Visualizing charges accumulated in an electric double layer by three-dimensional open-loop electric potential microscopy.
Charges accumulated in an electric double layer (EDL) play key roles in various interfacial phenomena and electronic devices. However, direct imaging of their spatial distribution has been a great challenge, which has hindered our nano-level understanding of the mechanisms of such interfacial phenomena and functions. In this study, we present direct imaging of charges accumulated at an electrode-electrolyte interface using three-dimensional open-loop electric potential microscopy (3D-OL-EPM). Conventional OL-EPM allows us to visualize two-dimensional potential distributions in liquid yet the zero of the measured potential is not well defined due to the influence of the long-range (LR) interaction between the cantilever and the sample. Here, we present practical ways to reduce such an influence by improving the equation for the potential calculation and subtracting the LR contribution estimated from a Z potential profile. These improvements enabled the calibration of the measured potential values with respect to the bulk solution potential. With these improvements, we visualized opposite charge accumulation behaviors on a polarizable and non-polarizable electrode with a varying electrode potential. Combining OL-EPM with a 3D tip scanning method, we also performed a 3D-OL-EPM measurement on a Cu fine wire and visualized the nanoscale distribution of the charges accumulated at the interface. Such real-space information on the charge distributions in an EDL should provide valuable insights into the mechanisms of interfacial phenomena and functions that are important in various academic and industrial research on electronic devices, electrochemistry, tribology and life sciences.


Alcohol and new onset atrial fibrillation: a case-control study of a current series.
The aetiological role of alcohol in new onset atrial fibrillation was evaluated in a case-control study of 100 consecutive patients aged 21-64 years. Clinical examination, routine diagnostic tests, and echocardiography revealed an underlying disease or other identifiable factor for atrial fibrillation in 65 patients (group 1); 35 patients had idiopathic atrial fibrillation (group 2). The most common diseases associated with atrial fibrillation were ischaemic heart disease (21%), hypertension (13%), and cardiomyopathy (8%). Data on alcohol consumption were obtained by interviewing the patients and their age and sex matched controls on admission. The mean daily alcohol intake of group 2 patients during the week preceding atrial fibrillation was significantly larger than that of either controls or group 1 patients. Compared with controls significantly more patients in both groups with atrial fibrillation had consumed alcohol within two days of the onset of the arrhythmia. Significantly more patients had onset of arrhythmia on Wednesday, Thursday, or Friday than on any other weekday, including patients with high alcohol intake. This study establishes alcohol as an important precipitating factor for new onset atrial fibrillation.


Regional and local environmental conditions do not shape the response to warming of a marine habitat-forming species.
The differential response of marine populations to climate change remains poorly understood. Here, we combine common garden thermotolerance experiments in aquaria and population genetics to disentangle the factors driving the population response to thermal stress in a temperate habitat-forming species: the octocoral Paramuricea clavata. Using eight populations separated from tens of meters to hundreds of kilometers, which were differentially impacted by recent mortality events, we identify 25 °C as a critical thermal threshold. After one week of exposure at this temperature, seven of the eight populations were affected by tissue necrosis and after 30 days of exposure at this temperature, the mean % of affected colonies increased gradually from 3 to 97%. We then demonstrate the weak relation between the observed differential phenotypic responses and the local temperature regimes experienced by each population. A significant correlation was observed between these responses and the extent of genetic drift impacting each population. Local adaptation may thus be hindered by genetic drift, which seems to be the main driver of the differential response. Accordingly, conservation measures should promote connectivity and control density erosion in order to limit the impact of genetic drift on marine populations facing climate change.


Characterization of aerosol optical properties, chemical composition and mixing states in the winter season in Shanghai, China.
Physical and chemical properties of ambient aerosols at the single particle level were studied in Shanghai from December 22 to 28, 2009. A Cavity-Ring-Down Aerosol Extinction Spectrometer (CRD-AES) and a nephelometer were deployed to measure aerosol light extinction and scattering properties, respectively. An Aerosol Time-of-Flight Mass Spectrometer (ATOFMS) was used to detect single particle sizes and chemical composition. Seven particle types were detected. Air parcels arrived at the sampling site from the vicinity of Shanghai until mid-day of December 25, when they started to originate from North China. The aerosol extinction, scattering, and absorption coefficients all dropped sharply when this cold, clean air arrived. Aerosol particles changed from a highly aged type before this meteorological shift to a relatively fresh type afterwards. The aerosol optical properties were dependent on the wind direction. Aerosols with high extinction coefficient and scattering Ångström exponent (SAE) were observed when the wind blew from the west and northwest, indicating that they were predominantly fine particles. Nitrate and ammonium correlated most strongly with the change in aerosol optical properties. In the elemental carbon/organic carbon (ECOC) particle type, the diurnal trends of single scattering albedo (SSA) and elemental carbon (EC) signal intensity had a negative correlation. We also found a negative correlation (r=-0.87) between high mass-OC particle number fraction and the SSA in a relatively clean period, suggesting that particulate aromatic components might play an important role in light absorption in urban areas.


Dissipation of Three Fungicides and Their Effects on Anthocyanins and Color of Monastrell Red Wines.
The effect of fungicides on fermentation is of paramount importance to control the quality and safety of wines. In this work, the quality (enological parameters, color, phenolic content, antioxidant activity, and fungicide residues) of wines from Monastrell grapes fortified with iprovalicarb, mepanipyrim, and tetraconazole fungicides was evaluated. Along the winemaking process, initial residues of mepanipyrim and tetraconazole were removed in more than 90% while the dissipation of iprovalicarb was around 73%. Significant statistical differences were found in the presence of iprovalicarb and mepanipyrim residues, especially at the highest concentration assayed. For both fungicides, increases in the volatile acidity (between 4 and 8.6 times), the lactic acid content (between 8.6 and 20.5 times), the percentage of polymeric anthocyanins (between 1.3 and 1.7 times), and also a slight increase of the total phenolic index and the total anthocyanin content determined by spectrophotometry were observed. On the contrary, the total monomeric anthocyanins content decreased about 16.3% and 28.6% in the presence of iprovalicarb and mepanipyrim, respectively. These results could be related to a higher development of acetic acid or lactic bacteria in the presence of these fungicides. The color of the final wines was also different in comparison with the control, with a higher yellow component, color intensity, tonality, and hue angle because of pH changes in the medium. Tetraconazole fermentations had a more similar trend to the control wine, probably due to the lower concentration of this fungicide in the grape must at the initial time. No effects on the antioxidant activity was observed for any of the target fungicides. A multivariate statistical analysis was done to view the interrelationships between different variables (color and anthocyanins profile). The obtained model allowed the wines to be separated according to the fungicide treatment applied.


Global transcriptome and physiological responses of Acinetobacter oleivorans DR1 exposed to distinct classes of antibiotics.
The effects of antibiotics on environment-originated nonpathogenic Acinetobacter species have been poorly explored. To understand the antibiotic-resistance mechanisms that function in nonpathogenic Acinetobacter species, we used an RNA-sequencing (RNA-seq) technique to perform global gene-expression profiling of soil-borne Acinetobacter oleivorans DR1 after exposing the bacteria to 4 classes of antibiotics (ampicillin, Amp; kanamycin, Km; tetracycline, Tc; norfloxacin, Nor). Interestingly, the well-known two global regulators, the soxR and the rpoE genes are present among 41 commonly upregulated genes under all 4 antibiotic-treatment conditions. We speculate that these common genes are essential for antibiotic resistance in DR1. Treatment with the 4 antibiotics produced diverse physiological and phenotypic changes. Km treatment induced the most dramatic phenotypic changes. Examination of mutation frequency and DNA-repair capability demonstrated the induction of the SOS response in Acinetobacter especially under Nor treatment. Based on the RNA-seq analysis, the glyoxylate-bypass genes of the citrate cycle were specifically upregulated under Amp treatment. We also identified newly recognized non-coding small RNAs of the DR1 strain, which were also confirmed by Northern blot analysis. These results reveal that treatment with antibiotics of distinct classes differentially affected the gene expression and physiology of DR1 cells. This study expands our understanding of the molecular mechanisms of antibiotic-stress response of environment-originated bacteria and provides a basis for future investigations.


Dispositional mindfulness and employment status as predictors of resilience in third year nursing students: a quantitative study.
Nursing students will graduate into stressful workplace environments and resilience is an essential acquired ability for surviving the workplace. Few studies have explored the relationship between resilience and the degree of innate dispositional mindfulness, compassion, compassion fatigue and burnout in nursing students, including those who find themselves in the position of needing to work in addition to their academic responsibilities. This paper investigates the predictors of resilience, including dispositional mindfulness and employment status of third year nursing students from three Australian universities. Participants were 240 undergraduate, third year, nursing students. Participants completed a resilience measure (Connor-Davidson Resilience Scale, CD-RISC), measures of dispositional mindfulness (Cognitive and Affective Mindfulness Scale Revised, CAMS-R) and professional quality of life (The Professional Quality of Life Scale version 5, PROQOL5), such as compassion satisfaction, compassion fatigue and burnout. An observational quantitative successive independent samples survey design was employed. A stepwise linear regression was used to evaluate the extent to which predictive variables were related each to resilience. The predictive model explained 57% of the variance in resilience. Dispositional mindfulness subset acceptance made the strongest contribution, followed by the expectation of a graduate nurse transition programme acceptance, with dispositional mindfulness total score and employment greater than 20 hours per week making the smallest contribution. This was a resilient group of nursing students who rated high with dispositional mindfulness and exhibited hopeful and positive aspirations for obtaining a position in a competitive graduate nurse transition programme after graduation.


Enzymic and molecular characterization of NADP-dependent glyceraldehyde-3-phosphate dehydrogenase from Synechococcus PCC 7942: resistance of the enzyme to hydrogen peroxide.
NADP-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been purified to electrophoretic homogeneity from Synechococcus PCC 7942 cells. The native enzyme had a molecular mass of 160 kDa and consisted of four subunits with a molecular mass of 41 kDa. The activity was 6-fold higher with NADPH than with NADH; the apparent Km values for NADPH and NADH were 62 +/- 4.5 and 420 +/- 10.5 microM respectively. The gene encoding NADP-dependent GAPDH was cloned from the chromosomal DNA of Synechococcus 7942. A 1140 bp open reading frame, encoding an enzyme of 380 amino acid residues (approx.molecular mass of 41.3 kDa) was observed. The deduced amino acid sequence of the gene had a greater sequence similarity to the NADP-dependent and chloroplastic form than to the NAD-dependent and cytosolic form. The Synechococcus 7942 enzyme lacked one of the cysteines involved in the light-dependent regulation of the chloroplast enzymes of higher plants. The recombinant enzyme expressed in Escherichia coli as well as the native enzyme purified from Synechococcus 7942 cells were resistant to 1 mM H2O2.


