Relevance of ammonium oxidation within biological soil crust communities.
Thin, vertically structured topsoil communities that become ecologically important in arid regions (biological soil crusts or BSCs) are responsible for much of the nitrogen inputs into pristine arid lands. We studied N(2) fixation and ammonium oxidation (AO) at subcentimetre resolution within BSCs from the Colorado Plateau. Pools of dissolved porewater nitrate/nitrite, ammonium and organic nitrogen in wetted BSCs were high in comparison with those typical of aridosoils. They remained stable during incubations, indicating that input and output processes were of similar magnitude. Areal N(2) fixation rates (6.5-48 micromol C(2)H(2) m(-2) h(-1)) were high, the vertical distribution of N(2) fixation peaking close to the surface if populations of heterocystous cyanobacteria were present, but in the subsurface if they were absent. Areal AO rates (19-46 micromol N m(-2) h(-1)) were commensurate with N(2) fixation inputs. When considering oxygen availability, AO activity invariably peaked 2-3 mm deep and was limited by oxygen (not ammonium) supply. Most probable number (MPN)-enumerated ammonia-oxidizing bacteria (6.7-7.9 x 10(3) cells g(-1) on average) clearly peaked at 2-3 mm depth. Thus, AO (hence nitrification) is a spatially restricted but important process in the nitrogen cycling of BSC, turning much of the biologically fixed nitrogen into oxidized forms, the fate of which remains to be determined.


Spastic paraplegia gene 7 in patients with spasticity and/or optic neuropathy.
Mutations in the spastic paraplegia 7 (SPG7) gene encoding paraplegin are responsible for autosomal recessive hereditary spasticity. We screened 135 unrelated index cases, selected in five different settings: SPG7-positive patients detected during SPG31 analysis using SPG31/SPG7 multiplex ligation-dependent probe amplification (n = 7); previously reported ambiguous SPG7 cases (n = 5); patients carefully selected on the basis of their phenotype (spasticity of the lower limbs with cerebellar signs and/or cerebellar atrophy on magnetic resonance imaging/computer tomography scan and/or optic neuropathy and without other signs) (n = 24); patients with hereditary spastic paraparesis referred consecutively from attending neurologists and the national reference centre in a diagnostic setting (n = 98); and the index case of a four-generation family with autosomal dominant optic neuropathy but no spasticity linked to the SPG7 locus. We identified two SPG7 mutations in 23/134 spastic patients, 21% of the patients selected according to phenotype but only 8% of those referred directly. Our results confirm the pathogenicity of Ala510Val, which was the most frequent mutation in our series (65%) and segregated at the homozygous state with spastic paraparesis in a large family with autosomal recessive inheritance. All SPG7-positive patients tested had optic neuropathy or abnormalities revealed by optical coherence tomography, indicating that abnormalities in optical coherence tomography could be a clinical biomarker for SPG7 testing. In addition, the presence of late-onset very slowly progressive spastic gait (median age 39 years, range 18-52 years) associated with cerebellar ataxia (39%) or cerebellar atrophy (47%) constitute, with abnormal optical coherence tomography, key features pointing towards SPG7-testing. Interestingly, three relatives of patients with heterozygote SPG7 mutations had cerebellar signs and atrophy, or peripheral neuropathy, but no spasticity of the lower limbs, suggesting that SPG7 mutations at the heterozygous state might predispose to late-onset neurodegenerative disorders, mimicking autosomal dominant inheritance. Finally, a novel missense SPG7 mutation at the heterozygous state (Asp411Ala) was identified as the cause of autosomal dominant optic neuropathy in a large family, indicating that some SPG7 mutations can occasionally be dominantly inherited and be an uncommon cause of isolated optic neuropathy. Altogether, these results emphasize the clinical variability associated with SPG7 mutations, ranging from optic neuropathy to spastic paraplegia, and support the view that SPG7 screening should be carried out in both conditions.


Presynaptic Calmodulin targets: lessons from structural proteomics.
Calmodulin (CaM) is a highly conserved Ca2+-binding protein that is exceptionally abundant in the brain. In the presynaptic compartment of neurons, CaM transduces changes in Ca2+ concentration into the regulation of synaptic transmission dynamics. Areas covered: We review selected literature including published CaM interactor screens and outline established and candidate presynaptic CaM targets. We present a workflow of biochemical and structural proteomic methods that were used to identify and characterize the interactions between CaM and Munc13 proteins. Finally, we outline the potential of ion mobility-mass spectrometry (IM-MS) for conformational screening and of protein-protein cross-linking for the structural characterization of CaM complexes. Expert commentary: Cross-linking/MS and native MS can be applied with considerable throughput to protein mixtures under near-physiological conditions, and thus effectively complement high-resolution structural biology techniques. Experimental distance constraints are applicable best when obtained by combining different cross-linking strategies, i.e. by using cross-linkers with different spacer length and reactivity, and by using the incorporation of unnatural photo-reactive amino acids. Insights from structural proteomics can be used to generate CaM-insensitive mutants of CaM targets for functional studies in vitro or ideally in vivo.


An evaluation of a specialist firesetting treatment programme for male and female mentally disordered offenders (the FIP-MO).
Individuals who set deliberate fires are frequently encountered by clinicians working in forensic mental health services. However, little attention has been paid to developing standardised treatment for this behaviour, and few evaluations of treatment have been conducted in forensic mental health services. This study evaluates a new standardised group cognitive behavioural treatment programme for individuals residing in forensic psychiatric hospitals who have engaged in deliberate firesetting (The Firesetting Intervention Programme for Mentally Disordered Offenders; FIP-MO). Sixty-three male and female patients with a history of deliberate firesetting commenced FIP-MO treatment. Patients who met the referral criteria for treatment but who resided at hospitals where FIP-MO treatment was not available were recruited as a treatment as usual comparison group. The treatment group completed a battery of psychometric assessments pre- and post-treatment, with the comparison group completing these at similar time points. Results showed that patients who completed the FIP-MO made significant improvements post-treatment, relative to the comparison group on fire-related measures (e.g., problematic interest and associations with fire) and anger expression. Further, effect size calculations showed that the treatment group made larger pre-post treatment shifts on the majority of outcome measures compared to the comparison group. These findings suggest that FIP-MO treatment is effective for reducing some of the key factors associated with deliberate firesetting.


