Tobacco smoking and somatic mutations in human bronchial epithelium.
Tobacco smoking causes lung cancer1-3, a process that is driven by more than 60 carcinogens in cigarette smoke that directly damage and mutate DNA4,5. The profound effects of tobacco on the genome of lung cancer cells are well-documented6-10, but equivalent data for normal bronchial cells are lacking. Here we sequenced whole genomes of 632 colonies derived from single bronchial epithelial cells across 16 subjects. Tobacco smoking was the major influence on mutational burden, typically adding from 1,000 to 10,000 mutations per cell; massively increasing the variance both within and between subjects; and generating several distinct mutational signatures of substitutions and of insertions and deletions. A population of cells in individuals with a history of smoking had mutational burdens that were equivalent to those expected for people who had never smoked: these cells had less damage from tobacco-specific mutational processes, were fourfold more frequent in ex-smokers than current smokers and had considerably longer telomeres than their more-mutated counterparts. Driver mutations increased in frequency with age, affecting 4-14% of cells in middle-aged subjects who had never smoked. In current smokers, at least 25% of cells carried driver mutations and 0-6% of cells had two or even three drivers. Thus, tobacco smoking increases mutational burden, cell-to-cell heterogeneity and driver mutations, but quitting promotes replenishment of the bronchial epithelium from mitotically quiescent cells that have avoided tobacco mutagenesis.


Ethchlorvynol-induced pulmonary edema in rats. An ultrastructural study.
Studies of ethchlorvynol (ECV)-induced pulmonary edema were undertaken for determination of the structural basis of increased microvascular permeability. Rats were administered an intravenous bolus dose of 15 mg/kg ECV and killed at time intervals between 5 minutes and 72 hours. Oyster glycogen and ferritin were used as permeability probes for identification of the sites of altered microvascular permeability. Edema fluid containing ferritin begins to accumulate in the alveolar interstitium 10 minutes after EVC . Thirty minutes after ECV, marked intersitial edema fluid is present containing both permeability probes. The absence of any appreciable transendothelial movement of either probe via vesicles and the presence of open endothelial junctions led the authors to propose the latter as the principal determinant of the increase in permeability. In addition to open endothelial junctions, prominent subendothelial blebs occur. These blebs develop in an otherwise intact endothelium and increase in frequency and size with time following their appearance at 10 minutes. Ferritin and glycogen progressively accumulate within the blebs. At 15 minutes the concentration of ferritin in blebs appears to equal that in plasma, whereas glycogen is absent or sparsely present in a few blebs. At 60 minutes both permeability probes have become concentrated in the blebs. The mechanism of formation of the blebs and concentration in them of the permeability probes cannot yet be specified. The lesion caused by ECV is completely reversible, so that by 72 hours after ECV there is complete resolution of interstitial edema, disappearance of the subendothelial blebs, and closure of endothelial junctions. A small amount of exudate remaining in the alveoli is cleared by 72 hours.


Neoadjuvant androgen ablation before radical prostatectomy in cT2bNxMo prostate cancer: 5-year results.
In the initial report of the Lupron Depot Neoadjuvant Prostate Cancer Study Group patients who received 3 months of androgen deprivation had a significant decrease in the positive margin rate. We monitored these patients for 5 years and to our knowledge present the longest followup of any neoadjuvant trial. A multi-institutional prospective randomized trial was performed between February 1992 and April 1994 involving patients with stage cT2b prostate cancer, including 138 who received 3 months of leuprolide plus flutamide before radical prostatectomy and 144 who underwent radical prostatectomy only. Patients were followed every 6 months with serum prostate specific antigen (PSA) testing for 5 years. Biochemical recurrence was defined as PSA greater than 0.4 ng./ml. At 5 years there was no difference in the biochemical recurrence rate. PSA was less than 0.4 ng./ml. in 64.8% of the patients in the neoadjuvant androgen ablation plus prostatectomy and 67.6% in the prostatectomy only group (p = 0.663). Although 3 months of androgen deprivation before radical prostatectomy resulted in an apparently significant decrease in positive surgical margins, a 5-year followup does not indicate any difference in the recurrence rate. Until studies document improvement in biochemical or clinical recurrence with longer periods of treatment, induction androgen deprivation before radical prostatectomy is not indicated.


Can safer-sex promotion leaflets change cognitive antecedents of condom use? An experimental evaluation.
An experimental evaluation of a safer sex promotion leaflet was undertaken to assess its capacity to change antecedent cognitions of condom use. The leaflet was identified in a previous study as addressing research-based cognitive antecedents of condom use. A pre-post-test experimental study including three conditions was conducted: (a) presentation of the leaflet; (b) presentation of the leaflet plus incentive for systematic processing; (c) no-leaflet control. The leaflet was evaluated in terms of its capacity to change eight cognitive correlates of condom use identified in a recent meta-analysis. The sample consisted of 230 tenth-grade students. Following baseline assessments, leaflet-induced change was measured immediately following the intervention and at a follow up 4 weeks post-intervention. The target leaflet alone did not result in significant changes in the cognitive antecedents of condom use compared with the control condition. However, in combination with an incentive for systematic processing, the target leaflet had a greater impact on cognitive antecedents than the no-leaflet control condition. The findings are discussed with regard to the development and evaluation of research-based health-promotion materials.


Primary graft dysfunction: definition, risk factors, short- and long-term outcomes.
Primary graft dysfunction (PGD) is a form of early lung graft dysfunction that is a major cause of early morbidity and mortality following lung transplantation. Survivors of PGD have decreased long-term lung function and an increased risk of developing bronchiolitis obliterans syndrome (BOS), a manifestation of chronic graft dysfunction. This article reviews the current definition, contributing factors, and guidelines for grading clinical PGD, as well as controversies surrounding them. The current literature examining clinical risk factors for PGD is summarized, as are studies exploring molecular and biochemical markers for PGD prediction and severity. Studies examining the short- and long-term effects of PGD on lung transplant outcomes are reviewed. Lastly, we highlight the emerging concepts in lung transplantation that will potentially impact PGD incidence and outcomes.


InterOrganizational practice committee guidance/recommendation for models of care during the novel coronavirus pandemic.
Objective: The Inter Organizational Practice Committee (IOPC) convened a workgroup to develop guidance on models to provide neuropsychological (NP) care during the COVID-19 pandemic while minimizing risks of novel coronavirus transmission as lockdown orders are lifted and ambulatory clinical services resume.Method: A collaborative panel of experts from major professional organizations developed provisional guidance for models of neuropsychological practice during the pandemic. The stakeholders included the American Academy of Clinical Neuropsychology/American Board of Clinical Neuropsychology, the National Academy of Neuropsychology, Society of Clinical neuropsychology (Division 40) of the American Psychological Association, the American Board of Professional Neuropsychology, and the American Psychological Association Services, Inc.Results: This guidance reviews the risks and benefits of conducting NP exams in several ways, including standard in-person, mitigated in-person, in-clinic teleneuropsychology (TeleNP), and in-home TeleNP. Strategies are provided for selecting the most appropriate model for a given patient, taking into account four levels of patient risk stratification, level of community risk and the concept of stepped models of care. Links are provided to governmental agency and professional organization resources as well as an outline and discussion of essential infection mitigation processes based on commonalities across recommendations from diverse federal, state, local, and professional organization recommendations.Conclusion: This document provides recommendations and guidance with analysis of the risks relative to the benefits of various models of neuropsychological care during the COVID-19 pandemic. These recommendations may be revised as circumstances evolve, with updates posted continuously on the IOPC website (https://iopc.online/).


Sticks and stones: investigating rude, dismissive and aggressive communication between doctors.
Destructive communication is a problem within the NHS; however previous research has focused on bullying. Rude, dismissive and aggressive (RDA) communication between doctors is a more widespread problem and underinvestigated. We conducted a mixed method study combining a survey and focus groups to describe the extent of RDA communication between doctors, its context and subsequent impact. In total, 606 doctors were surveyed across three teaching hospitals in England. Two structured focus groups were held with doctors at one teaching hospital. 31% of doctors described being subject to RDA communication multiple times per week or more often, with junior and registrar doctors affected twice as often as consultants. Rudeness was more commonly experienced from specific specialties: radiology, general surgery, neurosurgery and cardiology. 40% of respondents described that RDA moderately or severely affected their working day. The context for RDA communication was described in five themes: workload, lack of support, patient safety, hierarchy and culture. Impact of RDA communication was described as personal, including emotional distress and substance abuse, and professional, including demotivation. RDA communication between doctors is a widespread and damaging behaviour, occurring in contexts common in healthcare. Recognition of the impact on doctors and potentially patients is key to change.


Trust, continuity and agency: keys to understanding older patients' attitudes to general practice trainees.
Populations are ageing and therefore non-communicable diseases are becoming leading causes of global morbidity, which need to be the focus of primary care services and training. Some older patients are uncomfortable with general practitioner (GP) trainees managing their chronic conditions, reducing clinical experience opportunities for trainees. This Australian cross-sectional study explored the factors underlying patients' attitudes to trainees in an agency theory framework. Fifty patients aged 60 and over from each of 38 training practices were offered a questionnaire after their consultation. Principal component analysis of the results was undertaken. Factor scores were calculated. Binary logistic modelling was used to identify relationships between participant characteristics, behaviours, attitude items and factor scores. The response rate was 47.9% (n = 911). Three factors were identified: 'Interpersonal Trust' (IPT); 'Institution/system Trust' (ST); and 'Interpersonal Continuity' (IPC). Lower self-rated health (SRH) was associated with higher IPT factor scores (P = 0.023); higher SRH with higher ST scores (P = 0.001); and chronic illness with higher IPC scores (P = 0.005). Higher ST scores were associated with greater comfort with trainees' involvement in chronic care (P < 0.001) and frequency of trainee visits (P < 0.001), while higher IPC scores were negatively associated (P < 0.001 and P = 0.003, respectively). High IPT scores were associated with lower satisfaction with trainee visits (P = 0.001). These results indicate that better SRH, via higher institution/ST, is associated with favourable attitudes and attendance with trainees. In addition, chronic illness, via a higher need for IPC, is associated with lower comfort and attendance. These findings are consistent with agency theory, which shows potential as a framework for future interventions and research into older patient-trainee interactions.