Do children with autism learn to read more readily by computer assisted instruction or traditional book methods? A pilot study.
The study evaluates the progress of eight children aged 3-5 years with autism attending a specialist teaching unit in their development of reading skills in two conditions: computer instructed learning and book based learning. The authors developed a direct observation schedule to monitor autistic behaviours using computerized techniques. The children were matched by age, severity of autistic symptomatology and number of spoken words. They were initially randomly allocated to the computer or book condition and crossed over at 10 weeks. All of the children spent more time on task in the computer condition than in the book condition. By the end of the study after computer assisted learning, five of the eight children could reliably identify at least three words. It was found that children with autism spent more time on reading material when they accessed it through a computer and were less resistant to its use.


Osteo-chondroprogenitor cells are derived from Sox9 expressing precursors.
The transcription factor Sox9 is expressed in all chondroprogenitors and has an essential role in chondrogenesis. Sox9 is also expressed in other tissues, including central nervous system, neural crest, intestine, pancreas, testis, and endocardial cushions, and plays a crucial role in cell proliferation and differentiation in several of these tissues. To determine the cell fate of Sox9-expressing cells during mouse embryogenesis, we generated mice in which a Cre recombinase gene preceded by an internal ribosome entry site was inserted into the 3' untranslated region of the Sox9 gene (Sox9-Cre knock-in). In the developing skeleton, Sox9 was expressed before Runx2, an early osteoblast marker gene. Cell fate mapping by using Sox9-Cre;ROSA26 reporter (R26R) mice revealed that Sox9-expressing limb bud mesenchymal cells gave rise to both chondrocytes and osteoblasts. Furthermore, a mutant in which the Osterix gene was inactivated in Sox9-expressing cells exhibited a lack of endochondral and intramembranous ossification and a lack of mature osteoblasts comparable with Osterix-null mutants. In addition, Sox9-expressing limb bud mesenchymal cells also contributed to tendon and synovium formation. By using Sox9-Cre;R26R mice, we also were able to systematically follow Sox9-expressing cells from embryonic day 8.0 to 17.0. Our results showed that Sox9-expressing cells contributed to the formation of all cell types of the spinal cord, epithelium of the intestine, pancreas, and mesenchyme of the testis. Thus, our results strongly suggest that all osteo-chondroprogenitor cells, as well as progenitors in a variety of tissues, are derived from Sox9-expressing precursors during mouse embryogenesis.


Optical Coherence Tomography Used as a Modality to Delineate Basal Cell Carcinoma prior to Mohs Micrographic Surgery.
Optical coherence tomography (OCT) has potential as a modality for in vivo imaging of non-melanoma skin cancer (NMSC). By allowing identification of sub-surface margins of NMSC lesions, the use of OCT could improve the rate of complete excision and reduce the average number of stages during Mohs micrographic surgery (MMS). The objective of this study was to use OCT to delineate the apparent sub-surface margins of NMSC lesions prior to their excision by MMS. Lesions were scanned with reference to a physical marker on the skin, and the apparent margins were then identified from the OCT images and marked on the skin. Photographs of these margins and the Mohs defect were correlated and compared. OCT appears capable of visualizing the transition from lesional to normal tissue. In this case study, margins marked by use of the OCT system before surgery exhibit excellent correlation with the MMS defect. OCT offers the promise of better outcomes by enabling accurate margin mapping of NMSC in advance of MMS. Priorities now are to demonstrate this capability in a larger study, and to understand clearly indications and contraindications for use.


Visualization of local Ca2+ dynamics with genetically encoded bioluminescent reporters.
Measurements of local Ca2+ signalling at different developmental stages and/or in specific cell types is important for understanding aspects of brain functioning. The use of light excitation in fluorescence imaging can cause phototoxicity, photobleaching and auto-fluorescence. In contrast, bioluminescence does not require the input of radiative energy and can therefore be measured over long periods, with very high temporal resolution. Aequorin is a genetically encoded Ca(2+)-sensitive bioluminescent protein, however, its low quantum yield prevents dynamic measurements of Ca2+ responses in single cells. To overcome this limitation, we recently reported the bi-functional Ca2+ reporter gene, GFP-aequorin (GA), which was developed specifically to improve the light output and stability of aequorin chimeras [V. Baubet, et al., (2000) PNAS, 97, 7260-7265]. In the current study, we have genetically targeted GA to different microdomains important in synaptic transmission, including to the mitochondrial matrix, endoplasmic reticulum, synaptic vesicles and to the postsynaptic density. We demonstrate that these reporters enable 'real-time' measurements of subcellular Ca2+ changes in single mammalian neurons using bioluminescence. The high signal-to-noise ratio of these reporters is also important in that it affords the visualization of Ca2+ dynamics in cell-cell communication in neuronal cultures and tissue slices. Further, we demonstrate the utility of this approach in ex-vivo preparations of mammalian retina, a paradigm in which external light input should be controlled. This represents a novel molecular imaging approach for non-invasive monitoring of local Ca2+ dynamics and cellular communication in tissue or whole animal studies.


An extremely rare, remote intracerebral metastasis of oral cavity cancer: a case report.
Distant brain metastases from oral squamous cell carcinomas (OSCC) are extremely rare. Here we describe a case of a 53-year-old man with a primary OSCC who referred to the neurosurgical department because of epileptic seizures. MR imaging revealed an enhancing lesion in the right parietal lobe. A craniotomy with tumor removing was performed. Histopathological examination verified an invasive, minimally differentiated metastasis of the primary OSCC. The patient refused whole brain radiation therapy and died from pulmonary metastatic disease 10 months after the neurosurgical intervention without any cerebral recurrence. To the authors' knowledge, only two similar cases have been previously reported.


[Stapedo-Vestibular Ankylosis: Retrospective Study of Five Cases in São Tomé e Príncipe].
Otosclerosis is a common form of conductive hearing loss characterized by abnormal bone remodeling exclusively in the otic capsule. The prevalence of otosclerosis varies in racial populations and is described as being rare in black African populations. In this paper we aim to report five cases of clinical, and surgically confirmed, otosclerosis in black individuals, in São Tomé and Príncipe. Since February 2011, Ear, Nose and Throat consultations and surgeries specialty have been carried out at Dr. Ayres de Menezes Hospital in cooperation with the project 'Health for all'. A retrospective analysis was undertaken of the records of all patients subjected either to stapedectomy or partial stapedectomy until February 2014. Information regarding clinical presentation, audiometric data and surgery reports was recorded. Five adult patients underwent stapedectomy or partial stapedectomy. All of them presented with normal otoscopy, conductive or mixed hearing loss on audiogram and normal tympanometry with absent stapedial reflexes. None of the patients had signs of infection or history of head trauma. Three cases showed improvement in the air-bone gap after surgery. The other two were lost to follow-up. We documented and surgically confirmed five cases of clinical otosclerosis in this population. A thematic review was carried out and concluded that, despite being described as a rare event in this race, available literature on this topic is not enough to state that there is lower prevalence of otosclerosis amongst the African population. Even if not common, otosclerosis cannot be disregarded as a possible cause for conductive hearing loss among the population of São Tomé and Principe.


Participatory design in the development of an early therapy intervention for perinatal stroke.
Perinatal stroke is the leading cause of unilateral (hemiparetic) cerebral palsy, with life-long personal, social and financial consequences. Translational research findings indicate that early therapy intervention has the potential for significant improvements in long-term outcome in terms of motor function. By involving families and health professionals in the development and design stage, we aimed to produce a therapy intervention which they would engage with. Nine parents of children with hemiparesis and fourteen health professionals involved in the care of infants with perinatal stroke took part in peer review and focus groups to discuss evolving therapy materials, with revisions made iteratively. The materials and approach were also discussed at a meeting of the London Child Stroke Research Reference Group. Focus group data were coded using Normalisation Process Theory constructs to explore potential barriers and facilitators to routine uptake of the intervention. We developed the Early Therapy in Perinatal Stroke (eTIPS) program - a parent-delivered, home-based complex intervention addressing a current gap in practice for infants in the first 6 months of life after unilateral perinatal stroke and with the aim of improving motor outcome. Parents and health professionals saw the intervention as different from usual practice, and valuable (high coherence). They were keen to engage (high cognitive participation). They considered the tasks for parents to be achievable (high collective action). They demonstrated trust in the approach and felt that parents would undertake the recommended activities (high collective action). They saw the approach as flexible and adaptable (high reflexive monitoring). Following suggestions made, we added a section on involving the extended family, and obtained funding for a website and videos to supplement written materials. Focus groups with parents and health professionals provided meaningful feedback to iteratively improve the intervention materials prior to embarking on a pilot study. The intervention has a high potential to normalize and become a routine part of parents' interactions with their child following unilateral perinatal stroke.


Predictive Mortality Index for Community-Dwelling Elderly Koreans.
There are very few predictive indexes for long-term mortality among community-dwelling elderly Asian individuals, despite its importance, given the rapid and continuous increase in this population. We aimed to develop 10-year predictive mortality indexes for community-dwelling elderly Korean men and women based on routinely collected clinical data.We used data from 2244 elderly individuals (older than 60 years of age) from the southwest Seoul Study, a prospective cohort study, for the development of a prognostic index. An independent longitudinal cohort of 679 elderly participants was selected from the Korean Genome Epidemiology Study in Ansan City for validation.During a 10-year follow-up, 393 participants (17.5%) from the development cohort died. Nine risk factors were identified and weighed in the Cox proportional regression model to create a point scoring system: age, male sex, smoking, diabetes, systolic blood pressure, triglyceride, total cholesterol, white blood cell count, and hemoglobin. In the development cohort, the 10-year mortality risk was 6.6%, 14.8%, 18.2%, and 38.4% among subjects with 1 to 4, 5 to 7, 8 to 9, and ≥10 points, respectively. In the validation cohort, the 10-year mortality risk was 5.2%, 12.0%, 16.0%, and 16.0% according to these categories. The C-statistic for the point system was 0.73 and 0.67 in the development and validation cohorts, respectively.The present study provides valuable information for prognosis among elderly Koreans and may guide individualized approaches for appropriate care in a rapidly aging society.