A new chemotaxis device for cell migration studies.
This study presents the design and optimization for in vitro use of a new versatile chemotaxis device called the NANIVID (NANo IntraVital Imaging Device), developed using advanced nano/micro fabrication techniques. The device is fabricated using microphotolithographic techniques and two substrates are bonded together using a thin polymer layer creating a sealed device with one outlet. The main structure of the device consists of two Pyrex substrates: an etched chemoattractant reservoir and a top cover, with a final size of 0.2 × 2 × 3 mm. This reservoir contains a hydrogel blend with EGF which diffuses out through a small (∼9.10(3)μm(2)) outlet. This reservoir sustains a steady release of growth factor into the surrounding environment for several hours establishing a consistent concentration gradient from the device. The focus of this study was to design and optimize the new device for cell chemotaxis studies in breast cancer cells in cell culture. Our results show that we have created a flexible, cheap, miniature and autonomous chemotaxis device and demonstrate its usefulness in 2D and 3D cell culture. We also provide preliminary data for use of the device in vivo.


Ideal cardiovascular health and mortality: pooled results of three prospective cohorts in Chinese adults.
Evidence on the relations of the American Heart Association's ideal cardiovascular health (ICH) with mortality in Asians is sparse, and the interaction between behavioral and medical metrics remained unclear. We aimed to fill the gaps. A total of 198,164 participants without cancer and cardiovascular disease (CVD) were included from the China Kadoorie Biobank study (2004-2018), Dongfeng-Tongji cohort (2008-2018), and Kailuan study (2006-2019). Four behaviors (i.e., smoking, physical activity, diet, body mass index) and three medical factors (i.e., blood pressure, blood glucose, and blood lipid) were classified into poor, intermediate, and ideal levels (0, 1, and 2 points), which constituted 8-point behavioral, 6-point medical, and 14-point ICH scores. Results of Cox regression from three cohorts were pooled using random-effects models of meta-analysis. During about 2 million person-years, 20,176 deaths were recorded. After controlling for demographic characteristics and alcohol drinking, hazard ratios (95% confidence intervals) comparing ICH scores of 10-14 vs. 0-6 were 0.52 (0.41-0.67), 0.44 (0.37-0.53), 0.54 (0.45-0.66), and 0.86 (0.64-1.14) for all-cause, CVD, respiratory, and cancer mortality. A higher behavioral or medical score was independently associated with lower all-cause and CVD mortality among the total population and populations with different levels of behavioral or medical health equally, and no interaction was observed. ICH was associated with lower all-cause, CVD, and respiratory mortality among Chinese adults. Both behavioral and medical health should be improved to prevent premature deaths.


Politics, policies and processes: a multidisciplinary and multimethods research programme on policies on the social determinants of health inequity in Australia.
The development and implementation of multisectoral policy to improve health and reduce health inequities has been slow and uneven. Evidence is largely focused on the facts of health inequities rather than understanding the political and policy processes. This 5-year funded programme of research investigates how these processes could function more effectively to improve equitable population health. The programme of work is organised in four work packages using four themes (macroeconomics and infrastructure, land use and urban environments, health systems and racism) related to the structural drivers shaping the distribution of power, money and resources and daily living conditions. Policy case studies will use publicly available documents (policy documents, published evaluations, media coverage) and interviews with informants (policy-makers, former politicians, civil society, private sector) (~25 per case). NVIVO software will be used to analyse the documents to see how 'social and health equity' is included and conceptualised. The interview data will include qualitative descriptive and theory-driven critical discourse analysis. Our quantitative methodological work assessing the impact of public policy on health equity is experimental that is in its infancy but promises to provide the type of evidence demanded by policy-makers. Our programme is recognising the inherently political nature of the uptake, formulation and implementation of policy. The early stages of our work indicate its feasibility. Our work is aided by a Critical Policy Reference Group. Multiple ethics approvals have been obtained with the foundation approval from the Social and Behavioural Ethics Committee, Flinders University (Project No: 6786).The theoretical, methodological and policy engagement processes established will provide improved evidence for policy-makers who wish to reduce health inequities and inform a new generation of policy savvy knowledge on social determinants.


Loss of antigenicity in stored sections of breast cancer tissue microarrays.
Immunohistochemical characterization of tumor tissues in epidemiological studies is a promising approach to identify breast cancer subtypes with distinct etiology. The recent development of the tissue microarray (TMA) technique allows for standardized, rapid, and cost-effective immunohistochemical characterization of many cases, which is critical in epidemiological studies. Sectioning paraffin blocks at different times results in loss of material, which can be reduced by preparing many sections each time a block is cut. However, data suggest that staining intensity declines in whole sections prepared from conventional paraffin blocks with storage time, resulting in false-negative results. This problem would be accentuated in TMAs because of the limited tissue representation of each case. To evaluate this concern, we prepared a single TMA block from 125 invasive breast carcinomas collected in a population-based case-control study conducted in Poland and compared estrogen receptor (ER-alpha), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) expression in sections cut and stored for 6 months at room temperature with sections cut from the same TMA block and stained on the same day. Percentage of positive cases for stored versus fresh sections was similar for ER (59.0%) but significantly higher in fresh sections for PR (56.3% versus 64.1%, P = 0.01) and HER2 (45.5% versus 64.4%, P < 0.001). Among cases positive in both stored and fresh sections, the median percentage of immunoreactive cells was significantly reduced and the staining intensity was consistently lower in stored compared with fresh sections. We conclude that loss of immunoreactivity is an important problem in TMAs of breast cancer. Improved methods for sectioning TMAs and storing tissue sections aimed at reducing loss of immunoreactivity are critical for the use of TMAs in epidemiological studies.