Length scales and self-organization in dense suspension flows.
Dense non-Brownian suspension flows of hard particles display mystifying properties: As the jamming threshold is approached, the viscosity diverges, as well as a length scale that can be identified from velocity correlations. To unravel the microscopic mechanism governing dissipation and its connection to the observed correlation length, we develop an analogy between suspension flows and the rigidity transition occurring when floppy networks are pulled, a transition believed to be associated with the stress stiffening of certain gels. After deriving the critical properties near the rigidity transition, we show numerically that suspension flows lie close to it. We find that this proximity causes a decoupling between viscosity and the correlation length of velocities ξ, which scales as the length l(c) characterizing the response to a local perturbation, previously predicted to follow l(c)∼ 1/sqrt[z(c)-z] ∼ p(0.18), where p is the dimensionless particle pressure, z is the coordination of the contact network made by the particles, and z(c) is twice the spatial dimension. We confirm these predictions numerically and predict the existence of a larger length scale l(r)∼sqrt[p] with mild effects on velocity correlation and of a vanishing strain scale δγ ∼ 1/p that characterizes decorrelation in flow.


Induction of a salt appetite alters dendritic morphology in nucleus accumbens and sensitizes rats to amphetamine.
Sensitization to drugs, such as amphetamine, is associated with alterations in the morphology of neurons in the nucleus accumbens, a brain region critical to motivation and reward. The studies reported here indicate that a strong natural motivator, sodium depletion and associated salt appetite, also leads to alterations in neurons in nucleus accumbens. Medium spiny neurons in the shell of the nucleus accumbens of rats that had experienced sodium depletions had significantly more dendritic branches and spines than controls. In addition, a history of sodium depletions was found to have cross-sensitization effects, leading to enhanced psychostimulant responses to amphetamine. Thus, neuronal alterations common to salt and drug sensitization may provide a general mechanism for enhanced behavioral responses to subsequent exposures to these challenges.


Orthogonal representations for robust context-dependent task performance in brains and neural networks.
How do neural populations code for multiple, potentially conflicting tasks? Here we used computational simulations involving neural networks to define "lazy" and "rich" coding solutions to this context-dependent decision-making problem, which trade off learning speed for robustness. During lazy learning the input dimensionality is expanded by random projections to the network hidden layer, whereas in rich learning hidden units acquire structured representations that privilege relevant over irrelevant features. For context-dependent decision-making, one rich solution is to project task representations onto low-dimensional and orthogonal manifolds. Using behavioral testing and neuroimaging in humans and analysis of neural signals from macaque prefrontal cortex, we report evidence for neural coding patterns in biological brains whose dimensionality and neural geometry are consistent with the rich learning regime.


International Osteoporosis Foundation and International Federation of Clinical Chemistry and Laboratory Medicine position on bone marker standards in osteoporosis.
The International Osteoporosis Foundation (IOF) and the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) Working Group on Bone Marker Standards (WG-BMS) has evaluated the clinical potential of bone turnover markers (BTMs) in the prediction of fracture risk and for monitoring treatment. Research evidence suggests that BTMs may provide information on fracture risk independently from BMD, so that fracture risk prediction might be enhanced by their inclusion in assessment algorithms. The potential use of BTMs to predict the response to treatments for osteoporosis in the individual patient is also of great interest. Treatment-induced changes in specific markers account for a substantial proportion of fracture risk reduction. However, there is still a need for stronger evidence on which to base practice in both situations. IOF/IFCC recommends one bone formation marker (serum procollagen type I N propeptide, s-PINP) and one bone resorption marker (serum C-terminal cross-linking telopeptide of type I collagen, s-CTX) to be used as reference markers and measured by standardised assays in observational and intervention studies in order to enlarge the international experience of the application of markers to clinical medicine and to help resolve uncertainties over their clinical use.


Gender double standards in young people attending sexual health services in Northern Thailand.
Concern about the sexual and reproductive health of young people has been mounting recently in Thailand. Unequal gender relations have a considerable influence on the lives of young people, especially young women, yet few studies have explored the ways in which they have impacted on provision of sexual health care. Drawing upon data from a qualitative study in Northern Thailand, this paper explores the views and experiences of young people in seeking health care, highlighting the kinds of gender double standards and power imbalances that may pose obstacles to their use of sexual and reproductive health services. Findings reveal the vulnerability of sexually active young women in seeking support and care from partners, parents, and service providers. Those who experience adverse outcomes of sexual activity, such as unwanted pregnancy or infection, report facing indifference, victim blaming, or the threat of abandonment by their partners. Because of their fear of disclosure to their parents and communities, of their sexual activity, they opt for clandestine and unsafe abortion and seek the counsel of peers and drugstores rather than parents and providers. At the service provider level, young women report facing threatening and judgemental attitudes, indifferent counselling, and possible violation of confidentiality. This is in marked contrast to the treatment of young men, who generally meet with a more sympathetic and accepting response.


Immunogenicity and tolerability of recombinant serogroup B meningococcal vaccine administered with or without routine infant vaccinations according to different immunization schedules: a randomized controlled trial.
In the absence of an effective vaccine, serogroup B Neisseria meningitidis (MenB) remains a major cause of invasive disease in early childhood in developed countries. To determine the immunogenicity and reactogenicity of a multicomponent MenB vaccine (4CMenB) and routine infant vaccines when given either concomitantly or separately. Phase 2b, multicenter, open-label, parallel-group, randomized controlled study of 1885 infants enrolled at age 2 months from August 2008 to July 2010 in Europe. Participants were randomized 2:2:1:1 to receive (1) 4CMenB at 2, 4, and 6 months with routine vaccines (7-valent pneumococcal and combined diphtheria, tetanus, acellular pertussis, inactivated polio, hepatitis B, Haemophilus influenzae type b vaccines); (2) 4CMenB at 2, 4, and 6 months and routine vaccines at 3, 5, and 7 months; (3) 4CMenB with routine vaccines at 2, 3, and 4 months; or (4) routine vaccines alone at 2, 3, and 4 months. Percentage of participants with human complement serum bactericidal activity (hSBA) titer of 1:5 or greater against 3 MenB strains specific for vaccine antigens (NZ98/254, 44/76-SL, and 5/99). After three 4CMenB vaccinations, 99% or more of infants developed hSBA titers of 1:5 or greater against strains 44/76-SL and 5/99. For NZ98/254, this proportion was 79% (95% CI, 75.2%-82.4%) for vaccination at 2, 4, and 6 months with routine vaccines, 86.1% (95% CI, 82.9%-89.0%) for vaccination at 2, 4, and 6 months without routine vaccines, and 81.7% (95% CI, 76.6%-86.2%) for vaccination at 2, 3, and 4 months with routine vaccines. Responses to routine vaccines given with 4CMenB were noninferior to routine vaccines alone for all antigens, except for the responses to pertactin and serotype 6B pneumococcal polysaccharide. Fever was seen following 26% (158/602) to 41% (247/607) of 4CMenB doses when administered alone, compared with 23% (69/304) to 36% (109/306) after routine vaccines given alone and 51% (306/605) to 61% (380/624) after 4CMenB and routine vaccines administered together. A 4CMenB vaccine is immunogenic against reference strains when administered with routine vaccines at 2, 4, and 6 or at 2, 3, and 4 months of age, producing minimal interference with the response to routine infant vaccinations. clinicaltrials.gov Identifier: NCT00721396.


A novel, erythroid cell-specific murine transcription factor that binds to the CACCC element and is related to the Krüppel family of nuclear proteins.
We describe a novel erythroid cell-specific cDNA (EKLF [erythroid Krüppel-like factor]) isolated by enriching for genes expressed in a mouse erythroleukemia cell line but not expressed in a mouse monocyte-macrophage cell line. The complete cDNA sequence is predicted to encode a protein of approximately 38,000 Da that contains a proline-rich amino domain and three TFIIIA-like zinc fingers within the carboxy domain. Additional sequence analyses reveal that the EKLF zinc fingers are most homologous to the Krüppel family of transcription factors and also allow us to predict potential DNA-binding target sites for the EKLF protein. On the basis of this prediction, we show that EKLF is able to bind the sequence CCA CAC CCT, an essential element of the beta-globin promoter. Its tissue distribution establishes that the EKLF transcript is expressed only in bone marrow and spleen, the two hematopoietic organs of the mouse, and analysis of murine cell lines indicates that EKLF expression is limited to erythroid and mast cell lines. Cotransfection assays establish that EKLF transcriptionally activates a target promoter that contains its DNA-binding site. The tissue expression pattern of EKLF, in conjunction with its function as a transcriptional activator, strongly suggests that the EKLF protein may be intimately involved in establishment and/or maintenance of the erythroid cell phenotype.


Does the social context of early alcohol use affect risky drinking in adolescents? Prospective cohort study.
There are limited longitudinal data on the associations between different social contexts of alcohol use and risky adolescent drinking. Australian prospective longitudinal cohort of 1943 adolescents with 6 assessment waves at ages 14-17 years. Drinkers were asked where and how frequently they drank. Contexts were: at home with family, at home alone, at a party with friends, in a park/car, or at a bar/nightclub. The outcomes were prevalence and incidence of risky drinking (≥5 standard drinks (10g alcohol) on a day, past week) and very risky drinking (>20 standard drinks for males and >11 for females) in early (waves 1-2) and late (waves 3-6) adolescence. Forty-four percent (95 % CI: 41-46 %) reported past-week risky drinking on at least one wave during adolescence (waves 1-6). Drinking at a party was the most common repeated drinking context in early adolescence (28 %, 95 % CI 26-30 %); 15 % reported drinking repeatedly (3+ times) with their family in early adolescence (95 % CI: 14-17 %). For all contexts (including drinking with family), drinking 3+ times in a given context was associated with increased the risk of risky drinking in later adolescence. These effects remained apparent after adjustment for potential confounders (e.g. for drinking with family, adjusted RR 1.9; 95 % CI: 1.5-2.4). Similar patterns were observed for very risky drinking. Our results suggest that consumption with family does not protect against risky drinking. Furthermore, parents who wish to minimise high risk drinking by their adolescent children might also limit their children's opportunities to consume alcohol in unsupervised settings.


