considered at high risk of awareness and recall when undergoing general anaesthesia for caesarean section. In recent years the incidence may have been altered by developments in obstetric anaesthesia., Methods: A prospective observational study of general anaesthesia for caesarean section was conducted during 2005 and 2006 in 13 maternity hospitals dealing with approximately 49,500 deliveries per annum in Australia and New Zealand. As a component of this study the frequency of recall of intraoperative events was examined using a structured postoperative interview on two occasions., Results: There were 1095 general anaesthetics surveyed with 47% being performed for urgent fetal delivery. Thiopental was the most common induction agent (83%) with sevoflurane being used for maintenance in 63%. In 32% of cases a depth-of-anaesthesia monitor was used. In 763 cases (70%) there was least one postoperative interview enquiring into dreaming and awareness. There were two cases deemed to be consistent with awareness (incidence 0.26%, CI 0.03-0.9%, or 1 in 382) and three cases of possible awareness., Conclusion: Awareness with recall of intraoperative events remains a significant complication of obstetric general anaesthesia but was potentially avoidable in all cases detected in this study.

As medical schools across the nation consider the recent call made by the Association of American Medical Colleges to increase numbers of medical school students by 30% by 2015, it is important to explore the characteristics of the applicant pool. Understanding the make-up of the pool of recent applicants to the University of Kentucky College of Medicine can assist us in defining areas where the pool could be expanded in the future. Reviewing data from 2002-2006, we will examine the Kentucky county of origin of our applicants and matriculants. We will describe demographic characteristics of our applicants and matriculants with regard to gender, race and ethnicity, and international backgrounds. We will also look at factors that may discourage or dissuade prospective applicants from seeking admission to medical school including undergraduate grades, denial of the initial application to medical school, and cost considerations.

What Is Already Known About This Subject: There are presently no published data on tramadol transfer into breast milk or on its effects in the breastfed infant., What This Study Adds: We have provided quantitative data on the absolute and relative infant doses of rac-tramadol and it rac-O-desmethyl metabolite for the breastfed infant. We have also demonstrated a novel sparse sampling data collection method for investigating infant exposure via milk., Aims: To investigate the transfer of rac-tramadol and its rac-O-desmethyl metabolite into transitional milk, and assess unwanted effects in the breastfed infant., Methods: Tramadol HCl (100 mg six hourly) was administered to 75 breastfeeding mothers for postoperative analgesia on days 2-4 after Caesarian section. Milk and plasma samples were collected after administration of four or more doses. Rac-tramadol and rac-O-desmethyltramadol were measured by high performance liquid chromatography. Milk : plasma ratio (M : P) and infant doses were calculated by standard methods. The behavioural characteristics of the exposed breastfed infants and a matched control group of infants not exposed to tramadol were also studied. RESULTS At steady-state, mean (95% CI) M : P was 2.2 (2.0, 2.4) for rac-tramadol and 2.8 (2.5, 3.1) for rac-O-desmethyltramadol. The estimated absolute and relative infant doses were 112 (102, 122) microg kg(-1) day(-1) and 30 (28, 32) microg kg(-1) day(-1), and 2.24% (2.04, 2.44)% and 0.64% (0.59, 0.69)% for rac-tramadol and rac-O-desmethyltramadol, respectively. The exposed infants and control breastfed infants had similar characteristics, including Apgar scores at birth and Neurologic and Adaptive Capacity Scores., Conclusions: The combined relative infant dose of 2.88% at steady-state was low. The similarity of NACS in exposed infants and controls suggests that there were no significant behavioural adverse effects. We conclude that short-term maternal use of tramadol during establishment of lactation is compatible with breastfeeding.

Background: The diffusing capacity of the lungs for carbon monoxide (D(LCO)) is commonly measured during pulmonary function testing (PFT). Although adjustment of the measured D(LCO) for an elevated baseline carboxyhemoglobin level is recommended, carboxyhemoglobin is not routinely measured, which may reduce the accuracy of D(LCO) measurements. We sought to assess the utility of routine carboxyhemoglobin measurement and subsequent D(LCO) correction in patients referred for PFT., Methods: We retrospectively reviewed 100 consecutive PFT results, including D(LCO) assessment. We used a pulse CO-oximeter (recently approved by the Food and Drug Administration) to noninvasively measure baseline carboxyhemoglobin (S(pCO)). We used simple descriptive statistics to compare the S(pCO) values. In subjects with elevated S(pCO) (> 2%) we adjusted the percent-of-predicted D(LCO). Interpretation of D(LCO) was categorized according to the American Thoracic Society classification scheme for respiratory impairment., Results: The self-reported smokers had higher average S(pCO) than did self-reported nonsmokers (1.6% vs 3.5%, p < 0.001), although 14% of nonsmokers had an elevated S(pCO) and 26% of smokers had normal S(pCO). When the D(LCO) was corrected for elevated S(pCO), 2 patients moved from a category of moderate impairment to mild impairment. Both were smokers., Conclusions: The noninvasive measurement of carboxyhemoglobin is easy to perform during PFT. When precise measurement of D(LCO) is important, noninvasive measurement of carboxyhemoglobin may be of value. If routine S(pCO) measurement is considered, the highest yield is among current smokers.