Prediction of disease severity in young children presenting with acute febrile illness in resource-limited settings: a protocol for a prospective observational study.
In rural and difficult-to-access settings, early and accurate recognition of febrile children at risk of progressing to serious illness could contribute to improved patient outcomes and better resource allocation. This study aims to develop a prognostic clinical prediction tool to assist community healthcare providers identify febrile children who might benefit from referral or admission for facility-based medical care. This prospective observational study will recruit at least 4900 paediatric inpatients and outpatients under the age of 5 years presenting with an acute febrile illness to seven hospitals in six countries across Asia. A venous blood sample and nasopharyngeal swab is collected from each participant and detailed clinical data recorded at presentation, and each day for the first 48 hours of admission for inpatients. Multianalyte assays are performed at reference laboratories to measure a panel of host biomarkers, as well as targeted aetiological investigations for common bacterial and viral pathogens. Clinical outcome is ascertained on day 2 and day 28.Presenting syndromes, clinical outcomes and aetiology of acute febrile illness will be described and compared across sites. Following the latest guidance in prediction model building, a prognostic clinical prediction model, combining simple clinical features and measurements of host biomarkers, will be derived and geographically externally validated. The performance of the model will be evaluated in specific presenting clinical syndromes and fever aetiologies. The study has received approval from all relevant international, national and institutional ethics committees. Written informed consent is provided by the caretaker of all participants. Results will be shared with local and national stakeholders, and disseminated via peer-reviewed open-access journals and scientific meetings. NCT04285021.


Adsorption and diffusion characteristics of lithium on hydrogenated α- and β-silicene.
Using first-principles density functional theory calculations, we investigate adsorption properties and the diffusion mechanism of a Li atom on hydrogenated single-layer α- and β-silicene on a Ag(111) surface. It is found that a Li atom binds strongly on the surfaces of both α- and β-silicene, and it forms an ionic bond through the transfer of charge from the adsorbed atom to the surface. The binding energies of a Li atom on these surfaces are very similar. However, the diffusion barrier of a Li atom on H-α-Si is much higher than that on H-β-Si. The energy surface calculations show that a Li atom does not prefer to bind in the vicinity of the hydrogenated upper-Si atoms. Strong interaction between Li atoms and hydrogenated silicene phases and low diffusion barriers show that α- and β-silicene are promising platforms for Li-storage applications.


Anomalous twisting strength of tilt grain boundaries in armchair graphene nanoribbons.
The twisting response of armchair graphene nanoribbons with tilt grain boundaries is theoretically and numerically investigated. It is found that the critical instability twist rate of graphene nanoribbons with grain boundaries is generally about 10% higher than that of common armchair graphene nanoribbons when the width of nanoribbons is less than 4.0 nm. Our analytical analysis indicates that the strengthening effect is resulted from the rotation of the compressed direction, deflection of grain boundaries, and the reflexing of the creased angle in nanoribbons: the rotation of the compressed direction induced by grain boundaries improves the buckling strength of nanoribbons due to the chirality-dependent buckling in graphene; the deflection of grain boundaries leads to a nonzero strain in the axle wire of nanoribbons, which eventually decreases the compressed stress; grain boundaries induce a spontaneous creased angle in nanoribbons, which is reflexed under twist loading and impedes the propagation of instability in nanoribbons. Furthermore, we found and demonstrated that grain boundaries changed the transport properties of twisted graphene nanoribbons. It is expected that our findings would improve the fundamental understanding of the strain-engineering of graphene nanoribbons used in nanodevices.


Periodic health examination, 1995 update: 2. Prevention of dental caries. The Canadian Task Force on the Periodic Health Examination.
To make recommendations, based on current evidence, for practising physicians and dentists on interventions for the prevention of dental caries in their patients. Systemic fluoride administration, professionally administered fluoride, use of fluoride mouth rinses, fissure sealants, oral-hygiene practices, dietary practices, identification of groups at a high risk of dental caries, and early diagnosis and treatment. Reduced prevalence of dental caries and fluorosis, longer retention of teeth and lower treatment costs. Several MEDLINE searches were conducted for articles published from January 1980 to December 1992, including relevant review articles. Relevant clinical findings were evaluated and categorized with the use of the evidence-based methods and values of the Canadian Task Force on the Periodic Health Examination. Recommendations were developed for each method of caries prevention, with reduced incidence of dental caries and improved prevalence of caries-free teeth given high values. The potential benefits of these measures in the long-term are a lower incidence of tooth decay, longer retention of teeth and prevention of fluorosis. The cost saving can be considerable for patients and insurers; however, implementation of some recommendations will be difficult, since the traditional preventive practices of dentists and dental hygienists are not easily changed. There is good evidence that the following manoeuvres are effective in preventing dental caries: use of dentifrices containing fluoride, fluoridation of drinking water, fluoride supplements for patients in areas where there is a low level (0.3 ppm or less) of fluoride in the drinking water, professionally applied topical fluoride and the use of fluoride mouth rinses for patients with very active decay or at a high risk of dental caries and selective use of professionally applied fissure sealants on permanent molar teeth. There is poor evidence that the following manoeuvres are effective in preventing dental caries: professionally applied topical fluoride and the use of fluoride mouth rinses for patients with a low risk of caries, toothbrushing (without a dentifrice containing fluoride) and flossing, cleaning of teeth by a dentist or dental hygienist before topical application of fluoride or at a dental visit and dietary counselling for the general population. There is good evidence to recommend against the use of over-the-counter fluoride mouth rinses by the general population. These guidelines are compatible with those of the US Preventive Services Task Force. These guidelines were developed and endorsed by the task force, which is funded by Health Canada. Major funding was provided by the Faculty of Dentistry of the University of Toronto, Toronto and the Faculty of Dentistry of Dalhousie University, Halifax.