Breast cancer risk markedly lower with serum 25-hydroxyvitamin D concentrations ≥60 vs <20 ng/ml (150 vs 50 nmol/L): Pooled analysis of two randomized trials and a prospective cohort.
While numerous epidemiologic studies have found an association between higher serum 25-hydroxyvitamin D [25(OH)D] concentrations and lower breast cancer risk, few have assessed this association for concentrations >40 ng/ml. To investigate the relationship between 25(OH)D concentration and breast cancer risk across a broad range of 25(OH)D concentrations among women aged 55 years and older. Analyses used pooled data from two randomized clinical trials (N = 1129, N = 2196) and a prospective cohort (N = 1713) to examine a broad range of 25(OH)D concentrations. The outcome was diagnosis of breast cancer during the observation periods (median: 4.0 years). Three analyses were conducted: 1) Incidence rates were compared according to 25(OH)D concentration from <20 to ≥60 ng/ml (<50 to ≥150 nmol/L), 2) Kaplan-Meier plots were developed and 3) multivariate Cox regression was used to examine the association between 25(OH)D and breast cancer risk using multiple 25(OH)D measurements. Within the pooled cohort (N = 5038), 77 women were diagnosed with breast cancer (age-adjusted incidence: 512 cases per 100,000 person-years). Results were similar for the three analyses. First, comparing incidence rates, there was an 82% lower incidence rate of breast cancer for women with 25(OH)D concentrations ≥60 vs <20 ng/ml (Rate Ratio = 0.18, P = 0.006). Second, Kaplan-Meier curves for concentrations of <20, 20-39, 40-59 and ≥60 ng/ml were significantly different (P = 0.02), with the highest proportion breast cancer-free in the ≥60 ng/ml group (99.3%) and the lowest proportion breast cancer-free in the <20 ng/ml group (96.8%). The proportion with breast cancer was 78% lower for ≥60 vs <20 ng/ml (P = 0.02). Third, multivariate Cox regression revealed that women with 25(OH)D concentrations ≥60 ng/ml had an 80% lower risk of breast cancer than women with concentrations <20 ng/ml (HR = 0.20, P = 0.03), adjusting for age, BMI, smoking status, calcium supplement intake, and study of origin. Higher 25(OH)D concentrations were associated with a dose-response decrease in breast cancer risk with concentrations ≥60 ng/ml being most protective.


Gain of MYC underlies recurrent trisomy of the MYC chromosome in acute promyelocytic leukemia.
Gain of chromosome 8 is the most common chromosomal gain in human acute myeloid leukemia (AML). It has been hypothesized that gain of the MYC protooncogene is of central importance in trisomy 8, but the experimental data to support this are limited and controversial. In a mouse model of promyelocytic leukemia in which the MRP8 promoter drives expression of the PML-RARA fusion gene in myeloid cells, a Myc allele is gained in approximately two-thirds of cases as a result of trisomy for mouse chromosome 15. We used this model to test the idea that MYC underlies acquisition of trisomy in AML. We used a retroviral vector to drive expression of wild-type, hypermorphic, or hypomorphic MYC in bone marrow that expressed the PML-RARA transgene. MYC retroviruses cooperated in myeloid leukemogenesis and suppressed gain of chromosome 15. When the PML-RARA transgene was expressed in a Myc haploinsufficient background, we observed selection for increased copies of the wild-type Myc allele concomitant with leukemic transformation. In addition, we found that human myeloid leukemias with trisomy 8 have increased MYC. These data show that gain of MYC can contribute to the pathogenic effect of the most common trisomy of human AML.


Nucleolar organiser regions in adenocarcinoma in situ and invasive adenocarcinoma of the cervix.
Silver binding nucleolar regions (AgNORs) were evaluated in normal endocervix, adenocarcinoma, and its potential precursor, adenocarcinoma in situ (AIS), in an attempt to increase an understanding of the natural history of cervical adenocarcinoma and to identify a marker for abnormal endocervical (atypical glandular) cells which could aid diagnosis and follow up of endocervical lesions. For every 50 cells the mean AgNOR counts were as follows: normal endocervical cells (n = 15) 79.8 (95% Cl 68-91); AIS (n = 20) 200.7 (95% Cl 182-219); and invasive adenocarcinoma (n = 30) 299 (271-328). There was no overlap between the groups of normal endocervical cells and invasive adenocarcinoma, but there was significant overlap between cases of invasive adenocarcinoma and carcinoma in situ. In six out of 17 cases with AIS, NOR count in adjacent morphologically normal glandular cells ("internal" controls) was increased when compared with the "external" (normal endocervical) control group. This suggests the presence of wider field changes not previously identified using routine histological methods. The findings suggest that AIS is a potential premalignant precursor of invasive adenocarcinoma, but that assessment of NORs is of no practical use in discriminating between the histological types of cervical carcinoma.


Modality-dependent "what" and "where" preparatory processes in auditory and visual systems.
The present study examined the modality specificity and spatio-temporal dynamics of "what" and "where" preparatory processes in anticipation of auditory and visual targets using ERPs and a cue-target paradigm. Participants were presented with an auditory (Experiment 1) or a visual (Experiment 2) cue that signaled them to attend to the identity or location of an upcoming auditory or visual target. In both experiments, participants responded faster to the location compared to the identity conditions. Multivariate spatio-temporal partial least square (ST-PLS) analysis of the scalp-recorded data revealed supramodal "where" preparatory processes between 300-600 msec and 600-1200 msec at central and posterior parietal electrode sites in anticipation of both auditory and visual targets. Furthermore, preparation for pitch processing was captured at modality-specific temporal regions between 300 and 700 msec, and preparation for shape processing was detected at occipital electrode sites between 700 and 1150 msec. The spatio-temporal patterns noted above were replicated when a visual cue signaled the upcoming response (Experiment 2). Pitch or shape preparation exhibited modality-dependent spatio-temporal patterns, whereas preparation for target localization was associated with larger amplitude deflections at multimodal, centro-parietal sites preceding both auditory and visual targets. Using a novel paradigm, the study supports the notion of a division of labor in the auditory and visual pathways following both auditory and visual cues that signal identity or location response preparation to upcoming auditory or visual targets.


A Rab-E GTPase mutant acts downstream of the Rab-D subclass in biosynthetic membrane traffic to the plasma membrane in tobacco leaf epidermis.
The function of the Rab-E subclass of plant Rab GTPases in membrane traffic was investigated using a dominant-inhibitory mutant (RAB-E1(d)[NI]) of Arabidopsis thaliana RAB-E1(d) and in vivo imaging approaches that have been used to characterize similar mutants in the plant Rab-D2 and Rab-F2 subclasses. RAB-E1(d)[NI] inhibited the transport of a secreted green fluorescent protein marker, secGFP, but in contrast with dominant-inhibitory RAB-D2 or RAB-F2 mutants, it did not affect the transport of Golgi or vacuolar markers. Quantitative imaging revealed that RAB-E1(d)[NI] caused less intracellular secGFP accumulation than RAB-D2(a)[NI], a dominant-inhibitory mutant of a member of the Arabidopsis Rab-D2 subclass. Furthermore, whereas RAB-D2(a)[NI] caused secGFP to accumulate exclusively in the endoplasmic reticulum, RAB-E1(d)[NI] caused secGFP to accumulate additionally in the Golgi apparatus and a prevacuolar compartment that could be labeled by FM4-64 and yellow fluorescent protein (YFP)-tagged Arabidopsis RAB-F2(b). Using the vacuolar protease inhibitor E64-d, it was shown that some secGFP was transported to the vacuole in control cells and in the presence of RAB-E1(d)[NI]. Consistent with the hypothesis that secGFP carries a weak vacuolar-sorting determinant, it was shown that a secreted form of DsRed reaches the apoplast without appearing in the prevacuolar compartment. When fused to RAB-E1(d), YFP was targeted specifically to the Golgi via a saturable nucleotide- and prenylation-dependent mechanism but was never observed on the prevacuolar compartment. We propose that RAB-E1(d)[NI] inhibits the secretory pathway at or after the Golgi, causing an accumulation of secGFP in the upstream compartments and an increase in the quantity of secGFP that enters the vacuolar pathway.


Elective Nodal Irradiation Attenuates the Combinatorial Efficacy of Stereotactic Radiation Therapy and Immunotherapy.
Purpose: In the proper context, radiotherapy can promote antitumor immunity. It is unknown if elective nodal irradiation (ENI), a strategy that irradiates tumor-associated draining lymph nodes (DLN), affects adaptive immune responses and combinatorial efficacy of radiotherapy with immune checkpoint blockade (ICB).Experimental Design: We developed a preclinical model to compare stereotactic radiotherapy (Tumor RT) with or without ENI to examine immunologic differences between radiotherapy techniques that spare or irradiate the DLN.Results: Tumor RT was associated with upregulation of an intratumoral T-cell chemoattractant chemokine signature (CXCR3, CCR5-related) that resulted in robust infiltration of antigen-specific CD8+ effector T cells as well as FoxP3+ regulatory T cells (Tregs). The addition of ENI attenuated chemokine expression, restrained immune infiltration, and adversely affected survival when combined with ICB, especially with anti-CLTA4 therapy. The combination of stereotactic radiotherapy and ICB led to long-term survival in a subset of mice and was associated with favorable CD8 effector-to-Treg ratios and increased intratumoral density of antigen-specific CD8+ T cells. Although radiotherapy technique (Tumor RT vs. ENI) affected initial tumor control and survival, the ability to reject tumor upon rechallenge was partially dependent upon the mechanism of action of ICB; as radiotherapy/anti-CTLA4 was superior to radiotherapy/anti-PD-1.Conclusions: Our results highlight that irradiation of the DLN restrains adaptive immune responses through altered chemokine expression and CD8+ T-cell trafficking. These data have implications for combining radiotherapy and ICB, long-term survival, and induction of immunologic memory. Clinically, the immunomodulatory effect of the radiotherapy strategy should be considered when combining stereotactic radiotherapy with immunotherapy. Clin Cancer Res; 24(20); 5058-71. ©2018 AACR.