Perpetrators of abuse and violence against women and children are often reluctant participants in intervention programs. They frequently fail to attend scheduled appointments, are sometimes openly hostile to intervention staff, and often judge program materials as irrelevant to their situation. Recognizing this problem, researchers and practitioners have begun to develop models and tools to more appropriately assess and intervene with reluctant clients. Unfortunately, the resulting proliferation and inconsistent application of terms and theories have led to considerable confusion in characterizing reluctant clients and have significantly hampered research on strategies that may be helpful to better meet the needs of this client group. The purpose of this review is to help standardize the definition and measurement of treatment reluctance as it applies to violence perpetration and to review evidence for the importance of these aspects of client reluctance to intervention. Recommendations for assessing reluctance in research and clinical practice are also provided.

Child maltreatment constitutes significant risk for adolescent delinquency. Although an ecological model has been proposed to explain this relationship, most studies focus on individual risk factors. Prospective data from 1,788 students attending 23 schools were used to examine the additive influence of childhood maltreatment, individual-level risk factors, and school-level variables assessed at the beginning of Grade 9 on delinquency 4 to 6 months later. Individual-level results indicated that being male, experiencing childhood maltreatment, and poor parental nurturing were predictors of violent delinquency. School climate also played a significant role: Given the same individual risk profile, a student attending a school that was perceived by students as safe was less likely to engage in violent delinquency than was a student attending a school perceived to be unsafe. Moreover, the impact of childhood maltreatment on risk for engaging in violent delinquency was somewhat mitigated by schools' participation in a comprehensive violence prevention program.

The authors developed and tested a multilevel interactive model of the relationship between group undermining and individual undermining behavior in 2 multiwave studies of group members. Integrating the literature on group influences on individual behavior with the individual difference literature, the authors predicted a 3-way Group Undermining x Self-Esteem x Neuroticism interaction, such that the relationship between group and individual undermining would be strongest among those simultaneously high in self-esteem and neuroticism. The 3-way interaction was supported in Study 1 (457 participants in 103 groups) and replicated in Study 2 (415 participants in 93 groups) with additional controls and alternative measures of key constructs. The authors discuss the implications of the research and identify future research directions., ((c) 2006 APA, all rights reserved)

Multiple endocrine neoplasia type 1 (MEN1) is a cancer susceptibility syndrome affecting several endocrine tissues. Investigations of the biochemical function of the MEN1 protein, menin, have suggested a role as a transcriptional comodulator. The mechanism by which MEN1 inactivation leads to tumor formation is not fully understood. MEN1 was implicated to function in both regulation of cell proliferation and maintenance of genomic integrity. Here, we investigate the mechanism by which MEN1 affects DNA damage response. We found that Drosophila larval tissue and mouse embryonic fibroblasts mutant for the MEN1 homologue were deficient for a DNA damage-activated S-phase checkpoint. The forkhead transcription factor CHES1 (FOXN3) was identified as an interacting protein by a genetic screen, and overexpression of CHES1 restored both cell cycle arrest and viability of MEN1 mutant flies after ionizing radiation exposure. We showed a biochemical interaction between human menin and CHES1 and showed that the COOH terminus of menin, which is frequently mutated in MEN1 patients, is necessary for this interaction. Our data indicate that menin is involved in the activation of S-phase arrest in response to ionizing radiation. CHES1 is a component of a transcriptional repressor complex, that includes mSin3a, histone deacetylase (HDAC) 1, and HDAC2, and it interacts with menin in an S-phase checkpoint pathway related to DNA damage response.

Background: Though smokers are known to have elevated blood carboxyhemoglobin (COHb), due to inhalation of carbon monoxide (CO) in cigarette smoke, limited data exist regarding COHb levels in nonsmokers exposed to secondhand smoke., Methods: COHb was measured using a new noninvasive pulse CO-oximeter (Rad-57, Masimo, Irvine, California) in 38 subjects entering a bingo hall where smoking was allowed, then again as they exited 3 hours later., Results: The mean +/- SD baseline COHb for the entire group was 3.3 +/- 1.8%, for the 23 nonsmokers it was 2.2 +/- 0.7%, and for the 15 smokers it was 4.9 +/- 1.9%. The nonsmokers' mean COHb was unchanged, at 2.2 +/- 0.8%, whereas the smokers' mean COHb fell to 3.2 +/- 1.9%., Conclusions: The nonsmokers were not significantly exposed to CO from secondhand smoke in the setting we tested. The smokers probably consumed fewer cigarettes while playing bingo than they did prior to arrival. The Rad-57 pulse CO-oximeter is easy to use in the ambulatory setting.