Radiotheragnostics Paradigm for Radioactive Iodine (Iodide) Management of Differentiated Thyroid Cancer.
This review of radioactive iodide treatment (RAIT) extends from historical origins to its modern utilization in differentiated thyroid cancer (DTC). The principles embedded in the radiotheragnostics (RTGs) paradigm are detailed. The diverse approaches in current practice are addressed, and this broad variability represents a major weakness that erodes our specialty's trust-based relationship with patients and referring physicians. The currently developing inter-specialty collaboration should be hailed as a positive change. It promises to clarify the target-based terminology for RAIT. It defines RAIT of post total thyroidectomy (PTT), presumably benign thyroid as 'remnant ablation' (RA). 'Adjuvant treatment' (AT) referrers to RAIT of suspected microscopic DTC that is inherently occult on diagnostic imaging. RAIT directed at DTC lesion(s) overtly seen on diagnostic imaging is termed 'treatment of known disease' (TKD). It was recently recognized that a 'recurrent' DTC is actually occult residual DTC in the majority of cases. Thyroglobulin with remnant uptake concord (TRUC) method (aka Tulchinsky method) was developed to validate that a benign remnant in the post-thyroidectomy neck bed, as quantified by the RAI uptake, is concordant with a measured thyroglobulin (Tg) level at the time of the initial post-thyroidectomy evaluation. It allows recognition of occult residual DTC contribution to post-thyroidectomy Tg. Case examples demonstrate the application of the TRUC method for a logical selection of a specific RAIT category, using imaging-guided identification and management of RAI-avid versus RAI-nonavid residual DTC, i.e. the radiotheragnostics paradigm.


Emotions amplify speaker-listener neural alignment.
Individuals often align their emotional states during conversation. Here, we reveal how such emotional alignment is reflected in synchronization of brain activity across speakers and listeners. Two "speaker" subjects told emotional and neutral autobiographical stories while their hemodynamic brain activity was measured with functional magnetic resonance imaging (fMRI). The stories were recorded and played back to 16 "listener" subjects during fMRI. After scanning, both speakers and listeners rated the moment-to-moment valence and arousal of the stories. Time-varying similarity of the blood-oxygenation-level-dependent (BOLD) time series was quantified by intersubject phase synchronization (ISPS) between speaker-listener pairs. Telling and listening to the stories elicited similar emotions across speaker-listener pairs. Arousal was associated with increased speaker-listener neural synchronization in brain regions supporting attentional, auditory, somatosensory, and motor processing. Valence was associated with increased speaker-listener neural synchronization in brain regions involved in emotional processing, including amygdala, hippocampus, and temporal pole. Speaker-listener synchronization of subjective feelings of arousal was associated with increased neural synchronization in somatosensory and subcortical brain regions; synchronization of valence was associated with neural synchronization in parietal cortices and midline structures. We propose that emotion-dependent speaker-listener neural synchronization is associated with emotional contagion, thereby implying that listeners reproduce some aspects of the speaker's emotional state at the neural level.


Fluid shifts associated with gas-induced osmosis.
It has been proposed that equilibration of nitrous oxide with blood plasma increases osmotic pressure and thereby causes hemodilution. However, calculations show that the 250-torr osmotic gradient produced by 0.7 atmosphere of nitrous oxide dissolved in blood plasma (separated from the other body fluids by a membrane permeable only to water) would be eliminated by a water shift which would dilute the plasma by only 4 percent. Permeability of the membrane to nitrous oxide would further reduce the shift. In vivo measurements confirmed the smallness of any osmotically induced shifts by demonstrating no significant transient changes in hematocrit value when 0.7 atmosphere of nitrous oxide was added to or removed from an inhaled anesthetic mixture in man. These results cast doubt on the suggestions that gas-induced osmosis is an important factor in dysbarism or in clinical anesthesia.


Over 30 million psychedelic users in the United States.
We estimated lifetime prevalence of psychedelic use (lysergic acid diethylamide (LSD), psilocybin (magic mushrooms), mescaline, and peyote) by age category using data from a 2010 US population survey of 57,873 individuals aged 12 years and older. There were approximately 32 million lifetime psychedelic users in the US in 2010; including 17% of people aged 21 to 64 years (22% of males and 12% of females). Rate of lifetime psychedelic use was greatest among people aged 30 to 34 (total 20%, including 26% of males and 15% of females).


The influence of cigarette smoke on the epithelium of the vestibule: an electron microscopic study.
It is known that cigarette smoke induces cytological alterations on the respiratory and olfactory mucosa of the nasal cavity. We evaluated whether cigarette smoking had adverse effects on the epithelium of the vestibule, in the absence of any published ultrasutructural studies. We evaluated ten patients suffering from septum deviation, eight of whom were long-term smokers. While each layer of the epithelium obtained from the non-smokers consisted of a homogeneous cell population, each from the long-term smokers consisted of a heterogeneous cell population. The most prominent changes occurred in the shape and size of the cells and nuclei, the number and length of the cytoplasmic projections, the number and distribution pattern of the desmosomes, and the width of the intercellular spaces. We concluded that cigarette smoke produces hyperplastic and dysplastic changes, important factors related with cancer development, on the epithelium of the vestibule.