Identification and comparison of two sequence elements that confer cell-type specific transcription in yeast.
The MAT alpha 2 protein of budding yeast represses a set of genes; if the MATa1 protein is also present, a further set of genes is repressed. DNA sequence comparisons reveal a 20-base pair 'operator' sequence that is present in genes repressed by a1/alpha 2. A related, but distinct, sequence is found in genes repressed by alpha 2 alone.


Treatment of coronary artery disease from the inside: Light at the end of the tunnel?
Surgical treatment of coronary heart disease has shown its life saving benefits in millions of patients for more than half a century. Attempts to create less invasive ways to achieve similar results have attracted great attention since the introduction of balloon angioplasty in 1977. The fascination with such techniques was hampered by a 30% recurrence rate and a 5% rate of abrupt closure, requiring emergency bypass surgery in most instances. Angioplasty lost much of its unpredictability with the introduction of stents in 1986. The use of potent anti-platelet regimes added further safety, but recurrence remained a definite thread until the introduction of anti-mitotic coatings during the early years of this century. Very late thrombosis, however, continues to haunt interventionist and patients. Vanishing poly-lactic acid stents time still fail to exhibit comparable mechanical properties at present. So far bypass surgery has not yet lost its attraction in complicated and diffuse disease, despite its much higher patient discomfort.


Changing patterns of mobility as a response to climatic deterioration and aridification in the middle Holocene Southern Sahara.
The African Humid Period witnessed a rapid human re-occupation of the Sahara as numerous lakes formed during the Holocene climatic optimum circa 10-5 kya. Permanent waters attracted a variety of aquatic and terrestrial fauna allowing for long-term occupation of specific paleolake basins. The Gobero paleolake in central Niger was one such location that preserves a unique mortuary record from the southern Sahara. Here, we use radiogenic strontium isotope analysis to investigate how human communities adapted to aridification throughout the Holocene. In particular, we examine the effects of increasing climate instability on patterns of human mobility. Results of radiogenic strontium isotope analysis of enamel and bone samples from Middle Holocene burials (∼7.2-4.9 kya) indicate predominantly local values with no evidence for sex-based variation. Comparisons of radiogenic strontium isotope data with previously published (Stojanowski and Knudson: Am J Phys Anthropol 146 (2011) 49-61) Early Holocene burials (∼9.7-8.3 kya) indicate significant differences in both enamel and bone values. Middle Holocene individuals demonstrate a predominantly non-local signature for enamel values and a predominantly local signature for bone values. Those individuals with non-local bone values always demonstrated non-local enamel values; however, the opposite was not the case. This suggests a divergence of mobility strategies during the Middle Holocene with a minority of individuals maintaining a more mobile existence throughout their life and others maintaining a similar strategy as Early Holocene hunter-gatherers that was tied to the paleolake basin. The more mobile individuals likely lived during the terminal phase of the lake's occupation. One response to aridification by Saharan peoples, then, was increasing mobility.


Predicting vehicle belt fit for children ages 6-12.
To predict shoulder belt fit and lap belt fit as a function of child age, vehicle seat characteristics, and belt geometry. In a previous study, the lap belt and shoulder belt fit of 44 children aged 5-12 were measured in a simulated vehicle seat while varying cushion length, cushion angle, seatback angle, and belt anchorage geometry. A regression model was developed to predict lap belt fit and shoulder belt fit as a function of vehicle parameters and child stature. These regression models were applied to the stature distribution of 6- to 12-year-olds using a range of vehicle geometry data to predict the proportion of children expected to achieve good belt fit in the second-row, outboard seating positions of 46 vehicles when not using belt-positioning boosters. Across the ranges observed in vehicles, lap belt angle had the strongest effect on lap belt fit, although vehicle cushion length also contributed. Shoulder belt fit was most strongly affected by D-ring location. Vehicles with the geometric conditions most suitable for children are estimated to provide good lap belt fit for 25% of children aged 6 to 12. In 20% of vehicles, the shoulder belt is too far inboard for the target child population; 20% of vehicles are estimated to have shoulder belt fit too far outboard for children ages 6 to 12. Based on this geometric analysis, the rear seats of most vehicles are unlikely to provide good lap belt fit for up to 75% of children ages 6-12. Shoulder belt fit is outside the target range for 40% of children. Consequently, children under 12 years of age are likely to experience markedly poorer belt fit when transitioning out of a booster seat.


The evolution of sexual dimorphism in the house finch. I. Population divergence in morphological covariance structure.
Patterns of genetic variation and covariation strongly affect the rate and direction of evolutionary change by limiting the amount and form of genetic variation available to natural selection. We studied evolution of morphological variance-covariance structure among seven populations of house finches (Carpodacus mexicanus) with a known phylogenetic history. We examined the relationship between within- and among-population covariance structure and, in particular, tested the concordance between hierarchical changes in morphological variance-covariance structure and phylogenetic history of this species. We found that among-population morphological divergence in either males or females did not follow the within-population covariance patterns. Hierarchical patterns of similarity in morphological covariance matrices were not congruent with a priori defined historical pattern of population divergence. Both of these results point to the lack of proportionality in morphological covariance structure of finch populations, suggesting that random drift alone is unlikely to account for observed divergence. Furthermore, drift alone cannot explain the sex differences in within- and among-population covariance patterns or sex-specific patterns of evolution of covariance structure. Our results suggest that extensive among-population variation in sexual dimorphism in morphological covariance structure was produced by population differences in local selection pressures acting on each sex.


Single-Cell Sequencing of iPSC-Dopamine Neurons Reconstructs Disease Progression and Identifies HDAC4 as a Regulator of Parkinson Cell Phenotypes.
Induced pluripotent stem cell (iPSC)-derived dopamine neurons provide an opportunity to model Parkinson's disease (PD), but neuronal cultures are confounded by asynchronous and heterogeneous appearance of disease phenotypes in vitro. Using high-resolution, single-cell transcriptomic analyses of iPSC-derived dopamine neurons carrying the GBA-N370S PD risk variant, we identified a progressive axis of gene expression variation leading to endoplasmic reticulum stress. Pseudotime analysis of genes differentially expressed (DE) along this axis identified the transcriptional repressor histone deacetylase 4 (HDAC4) as an upstream regulator of disease progression. HDAC4 was mislocalized to the nucleus in PD iPSC-derived dopamine neurons and repressed genes early in the disease axis, leading to late deficits in protein homeostasis. Treatment of iPSC-derived dopamine neurons with HDAC4-modulating compounds upregulated genes early in the DE axis and corrected PD-related cellular phenotypes. Our study demonstrates how single-cell transcriptomics can exploit cellular heterogeneity to reveal disease mechanisms and identify therapeutic targets.


Overcoming challenges to dissemination and implementation of research findings in under-resourced countries.
Louis Pasteur once commented on the happiness that a scientist finds when, besides making a discovery, study results find practical application. Where health status is poor and resources are limited, finding such applications is a necessity, not merely a joy.Dissemination, or the distribution of new knowledge gained through research, is essential to the ethical conduct of research. Further, when research is designed to improve health, dissemination is critical to the development of evidence-based medicine and the adoption of evidence-supported interventions and improved practice patterns within specific settings. When dissemination is lacking, research may be considered a waste of resources and a useless pursuit unable to influence positive health outcomes.Effective translation of the findings of health research into policy and the practice of medicine has been slow in many countries considered low or lower middle-income (as defined by the World Bank). This is because such countries often have health care systems that are under-resourced (e.g., lacking personnel or facilities) and thus insufficiently responsive to health needs of their populations. However, implementation research has produced many tools and strategies that can prompt more effective and timelier application of research findings to real world situations.A conscientious researcher can find many suggestions for improving the integration of research evidence into practice. First and foremost, the truthful reporting of results is emphasized as essential because both studies with desirable findings as well those with less than ideal results can provide new and valuable knowledge. Consideration in advance of the audience likely to be interested in study findings can result in suitable packaging and targeted communication of results. Other strategies for avoiding the barriers that can negatively impact implementation of research evidence include the early involvement of stakeholders as research is being designed and discussion before initiation of proposed research with those who will be affected by it. It is also important to recognize the role of education and training for ensuring the skills and knowledge needed for not only the conduct of high quality research but also for the meaningful promotion of results and application of research findings to achieve intended purposes.


Inferring yeast cell cycle regulators and interactions using transcription factor activities.
Since transcription factors are often regulated at the post-transcriptional level, their activities, rather than expression levels may provide valuable information for investigating functions and their interactions. The recently developed Network Component Analysis (NCA) and its generalized form (gNCA) provide a robust framework for deducing the transcription factor activities (TFAs) from various types of DNA microarray data and transcription factor-gene connectivity. The goal of this work is to demonstrate the utility of TFAs in inferring transcription factor functions and interactions in Saccharomyces cerevisiae cell cycle regulation. Using gNCA, we determined 74 TFAs from both wild type and fkh1 fkh2 deletion mutant microarray data encompassing 1529 ORFs. We hypothesized that transcription factors participating in the cell cycle regulation exhibit cyclic activity profiles. This hypothesis was supported by the TFA profiles of known cell cycle factors and was used as a basis to uncover other potential cell cycle factors. By combining the results from both cluster analysis and periodicity analysis, we recovered nearly 90% of the known cell cycle regulators, and identified 5 putative cell cycle-related transcription factors (Dal81, Hap2, Hir2, Mss11, and Rlm1). In addition, by analyzing expression data from transcription factor knockout strains, we determined 3 verified (Ace2, Ndd1, and Swi5) and 4 putative interaction partners (Cha4, Hap2, Fhl1, and Rts2) of the forkhead transcription factors. Sensitivity of TFAs to connectivity errors was determined to provide confidence level of these predictions. By subjecting TFA profiles to analyses based upon physiological signatures we were able to identify cell cycle related transcription factors consistent with current literature, transcription factors with potential cell cycle dependent roles, and interactions between transcription factors.