Recent advances in the synthesis of pyrroles by multicomponent reactions.
Pyrrole is one of the most important one-ring heterocycles. The ready availability of suitably substituted and functionalized pyrrole derivatives is essential for the progress of many branches of science, including biology and materials science. Access to this key heterocycle by multicomponent routes is particularly attractive in terms of synthetic efficiency, and also from the environmental point of view. We update here our previous review on this topic by describing the progress made in this area in the period between mid-2009 and the end of 2013.


The primary cilium as the cell's antenna: signaling at a sensory organelle.
Almost every vertebrate cell has a specialized cell surface projection called a primary cilium. Although these structures were first described more than a century ago, the full scope of their functions remains poorly understood. Here, we review emerging evidence that in addition to their well-established roles in sight, smell, and mechanosensation, primary cilia are key participants in intercellular signaling. This new appreciation of primary cilia as cellular antennae that sense a wide variety of signals could help explain why ciliary defects underlie such a wide range of human disorders, including retinal degeneration, polycystic kidney disease, Bardet-Biedl syndrome, and neural tube defects.


Changes in gene expression within the nucleus accumbens and striatum following sexual experience.
Sexual experience, like repeated drug use, produces long-term changes including sensitization in the nucleus accumbens and dorsal striatum. To better understand the molecular mechanisms underlying the neuroadaptations following sexual experience, we employed a DNA microarray approach to identify genes differentially expressed between sexually experienced and sexually naive female hamsters within the nucleus accumbens and dorsal striatum. For 6 weeks, a stimulus male was placed in the home cage of one-half of the hormonally primed, ovariectomized female hamsters. On the seventh week, the two experimental groups were subdivided, with one half paired with a stimulus male. In comparison with sexually naive animals, sexually experienced hamsters receiving a stimulus male on week 7 exhibited an increase in a large number of genes. Conversely, sexually experienced female hamsters not receiving a stimulus male on week 7 exhibited a reduction in the expression of many genes. For directional changes and the categories of genes regulated by the experimental conditions, data were consistent across the nucleus accumbens and dorsal striatum. However, the specific genes exhibiting changes in expression were disparate. These experiments, among the first to profile genes regulated by female sexual behavior, will provide insight into the mechanisms by which both motivated behaviors and drugs of abuse induce long-term changes in the mesolimbic and nigrostriatal dopamine pathways.


Coagulation of human prostate volumes with MRI-controlled transurethral ultrasound therapy: results in gel phantoms.
The feasibility and safety of magnetic resonance imaging (MRI)-controlled transurethral ultrasound therapy were demonstrated recently in a preliminary human study in which a small subvolume of prostate tissue was treated prior to radical prostatectomy. Translation of this technology to full clinical use, however, requires the capability to generate thermal coagulation in a volume up to that of the prostate gland itself. The aim of this study was to investigate the parameters required to treat a full 3D human prostate accurately with a multi-element transurethral applicator and multiplanar MR temperature control. The approach was a combination of simulations (to select appropriate parameters) followed by experimental confirmation in tissue-mimicking phantoms. A ten-channel, MRI-compatible transurethral ultrasound therapy system was evaluated using six human prostate models (average volume: 36 cm(3)) obtained from the preliminary human feasibility study. Real-time multiplanar MR thermometry at 3 T was used to control the spatial heating pattern in up to nine planes simultaneously. Treatment strategies incorporated both single (4.6 or 8.1 MHz) and dual (4.6 and 14.4 MHz) frequencies, as well as maximum acoustic surface powers of 10 or 20 W cm(-2). Treatments at 4.6 MHz were capable of coagulating a volume equivalent to 97% of the prostate. Increasing power from 10 to 20 W cm(-2) reduced treatment times by approximately 50% with full treatments taking 26 ± 3 min at a coagulation rate of 1.8 ± 0.4 cm(3) min(-1). A dual-frequency 4.6∕14.4 MHz treatment strategy was shown to be the most effective configuration for achieving full human prostate treatment while maintaining good treatment accuracy for small treatment radii. The dual-frequency approach reduced overtreatment close to the prostate base and apex, confirming the simulations. This study reinforces the capability of MRI-controlled transurethral ultrasound therapy to treat full prostate volumes in a short treatment time with good spatial targeting accuracy and provides key parameters necessary for the next clinical trial.


The Cdc14 phosphatase and the FEAR network control meiotic spindle disassembly and chromosome segregation.
During meiosis, DNA replication is followed by two consecutive rounds of chromosome segregation. Cells lacking the protein phosphatase CDC14 or its regulators, SPO12 and SLK19, undergo only a single meiotic division, with some chromosomes segregating reductionally and others equationally. We find that this abnormal chromosome behavior is due to an uncoupling of meiotic events. Anaphase I spindle disassembly is delayed in cdc14-1, slk19Delta, or spo12Delta mutants, but the chromosome segregation cycle continues, so that both meiotic chromosome segregation phases take place on the persisting meiosis I spindle. Our results show that Cdc14, Slk19, and Spo12 are not only required for meiosis I spindle disassembly but also play a pivotal role in establishing two consecutive chromosome segregation phases, a key feature of the meiotic cell cycle.


Partitioning and correlating subgroup characteristics from Aligned Pattern Clusters.
Evolutionarily conserved amino acids within proteins characterize functional or structural regions. Conversely, less conserved amino acids within these regions are generally areas of evolutionary divergence. A priori knowledge of biological function and species can help interpret the amino acid differences between sequences. However, this information is often erroneous or unavailable, hampering discovery with supervised algorithms. Also, most of the current unsupervised methods depend on full sequence similarity, which become inaccurate when proteins diverge (e.g. inversions, deletions, insertions). Due to these and other shortcomings, we developed a novel unsupervised algorithm which discovers highly conserved regions and uses two types of information measures: (i) data measures computed from input sequences; and (ii) class measures computed using a priori class groupings in order to reveal subgroups (i.e. classes) or functional characteristics. Using known and putative sequences of two proteins belonging to a relatively uncharacterized protein family we were able to group evolutionarily related sequences and identify conserved regions, which are strong homologous association patterns called Aligned Pattern Clusters, within individual proteins and across the members of this family. An initial synthetic demonstration and in silico results reveal that (i) the data measures are unbiased and (ii) our class measures can accurately rank the quality of the evolutionarily relevant groupings. Furthermore, combining our data and class measures allowed us to interpret the results by inferring regions of biological importance within the binding domain of these proteins. Compared to popular supervised methods, our algorithm has a superior runtime and comparable accuracy. The dataset and results are available at www.pami.uwaterloo.ca/∼ealee/files/classification2015 CONTACT: email@example.com Supplementary data are available at Bioinformatics online.


The pains and reliefs of the transitions into and out of spousal caregiving. A cross-national comparison of the health consequences of caregiving by gender.
Spousal caregiving offers a unique opportunity to investigate how gender shapes the influence of care responsibilities on health at older ages. However, empirical evidence supporting a causal link between the transitions into and out of caregiving and health is mixed. This study investigates the influence of spousal care transitions on the health of older men and women living in 17 European countries. We use five waves of the Survey of Health, Ageing and Retirement in Europe (SHARE) between the years 2004 and 2015 for a total of 43,435 individuals and 117,831 observations. Health is defined as a Frailty Index calculated from 40 items. Caregiving is defined as intensive help with personal care provided to spouses. Results from asymmetric fixed-effects linear regression models show that the transitions into caregiving have a detrimental effect on health. On the contrary, the transitions out of caregiving have in most cases no beneficial consequences on health. Most importantly, we found evidence supporting differential effects of caregiving transitions by gender and welfare arrangement: the transitions out of caregiving are associated with better health conditions only for Southern and Eastern European women. Our study highlights the asymmetric and gendered nature of care transitions and suggests that the impact of caregiving is somewhat permanent and has long lasting effects for the caregiver. Policies should account for this asymmetry when assessing the impact and consequences of caregiving.


A survey of the complex transcriptome from the highly polyploid sugarcane genome using full-length isoform sequencing and de novo assembly from short read sequencing.
Despite the economic importance of sugarcane in sugar and bioenergy production, there is not yet a reference genome available. Most of the sugarcane transcriptomic studies have been based on Saccharum officinarum gene indices (SoGI), expressed sequence tags (ESTs) and de novo assembled transcript contigs from short-reads; hence knowledge of the sugarcane transcriptome is limited in relation to transcript length and number of transcript isoforms. The sugarcane transcriptome was sequenced using PacBio isoform sequencing (Iso-Seq) of a pooled RNA sample derived from leaf, internode and root tissues, of different developmental stages, from 22 varieties, to explore the potential for capturing full-length transcript isoforms. A total of 107,598 unique transcript isoforms were obtained, representing about 71% of the total number of predicted sugarcane genes. The majority of this dataset (92%) matched the plant protein database, while just over 2% was novel transcripts, and over 2% was putative long non-coding RNAs. About 56% and 23% of total sequences were annotated against the gene ontology and KEGG pathway databases, respectively. Comparison with de novo contigs from Illumina RNA-Sequencing (RNA-Seq) of the internode samples from the same experiment and public databases showed that the Iso-Seq method recovered more full-length transcript isoforms, had a higher N50 and average length of largest 1,000 proteins; whereas a greater representation of the gene content and RNA diversity was captured in RNA-Seq. Only 62% of PacBio transcript isoforms matched 67% of de novo contigs, while the non-matched proportions were attributed to the inclusion of leaf/root tissues and the normalization in PacBio, and the representation of more gene content and RNA classes in the de novo assembly, respectively. About 69% of PacBio transcript isoforms and 41% of de novo contigs aligned with the sorghum genome, indicating the high conservation of orthologs in the genic regions of the two genomes. The transcriptome dataset should contribute to improved sugarcane gene models and sugarcane protein predictions; and will serve as a reference database for analysis of transcript expression in sugarcane.