Studies on composites of collagen and a synthetic polymer. Second report - mode of reaction of a laminar composite with living tissue, and results of long-term implantation.
We have been studying composites of synthetic polymers and proctase-treated bovine collagen (telopeptide-poor collagen). In a previous paper, it is described that one of our composites appears to have a high tissue affinity, because it was found under a scanning electron microscope, well bonding the living tissue 6 weeks after implantation in the rabbit subcutaneous tissue. In the present study, the composites were also implanted in the subcutaneous tissue in rabbits to observe the mode of its bonding to the living tissue, and at the same time the bonding force was measured at various intervals during a period of 1.5 years, to observe the degree and duration of the effect of the composition, and further to study the optimal conditions for preparation of such a composite. As a method of composition, ultraviolet radiation and glutaraldehyde treatment were also comparatively examined, besides gamma-radiation that had been employed in a previous study. As the results, it is revealed that the composite bonds to the living tissue in such a manner that a portion of the collagen part is digested and absorbed to be replaced by the invading connective tissue and a part of collagen near the surface of the synthetic polymers had escaped digestion to combine with the rabbit's own collagen fiber. From these results we like to insist that the mode of reaction of our composites may be similar to real organization and be different in natur from mere encapsulation which was shown by the artificial materials hitherto in use to be considered to have a good tissue affinity. Therefor we think now the encapsulation is pseudoorganization. It is also shown that the bonding force of the composite to the living tissue and its duration depend chiefly on the degree of swelling, that is, the degree of intermolecular cross-linking, of the composed collagen part. Out of the gamma-radiated composites, those radiated 1--3 Mrads proved desirable in both tissue affinity and duration of tissue bonding, with a potent bonding force remaining effective even 1.5 years later, while the composite radiated with 5 Mrads or more proved poorer in these properties because of destruction of the collagen part, vastly varying in the properties. By ultraviolet radiation, it was easy to control the degree of swelling of collagen part with the radiation dose, however, this method gave a lower bonding force and a shorter duration of the force than gamma-radiation. The glutaraldehyde method allowed the collagen to remain over a long time, but gave the composite less tissue affinity than the other two methods.


Formation of Geopolymers Using Sodium Silicate Solution and Aluminum Orthophosphate.
This paper reports the formation and structure of fast setting geopolymers activated by using three sodium silicate solutions with different modules (1.6, 2.0 and 2.4) and a berlinite-type aluminum orthophosphate. By varying the concentration of the aluminum orthophosphate, different Si/Al-ratios were established (6, 3 and 2). Reaction kinetics of binders were determined by isothermal calorimetric measurements at 20 °C. X-ray diffraction analysis as well as nuclear magnetic resonance (NMR) measurements were performed on binders to determine differences in structure by varying the alkalinity of the sodium silicate solutions and the Si/Al-ratio. The calorimetric results indicated that the higher the alkalinity of the sodium silicate solution, the higher the solubility and degree of conversion of the aluminum orthophosphate. The results of X-ray diffraction and Rietveldt analysis, as well as the NMR measurements, confirmed the assumption of the calorimetric experiments that first the aluminum orthophosphate was dissolved and then a polycondensation to an amorphous aluminosilicate network occurred. The different amounts of amorphous phases formed as a function of the alkalinity of the sodium silicate solution, indicate that tetrahydroxoaluminate species were formed during the dissolution of the aluminum orthophosphate, which reduce the pH value. This led to no further dissolution of the aluminum orthophosphate, which remained unreacted.


Grandmothers and caregiving to grandchildren: continuity, change, and outcomes over 24 months.
Transitions in caregiving, such as becoming a primary caregiver to grandchildren or having adult children and grandchildren move in or out, may affect the well-being of the grandmother. This report describes caregiving patterns at 3 time points over 24 months in a sample of 485 Ohio grandmothers and examines the effects of stability and change in grandmother caregiving roles (raising a grandchild, living in a multigenerational home, or not caregiving to grandchildren). Drawing on the Resiliency Model of Family Stress, the study examined caregiving stress and reward, intrafamily strain, social support, resourcefulness, depressive symptoms, mental and physical health, and perceived family functioning. Caregiver group, time of measurement, switching between caregiver groups, and baseline age, race, education, work status, and marital status were considered as independent variables within the context of a one-way treatment structure in a mixed-model multivariate analysis. There were significant caregiver group effects for all variables, except mental health and resourcefulness. Grandmothers raising grandchildren reported the most stress, intrafamily strain, and perceived problems in family functioning, the worst physical health and more depressive symptoms, and the least reward and subjective support. Across groups, there were significant time effects, with worsening physical health and increased stress over time. Switching to higher levels of caregiving was associated with worsening physical health and increases in stress, intrafamily strain, and perceived problems in family functioning. Recommendations for research and for practice, especially during times of caregiving transition or for grandmothers raising grandchildren, are discussed.


Imprinted micelles for chiral recognition in water: shape, depth, and number of recognition sites.
Chiral molecular recognition is important to biology, separation, and asymmetric catalysis. Because there is no direct correlation between the chiralities of the host and the guest, it is difficult to design a molecular receptor for a chiral guest in a rational manner. By cross-linking surfactant micelles containing chiral template molecules, we obtained chiral nanoparticle receptors for a number of 4-hydroxyproline derivatives. Molecular imprinting allowed us to transfer the chiral information directly from the guest to host, making the molecular recognition between the two highly predictable. Hydrophobic interactions between the host and the guest contributed strongly to the enantio- and diastereoselective differentiation of these compounds in water, whereas ion-pair interactions, which happened near the surface of the micelle, were less discriminating. The chiral recognition could be modulated by tuning the size and shape of the binding pockets.


A zebra or a painted horse? Are hospital PPPs infrastructure partnerships with stripes or a separate species?
Public Private Partnerships (PPP) have been common in infrastructure for many years and are increasingly being considered as a means to finance, build, and manage hospitals. However, the growth of hospital PPPs in the past two decades has led to confusion about what sorts of contractual arrangements between public and private partners consititute a PPP, and what key differences distinguish public private partnership for hospitals from PPPs for infrastructure. Based on experiences from around the world we indentify six key areas where hospital PPPs differ from infrastructure partnerships. We draw upon the hospital partnerships that have been documented in OECD countries and a growing number of middle-income countries to identify four distinct types of hospital PPPs: service focused partnerships in which private partners manage operations within publicly constructed facilities; facilities and finance PPPs, focused on mobilizing capital and creating new hospitals; combined PPPs, involving both facility and clinical operations; and co-located PPPs where privately operated services are developed within the grounds of a public hospital. These four types of hospital PPPs have differing goals, and therefore different contractual and functional aspects, as well as differing risks to both public and private partners. By clarifying these, we provide a base upon which hospital PPPs can be assessed against appropriate goals and benchmarks.