Manipulating reproductive effort leads to changes in female reproductive scheduling but not oxidative stress.
The trade-off between reproductive investment and lifespan is the single most important concept in life-history theory. A variety of sources of evidence support the existence of this trade-off, but the physiological costs of reproduction that underlie this relationship remain poorly understood. The Free Radical Theory of Ageing suggests that oxidative stress, which occurs when there is an imbalance between the production of damaging Reactive Oxygen Species (ROS) and protective antioxidants, may be an important mediator of this trade-off. We sought to test this theory by manipulating the reproductive investment of female mice (Mus musculus domesticus) and measuring the effects on a number of life history and oxidative stress variables. Females with a greater reproductive load showed no consistent increase in oxidative damage above females who had a smaller reproductive load. The groups differed, however, in their food consumption, reproductive scheduling and mean offspring mass. Of particular note, females with a very high reproductive load delayed blastocyst implantation of their second litter, potentially mitigating the costs of energetically costly reproductive periods. Our results highlight that females use strategies to offset particularly costly periods of reproduction and illustrate the absence of a simple relationship between oxidative stress and reproduction.


Nitric oxide-releasing nanoparticles improve doxorubicin anticancer activity.
Anticancer drug delivery systems are often limited by hurdles, such as off-target distribution, slow cellular internalization, limited lysosomal escape, and drug resistance. To overcome these limitations, we have developed a stable nitric oxide (NO)-releasing nanoparticle (polystyrene-maleic acid [SMA]-tert-dodecane S-nitrosothiol [tDodSNO]) with the aim of enhancing the anticancer properties of doxorubicin (Dox) and a Dox-loaded nanoparticle (SMA-Dox) carrier. Effects of SMA-tDodSNO and/or in combination with Dox or SMA-Dox on cell viability, apoptosis, mitochondrial membrane potential, lysosomal membrane permeability, tumor tissue, and tumor growth were studied using in vitro and in vivo model of triple-negative breast cancer (TNBC). In addition, the concentrations of SMA-Dox and Dox in combination with SMA-tDodSNO were measured in cells and tumor tissues. Combination of SMA-tDodSNO and Dox synergistically decreased cell viability and induced apoptosis in 4T1 (TNBC cells). Incubation of 4T1 cells with SMA-tDodSNO (40 µM) significantly enhanced the cellular uptake of SMA-Dox and increased Dox concentration in the cells resulting in a twofold increase (P<0.001). Lysosomal membrane integrity, evaluated by acridine orange (AO) staining, was impaired by 40 µM SMA-tDodSNO (P<0.05 vs control) and when combined with SMA-Dox, this effect was significantly potentiated (P<0.001 vs SMA-Dox). Subcutaneous administration of SMA-tDodSNO (1 mg/kg) to xenografted mice bearing 4T1 cells showed that SMA-tDodSNO alone caused a twofold decrease in the tumor size compared to the control group. SMA-tDodSNO in combination with SMA-Dox resulted in a statistically significant 4.7-fold reduction in the tumor volume (P<0.001 vs control), without causing significant toxicity as monitored through body weight loss. Taken together, these results suggest that SMA-tDodSNO can be used as a successful strategy to increase the efficacy of Dox and SMA-Dox in a model of TNBC.


Linguistic analysis of communication in therapist-assisted internet-delivered cognitive behavior therapy for generalized anxiety disorder.
Therapist-assisted Internet-delivered cognitive behavior therapy (ICBT) involves elements of expressive writing through secure messaging with a therapist. Expressive writing has been associated with psychological and physical health benefits in past research; furthermore, certain linguistic dimensions in expressive writing have been identified as particularly beneficial to health, such as less frequent use of negative emotion words and greater use of positive emotion words. No research, to date, has analyzed linguistic dimensions in client communication over the course of therapist-assisted ICBT for individuals with symptoms of generalized anxiety. This naturalistic study examined messages sent to therapists during the course of ICBT using linguistic analysis, and explored covariation of word use with symptom improvement. Data were obtained from patients with symptoms of generalized anxiety (N = 59) who completed 12 modules of therapist-assisted ICBT and rated symptoms of anxiety, depression, and panic at the beginning of each module. Linguistic analysis categorized text submitted to therapists into different word categories. Results found that patients' use of negative emotion, anxiety, causation, and insight words reduced over the course of treatment, while past tense words increased. Furthermore, negative emotion words significantly covaried with symptom ratings over the course of treatment. While causal statements cannot be made, findings improve our understanding of patient communication in ICBT and suggest that the further study of linguistic dimensions as psychological indicators and the potential utility of expressive writing strategies in therapist-assisted ICBT may be worthwhile.


Physical activity levels before and after a diagnosis of breast carcinoma: the Health, Eating, Activity, and Lifestyle (HEAL) study.
Increased body weight at the time patients are diagnosed with breast carcinoma has been associated with an increased risk of recurrence and reduced survival. Weight gain also is common after diagnosis. Increasing physical activity (PA) after diagnosis may minimize these adverse outcomes. In this population-based study, the authors investigated whether PA levels after diagnosis declined from prediagnosis levels and whether any changes in PA varied by disease stage, adjuvant treatment, patient age, or body mass index (BMI) in 812 patients with incident breast carcinoma (from in situ to Stage IIIa). Types of sports and household activities and their frequency and duration for the year prior to diagnosis and for the month prior to the interview (i.e., 4-12 months postdiagnosis) were assessed during a baseline interview. Patients decreased their total PA by an estimated 2.0 hours per week from prediagnosis to postdiagnosis, an 11% decrease (P < 0.05). Greater decreases in sports PA were observed among women who were treated with radiation and chemotherapy (50% decrease) compared with women who underwent surgery only (24% decrease) or who were treated with radiation only (23%; (P < 0.05). Greater decreases in sports PA were observed among obese patients (41% decrease) compared with patients of normal weight (24% decrease; P < 0.05). PA levels were reduced significantly after patients were diagnosed with breast carcinoma. Greater decreases in PA observed among heavier patients implied a potential for greater weight gain among women who already were overweight. Randomized, controlled trials are needed to evaluate how PA may improve the prognosis for patients with breast carcinoma.


Assessing the Taxonomic Validity of Austrodiplostomum SPP. (Digenea: Diplostomidae) Through Nuclear and Mitochondrial Data.
Adults of the genus Austrodiplostomum are parasites in cormorants of the New World, whereas metacercariae are parasites from eye globe and brain of freshwater and brackish water fishes. In this study, specimens of Austrodiplostomum mordax from South America (type-species) were analyzed together with other specimens of Austrodiplostomum spp. collected from several locations across Middle America and North America. Partial DNA sequences of the mitochondrial gene cytochrome c oxidase subunit I ( COI), the internal transcribed spacers ( ITS1, ITS2, and 5.8S gene), and the D2-D3, domains of the large subunit ( LSU) of nuclear ribosomal DNA, were generated for both developmental stages and compared with available sequences of Austrodiplostomum spp. Phylogenetic analyses inferred with each molecular marker using maximum likelihood and Bayesian inference revealed the existence of 4 lineages representing 2 described species, A. mordax and Austrodiplostomum compactum (syn. Austrodiplostomum ostrowskiae) and 2 undescribed species of Austrodiplostomum recognized in previous studies. The COI haplotype network inferred with 172 sequences detected 28 haplotypes divided into 4 clusters, separating each other by 33 and 40 substitutions and with a genetic divergence ranging from 9 to 12%. The largest group included specimens identified as A. compactum plus those identified as A. ostrowskiae, supporting the synonymy of both species. As a result, we conclude that A. compactum is widely distributed across the Americas, in locations of the United States, Mexico, El Salvador, Honduras, Costa Rica, Venezuela, Peru, and Brazil. The other 2 undescribed species of the genus Austrodiplostomum were previously recorded in the United States and now are reported in Mexico. These 2 species cannot be described because adult forms have not been found in their definitive hosts. Finally, the species A. mordax has been found only in some lakes from Argentina, and it was validated in this study through molecular analyses.


Gabapentin for postoperative pain after photorefractive keratectomy: a prospective, randomized, double-blind, placebo-controlled trial.
To determine whether treatment with oral gabapentin reduces postoperative pain after photorefractive keratectomy (PRK). This prospective, randomized, double-blind, placebo-controlled study comprised 40 patients scheduled for bilateral PRK. Exclusion criteria were previous refractive surgery; diseases that could affect epithelial healing; use of antihistamines, nonsteroidal anti-inflammatory drugs, or steroids; and use of mitomycin C during PRK. Patients were divided into two groups: 20 patients received gabapentin capsules (300 mg) and 20 patients received identical placebo capsules. Visual acuity, slit-lamp examination, and a 12-question, 10-point visual numerical scale questionnaire were evaluated at each postoperative follow-up. Main outcome measures were the severity of pain at each follow-up and during the previous 24 hours. Secondary outcome measures were maximum and minimum severity of pain, healing time, uncorrected distance visual acuity, and the rating of other symptoms and potential side effects. The gabapentin group had less pain during the first 72 hours (P=.024 to .001). A significant difference in favor of gabapentin was found for the first 48 hours regarding minimum pain severity (P=.005 and .01 for 24 and 48 hours, respectively) and for the first 72 hours for maximum pain severity (P=.014, .007, and .001 for 24, 48, and 72 hours, respectively). No significant differences were observed for side effects, symptoms, or healing time except discomfort in favor of gabapentin during the first 24 hours (P=.043). Gabapentin significantly reduced postoperative pain after PRK compared to placebo, with no increase in reported side effects.


Cardiac reflexes in a warming world: thermal plasticity of barostatic control and autonomic tones in a temperate fish.
Thermal plasticity of cardiorespiratory function allows ectotherms like fish to cope with seasonal temperature changes and is critical for resilience to climate change. Yet, the chronic thermal effects on cardiovascular homeostatic reflexes in fish are little understood although this may have important implications for physiological performance and overall resilience to climate warming. We compared cardiac autonomic control and baroreflex regulation of heart rate in perch (Perca fluviatilis L.) from a reference area in the Baltic Sea at 18-19°C with conspecifics from the Biotest enclosure, a chronically heated ecosystem receiving warmed effluent water (24-25°C) from a nuclear power plant. Resting heart rate of Biotest fish displayed clear thermal compensation and was 58.3±2.3 beats min-1 compared with 52.4±2.6 beats min-1 in reference fish at their respective environmental temperatures (Q10=1.2). The thermally compensated heart rate of Biotest fish was a combined effect of elevated inhibitory cholinergic tone (105% in Biotest fish versus 70% in reference fish) and reduced intrinsic cardiac pacemaker rate. A barostatic response was evident in both groups, as pharmacologically induced increases and decreases in blood pressure resulted in atropine-sensitive bradycardia and tachycardia, respectively. Yet, the tachycardia in Biotest fish was significantly greater, presumably due to the larger scope for vagal release. Acclimation of Biotest fish to 18°C for 3 weeks abolished differences in intrinsic heart rate and autonomic tone, suggesting considerable short-term thermal plasticity of cardiovascular control in this species. The heightened hypotensive tachycardia in Biotest perch may represent an important mechanism of ectothermic vertebrates that safeguards tissue perfusion pressure when tissue oxygen demand is elevated by environmental warming.