Fibromyalgia is associated with impaired balance and falls.
The purpose of this study was to determine whether fibromyalgia (FM) patients differ from matched healthy controls in clinical tests of balance ability and fall frequency. Thirty-four FM patients and 32 age-matched controls were administered the Balance Evaluation-Systems Test (BESTest), rated their balance confidence with the Activities-Specific Balance Confidence (ABC) Scale, and reported the number of falls in the last 6 months. The Fibromyalgia Impact Questionnaire was used to assess FM severity. FM patients had significantly impaired balance in all components of the BESTest compared with controls. They also scored more poorly on balance confidence. Overall FM severity (Fibromyalgia Impact Questionnaire) correlated significantly with the BESTest and the ABC scale. The BESTest and ABC correlated significantly with 6 commonly reported FM symptoms (excluding pain). FM patients reported a total of 37 falls over the last 6-months compared with 6 falls in healthy controls. FM is associated with balance problems and increased fall frequency. Patients were aware of their balance problems. These results suggest that FM may affect peripheral and/or central mechanisms of postural control. Further objective study is needed to identify the relative contributions of various neural and musculoskeletal and other impairments to postural stability in FM to provide clinicians with methods to maximize postural stability and help fall prevention.


Significance of HE4 estimation in comparison with CA125 in diagnosis of ovarian cancer and assessment of treatment response.
Human epididymis protein 4 (HE4) is a novel and specific biomarker for ovarian cancer. The aim of this study is to evaluate a new tumor marker, HE4, in comparison with CA125 in diagnosis of epithelial ovarian cancer (EOC) and benign gynecological diseases. CA125 and HE4 serum levels were determined in 30 patients with epithelial ovarian cancer (21 serous, 6 endometrioid and 3 mucinous tumors), 20 patients with benign gynecological diseases (8 patients with ovarian cyst, 5 patients with endometriosis, 4 patients with fibroid and 3 patients with pelvic inflammatory disease) and 20 healthy women. CA125 and HE4 cut-offs were 35 U/ml and 150 pmol/l, respectively. Serum HE4 and CA125 concentrations were significantly higher in the ovarian cancer patients compared with those seen in patients with benign disease or in the healthy controls (p = 0.001 and p < 0.001 respectively). In the receiver operating characteristic analysis (ROC), the area under the curve (AUC) values for HE4 was 0.96 (95% confidence interval, 0.9-1.0) and CA125 was 0.82 (95% confidence interval, 0.7-0.94). Compared to CA125, HE4 had higher sensitivity (90% vs. 83.3%), specificity (95% vs. 85%), PPV (93.1% vs. 80.7%) and NPV (92.7% vs. 87.2%), the combination of HE4 + CA125 the sensitivity and PPV reached 96.7% and 97% respectively. Measuring serum HE4 concentrations along with CA125 concentrations may provide higher accuracy for detecting epithelial ovarian cancer. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1060413168685759.


White matter microstructure in female to male transsexuals before cross-sex hormonal treatment. A diffusion tensor imaging study.
Some gray and white matter regions of the brain are sexually dimorphic. The best MRI technique for identifying subtle differences in white matter is diffusion tensor imaging (DTI). The purpose of this paper is to investigate whether white matter patterns in female to male (FtM) transsexuals before commencing cross-sex hormone treatment are more similar to that of their biological sex or to that of their gender identity. DTI was performed in 18 FtM transsexuals and 24 male and 19 female heterosexual controls scanned with a 3 T Trio Tim Magneton. Fractional anisotropy (FA) was performed on white matter fibers of the whole brain, which was spatially analyzed using Tract-Based Spatial Statistics. In controls, males have significantly higher FA values than females in the medial and posterior parts of the right superior longitudinal fasciculus (SLF), the forceps minor, and the corticospinal tract. Compared to control females, FtM showed higher FA values in posterior part of the right SLF, the forceps minor and corticospinal tract. Compared to control males, FtM showed only lower FA values in the corticospinal tract. Our results show that the white matter microstructure pattern in untreated FtM transsexuals is closer to the pattern of subjects who share their gender identity (males) than those who share their biological sex (females). Our results provide evidence for an inherent difference in the brain structure of FtM transsexuals.


Feasibility and Acceptability of a Task-Shifted Intervention to Enhance Adherence to HIV Medication and Improve Depression in People Living with HIV in Zimbabwe, a Low Income Country in Sub-Saharan Africa.
Using a pilot trial design in an HIV care clinic in Zimbabwe, we randomised 32 adults with poor adherence to antiretroviral therapy and at least mild depression to either six sessions of Problem-Solving Therapy for adherence and depression (PST-AD) delivered by an adherence counsellor, or to Enhanced Usual Care (Control). Acceptability of PST-AD was high, as indicated by frequency of session attendance and through qualitative analyses of exit interviews. Fidelity was >80% for the first two sessions of PST-AD but fidelity to the adherence component of PST-AD dropped by session 4. Contamination occurred, in that seven patients in the control arm received one or two PST-AD sessions before follow-up assessment. Routine health records proved unreliable for measuring HIV viral load at follow-up. Barriers to measuring adherence electronically included device failure and participant perception of being helped by the research device. The study was not powered to detect clinical differences, however, promising change at 6-months follow-up was seen in electronic adherence, viral load suppression (PST-AD arm 9/12 suppressed; control arm 4/8 suppressed) and depression (Patient Health Questionnaire-4.7 points in PST-AD arm vs. control, adjusted p value = 0.01). Results inform and justify a future randomised controlled trial of task-shifted PST-AD.


Physiologic childbirth and active management of the third stage of labor: A latent class model of risk for postpartum hemorrhage.
Postpartum hemorrhage (PPH) is a threat to maternal mortality worldwide. Evidence supports active management of third stage labor (AMTSL) for preventing PPH. However, trials of AMTSL include women at varying risk levels, such as women undergoing physiologic labor and those with labor complications. Counseling women about their risk for PPH and AMTSL is difficult as many women who appear low-risk can still have PPH. This study uses outcomes of 2322 vaginal births from a hospital midwifery service in the United States to examine risks for PPH and effectiveness of AMTSL. Using a latent class analysis approach, physiologic birth practices and other risk factors for PPH were analyzed to understand if discrete classes of clinical characteristics would emerge. The effect of AMTSL on the PPH outcome was also considered by class. A four-class solution best fit the data; each class was clinically distinct. The two largest Classes (A and B) represented women with term births and lower average parity, with higher rates of nulliparity in Class B. Class A women had more physiologic birth elements and less labor induction or labor dysfunction compared with Class B. PPH and AMTSL use was higher in Class B. In Class B, AMTSL lowered risk for PPH. However, in Class A, AMTSL was associated with higher risk for PPH and delayed placental delivery (>30 minutes). AMTSL may not be as beneficial to women undergoing physiologic birth. Further study of the etiology of PPH in these women is indicated to inform preventive care.


Cardiovascular Magnetic Resonance in Nonischemic Myocardial Inflammation: Expert Recommendations.
This JACC Scientific Expert Panel provides consensus recommendations for an update of the cardiovascular magnetic resonance (CMR) diagnostic criteria for myocardial inflammation in patients with suspected acute or active myocardial inflammation (Lake Louise Criteria) that include options to use parametric mapping techniques. While each parameter may indicate myocardial inflammation, the authors propose that CMR provides strong evidence for myocardial inflammation, with increasing specificity, if the CMR scan demonstrates the combination of myocardial edema with other CMR markers of inflammatory myocardial injury. This is based on at least one T2-based criterion (global or regional increase of myocardial T2 relaxation time or an increased signal intensity in T2-weighted CMR images), with at least one T1-based criterion (increased myocardial T1, extracellular volume, or late gadolinium enhancement). While having both a positive T2-based marker and a T1-based marker will increase specificity for diagnosing acute myocardial inflammation, having only one (i.e., T2-based OR T1-based) marker may still support a diagnosis of acute myocardial inflammation in an appropriate clinical scenario, albeit with less specificity. The update is expected to improve the diagnostic accuracy of CMR further in detecting myocardial inflammation.


Validation of the French translation of the Dutch residency educational climate test.
The learning environment is one of the most influential factors in training of medical residents. The Dutch Residency Educational Climate Test (D-RECT) is one of the strongest instruments for measuring the learning environment. However, it has not been translated in French. The objective of this study is the psychometric validation of the DRECT French version. After translation of the D-RECT questionnaire into French, residents of five Moroccan hospitals were invited to complete the questionnaire between July and September 2018. Confirmatory factor analysis was used to evaluate the validity of the construct using the standardized root mean square residual (SRMR), the root mean square error approximation (RMSEA), the Comparative Fit Index (CFI) and the Tucker- Lewis Index (TLI). Reliability analysis was analysed using Internal consistency and Test-retest. During the study period, 211 residents completed the questionnaire. Confirmatory factor analysis showed an adequate model fit with the following indicators: SRMR = 0.058 / RMSEA = 0.07 / CFI = 0.88 / TLI = 0.87. The French translation had a good internal consistency (Cronbach alpha score > 0.7 for all subscales) and a good temporal stability (correlation score between two measurements = 0.89). This French version has an acceptable validity of the construct, a good internal consistency and good temporal reliability, and may be used to evaluate the learning climate. Additional research is necessary in other French-speaking contexts, in order to confirm these results.


[Pilot study to resolve problems of visual acuity assessment in grading vision of the physically handicapped--visual acuity and difficulties in daily life of age-related macular degeneration-].
To resolve the problems of visual acuity assessment in grading the vision of the physically handicapped as proposed by the Subcommittee for Promoting the Realization of a Cohesive Society with the Visually Disabled, Science Council of Japan, a method suitable for assessing visual disturbances, and the relationship between the degree of visual disturbances and the degree of difficulty in activities of daily life are clarified. 151 persons with age-related macular degeneration were studied. Examination methods for measuring visual acuity and reading performance were studied, and interviews using the daily living task dependent on vision (DLTV) questionnaire were performed. The correlations between total DLTV score and each examination method were analyzed. The median total DLTV score for each grade of visual acuity of the better eye was calculated. Spearman's correlation coefficient between distance corrected visual acuity of the better eye and total DLTV score was 0.76. Median DLTV scores for visual acuities (better eye) of 0.2, 0.3, 0.4, 0.5 were 65, 73.5, 62, 79 respectively. Visual acuity can be assessed by measuring distant corrected visual acuity of the better eye and setting the upper limit of visual disturbance at either 0.3 or 0.4.