The relationship between depressive symptoms and shoulder mobility among older women: assessment at one year after breast cancer diagnosis.
To examine the association between depressive symptoms and shoulder range of motion at one year after breast cancer diagnosis, controlling for patient characteristics, tumour stage and cancer therapy (surgery, axillary node dissection and radiation). Prospective trial of nurse case management involving 187 older women with complete data, age 60 years and older, newly diagnosed with breast cancer, from 1 November, 1993 to 31 October, 1996 in south-eastern Texas. Depressive symptoms, sociodemographic characteristics and breast cancer treatment were measured at two months and shoulder range of motion at 12 months. The relationship among the variables was evaluated with bivariate chi-square statistics and logistic regression analysis. All logistic models also included a variable indicating whether or not the woman received nurse case management, to control for intervention status. Increasing depressive symptoms at baseline were associated with lower arm mobility at 12 months following breast cancer diagnosis. Each unit increase in depressive symptoms at baseline was associated with an 8% decreased odds of having full range of motion of the shoulder (odds ratio (OR) 0.92, 95% confidence interval (CI) 0.87, 0.97), after controlling for relevant patient and treatment factors. Older women with depressive symptoms have an elevated risk of not fully recovering shoulder mobility after being treated for breast cancer. Future studies are needed to assess benefits from early intervention with psychological and or physical interventions in the presence of depressive symptoms on shoulder mobility.


A Case of Sudden Death in Decameron IV.6: Aortic Dissection or Atrial Myxoma?
Giovanni Boccaccio's Decameron contains a novella that details the sudden death of a young man called Gabriotto, including a portrayal of the discomfort that the protagonist experienced and a rudimentary autopsy performed by local physicians. The intriguing description of symptoms and pathologies has made it possible to read a 7-century-old case through the modern clinical lens. Thanks to the medical and philological analysis of the text-despite the vast difference between modern and medieval medicine-2 hypothetical diagnoses have emerged: either an aortic dissection or an atrial myxoma.


Placental expression of VEGF, PlGF and their receptors in a model of placental insufficiency-intrauterine growth restriction (PI-IUGR).
Placental development requires adequate and organized interaction of vascular growth factors and their receptors, including vascular endothelial growth factor (VEGF) and placental growth factor (PlGF). Both VEGF and PlGF, acting through the tyrosine kinase receptors VEGFR-1 and VEGFR-2, have been implicated in playing a role in ovine placental vascular development. The present studies describe the placental expression of components of the VEGF family at two maturational time points (55 and 90 days post coitus, dpc) in a hyperthermic-induced ovine model of placental insufficiency-intrauterine growth restriction (PI-IUGR). Both caruncular and cotyledonary VEGF and PlGF mRNA concentration increased with gestational age (P< 0.05), whereas only cotyledonary VEGF and PlGF protein concentration increased over gestation (P< 0.002). At 55 dpc, VEGF mRNA concentration was elevated in hyperthermic (HT) ewes, compared to control thermoneutral (TN) animals (TN; 0.52+/-0.08 vs HT; 1.27+/-0.17 VEGF/GAPDH, P< 0.001). At 90 dpc, expression of PlGF and VEGF mRNA was not altered by the HT treatment. Both TN cotyledonary VEGFR-1 and VEGFR-2 mRNA expression levels rose significantly over the period studied (P< 0.05 and P< 0.01 respectively). Receptor mRNA concentration in HT cotyledonary tissue was significantly reduced at 90 dpc (VEGFR-1; TN 0.21+/-0.02 vs HT 0.11+/-0.01 VEGFR-1/actin, P< 0.05, VEGFR-2; TN 0.18+/-0.05 vs HT 0.07+/-0.01 VEGFR-2/actin, P< 0.01). Soluble VEGFR-1 (sVEGFR-1) mRNA was not detected in these tissues. These alterations in growth factor and growth factor receptor mRNA expression, as a result of environmental heat stress early in placental development, could impair normal placental vascular development. Furthermore, alterations in VEGF, VEGFR-1 and VEGFR-2 mRNA expression, during the period of maximal placental growth, may contribute to the development of placental insufficiency, and ultimately intrauterine growth restriction.


Single-cell transcriptomics uncovers distinct molecular signatures of stem cells in chronic myeloid leukemia.
Recent advances in single-cell transcriptomics are ideally placed to unravel intratumoral heterogeneity and selective resistance of cancer stem cell (SC) subpopulations to molecularly targeted cancer therapies. However, current single-cell RNA-sequencing approaches lack the sensitivity required to reliably detect somatic mutations. We developed a method that combines high-sensitivity mutation detection with whole-transcriptome analysis of the same single cell. We applied this technique to analyze more than 2,000 SCs from patients with chronic myeloid leukemia (CML) throughout the disease course, revealing heterogeneity of CML-SCs, including the identification of a subgroup of CML-SCs with a distinct molecular signature that selectively persisted during prolonged therapy. Analysis of nonleukemic SCs from patients with CML also provided new insights into cell-extrinsic disruption of hematopoiesis in CML associated with clinical outcome. Furthermore, we used this single-cell approach to identify a blast-crisis-specific SC population, which was also present in a subclone of CML-SCs during the chronic phase in a patient who subsequently developed blast crisis. This approach, which might be broadly applied to any malignancy, illustrates how single-cell analysis can identify subpopulations of therapy-resistant SCs that are not apparent through cell-population analysis.


[Expand newborn screening using tandem mass spectrometry: two years' experience in Nuevo León, Mexico].
To initiate a statewide expanded metabolic screening program in neonates with the purpose of identifying the most common inborn errors of metabolism. From March 2002 through February 2004, a blood sample was obtained between 24 and 48 hours after delivery from every consecutive child born in public hospitals in Nuevo León. It was spotted on filter paper and analyzed by tandem mass spectrometry for expanded metabolic screening. A total of 42 264 samples were analyzed. Were obtained seven positive results, one for each disorder: homocystinuria, hyperphenylalaninemia, citrulinemia, transient tyrosinemia, 3-methylcrotonyl CoA carboxylase deficiency, 3-hydroxy-3-methylglutaryl CoA deficiency, and classic galactosemia. The estimated incidence of inborn errors of metabolism is 1:5 000, with a false positive rate of 0.22%. The program permitted the identification of metabolic disorders in the newborn, allowing an early intervention and prevention of life-threatening events and permanent neurological damage.