Self-other organization: why early life did not evolve through natural selection.
The improbability of a spontaneously generated self-assembling molecule has suggested that life began with a set of simpler, collectively replicating elements, such as an enclosed autocatalytic set of polymers (or protocell). Since replication occurs without a self-assembly code, acquired characteristics are inherited. Moreover, there is no strict distinction between alive and dead; one can only infer that a protocell was alive if it replicates. These features of early life render natural selection inapplicable to the description of its change-of-state because they defy its underlying assumptions. Moreover, natural selection describes only randomly generated novelty; it cannot describe the emergence of form at the interface between organism and environment. Self-organization is also inadequate because it is restricted to interactions amongst parts; it too cannot account for context-driven change. A modified version of selection theory or self-organization would not work because the description of change-of-state through interaction with an incompletely specified context has a completely different mathematical structure, i.e. entails a non-Kolmogorovian probability model. It is proposed that the evolution of early life is appropriately described as lineage transformation through context-driven actualization of potential (CAP), with self-organized change-of-state being a special case of no contextual influence, and competitive exclusion of less fit individuals through a selection-like process possibly (but not necessarily) playing a secondary role. It is argued that natural selection played an important role in evolution only after genetically mediated replication was established.


A biogeographic and ecological perspective to the evolution of reproductive behaviour in the family Salamandridae.
Amphibians have a complex reproductive behaviour, which shows the highest diversity among tetrapodes. The family Salamandridae, distributed across the entire Holarctic, is one of the most diverse groups of extant salamanders comprising 114 species in 21 genera. The family has a remarkable diversity of courtship modes, amplexus and sperm transfer. It is often hypothesised that this diversity has evolved in adaptation to a specific mating and/or breeding habitat. We test this hypothesis based upon a phylogenetic reconstruction using the complete mitochondrial genome sequences of 45 Salamandridae species, representing all existing genera. We used ancestral character state reconstruction methods and geographic range models and applied relaxed Bayesian molecular clock models to discuss the results in a temporal framework of Salamandridae evolution. Our results show that the family Salamandridae started to diversify in the Late Cretaceous (ca. 87 mya) and is of Western Palearctic origin. Ancestral character state reconstruction predicts that its common ancestor was oviparous, mated on land without amplexus and probably showed a pin wheel spermatophore transfer, which is still found in the Italian endemic Salamandrina terdigidata. Our results suggest that several colonization of continents with subsequent radiations took place, once to the Nearctic and twice into Eastern Asian realms. However, these events were only in one case associated with a change in mating behaviour (dorsal amplexus in Nearctic newts). Around the Cretaceous-Paleogene boundary (K-Pg boundary) several Salamandridae lineages further diverged, again with no obvious changes in mating behaviour. Overall, there is no significant signal for mating character evolution being caused by changes in habitat type, with only a slight tendency that changes in mating habitat might have led to changes in the type of sperm transfer which in turn was associated with changes in the presence or absence of amplexus.


Retroviral invasion of the koala genome.
Endogenous retroviruses are a common ancestral feature of mammalian genomes with most having been inactivated over time through mutation and deletion. A group of more intact endogenous retroviruses are considered to have entered the genomes of some species more recently, through infection by exogenous viruses, but this event has never been directly proved. We have previously reported koala retrovirus (KoRV) to be a functional virus that is associated with neoplasia. Here we show that KoRV also shows features of a recently inserted endogenous retrovirus that is vertically transmitted. The finding that some isolated koala populations have not yet incorporated KoRV into their genomes, combined with its high level of activity and variability in individual koalas, suggests that KoRV is a virus in transition between an exogenous and endogenous element. This ongoing dynamic interaction with a wild species provides an exciting opportunity to study the process and consequences of retroviral endogenization in action, and is an attractive model for studying the evolutionary event in which a retrovirus invades a mammalian genome.


[Quality of life and alexithymia in somatoform pain disorder].
Patients with somatoform pain disorders (SPD) frequently display reduced quality of life (QoL) and increased levels of alexithymia. This study investigated the association of QoL and alexithymia in a sample of SPD. Fifty-one patients with SPD (average time since onset: 11.6 years) were assessed in terms of alexithymia (TAS-20), QoL (WHOQOL-BREF), psychological distress and somatisation (SCL-90-R), and depression (MADRS). In SPD patients a significant negative correlation was observed between QoL and alexithymia, particularly the psychological domain of QoL and the TAS-20 total score (r=-.63, p<.001). The TAS-20 subscale "Difficulty Describing Feelings" was revealed to be a significant predictor of the psychological domain of QoL (beta=-.34, p<.01), even after controlling for depression, somatisation and gender. Patients with SPD show a remarkably reduced QoL and alexithymia appears to play a significant role for low QoL. Clinicians need to pay careful attention to alexithymia with regard to diagnosis and treatment planning in SPD patients.


Which components of medical homes reduce the time burden on families of children with special health care needs?
To examine which components of medical homes affect time families spend arranging/coordinating health care for their children with special health care needs (CSHCNs) and providing health care at home. 2009-2010 National Survey of Children with Special Health Care Needs (NS-CSHCN), a population-based survey of 40,242 CSHCNs. NS-CSHCN is a cross-sectional, observational study. We used generalized ordered logistic regression, testing for nonproportional odds in the associations between each of five medical home components and time burden, controlling for insurance, child health, and sociodemographics. Medical home components were collected using Child and Adolescent Health Measurement Initiative definitions. Family-centered care, care coordination, and obtaining needed referrals were associated with 15-32 percent lower odds of time burdens arranging/coordinating and 16-19 percent lower odds providing health care. All five components together were associated with lower odds of time burdens, with greater reductions for higher burdens providing care. Three of the five medical home components were associated with lower family time burdens arranging/coordinating and providing health care for children with chronic conditions. If the 55 percent of CSHCNs lacking medical homes had one, the share of families with time burdens arranging care could be reduced by 13 percent.


Using scenarios to capture work processes in shared home care.
Shared home care is increasingly common, and in order to develop ICT that support such complex cooperative work it is crucial obtain an understanding of the work routines, information demands, and other central preconditions at the clinical level before the development is initiated. Scenarios are proposed as a technique that can be useful for capturing work processes in shared home care and experiences from the Old@Home project are presented. The scenarios are useful not only in the initial phases of the development project but throughout the development process, improving the accessibility of end user requirements and usability issues for the design team, and as a basis for use cases and further design.


Soluble epidermal growth factor receptor (sEGFR) [corrected] and cancer antigen 125 (CA125) as screening and diagnostic tests for epithelial ovarian cancer.
Epithelial ovarian cancer (EOC) is the leading cause of death among all gynecologic cancers in the United States. Because women who are diagnosed with early stage disease have a better prognosis than women diagnosed with late stage disease, early detection represents a potentially practical approach to reduce the mortality associated with EOC. Unfortunately, no single screening test has proven to be effective for this purpose, and a valid and feasible screening program to detect early stage EOC in the general population has not yet been devised. Consequently, research has focused on coupling two or more screening modalities to improve program validity and feasibility. Serum cancer antigen 125 (CA125) and a soluble isoform of the epidermal growth factor receptor (p110 sEGFR) have been studied individually as biomarkers of ovarian cancer. In this study, we compare serum CA125 levels and sEGFR concentrations in women with EOC to women with benign gynecologic conditions of ovarian and non-ovarian origin. We show that serum sEGFR concentrations are lower in patients with EOC than in women with benign gynecologic conditions, whereas serum CA125 levels are higher in patients to EOC compared with women with benign gynecologic conditions. These data also reveal that age and serum sEGFR concentrations modify the association between CA125 levels and EOC versus benign gynecologic disease. Hence, age- and sEGFR-dependent CA125 cutoff thresholds improve the ability of CA125 to discern EOC patients from women with benign ovarian tumors and non-ovarian gynecologic conditions. Our analyses show that parallel testing with fixed sEGFR and CA125 cutoff thresholds optimizes sensitivity to detect EOC, whereas serial testing with age- and sEGFR-dependent CA125 cutoff thresholds optimizes test specificity, and overall accuracy to discern patients with EOC from women with benign ovarian and non-ovarian gynecologic conditions. The combined use of serologic sEGFR and CA125, thus, has improved utility for screening and diagnosing EOC, which may increase the positive predictive value of a multimodal screening program that incorporates these biomarkers to detect and subsequently differentiate benign from malignant ovarian tumors.


Conversion: Simulated Method of Exchanging Tourniquet Use for Pressure Dressing Use.
Given little data to assess guidelines, we sought a way to exchange one type of intervention, field tourniquet use, for another, use of a pressure dressing. The study purpose was to test performance of controlling simulated bleeding with a stepwise procedure of tourniquet conversion. An experiment was designed to assess 15 tests of a caregiver making tourniquet-dressing conversions. Tests were divided into trials: tourniquet use and its conversion. In laboratory conditions, the tourniquet trial was care under gunfire; then, the conversion trial was emergency healthcare. A HapMed Leg Tourniquet Trainer simulated a limb amputation. An investigator provided healthcare. Mean (± standard deviation [SD]) test time and blood loss were 9 ± 3.6 minutes and 334 ± 353.9mL, respectively. The first test took 17 minutes. By test number, times decreased; the last six took ≤7 minutes. All tourniquet trials controlled bleeding. Mean (±SD) tourniquet pressure and blood loss were 222 ± 18.0mmHg and 146 ± 40.9mL, respectively. Bleeding remained uncontrolled in one conversion. Initial attempts to wrap a dressing were effective in 73% of tries (n = 11 of 15). Four of 15 wrap attempts (27%) were repeated to troubleshoot bleeding recurrence, and the first three tests required a repetition. Mean (±SD) dressing pressures and blood losses were 141 ± 17.6mmHg and 188 ± 327.4mL, respectively. Unsatisfactory conversion trials had a dressing pressure <137mmHg. Dressings and wraps hid the wound to impair assessment of bleeding. In testing a method of converting a limb tourniquet to a pressure dressing, the caregiver performed faster with experience accrual. The tourniquet results were uniformly good, but conversion results were worse and more varied. Simulating conversion was disappointing on a manikin and indicated that its redesign might be needed to suit this method. The procedural method constituted a start for further development.