Advancing an interdisciplinary framework to study seed dispersal ecology.
Although dispersal is generally viewed as a crucial determinant for the fitness of any organism, our understanding of its role in the persistence and spread of plant populations remains incomplete. Generalizing and predicting dispersal processes are challenging due to context dependence of seed dispersal, environmental heterogeneity and interdependent processes occurring over multiple spatial and temporal scales. Current population models often use simple phenomenological descriptions of dispersal processes, limiting their ability to examine the role of population persistence and spread, especially under global change. To move seed dispersal ecology forward, we need to evaluate the impact of any single seed dispersal event within the full spatial and temporal context of a plant's life history and environmental variability that ultimately influences a population's ability to persist and spread. In this perspective, we provide guidance on integrating empirical and theoretical approaches that account for the context dependency of seed dispersal to improve our ability to generalize and predict the consequences of dispersal, and its anthropogenic alteration, across systems. We synthesize suitable theoretical frameworks for this work and discuss concepts, approaches and available data from diverse subdisciplines to help operationalize concepts, highlight recent breakthroughs across research areas and discuss ongoing challenges and open questions. We address knowledge gaps in the movement ecology of seeds and the integration of dispersal and demography that could benefit from such a synthesis. With an interdisciplinary perspective, we will be able to better understand how global change will impact seed dispersal processes, and potential cascading effects on plant population persistence, spread and biodiversity.


Epidemiology and control of dengue virus infections in Thai villages in 1987.
A severe epidemic of dengue hemorrhagic fever (DHF) in Nakhon Ratchasima, Thailand in August of 1987 prompted a field investigation. DHF rates of 0.4-6.5 cases per 1,000 residents in subdistricts and 2-15 cases per 1,000 residents in 10 villages investigated were reported. Epidemics peaked in neighboring villages at different times; in June and July, and in August before the rainy season began late in the month. In 4 primary schools representing 6 villages, sera from groups of randomly selected children were tested for dengue IgM with the antibody capture ELISA test. Rates of recent dengue infection were 10-65% in the schools and correlated closely with reported rates of DHF. In an effort to control vectors, malathion fog and temephos (1% abate sand granules) were applied. Villagers were educated in prevention and were urged to cover water receptacles. The percentage of houses with larvae dropped from 67 to 20, the percentage of containers with larvae decreased from 30 to 5, and the number of containers with larvae per 100 households decreased from 221 to 33. This was a serious epidemic in which conventional control measures were only moderately effective.


SU-E-T-455: Novel Planning Techniques for Lung SBRT.
Two new planning techniques in lung SBRT are presented. The first technique involves a differential dosing strategy for patients with compromised pulmonary function. The second technique compares supine vs prone setup for extreme posterior lesions. CyberKnife™(CK) prone setup is complicated due to patient discomfort from longer treatment times and the inability to track without fiducials. CK(G.3) Multiplan™(V3.5). Synchrony™, XSight™ Lung Tracking Systems. Our lung SBRT patients are tracked using fiducials with Synchrony or without fiducials with XSight using a 60 Gy in three fractions scheme. If the ray-tracing optimized plan results in V15Gy greater than 10% we employ the differential dosing technique. A new PTV is created to encompass the 54 Gy isodose surface. A more conformal plan with a dose constraint of 60Gy to CTV and 54Gy to PTV is created appreciably lowering the V15Gy. Alternatively a 54Gy PTV can be created from measured respiration excursions. For the prone/supine comparison, plans were generated for both positions then evaluated. Patients planned with the differential dose technique have a good tumor response as evidenced by a follow-up PET. Although prone setup is preferred for extremely posterior tumors, a dosimetrically comparable supine plan can be created to replicate the prone plan. The supine low dose spillage was only slightly greater than in the prone case. We conclude that the 54Gy/60Gy dosed patients have almost the same tumor response as the standard 60 Gy treatment. A dosimetrically similar supine plan obviates the need for a prone delivery.


Curiosity improves coping efficacy and reduces suicidal ideation severity among military veterans at risk for suicide.
Curiosity, the tendency to engage in novel and challenging opportunities, may be an important source of resilience for those at risk for suicide. We hypothesized that curiosity would have a buffering effect against risk conferred by multiple sources of distress, whereby curiosity would be associated with reduced suicidal ideation and increased coping efficacy. As part of a larger intervention trial designed to improve coping skills and reduce suicidal ideation, 117 military veterans with suicidal ideation completed measures of curiosity and distress (perceived stress, depression, anxiety, and sleep disturbances) at baseline, and completed measures of suicidal ideation and coping efficacy (to stop negative thoughts, to enlist support from friends and family) at baseline and 3-, 6-, and 12-week follow up. Growth curve models showed that curiosity moderated the association between distress and suicidal ideation at baseline and that curiosity moderated the association between distress and increased coping efficacy to stop negative thoughts over time. Findings suggest that curiosity may buffer against the effect of heightened levels of distress on suicidal ideation and help facilitate stronger gains in coping efficacy over time. Additional work should further examine the role of curiosity as a protective factor for veterans with suicidal ideation.


Behavior change evaluation of a culturally consistent reproductive health program for young Kenyans.
Few rigorous evaluations have been conducted of locally designed, culturally consistent adolescent reproductive health programs. A quasi-experimental research design was used to measure behavioral changes associated with a culturally consistent reproductive health program for young people in Kenya. Baseline and endline surveys were conducted in 1997 and 2001, respectively, in the project and control areas. Multivariate analysis was used to assess whether the project was associated with changes in young people's sexual initiation, safer-sex behavior and discussion of reproductive health issues with adults. The 36-month project was associated with considerable changes in young people's sexual and reproductive health-related behavior, but behavior change differed by gender. Females in the project site were significantly more likely than those in the control site to adopt secondary abstinence (odds ratio, 3.3) and less likely to have had three or more sex partners (0.1). Males in the project site were more likely to use condoms than those in the control site (3.7). Both males and females in the project site were more likely to discuss sexual and reproductive health issues with a nonparent adult than were young people in the control site (1.9 and 5.5, respectively). Interventions that adapt to indigenous traditions can be both acceptable to communities and associated with significant changes in young people's behavior.


The Differences in Local Translatome across Distinct Neuron Types Is Mediated by Both Baseline Cellular Differences and Post-transcriptional Mechanisms.
Local translation in neurites is a phenomenon that enhances the spatial segregation of proteins and their functions away from the cell body, yet it is unclear how local translation varies across neuronal cell types. Further, it is unclear whether differences in local translation across cell types simply reflect differences in transcription or whether there is also a cell type-specific post-transcriptional regulation of the location and translation of specific mRNAs. Most of the mRNAs discovered as being locally translated have been identified from hippocampal neurons because their laminar organization facilitates neurite-specific dissection and microscopy methods. Given the diversity of neurons across the brain, studies have not yet analyzed how locally translated mRNAs differ across cell types. Here, we used the SynapTRAP method to harvest two broad cell types in the mouse forebrain: GABAergic neurons and layer 5 projection neurons. While some transcripts overlap, the majority of the local translatome is not shared across these cell types. In addition to differences driven by baseline expression levels, some transcripts also exhibit cell type-specific post-transcriptional regulation. Finally, we provide evidence that GABAergic neurons specifically localize mRNAs for peptide neurotransmitters, including somatostatin and cortistatin, suggesting localized production of these key signaling molecules in the neurites of GABAergic neurons. Overall, this work suggests that differences in local translation in neurites across neuronal cell types are poised to contribute substantially to the heterogeneity in neuronal phenotypes.


The influence of protein provision in the early phase of intensive care on clinical outcomes for critically ill patients on mechanical ventilation.
Mechanically ventilated patients often face progressive and rapid losses of body mass and muscle because of hypermetabolism and increased protein catabolism. To investigate the impact of adequate nutritional provision during the early phase of intensive care unit (ICU) admission on the clinical outcomes in patients with medical illnesses receiving mechanical ventilation support. Two hundred and eleven mechanically ventilated patients admitted to a 30-bed medical ICU were included. Three groups, based on nutrition intake, were examined: adequate protein intake (aPI), n=34; insufficient protein intake/ adequate energy intake (iPI/aEI), n=25; insufficient protein and energy intake (iPI/iEI), n=152. Patients' mean age was 65±14 years; body mass index, 22±4; Acute Physiology and Chronic Health Evaluation II score, 24±7. The aPI group had significantly lower rates of in-ICU (14.7%) and in-hospital (23.5%) mortality than patients with insufficient protein intake: in-ICU mortality, iPI/aEI, 36%; iPI/iEI, 44.1% (p=0.006); in-hospital mortality, iPI/aEI, 56.0%; iPI/iEI, 52.0% (p=0.008). In the multivariate analysis, the hazard ratios (95% confidence intervals) for 60-day survival were 2.59 (1.02-6.59; p=0.046) and 2.88 (1.33-6.26; p=0.008) for the iPI/aEI and iPI/iEI groups, respectively. Despite possible selection bias owing to the retrospective nature of the study, achievement of >90% of target protein intake was associated with improved ICU outcomes in mechanically ventilated critically ill patients, based on real-world clinical circumstances.


Two-dimensional subwavelength gratings with different frontside/backside reflectance.
A concept of two-dimensional periodic subwavelength gratings exhibiting different reflectance on the front and back sides is presented. This structure incorporates an arrangement of a metalized hole array and a nanodot array on top. We exemplify the optical properties for an aluminum grating with 243 nm period, and we demonstrate its color filtering properties for unpolarized light in reflectance as well as in transmittance yielding different colors for frontside and backside illumination. These findings have been confirmed by a numerical study. The gratings may be replicated by a simple nanoimprint process on plastic foils predestined for industrial mass production.