Identification of cystic fibrosis transmembrane conductance regulator channel-lining residues in and flanking the M6 membrane-spanning segment.
The cystic fibrosis transmembrane conductance regulator (CFTR) forms a chloride channel that is regulated by phosphorylation and ATP binding. Work by others suggested that some residues in the sixth transmembrane segment (M6) might be exposed in the channel and play a role in ion conduction and selectivity. To identify the residues in M6 that are exposed in the channel and the secondary structure of M6, we used the substituted cysteine accessibility method. We mutated to cysteine, one at a time, 24 consecutive residues in and flanking the M6 segment and expressed these mutants in Xenopus oocytes. We determined the accessibility of the engineered cysteines to charged, lipophobic, sulfhydryl-specific methanethiosulfonate (MTS) reagents applied extracellularly. The cysteines substituted for Ile331, Leu333, Arg334, Lys335, Phe337, Ser341, Ile344, Arg347, Thr351, Arg352, and Gln353 reacted with the MTS reagents, and we infer that they are exposed on the water-accessible surface of the protein. From the pattern of the exposed residues we infer that the secondary structure of the M6 segment includes both alpha-helical and extended regions. The diameter of the channel from the extracellular end to the level of Gln353 must be at least 6 A to allow the MTS reagents to reach these residues.


Evaluation of a pilot project: preparenthood and pregrandparenthood education.
A pilot project in expanding perinatal education is presented in this article. The author has experimented with a series of group sessions on the introduction to the transition to parenthood for couples who are either engaged or recently married. Such couples, if introduced to the changes associated with the transition to parenthood at a point of development in the commitment process, may increase their chances of weathering the turbulence that is often a part of the transition. The potential grandparents are part of the process since they are usually an important source of social support. Preparenthood and pregrandparenthood education is a useful extension of perinatal education for the beginning of this century.


Vasopressin and oxytocin release during prolonged environmental hypoxia in the rat.
The mechanism causing peripheral oedema in hypoxaemic chronic obstructive pulmonary disease has not been established. Vasopressin, a powerful antidiuretic hormone involved in salt and water homeostasis, is released in response to acute hypoxia. However, the effect of prolonged hypoxaemia on hypothalamic and pituitary release of the magnocellular hypothalamic hormones, vasopressin and oxytocin, has not previously been studied. Male Wistar rats were randomly allocated to either normobaric, hypoxic (10% O2) or control (21% O2) environmental chambers. An initial series of experiments examined plasma vasopressin concentration, osmolality, sodium concentration, packed cell volume (PCV), and weight gain at weekly intervals (n = 4-6) for six weeks. The maximum increase in plasma vasopressin concentration and PCV occurred after five weeks. In a second experiment vasopressin and oxytocin concentrations in the hypothalamus, pituitary gland, and plasma were measured in eight control and eight hypoxic rats after five weeks in the environmental chambers. In rats exposed to environmental hypoxia PCV increased (p < 0.001) and weight gain decreased (p < 0.05) compared with controls. The plasma vasopressin concentration increased progressively from a baseline of 1.36 (0.2) pmol/l (n = 6) to a maximum of 4.38 (0.8) pmol/l (n = 6; p < 0.01) during the first five weeks of environmental hypoxia (difference 3.02 (95% CI 1.18 to 4.86)). Plasma osmolality and sodium concentration were unchanged in hypoxic rats compared with controls during the six week period. The hypothalamic vasopressin concentration was increased (p < 0.001) after five weeks of environmental hypoxia (91.6 (4.8) pmol/ hypothalamus) compared with controls (57.4 (5.1) pmol/hypothalamus), the difference being 34.2 pmol/hypothalamus (95% CI 21.6 to 46.5). The pituitary vasopressin concentration was unchanged. In hypoxic rats hypothalamic oxytocin (59.6 (3.2) pmol/hypothalamus) was greater (p < 0.01) than in controls (42 (3.8) pmol/hypothalamus), a difference of 17.6 pmol/ hypothalamus (95% CI 8.7 to 26.5). Similarly, the plasma oxytocin concentration was increased (p < 0.05) in hypoxic rats (6.78 (1.2) pmol/l) compared with controls (3.3 (0.8) pmol/l), a difference of 3.48 pmol/l (95% CI 0.89 to 6.07). The pituitary oxytocin concentration was unchanged in the two groups. These results demonstrate an increase in hypothalamic production of vasopressin and oxytocin in rats during prolonged hypoxaemia. Increased plasma concentrations of neurohypophysial hormones would be expected to impair sodium and water homeostasis in patients with hypoxaemia. However, the absence of change in the plasma osmolality and sodium concentrations in this study and previous clinical investigations suggests that compensatory mechanisms modulate the actions of both vasopressin and oxytocin. A reduction in renal blood flow or decreased renal responsiveness to the neurohypophyseal hormones may be involved.