Morbidity and mortality of carotid endarterectomy: rates of occurrence in asymptomatic and symptomatic patients.
During a six-year period ending December 1979, a retrospective study of 252 carotid endarterectomies in 211 patients was undertaken to review morbidity and mortality, and factors related to them. For purposes of the study, patients were categorized into asymptomatic and symptomatic groups. Indications for operation were internal carotid artery stenosis that occluded 50% or more of the vessel, an ulcerated plaque proved by angiography, or both. There were no postoperative deaths or neurological complications in the asymptomatic group. In the symptomatic group, there was an overall postoperative mortality of 2% and a stroke rate of 2%. All postoperative CNS deaths and strokes occurred in patients in whom there was difficulty in controlling blood pressure postoperatively. We conclude that with regard to carotid endarterectomy for treatment of extracranial occlusive disease (1) there is an acceptable morbidity and mortality in the symptomatic patient, (2) in the asymptomatic patient the procedure is associated with minimal complications, and (3) strict control of blood pressure in the immediate postoperative period is a critical factor in preventing neurological morbidity and mortality.


Alterations of ionized and total magnesium levels in pregnant women with gestational diabetes mellitus.
The aim of this prospective study was to determine ionized and total magnesium (Mg) levels in pregnant subjects with and without gestational diabetes mellitus (GDM). Eighty-five women, 26-28 weeks pregnant, were recruited for routine oral glucose tolerance tests (OGTT); 45 had normal OGTT results and 40 were diagnosed with GDM. Electrolyte levels, including ionized and total Mg, were analyzed. Gestational age and BMI were similar between the two groups (p = 0.800, p = 0.025). Multivitamin use was higher in the control group (p = 0.036). Fasting blood glucose was higher in the GDM group (p < 0.001). The median total Mg levels were 1.9 mg/dl (range 1.6-2.2) in the control group and 1.8 mg/dl (range 1.2-2.1) in the GDM group (p < 0.001). The median ionized Mg levels were 0.5 mmol/l (range 0.4-0.6) in the control group and 0.4 mmol/l (range 0.4-0.5) in the GDM group (p < 0.001). Our study revealed a relationship between low total and ionized Mg levels and GDM, as in type 2 diabetes mellitus (DM). The literature regarding type 2 DM and our findings suggest that Mg is the key ion in the pathophysiology of GDM. Low-dose Mg supplementation was not related to GDM; however, pharmacological doses in the various stages of pregnancy could be beneficial and should be investigated.


Interchannel pulse collision in a wavelength-division-multiplexed system with strong dispersion management.
We develop a perturbation theory to analytically calculate the effects of complete and incomplete interchannel collisions of Gaussian pulses in a wavelength-division-multiplexed system with strong dispersion management. We show that, for complete collisions, the collision-induced frequency shift of a Gaussian pulse is negligible, whereas its position shift is significant and can be found in a simple analytical form. For strong dispersion management we find that incomplete collisions can be neglected, whereas for dispersion management of moderate strength the contribution of the incomplete collisions can be significant. The analytical predictions are in satisfactory agreement with numerical results. We also give an estimate of the limit imposed on the transmission distance by such collisions.


Release-Modulated Antioxidant Activity of a Composite Curcumin-Chitosan Polymer.
Curcumin is known to have immense therapeutic potential but is hindered by poor solubility and rapid degradation in solution. To overcome these shortcomings, curcumin has been conjugated to chitosan through a pendant glutaric anhydride linker using amide bond coupling chemistry. The hybrid polymer has been characterized by UV-visible, fluorescence, and infrared spectroscopies as well as zeta potential measurements and SEM imaging. The conjugation reactivity was confirmed through gel permeation chromatography and quantification of unconjugated curcumin. An analogous reaction of curcumin with glucosamine, a small molecule analogue for chitosan, was performed and the purified product characterized by mass spectrometry, UV-visible, fluorescence, and infrared spectroscopies. Conjugation of curcumin to chitosan has greatly improved curcumin aqueous solubility and stability, with no significant curcumin degradation detected after one month in solution. The absorbance and fluorescence properties of curcumin are minimally perturbed (λmax shifts of 2 and 5 nm, respectively) by the conjugation reaction. This conjugation strategy required use of one out of two curcumin phenols (one of the main antioxidant functional groups) for covalent linkage to chitosan, thus temporarily attenuating its antioxidant capacity. Hydrolysis-based release of curcumin from the polymer, however, is accompanied by full restoration of curcumin's antioxidant potential. Antioxidant assays show that curcumin radical scavenging potential is reduced by 40% after conjugation, but that full antioxidant potential is restored upon hydrolytic release from chitosan. Release studies show that curcumin is released over 19 days from the polymer and maintains a concentration of 0.23 ± 0.12 μM curcumin/mg polymer/mL solution based on 1% curcumin loading on the polymer. Release studies in the presence of carbonic anhydrase, an enzyme with known phenolic esterase activity, show no significant difference from nonenzymatic release studies, implying that simple ester hydrolysis is the dominant release mechanism. Conjugation of curcumin to chitosan through a phenol ester modification provides improved stability and solubility to curcumin, with ester hydrolysis restoring the full antioxidant potential of curcumin.


A real-time in vivo dosimetric verification method for high-dose rate intracavitary brachytherapy of nasopharyngeal carcinoma.
A real-time in vivo dosimetric verification method using metal-oxide-semiconductor field effect transistor (MOSFET) dosimeters has been developed for patient dosimetry in high-dose rate (HDR) intracavitary brachytherapy of nasopharyngeal carcinoma (NPC). The necessary calibration and correction factors for MOSFET measurements in (192)Iridium source were determined in a water phantom. With the detector placed inside a custom-made nasopharyngeal applicator, the actual dose delivered to the tumor was measured in vivo and compared to the calculated values using a commercial brachytherapy planning system. Five MOSFETs were independently calibrated with the HDR source, yielding calibration factors of 0.48 ± 0.007 cGy∕mV. The maximum sensitivity variation was no more than 7% in the clinically relevant distance range of 1-5 cm from the source. A total of 70 in vivo measurements in 11 NPC patients demonstrated good agreement with the treatment planning. The mean differences between the planned and the actually delivered dose within a single treatment fraction were -0.1% ± 3.8% and -0.1% ± 3.7%, respectively, for right and left side assessments. The maximum dose deviation was less than 8.5%. In vivo measurement using the real-time MOSFET dosimetry system is possible to evaluate the actual dose to the tumor received by the patient during a treatment fraction and thus can offer another line of security to detect and prevent large errors.


Using Scent Detection Dogs in Conservation Settings: A Review of Scientific Literature Regarding Their Selection.
Dogs are widely used for scent detection work, assisting in searches for, among other things, missing persons, explosives, and even cancers. They are also increasingly used in conservation settings, being deployed for a range of diverse purposes. Although scent detecting dogs have been used in conservation roles for over 100 years, it is only recently that the scientific literature has begun to document their effectiveness and, importantly, how suitable dogs should initially be selected by organizations wanting to develop a detection program. In this paper, we review this literature, with the aim of extracting information that might be of value to conservation groups considering whether to invest in the use of dogs. We conclude that selection of appropriate dogs is no easy task. While olfactory ability is critical, so also are a range of other characteristics. These include biological, psychological, and social traits. At present, no validated selection tools have been published. Existing organizations have adapted selection instruments from other contexts for their use, but very little published information is available regarding the effectiveness of these instruments in a conservation setting. In the absence of clear guidelines, we urge those wanting to invest in one or more dogs for conservation purposes to proceed with extreme caution and, preferably, under the watchful eyes of an experienced professional.


Rehabilitation after lumbar disc surgery.
Several rehabilitation programs are available for individuals after lumbar disc surgery. To evaluate the effects of active rehabilitation for adults after first-time lumbar disc surgery. We searched CENTRAL (The Cochrane Library 2007, Issue 2) and MEDLINE, EMBASE, CINAHL and PsycINFO to May 2007. We only included randomised controlled trials (RCTs). Pairs of review authors independently assessed studies for eligibility and risk of bias. A meta-analysis was performed with clinically homogeneous studies. The GRADE approach was used to determine the quality of evidence. Fourteen studies were included, seven of which had a low risk of bias. Most programs were only assessed in one study. Statistical pooling was only completed for three comparisons in which exercises were started four to six weeks post-surgery: exercise programs versus no treatment, high versus low intensity exercise programs, and supervised versus home exercises.There is low quality evidence (three RCTS, N = 156) that exercises are more effective than no treatment for pain at short-term follow-up (WMD -11.13; 95% CI -18.44 to -3.82) and moderate evidence (two RCTs, N = 136) that they are more effective for functional status on short-term follow-up (WMD -6.50; 95% CI -9.26 to -3.74). None of the studies reported that exercises increased the re-operation rate.There is low quality evidence (two RCTs, N =103) that high intensity are slightly more effective than low intensity exercise programs for pain in the short term (WMD -10.67; 95% CI -17.04 to -4.30) and moderate evidence (two RCTs, N = 103) that they are more effective for functional status in the short term (SMD -0.77; 95% CI -1.17 to -0.36).There is low quality evidence (three RCTS, N = 95) that there were no significant differences between supervised and home exercises for short-term pain relief (SMD -1.12; 95% CI -2.77 to 0.53) or functional status (three RCTs, N = 88; SMD -1.18; 95% CI -2.63 to 0.26). Exercise programs starting four to six weeks post-surgery seem to lead to a faster decrease in pain and disability than no treatment. High intensity exercise programs seem to lead to a faster decrease in pain and disability than low intensity programs. There were no significant differences between supervised and home exercises for pain relief, disability, or global perceived effect. There is no evidence that active programs increase the re-operation rate after first-time lumbar surgery.