The empirical status of psychodynamic psychotherapy - an update: Bambi's alive and kicking.
The Task Force on Promotion and Dissemination of Psychological Procedures proposed rigorous criteria to define empirically supported psychotherapies. According to these criteria, 2 randomized controlled trials (RCTs) showing efficacy are required for a treatment to be designated as 'efficacious' and 1 RCT for a designation as 'possibly efficacious'. Applying these criteria modified by Chambless and Hollon, this article presents an update on the evidence for psychodynamic therapy (PDT) in specific mental disorders. A systematic search was performed using the criteria by Chambless and Hollon for study selection, as follows: (1) RCT of PDT in adults, (2) use of reliable and valid measures for diagnosis and outcome, (3) use of treatment manuals or manual-like guidelines, (4) adult population treated for specific problems and (5) PDT superior to no treatment, placebo or alternative treatment or equivalent to an established treatment. A total of 39 RCTs were included. Following Chambless and Hollon, PDT can presently be designated as efficacious in major depressive disorder (MDD), social anxiety disorder, borderline and heterogeneous personality disorders, somatoform pain disorder, and anorexia nervosa. For MDD, this also applies to the combination with pharmacotherapy. PDT can be considered as possibly efficacious in dysthymia, complicated grief, panic disorder, generalized anxiety disorder, and substance abuse/dependence. Evidence is lacking for obsessive-compulsive, posttraumatic stress, bipolar and schizophrenia spectrum disorder(s). Evidence has emerged that PDT is efficacious or possibly efficacious in a wide range of common mental disorders. Further research is required for those disorders for which sufficient evidence does not yet exist.


A coalition partnership of vision health through a health-promoting school program for primary school students in Taiwan.
Myopia, the most common refractive error, is the most common cause of avoidable visual impairment among children and has reached epidemic proportions among children and young adults in urban areas of East and Southeast Asia that contain populations of Chinese ancestry. Moreover, vision health is an important theme of the health-promoting school program issued by the Ministry of Education in Taiwan. The aim of this study is to assess the impact of pre- and post-intervention proposed by the health-promoting school (HPS) model. The objectives are to understand whether the HPS model influenced the vision screening results and the attitude, knowledge level, and vision care behavior of the students involved. A prospective cohort study design was used to evaluate a vision health program. Four elementary schools, local education authorities, and one university in northern Taiwan established a coalition partnership to design a six-month program to combat myopia among students. The target population was 6668 school children from local elementary schools. For the purpose of this study, the outcome of visual acuity testing (in logMAR) was analyzed with a sampling of 373 school children (aged 11-12 years old) who were chosen from high prevalence of poor vision classes. After the HPS program, the attitudes, behaviors, and knowledge levels of the school children regarding vision health were significantly improved. The pre-intervention mean logMAR of all participating students ( N = 373) was -.10, which increased to -.19 after the intervention. Analysis using the Wilcoxon signed-rank test showed that the logMAR value was significantly improved after the intervention ( t = 2.13, p < 0.05). Our findings highlight the relevance and effectiveness of the coalition's efforts, which reinforces the usefulness of co-operatively implementing the HPS program.


Neulasta (pegfilgrastim): a once-per-cycle option for the management of chemotherapy-induced neutropenia.
Neutropenia is a significant hematologic complication induced by cytotoxic chemotherapy. The clinical consequences of chemotherapy-induced neutropenia are often severe and can be potentially life-threatening. Patients who develop febrile neutropenia often need to be hospitalized, reducing their quality of life and increasing costs. Neutropenia can also compromise the ability to deliver chemotherapy at the full dose and on schedule. To help prevent the occurrence of neutropenia, patients with a high risk of developing chemotherapy-related infections may be given prophylactic colony-stimulating factors. Filgrastim is a recombinant human granulocyte colony-stimulating factor that has been widely used (in over 3 million patients) for over 12 years in the management of neutropenia. Pegfilgrastim is an approved, long-acting, next generation of granulocyte colony-stimulating factor that has similar clinical benefits to filgrastim but has novel pharmacokinetic properties. Pegfilgrastim shows at least comparable safety and efficacy to filgrastim, with the added benefit of simplified once-per-chemotherapy-cycle dosing. In addition, two randomized, controlled pivotal trials have shown that a single dose of pegfilgrastim given once per cycle led to a lower observed incidence of febrile neutropenia following myelosuppressive chemotherapy, compared with daily injections of filgrastim. Clinical trials are currently expanding the clinical experience with pegfilgrastim in a variety of solid tumors and hematologic malignancies. In addition to prevention of chemotherapy-induced neutropenia in 21- and 28-day regimens, future studies are examining the suitability of pegfilgrastim in dose-dense therapy and other cancer settings.


Statistical analysis of nonlinear parameter estimation for Monod biodegradation kinetics using bivariate data.
A nonlinear regression technique for estimating the Monod parameters describing biodegradation kinetics is presented and analyzed. Two model data sets were taken from a study of aerobic biodegradation of the polycyclic aromatic hydrocarbons (PAHs), naphthalene and 2-methylnaphthalene, as the growth-limiting substrates, where substrate and biomass concentrations were measured with time. For each PAH, the parameters estimated were: q(max), the maximum substrate utilization rate per unit biomass; K(S), the half-saturation coefficient; and Y, the stoichiometric yield coefficient. Estimating parameters when measurements have been made for two variables with different error structures requires a technique more rigorous than least squares regression. An optimization function is derived from the maximumlikelihood equation assuming an unknown, nondiagonal covariance matrix for the measured variables. Because the derivation is based on an assumption of normally distributed errors in the observations, the error structures of the regression variables were examined. Through residual analysis, the errors in the substrate concentration data were found to be distributed log-normally, demonstrating a need for log transformation of this variable. The covariance between ln C and X was found to be small but significantly nonzero at the 67% confidence level for NPH and at the 94% confidence level for 2MN. The nonlinear parameter estimation yielded unique values for q(max), K(S), and Y for naphthalene. Thus, despite the low concentrations of this sparingly soluble compound, the data contained sufficient information for parameter estimation. For 2-methylnaphthalene, the values of q(max) and K(S) could not be estimated uniquely; however, q(max)/K(S) was estimated. To assess the value of including the relatively imprecise biomass concentration data, the results from the bivariate method were compared with a univariate method using only the substrate concentration data. The results demonstrated that the bivariate data yielded a better confidence in the estimates and provided additional information about the model fit and model adequacy. The combination of the value of the bivariate data set and their nonzero covariance justifies the need for maximum likelihood estimation over the simpler nonlinear least squares regression.


Detection of plasmid-mediated beta-lactamases with DNA probes.
beta-Lactamase identification by colony hybridization with 32P-labeled DNA probes for TEM-1, SHV-1, OXA-1, OXA-2, PSE-1, PSE-2, and PSE-4 was compared with isoelectric focusing in 122 clinical isolates making a variety of enzyme types. All strains producing a probe-type enzyme gave a positive hybridization reaction. Cross-hybridization was observed between TEM-1 and TEM-2 or TLE-1, between SHV-1 and SHV-2, between OXA-1 and OXA-4, between OXA-2 and OXA-3 (weak), between PSE-2 and OXA-6 or OXA-5 (weak), and among PSE-1, PSE-4, and CARB-3. With allowance for such cross-hybridization, only six strains gave false-positive reactions, and the procedure was 99% specific.


A novel UGGT1 and p97-dependent checkpoint for native ectodomains with ionizable intramembrane residue.
Only native polypeptides are released from the endoplasmic reticulum (ER) to be transported at the site of activity. Persistently misfolded proteins are retained and eventually selected for ER-associated degradation (ERAD). The paradox of a structure-based protein quality control is that functional polypeptides may be destroyed if they are architecturally unfit. This has health-threatening implications, as shown by the numerous "loss-of-function" proteopathies, but also offers chances to intervene pharmacologically to promote bypassing of the quality control inspection and export of the mutant, yet functional protein. Here we challenged the ER of human cells with four modular glycopolypeptides designed to alert luminal and membrane protein quality checkpoints. Our analysis reveals the unexpected collaboration of the cytosolic AAA-ATPase p97 and the luminal quality control factor UDP-glucose:glycoprotein glucosyltransferase (UGGT1) in a novel, BiP- and CNX-independent checkpoint. This prevents Golgi transport of a chimera with a native ectodomain that passes the luminal quality control scrutiny but displays an intramembrane defect. Given that human proteopathies may result from impaired transport of functional polypeptides with minor structural defects, identification of quality checkpoints and treatments to bypass them as shown here upon silencing or pharmacologic inhibition of UGGT1 or p97 may have important clinical implications.


Structure-activity relationship study of 16 a-thiocamptothecins: an integrated in vitro and in silico approach.
The instability of the hydroxylactone E ring represents a critical drawback of camptothecins, because the lactone ring is recognized to be essential for stabilization of topoisomerase I-mediated DNA cleavage. In an attempt to investigate the effect of the thiopyridone pharmacophofore on the molecular and pharmacological features of the drug, we prepared a series of novel 16 a-thiocamptothecin analogues. Due to the sulfur atom, a destabilization of the hydrogen bond between the hydroxy group in position 17 of the opened E ring and the carbonyl of the pyridone moiety is predicted, thus shifting the equilibrium toward the closed lactone form and increasing the lipophilic properties of the compounds. This feature was associated with superior antiproliferative potency, with reduced interaction with the human serum albumin and with substantial increase of the persistence of the topoisomerase I-DNA cleavable complex. These effects were prominent for thio-SN38, the most active compound of the series. The favorable interactions at the molecular and cellular level of the reported thiocamptothecins confer promising features, and these compounds warrant preclinical development.


The art of maintaining everyday life: collaboration among older parents, their adult children, and health care professionals in reablement.
Background: A shift in the work-divide among generations and an ageing population have altered the balance of care and support between families and welfare states. Although state policy has increasingly acknowledged that older adults ageing in place receive support from family members, how adult children perceive their collaboration with their parents and health care professionals in reablement services remains unclear. The aim of this study is to identify how adult children perceive the collaboration between older parents, family members, and health care professionals in reablement services. Methods: This study has a qualitative research design with a constructivist grounded theory approach. In total, 15 adult children - 6 sons, 8 daughters, and a daughter-in-law, aged 47-64 years - whose parents had received reablement services, participated in in-depth interviews. Results: Our findings clarify how children and their older parents' reablement services can collaborate to support how the adult children manage and maintain both their own and their parents' everyday lives. The core category derived from our data analysis was the art of maintaining everyday life, with four subcategories indicating the different dimensions of that process: doing what is best for one's parents, negotiating the dilemmas of everyday life, managing parents' reablement, and ensuring the flow of everyday life. Conclusion: To promote collaboration among older adults, their children, and health care professionals in reablement, health care professionals need to proactively involve older adults' family members in the reablement processes, particularly because older adults and their children do not always express all of their care-related needs to reablement services.