Most case definitions for carbon monoxide (CO) poisoning include demonstration of an elevated blood carboxyhemoglobin (COHb) concentration. Further, it is generally believed that treatment of CO poisoning is more effective when performed as soon as possible after the exposure. This suggests that a hospital's inability to measure blood COHb could lead to delayed or missed diagnosis or treatment. This study evaluated the ability of hospitals in the Pacific Northwest to measure COHb levels. The clinical laboratory of every acute care hospital in Washington, Idaho, Montana, and Alaska was surveyed regarding the ability to measure COHb levels, the method utilized and the time required. If they could not measure COHb, they were asked whether samples are sent elsewhere, the location of the referral laboratory, and time required. Results were then compared to the list of hospitals referring CO-poisoned patients to a regional center for hyperbaric oxygen therapy from 2003-2004. In the four states, only 44% of acute care hospitals have the capability to measure COHb. The remaining 56% send blood samples to other laboratories. The average time to get a result is 10 +/- 10 min in hospitals with co-oximetry and 904 +/- 1360 min in those without, a difference of 15 h (p < 0.0001). When samples are sent out, the average distance is 121 miles, often bypassing a hospital with CO-oximetry capability. Over 90% of CO-poisoned patients referred for hyperbaric treatment came from hospitals able to measure COHb. Fewer than one-half of acute care hospitals in a four-state region have the capability to measure COHb levels. This has the potential to significantly impact diagnosis or treatment of patients with acute CO poisoning.

Checkpoint Suppressor 1 (CHES1; FOXN3) encodes a member of the forkhead/winged-helix transcription factor family. The human CHES1 cDNA was originally identified by its ability to function as a high-copy suppressor of multiple checkpoint mutants of Saccharomyces cerevisiae. Accumulating expression profile data suggest that CHES1 plays a role in tumorigenicity and responses to cancer treatments, though nothing is known regarding the transcriptional function of CHES1 or other FOXN proteins in human cells. In this report, we find that the carboxyl terminus of CHES1 fused to a heterologous DNA binding domain consistently represses reporter gene transcription in cell lines derived from tumor tissues. Using a cytoplasmic two-hybrid screening approach, we find that this portion of CHES1 interacts with Ski-interacting protein (SKIP; NCoA-62), which is a transcriptional co-regulator known to associate with repressor complexes. We verify this interaction through co-immunoprecipitation experiments performed in mammalian cells. Further analysis of the CHES1/SKIP interaction indicates that CHES1 binds to a region within the final 66 hydrophobic residues of SKIP thus defining a new protein-protein interaction domain of SKIP. These data suggest that CHES1 recruits SKIP to repress genes important for tumorigenesis and the response to cancer treatments.

Maltose-limited, continuous growth of the hyperthermophile Thermotoga maritima at different temperatures and dilution rates (80 degrees C/0.25 h(-1), 80 degrees C/0.17 h(-1), and 85 degrees C/0.25 h(-1)) showed that transcriptome-wide variation in gene expression within mechanical steady states was minimal compared to that between steady states, supporting the efficacy of chemostat-based approaches for functional genomics studies.

Preservation and long-term storage of red blood cells (RBCs) is needed to ensure a readily available, safe blood supply for transfusion medicine. Effective preservation procedures are required at various steps in the production of a RBC product including testing, inventory, quality control, and product distribution. Biopreservation is the process of maintaining the integrity and functionality of cells held outside the native environment for extended storage times. The biopreservation of RBCs for clinical use can be categorized based on the techniques used to achieve biologic stability and ensure a viable state after long-term storage. This paper will review the history, science, current practices, and emerging technologies of current RBC biopreservation approaches: hypothermic storage, cryopreservation, and lyophilization.

Objective: Obese patients with sepsis have higher morbidity and mortality than lean counterparts, but the mechanisms involved are unknown. The authors examined the inflammatory and thrombogenic responses of the cerebral microvasculature to sepsis induced by cecal ligation and perforation in obese and lean wild-type mice., Methods: Leukocyte and platelet adhesion in cerebral microvasculature and behavioral responses were measured in wild-type and obese mice 4 h postperforation. P-selectin expression in different vascular beds was assessed 6 h postperforation. The effects of immunoblockade of P-selectin, ICAM-1, and CD18 on leukocyte and platelet recruitment were evaluated in obese septic animals., Results: Cerebral venules of obese and wild-type mice assumed a proinflammatory and prothrombogenic phenotype 4 h post-perforation, with greatly exaggerated responses in obese mice compared to the lean counterparts. These enhanced responses were attenuated by blocking P-selectin, CD18, or ICAM-1. Obese mice also exhibited a more profound behavioral deficit after sepsis, which appears to be unrelated to the recruitment of leukocytes and platelets. Cecal ligation and perforation-induced P-selectin expression was greater in obese mice compared with lean counterparts., Conclusions: These findings suggest that the increased morbidity to sepsis in obesity may result from exaggerated microvascular inflammatory and thrombogenic responses that include the activation of endothelial cells with subsequent expression of adhesion molecules, such as P-selectin.