Multi-omics profiling of calcium-induced human keratinocytes differentiation reveals modulation of unfolded protein response signaling pathways.
By proteomic, metabolomic and transcriptomic approaches we shed light on the molecular mechanism by which human keratinocytes undergo to terminal differentiation upon in vitro calcium treatment. Proteomic analysis revealed a selective induction of the ribosomal proteins RSSA, an inhibitor of cell proliferation and inducer of differentiation, HSP 60, a protein folding chaperone and GRP78, an unfolding protein response signal. Additionally, we observed an induction of EF1D, a transcription factor for genes that contain heat-shock responsive elements. Conversely, RAD23, a protein involved in regulating ER-associated protein degradation was down-regulated. All these modifications indicated an ER stress response, which in turn activated the unfolded protein response signaling pathway through ATF4, as confirmed both by the modulation of amino acids metabolism genes, such as XBP1, PDI and GPR78, and by the metabolomic analysis. Finally, we detected a reduction of PDI protein, as confirmed by the increase of oxidized glutathione. Metabolome analysis indicated that glycolysis failed to fuel the Krebs cycle, which continued to decrease during differentiation, at glance with the PPP pathway, allowing NADH production and glutathione reduction. Since unfolded protein response is linked to keratinization, these results may be useful for studying pathological mechanisms as well as potential treatments for different pathological conditions. Abbreviation: UPR, unfolded protein response; HEK, human epidermal keratinocytes; HKGS, human keratinocytes growth factor.


Resonant Mie scattering (RMieS) correction of infrared spectra from highly scattering biological samples.
Infrared spectra of single biological cells often exhibit the 'dispersion artefact' observed as a sharp decrease in intensity on the high wavenumber side of absorption bands, in particular the Amide I band at approximately 1655 cm(-1), causing a downward shift of the true peak position. The presence of this effect makes any biochemical interpretation of the spectra unreliable. Recent theory has shed light on the origins of the 'dispersion artefact' which has been attributed to resonant Mie scattering (RMieS). In this paper a preliminary algorithm for correcting RMieS is presented and evaluated using simulated data. Results show that the 'dispersion artefact' appears to be removed; however, the correction is not perfect. An iterative approach was subsequently implemented whereby the reference spectrum is improved after each iteration, resulting in a more accurate correction. Consequently the corrected spectra become increasingly more representative of the pure absorbance spectra. Using this correction method reliable peak positions can be obtained.


Bisphosphonates: from the laboratory to the clinic and back again.
Bisphosphonates (BPs) used as inhibitors of bone resorption all contain two phosphonate groups attached to a single carbon atom, forming a "P-C-P" structure. The bisphosphonates are therefore stable analogues of naturally occuring pyrophosphate-containing compounds, which now helps to explain their intracellular as well as their extracellular modes of action. Bisphosphonates adsorb to bone mineral and inhibit bone resorption. The mode of action of bisphosphonates was originally ascribed to physico-chemical effects on hydroxyapatite crystals, but it has gradually become clear that cellular effects must also be involved. The marked structure-activity relationships observed among more complex compounds indicate that the pharmacophore required for maximal activity not only depends upon the bisphosphonate moiety but also on key features, e.g., nitrogen substitution in alkyl or heterocyclic side chains. Several bisphosphonates (e.g., etidronate, clodronate, pamidronate, alendronate, tiludronate, risedronate, and ibandronate) are established as effective treatments in clinical disorders such as Paget's disease of bone, myeloma, and bone metastases. Bisphosphonates are also now well established as successful antiresorptive agents for the prevention and treatment of osteoporosis. In particular, etidronate and alendronate are approved as therapies in many countries, and both can increase bone mass and produce a reduction in fracture rates to approximately half of control rates at the spine, hip, and other sites in postmenopausal women. In addition to inhibition of osteoclasts, the ability of bisphosphonates to reduce the activation frequency and birth rates of new bone remodeling units, and possibly to enhance osteon mineralisation, may also contribute to the reduction in fractures. The clinical pharmacology of bisphosphonates is characterized by low intestinal absorption, but highly selective localization and retention in bone. Significant side effects are minimal. Current issues with bisphosphonates include the introduction of new compounds, the choice of therapeutic regimen (e.g., the use of intermittent dosing rather than continuous), intravenous vs. oral therapy, the optimal duration of therapy, the combination with other drugs, and extension of their use to other conditions, including steroid-associated osteoporosis, male osteoporosis, arthritis, and osteopenic disorders in childhood. Bisphosphonates inhibit bone resorption by being selectively taken up and adsorbed to mineral surfaces in bone, where they interfere with the action of osteoclasts. It is likely that bisphosphonates are internalized by osteoclasts and interfere with specific biochemical processes and induce apoptosis. The molecular mechanisms by which these effects are brought about are becoming clearer. Recent studies show that bisphosphonates can be classified into at least two groups with different modes of action. Bisphosphonates that closely resemble pyrophosphate (such as clodronate and etidronate) can be metabolically incorporated into nonhydrolysable analogues of ATP that may inhibit ATP-dependent intracellular enzymes. The more potent, nitrogen-containing bisphosphonates (such as pamidronate, alendronate, risedronate, and ibandronate) are not metabolized in this way but can inhibit enzymes of the mevalonate pathway, thereby preventing the biosynthesis of isoprenoid compounds that are essential for the posttranslational modification of small GTPases. The inhibition of protein prenylation and the disruption of the function of these key regulatory proteins explains the loss of osteoclast activity and induction of apoptosis. These different modes of action might account for subtle differences between compounds in terms of their clinical effects. In conclusion, bisphosphonates are now established as an important class of drugs for the treatment of bone diseases, and their mode of action is being unravelled. As a result, their full therapeutic potential is gradual


Modelling age and secular differences in fitness between basketball players.
Concerns about the value of physical testing and apparently declining test performance in junior basketball players prompted this retrospective study of trends in anthropometric and fitness test scores related to recruitment age and recruitment year. The participants were 1011 females and 1087 males entering Basketball Australia's State and National programmes (1862 and 236 players, respectively). Players were tested on 2.6 +/- 2.0 (mean +/- s) occasions over 0.8 +/- 1.0 year. Test scores were adjusted to recruitment age (14-19 years) and recruitment year (1996-2003) using mixed modelling. Effects were estimated by log transformation and expressed as standardized (Cohen) differences in means. National players scored more favourably than State players on all tests, with the differences being generally small (standardized differences, 0.2-0.6) or moderate (0.6-1.2). On all tests, males scored more favourably than females, with large standardized differences (>1.2). Athletes entering at age 16 performed at least moderately better than athletes entering at age 14 on most tests (standardized differences, 0.7-2.1), but test scores often plateaued or began to deteriorate at around 17 years. Some fitness scores deteriorated over the 8-year period, most notably a moderate increase in sprint time and moderate (National male) to large (National female) declines in shuttle run performance. Variation in test scores between National players was generally less than that between State players (ratio of standard deviations, 0.83-1.18). More favourable means and lower variability in athletes of a higher standard highlight the potential utility of these tests in junior basketball programmes, although secular declines should be a major concern of Australian basketball coaches.


Breast Density and Benign Breast Disease: Risk Assessment to Identify Women at High Risk of Breast Cancer.
Women with proliferative breast lesions are candidates for primary prevention, but few risk models incorporate benign findings to assess breast cancer risk. We incorporated benign breast disease (BBD) diagnoses into the Breast Cancer Surveillance Consortium (BCSC) risk model, the only breast cancer risk assessment tool that uses breast density. We developed and validated a competing-risk model using 2000 to 2010 SEER data for breast cancer incidence and 2010 vital statistics to adjust for the competing risk of death. We used Cox proportional hazards regression to estimate the relative hazards for age, race/ethnicity, family history of breast cancer, history of breast biopsy, BBD diagnoses, and breast density in the BCSC. We included 1,135,977 women age 35 to 74 years undergoing mammography with no history of breast cancer; 17% of the women had a prior breast biopsy. During a mean follow-up of 6.9 years, 17,908 women were diagnosed with invasive breast cancer. The BCSC BBD model slightly overpredicted risk (expected-to-observed ratio, 1.04; 95% CI, 1.03 to 1.06) and had modest discriminatory accuracy (area under the receiver operator characteristic curve, 0.665). Among women with proliferative findings, adding BBD to the model increased the proportion of women with an estimated 5-year risk of 3% or higher from 9.3% to 27.8% (P<.001). The BCSC BBD model accurately estimates women's risk for breast cancer using breast density and BBD diagnoses. Greater numbers of high-risk women eligible for primary prevention after BBD diagnosis are identified using the BCSC BBD model.


Do lean practices lead to more time at the bedside?
The aim of this review is to evaluate the application of value-added processes in healthcare, with an emphasis on their effects on bedside nursing. Literature relevant to Lean methodology and inpatient care was reviewed, excluding all research related to other service lines (i.e., surgical services, emergency services, laboratory, radiology, etc.). Increased value is also an important tenet of transforming care at the bedside (TCAB), an initiative launched by the Institute for Healthcare Improvement (IHI) and the Robert Wood Johnson Foundation (RWJF). Therefore, articles concerning TCAB were also included in this review. A systematic study of the literature revealed varied applications of Lean principles in practice, ranging from the implementation of a single tool, to full organizational restructuring. All articles reviewed reported positive results, although the majority lacked strong supporting evidence for claims of improvement. Even though there is some indication that the application of Lean principles to nursing processes is successful in improving specific outcomes, the authors cannot conclude that the implementation of Lean methodology or TCAB greatly influences direct patient care, or increases time spent at the bedside.


Molecular and biological actions of cyclosporin A and FK506 on T cell development and function.
The microbial products cyclosporin A (CsA) and FK506 are potent immunosuppressive agents in experimental and clinical transplantation. Binding of both drugs to intracellular immunophilins results in the inhibition of calcineurin activity and the prevention of interleukin-2 gene transcription and, thus, in the inhibition of T cell receptor-dependent T cell activation. CsA and FK506 also have a profound effect on the structure and function of the thymus with disruption of positive and negative selection of T cells. These influences on thymic microenvironment and T cell ontogeny disrupt the induction or maintenance of self-tolerance (or both) and are thus of relevance to clinical transplantation immunology.