Proliferative and anti-proliferative effects of retinoic acid at doses similar to endogenous levels in Leydig MLTC-1/R2C/TM-3 cells.
Vitamin A is suggested to be protective against oxidative stress. However, different authors observed pro-oxidant effects of retinoids both in experimental works and clinical trials. These discordances are the bases for the investigation of the proliferative and anti-proliferative properties of retinoic acid (RA) in biological systems. Cell viability is determined with the MTT assay. Oxidative stress parameters are detected measuring catalase (CAT) and glutathione S-transferase (GST) enzymatic activities. FABP5 mRNA levels are measured by RT-PCR. Autophagy and apoptosis are analyzed by Monodansylcadaverine (MDC) staining and TUNEL assay, respectively. RA, at nutraceutic/endogenous doses (10-200 nM), increases cell viability of testes tumor Leydig cell lines (MLTC-1 and R2C) and modulates antioxidant enzyme activities, as CAT and GST. RA is able to induce proliferation through non-classical and redox-dependent mechanisms accompanied by increased levels of FABP5 mRNA. The redox environment of the cell is currently thought to be extremely important for controlling either apoptosis or autophagy. Apoptosis occurs at pharmacological doses, while autophagy, which plays a critical role in removing damaged or surplus organelles in order to maintain cellular homeostasis, is triggered at the critical concentration of 500 nM RA, both in normal and tumoral cells. Slight variations of RA concentrations are evaluated as a threshold value to distinguish between the proliferative or anti-proliferative effects. Although retinoids have a promising role as antineoplastic agents, physiological levels of RA play a key role in Leydig cancer progression, fostering proliferation and growth of testicular tumoral mass.


Review of Valuation Methods of Preference-Based Measures of Health for Economic Evaluation in Child and Adolescent Populations: Where are We Now and Where are We Going?
Methods for measuring and valuing health benefits for economic evaluation and health technology assessment in adult populations are well developed. In contrast, methods for assessing interventions for child and adolescent populations lack detailed guidelines, particularly regarding the valuation of health and quality of life in these age groups. This paper critically examines the methodological considerations involved in the valuation of child- and adolescent-specific health-related quality of life by existing preference-based measures. It also describes the methodological choices made in the valuation of existing generic preference-based measures developed with and/or applied in child and adolescent populations: AHUM, AQoL-6D, CHU9D, EQ-5D-Y, HUI2, HUI3, QWB, 16D and 17D. The approaches used to value existing child- and adolescent-specific generic preference-based measures vary considerably. While the choice of whose preferences and which perspective to use is a matter of normative debate and ultimately for decision by reimbursement agencies and policy makers, greater research around these issues would be informative and would enrich these discussions. Research can also inform the other methodological choices required in the valuation of child and adolescent health states. Gaps in research evidence are identified around the impact of the child described in health state valuation exercises undertaken by adults, including the possibility of informed preferences; the appropriateness and acceptability of valuation tasks for adolescents, in particular tasks involving the state 'dead'; anchoring of adolescent preferences; and the generation and use of combined adult and adolescent preferences.


Testing for human immunodeficiency virus among cancer survivors under age 65 in the United States.
Knowing the human immunodeficiency virus (HIV) serostatus of patients at the time of cancer diagnosis or cancer recurrence is prerequisite to coordinating HIV and cancer treatments and improving treatment outcomes. However, there are no published data about HIV testing among cancer survivors in the United States. We sought to provide estimates of the proportion of cancer survivors tested for HIV and to characterize factors associated with having had HIV testing. We used data from the 2009 Behavioral Risk Factor Surveillance System to calculate the proportion of cancer survivors under age 65 who had undergone HIV testing, by demographic and health-related factors and by state. Adjusted proportion estimates were calculated by multivariable logistic regression. Only 41% of cancer survivors in the United States under the age of 65 reported ever having had an HIV test. The highest proportion of survivors tested was among patients aged 25 to 34 years (72.2%), non-Hispanic blacks (59.5%), and cervical cancer survivors (51.2%). The proportion tested was highest in the District of Columbia (68.3%) and lowest in Nebraska (24.1%). Multivariable analysis showed that factors associated with HIV testing included being non-Hispanic black or Hispanic, being younger, having higher education, not being married or living with a partner, not being disabled, and having medical cost concerns. Having an AIDS-related cancer was associated with HIV testing only among females. The proportions of HIV testing varied substantially by demographic and health-related factors and by state. Our study points to the need for public health interventions to promote HIV testing among cancer survivors.


Challenges in developing national HIV guidelines: experience from the eastern Mediterranean.
To appraise the process of development and clinical content of national human immunodeficiency virus (HIV) clinical practice guidelines of countries in the eastern Mediterranean and to formulate recommendations for future guideline development and adaptation. Twenty-three countries in the World Health Organization (WHO) Eastern Mediterranean and United Nations Children's Fund Middle East and North Africa regions were invited to submit national HIV clinical practice guidelines for review. The guideline development methodology was assessed using an adaptation of the Appraisal of Guidelines Research and Evaluation (AGREE) instrument and guideline content, using a checklist to evaluate concordance with WHO 2006 generic guidelines. Twelve countries submitted 20 guidelines developed between 2004 and 2009. Median scores were poor (i.e. < 0.6) for the methodological quality domains of rigour of development, stakeholder involvement and applicability and flexibility. Scores were better for the domains of scope and purpose (median: 0.82, interquartile range, IQR: 0.58-0.89) and clarity and presentation (median: 0.67, IQR: 0.50-0.78). Concerning guideline content, recommended first-line treatment and eligibility criteria for antiretroviral therapy (ART) in adults were in line with WHO recommendations in most guidelines. However, recommendations on antiretroviral prophylaxis for the prevention of vertical HIV transmission, diagnosis and treatment of HIV infection in infants, monitoring patients on ART, treatment failure and co-morbidities were often lacking. The large majority of national HIV clinical practice guidelines had methodological weaknesses and content inaccuracies. Countries require assistance with the adaptation process to ensure that guidelines are valid and up to date and accurately reflect WHO global clinical care recommendations for patients with HIV.


Insomnia and urban neighbourhood contexts--are associations modified by individual social characteristics and change of residence? Results from a population-based study using residential histories.
Until now, insomnia has not been much of interest in epidemiological neighbourhood studies, although literature provides evidence enough for insomnia-related mechanisms being potentially dependent on neighbourhood contexts. Besides, studies have shown differences in sleep along individual social characteristics that might render residents more vulnerable to neighbourhood contextual exposures. Given the role of exposure duration and changes in the relationship between neighbourhoods and health, we studied associations of neighbourhood unemployment and months under residential turnover with insomnia by covering ten years of residential history of nearly 3,000 urban residents in the Ruhr Area, Germany. Individual data were retrieved from the Heinz Nixdorf Recall Study, a population-based study of randomly chosen participants from adjacent cities, which contains self-rated insomnia symptoms and individual social characteristics. Participants' residential addresses were retrospectively assessed using public registries. We built individually derived exposure measures informing about mean neighbourhood unemployment rates and months under high residential turnover. These measures were major predictors in multivariate logistic regressions modelling the association between social neighbourhood characteristics and insomnia in the whole sample and subgroups defined by low income, low education, social isolation, and change of residence. Traffic-related noise, age, gender, economic activity, and education were considered as covariates. Nearly 12 per cent of the participants complained about insomnia. Associations of neighbourhood unemployment with insomnia were more consistent than those of residential turnover in the whole sample (adjusted OR 1.42, 95% CI 1.00-2.03 for neighbourhood unemployment and OR 1.33, 95% CI 0.78-2.25 for residential turnover in the highest exposure categories). In low-income and socially isolated participants, neighbourhood unemployment odds of reporting insomnia were particularly elevated (adjusted OR 2.90, 95% CI 1.39-6.02 and OR 3.32, 95% CI 1.11-9.96, respectively). Less educated participants displayed relatively high odds of reporting insomnia throughout all upper neighbourhood unemployment exposure categories. Change of residence weakened associations, whereas undisrupted exposure sharpened them by trend. Our findings hint at multiple stressors being effective in both the neighbourhood context and individual resident, possibly reflecting precarious life situations undermining residents' sleep and health chances. Moreover, our results suggest a temporal dependency in the association between neighbourhood and insomnia.


Co-adjustment of yolk antioxidants and androgens in birds.
Mothers can shape the developmental trajectory of their offspring through the transmission of resources such as hormones, antioxidants or immunoglobulins. Over the last two decades, an abundant literature on maternal effects in birds has shown that several of these compounds (i.e. androgens, glucocorticoids and antioxidants) often influence the same offspring phenotypic traits (i.e. growth, immunity or oxidative stress levels), making interaction effects between egg components a likely scenario. However, the potential interactive effects of maternally transmitted compounds on offspring development and potential co-adjustment of these compounds within an egg are still poorly understood. Here, we report the results of an interspecific comparative analysis on birds' egg yolk composition (i.e. androgens and antioxidants) where we found that yolk carotenoid and vitamin E concentrations are positively associated, supporting the hypothesis that these two antioxidants act in synergy. The concentrations of vitamin E also increased with increasing concentrations of testosterone. This last result confirms the emerging idea that androgens and antioxidants are co-adjusted within eggs and that maternally transmitted antioxidants might limit the potential direct and indirect effects of prenatal exposure to high testosterone levels on oxidative stress.