The current study examined stage of change as a predictor of outcome in batterer treatment. Men (N=119) were classified into the transtheoretical model's stages of change and assessed 3 times over treatment. Hierarchical linear modeling revealed significant variation in men's progress, predictable from their stage of change. As hypothesized, men in the precontemplation stage showed little positive change in empathy, communication, or abusive behavior, whereas men in the contemplation and action stages showed positive growth in all of these domains. These effects occurred in the initial 10 weeks of treatment, after which men progressed at a more homogeneous rate. Interpretation is complicated by pretreatment differences that draw into question stage-related patterns in final outcome. Implications for general models of abuse cessation and for stage-specific trajectories are discussed.

The purpose of this study was to develop a tape measure method to assess the size of the hand and wrist. This tape measure method was designed to be readily performed, easily administered, reliable, and valid compared with water volumetry. The study sample included 50 normal consenting volunteers. Two examiners measured each hand of each subject three times, alternating hands between measurements. In the second phase of the study, the same two examiners measured both hands of the subjects using the traditional water volumetry method. The intraclass correlation coefficients for intra- and interater reliability obtained were 0.99. Pearson product-moment correlation values between the figure-of-eight tape measure and the water volumetry method for the two examiners were 0.94 and 0.95, respectively. The figure-of-eight method demonstrated excellent reliability and concurrent validity compared with the water volumetry method. These results support the application of the figure-of-eight method as a reliable and valid evaluation tool for the assessment of hand and wrist size.

We previously reported that expression of the human forkhead/winged helix transcription factor, CHES1 (checkpoint suppressor 1; FOXN3), suppresses sensitivity to DNA damage and restores damage-induced G(2)/M arrest in checkpoint-deficient strains of Saccharomyces cerevisiae. We find that a functional glutathione S-transferase-Ches1 fusion protein binds in vivo to Sin3, a component of the S. cerevisiae Sin3/Rpd3 histone deacetylase complex. Checkpoint mutant strains with SIN3 deleted show increased resistance to UV irradiation, which is not further enhanced by CHES1 expression. Conversely, overexpression of SIN3 blocks the Ches1-mediated G(2)/M delay in response to DNA damage, which is consistent with Ches1 acting by inhibiting the Sin3/Rpd3 complex. Deletion of either SIN3 or RPD3 in rad9 or mec1 checkpoint mutant strains suppresses sensitivity to replication blocks and DNA damage resulting from Cdc9 ligase deficiency and UV irradiation. SIN3 or RPD3 deletions also restored G(2)/M arrest after DNA damage without concomitant Rad53 phosphorylation in mec1 mutant strains. This DNA damage response is absent in mad1 spindle checkpoint mutants. These data suggest that modulation of chromatin structure may regulate checkpoint responses in S. cerevisiae. Inhibition of histone deacetylation results in a DNA damage checkpoint response mediated by the spindle checkpoint pathway that compensates for loss of the primary DNA damage checkpoint pathway.

This paper addresses fluctuations in the applicant and matriculant pools both across United States medical schools and at the University of Kentucky College of Medicine (UKCOM) for 1992-2002. It also presents data regarding the increasing costs of a medical education. Over the past decade, both nationally and at the UKCOM, there has been an over-all reduction in the number of applicants to medical school. In this changing applicant pool, the percentage of female matriculants has increased both nationally and at the UKCOM. However, the number of underrepresented minorities applying to and matriculating in the US and at the UKCOM has dropped since the mid-1990s. Although the applicant pool has decreased in size over the time period examined, the academic quality of applicants as measured by the undergraduate grade point average and Medical College Admission Test scores has increased both nationally and at UKCOM. Costs of a medical education have risen over time, as has the debt burden of medical school graduates due to increasing undergraduate debt, consumer debt, and medical school tuition. Potential causes for and implication of these changing trends are discussed.

Posttraumatic stress disorder is one of the most common and often prolonged consequences of childhood maltreatment. In this article the authors consider theories of trauma continuity, with emphasis on a relational path to maladjustment that links childhood maltreatment to elevated trauma symptomatology and intimate victimization in adolescent dating relationships.