MicroRNA Signature of Traumatic Brain Injury: From the Biomarker Discovery to the Point-of-Care.
Traumatic brain injury (TBI) is a serious problem that causes high morbidity and mortality around the world. Currently, no reliable biomarkers are used to assess the severity and predict the recovery. Many protein biomarkers were extensively studied for diagnosis and prognosis of different TBI severities such as S-100β, glial fibrillary acidic protein (GFAP), neuron-specific enolase (NSE), neurofilament light chain (NFL), cleaved tau protein (C-tau), and ubiquitin C-terminal hydrolase-L1 (UCH-L1). However, none of these candidates is currently used in the clinical practice, due to relatively low sensitivity, for the diagnosis of mild TBI (mTBI) or mild to moderate TBI (MMTBI) patients who are clinically well and do not have a detectable intracranial pathology on the scans. MicroRNAs (miRNAs or miRs) are a class of small endogenous molecular regulators, which showed to be altered in different pathologies, including TBI and for this reason, their potential role in diagnosis, prognosis and therapeutic applications, is explored. Promising miRNAs such as miR-21, miR-16 or let-7i were identified as suitable candidate biomarkers for TBI and can differentiate mild from severe TBI. Also, they might represent new potential therapeutic targets. Identification of miRNA signature in tissue or biofluids, for several pathological conditions, is now possible thanks to the introduction of new high-throughput technologies such as microarray platform, Nanostring technologies or Next Generation Sequencing. This review has the aim to describe the role of microRNA in TBI and to explore the most commonly used techniques to identify microRNA profile. Understanding the strengths and limitations of the different methods can aid in the practical use of miRNA profiling for diverse clinical applications, including the development of a point-of-care device.


Transposon-associated lincosamide resistance lnu(C) gene identified in Brachyspira hyodysenteriae ST83.
Treatment of Swine Dysentery (SD) caused by Brachyspira hyodysenteriae (B. hyodysenteriae) is carried out using antimicrobials such as macrolides, lincosamides and pleuromutilins leading to the selection of resistant strains. Whole genome sequencing of a multidrug-resistant B. hyodysenteriae strain called BH718 belonging to sequence type (ST) 83 revealed the presence of the lincosamide resistance gene lnu(C) on the small 1724-bp transposon MTnSag1. The strain also contains an A to T substitution at position 2058 (A2058T) in the 23S rRNA gene which is known to be associated with macrolide and lincosamide resistance in B. hyodysenteriae. Testing of additional strains showed that those containing lnu(C) exhibited a higher minimal inhibitory concentration (MIC) of lincomycin (MIC ≥ 64 mg/L) compared to strains lacking lnu(C), even if they also harbor the A2058T mutation. Resistance to pleuromutilins could not be explained by the presence of already reported mutations in the 23S rRNA gene and in the ribosomal protein L3. This study shows that B. hyodysenteriae has the ability to acquire mobile genetic elements conferring resistance to antibiotics.


Solute exchange through gap junctions lessens the adverse effects of inactivating mutations in metabolite-handling genes.
Growth of cancer cells in vitro can be attenuated by genetically inactivating selected metabolic pathways. However, loss-of-function mutations in metabolic pathways are not negatively selected in human cancers, indicating that these genes are not essential in vivo. We hypothesize that spontaneous mutations in 'metabolic genes' will not necessarily produce functional defects because mutation-bearing cells may be rescued by metabolite exchange with neighboring wild-type cells via gap junctions. Using fluorescent substances to probe intercellular diffusion, we show that colorectal cancer (CRC) cells are coupled by gap junctions assembled from connexins, particularly Cx26. Cells with genetically inactivated components of pH regulation (SLC9A1), glycolysis (ALDOA), or mitochondrial respiration (NDUFS1) could be rescued through access to functional proteins in co-cultured wild-type cells. The effect of diffusive coupling was also observed in co-culture xenografts. Rescue was largely dependent on solute exchange via Cx26 channels, a uniformly and constitutively expressed isoform in CRCs. Due to diffusive coupling, the emergent phenotype is less heterogenous than its genotype, and thus an individual cell should not be considered as the unit under selection, at least for metabolite-handling processes. Our findings can explain why certain loss-of-function mutations in genes ascribed as 'essential' do not influence the growth of human cancers.


Mcp6, a meiosis-specific coiled-coil protein of Schizosaccharomyces pombe, localizes to the spindle pole body and is required for horsetail movement and recombination.
We report here that a meiosis-specific gene of Schizosaccharomyces pombe denoted mcp6+ (meiotic coiled-coil protein) encodes a protein that is required for the horsetail movement of chromosomes at meiosis I. The mcp6+ gene is specifically transcribed during the horsetail phase. Green fluorescent protein (GFP)-tagged Mcp6 appears at the start of karyogamy, localizes to the spindle-pole body (SPB) and then disappears before chromosome segregation at meiosis I. In the mcp6Delta strain, the horsetail movement was either hampered (zygotic meiosis) or abolished (azygotic meiosis) and the pairing of homologous chromosomes was impaired. Accordingly, the allelic recombination rates of the mcp6Delta strain were only 10-40% of the wild-type rates. By contrast, the ectopic recombination rate of the mcp6Delta strain was twice the wild-type rate. This is probably caused by abnormal homologous pairing in mcp6Delta cells because of aberrant horsetail movement. Fluorescent microscopy indicates that SPB components such as Sad1, Kms1 and Spo15 localize normally in mcp6Delta cells. Because Taz1 and Swi6 also localized with Sad1 in mcp6Delta cells, Mcp6 is not required for telomere clustering. In a taz1Delta strain, which does not display telomere clustering, and the dhc1-d3 mutant, which lacks horsetail movement, Mcp6 localized with Sad1 normally. However, we observed abnormal astral microtubule organization in mcp6Delta cells. From these results, we conclude that Mcp6 is necessary for neither SPB organization nor telomere clustering, but is required for proper astral microtubule positioning to maintain horsetail movement.


Elevated glutathione levels confer cellular sensitization to cisplatin toxicity by up-regulation of copper transporter hCtr1.
Previous studies have demonstrated that treating cultured cells with cisplatin (CDDP) up-regulated the expression of glutathione (GSH) and its de novo rate-limiting enzyme glutamate-cysteine ligase (GCL), which consists of a catalytic (GCLC) and a modifier (GCLM) subunit. It has also been shown that many CDDP-resistant cell lines exhibit high levels of GCLC/GCLM and GSH. Because the GSH system is the major intracellular regulator of redox conditions that serve as an important detoxification cytoprotector, these results have been taken into consideration that elevated levels of GCL/GSH are responsible for the CDDP resistance. In contrast to this context, we demonstrated here that overexpression of GSH by transfection with an expression plasmid containing the GCLC cDNA conferred sensitization to CDDP through up-regulation of human copper transporter (hCtr) 1, which is also a transporter for CDDP. Depleting GSH levels in these transfected cells reversed CDDP sensitivity with concomitant reduction of hCtr1 expression. Although rates of copper transport were also up-regulated in the transfected cells, these cells exhibited biochemical signature of copper deficiency, suggesting that GSH functions as an intracellular copper-chelator and that overexpression of GSH can alter copper metabolism. More importantly, our results reveal a new role of GSH in the regulation of CDDP sensitivity. Overproduction of GSH depletes the bioavailable copper pool, leading to up-regulation of hCtr1 and sensitization of CDDP transport and cell killing. These findings also have important implications in that modulation of the intracellular copper pool may be a novel strategy for improving chemotherapeutic efficacy of platinum-based antitumor agents.


CDA: combinatorial drug discovery using transcriptional response modules.
Anticancer therapies that target single signal transduction pathways often fail to prevent proliferation of cancer cells because of overlapping functions and cross-talk between different signaling pathways. Recent research has identified that balanced multi-component therapies might be more efficacious than highly specific single component therapies in certain cases. Ideally, synergistic combinations can provide 1) increased efficacy of the therapeutic effect 2) reduced toxicity as a result of decreased dosage providing equivalent or increased efficacy 3) the avoidance or delayed onset of drug resistance. Therefore, the interest in combinatorial drug discovery based on systems-oriented approaches has been increasing steadily in recent years. Here we describe the development of Combinatorial Drug Assembler (CDA), a genomics and bioinformatics system, whereby using gene expression profiling, multiple signaling pathways are targeted for combinatorial drug discovery. CDA performs expression pattern matching of signaling pathway components to compare genes expressed in an input cell line (or patient sample data), with expression patterns in cell lines treated with different small molecules. Then it detects best pattern matching combinatorial drug pairs across the input gene set-related signaling pathways to detect where gene expression patterns overlap and those predicted drug pairs could likely be applied as combination therapy. We carried out in vitro validations on non-small cell lung cancer cells and triple-negative breast cancer (TNBC) cells. We found two combinatorial drug pairs that showed synergistic effect on lung cancer cells. Furthermore, we also observed that halofantrine and vinblastine were synergistic on TNBC cells. CDA provides a new way for rational drug combination. Together with phExplorer, CDA also provides functional insights into combinatorial drugs. CDA is freely available at http://cda.i-pharm.org.


Hypoperfusion and T1-hypointense lesions in white matter in multiple sclerosis.
Longitudinal magnetic resonance imaging (MRI) studies show that a fraction of the multiple sclerosis (MS) T2-lesions contain T1-hypointense components that may persist to represent severe, irreversible tissue damage. It is not known why certain lesions convert to persistent T1-hypointense lesions. We hypothesized that the T1-hypointense lesions disproportionately distribute in the more hypoperfused areas of the brain. Here we investigated the association between hypoperfusion and T1-hypointense lesion distributions. MRI and cerebral blood flow (CBF) data were acquired on 45 multiple sclerosis (MS) patients and 20 healthy controls. CBF maps were generated using pseudo-continuous arterial spin labeling technique. The lesion probability distribution maps were superimposed on the CBF maps. Two distinct CBF clusters were observed in the white matter (WM) both in healthy controls and MS patients. An overall reduction in CBF was observed in MS patients compared to healthy controls. The majority of the T1-hypointense lesions were concentrated almost exclusively in the WM regions with lower CBF. The T2-hyperintense lesions were more generally distributed in both higher and lower perfused WM. This study suggests an association between hypoperfusion and T1-hypointense lesions.