ADAM gene expression and regulation during human osteoclast formation.
In this study, we identified the expression and the regulation of ADAM members (a disintegrin and metalloprotease) at both gene and protein levels during human osteoclast differentiation and activity. Human peripheral blood monocytes (HPBMC) treated with M-CSF and RANKL were used as an in vitro fusion model. In parallel, we used human osteoclastoma (OCL) tumor as a source of mature osteoclasts, and human osteoblastic cells as a control representing nonfusing and non-resorbing bone cells. RT-PCR using ADAM-specific primers enabled us to identify the expression of ADAM 8, 9, 10, 15, 17, and 28 in both osteoclasts and osteoblasts. Using primers specific for each ADAM 12 isoform (L and S), we observed a strong signal for both forms (ADAM 12L and ADAM 12S) in osteoblastic cells, while only ADAM 12S was detectable in HPBMC-derived osteoclasts and osteoclastoma. Gene regulation was studied using real-time PCR analysis performed during HPBMC differentiation; this showed a progressive increase of ADAM 12 mRNA level from day 1 to 8 of the culture, while at around day 9, ADAM 12 mRNA level decreased 2-fold. We also showed that ADAM 8, ADAM 17, and ADAM 28 decreased according to the stage of HPBMC differentiation or fusion. ADAM 10 was unaltered during cell fusion. However, confocal immunolocalization showed that ADAM 10 protein re-localized from the nuclei and cytoplasm to the plasma membrane during culture and to the ruffled border in resorbing cells. The same re-localization process was observed using an ADAM 12S-specific antibody during HPBMC differentiation. Between days 12 and 14, ADAM 12 co-localized with the F-actin ring, and at day 15, a strong signal was also present in ruffled border or sealing zone area of osteoclasts. Our results describe the expression and regulation of various ADAMs in human bone cells and the selective expression of ADAM 12L in osteoblasts. Our gene regulation and protein localization studies suggest a function for ADAM 10 and ADAM 12S in the formation of osteoclasts from HPBMC and resorption activity.


No effect of classroom sharing on educational achievement in twins: a prospective, longitudinal cohort study.
A returning dilemma for families with multiple births is whether twins should share the same, or a parallel classroom, or in other words, whether they should be separated at school or not. This study investigated the effects of sharing a classroom during primary school on cognitive achievement in twins. Subjects were 839 monozygotic and 1164 dizygotic twin pairs who were registered at birth at The Netherlands Twin Register. A prospective, longitudinal study design was used with educational achievement at age 12 years, measured with a standardised test (CITO test), as outcome measure. Most twin pairs (72%) shared a classroom during their schooling, 19% were in separate, but parallel, classes, and 9% "partly" shared a classroom. Twins who were in parallel classrooms had higher CITO scores (mean 539.51; SD 8.12), compared to twins who shared a classroom (537.99; SD 8.52). When controlling for socioeconomic status, and externalising problems before starting primary school (age 3), there was no significant difference in educational achievement between separated and non-separated twin pairs (p = 0.138). In addition, there was no interaction with sex or zygosity of the twins (p = 0.798). There is no difference in educational achievement between twins who share a classroom and twins who do not share a classroom during their primary school time. The choice of separation should be made by teachers, parents and their twin children, based on individual characteristics of a twin pair.


New first-trimester crown-rump length equations from a French general population.
The purpose of this study was to obtain precise knowledge of fetal biometric measurements, in particular crown-rump length (CRL). Our results have been carefully compared to equations found in the literature. Single-operator measurements of 2,123 spontaneous pregnancies from a general French population provided new statistical relationships between fetal age (FA) and CRL. Comparisons were made with measurements obtained from 402 in vitro fertilizations (IVFs) for which FA were known. Heteroskedastic and robust regressions were compared by cross-validation, and prediction errors were studied. All ultrasound measurements were taken during standard follow-ups of pregnancies, without any additional features. From a cleaned subsample of 513 spontaneous pregnancies, we reported good modeling of first-term embryonic growth, with equations and predictions of standard deviations agreeing with objective datations for IVFs. Most precise CRL measurements were predicted for FA of 49 days. Our results allow future detection of fetal growth abnormalities using Z-scores throughout the first trimester.


Perpetual landfilling through aeration of the waste mass; lessons from test cells in Georgia (USA).
Municipal solid waste (MSW) landfills worldwide are experiencing the consequences of conventional landfilling techniques, whereby anaerobic conditions are created within the landfilled waste. Under anaerobic conditions within a landfill site slow stabilization of the waste mass occurs, producing methane, (an explosive 'green house' gas) and leachate (which can pollute groundwater) over long periods of time. As a potential solution, it was demonstrated that the aerobic degradation of MSW within a landfill can significantly increase the rate of waste decomposition and settlement, decrease the methane production and leachate leaving the system, and potentially increase the operational life of the site. Readily integrated into the existing landfill infrastructure, this approach can safely and cost-effectively convert a MSW landfill from anaerobic to aerobic degradation processes, thereby effectively composting much of the organic portions (one of the potentially polluting elements in a conventional landfill site) of the waste. This paper summarizes the successful results of two separate aerobic landfill projects located in Georgia (USA) and discusses the potential economic and environmental impacts to worldwide solid waste management practices.


Characterization of synthetic Chikungunya viruses based on the consensus sequence of recent E1-226V isolates.
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that re-emerged in 2004 and has caused massive outbreaks in recent years. The lack of a licensed vaccine or treatment options emphasize the need to obtain more insight into the viral life cycle and CHIKV-host interactions. Infectious cDNA clones are important tools for such studies, and for mechanism of action studies on antiviral compounds. Existing CHIKV cDNA clones are based on a single genome from an individual clinical isolate, which is expected to have evolved specific characteristics in response to the host environment, and possibly also during subsequent cell culture passaging. To obtain a virus expected to have the general characteristics of the recent E1-226V CHIKV isolates, we have constructed a new CHIKV full-length cDNA clone, CHIKV LS3, based on the consensus sequence of their aligned genomes. Here we report the characterization of this synthetic virus and a green fluorescent protein-expressing variant (CHIKV LS3-GFP). Their characteristics were compared to those of natural strain ITA07-RA1, which was isolated during the 2007 outbreak in Italy. In cell culture the synthetic viruses displayed phenotypes comparable to the natural isolate, and in a mouse model they caused lethal infections that were indistinguishable from infections with a natural strain. Compared to ITA07-RA1 and clinical isolate NL10/152, the synthetic viruses displayed similar sensitivities to several antiviral compounds. 3-deaza-adenosine was identified as a new inhibitor of CHIKV replication. Cyclosporin A had no effect on CHIKV replication, suggesting that cyclophilins -opposite to what was found for other +RNA viruses- do not play an essential role in CHIKV replication. The characterization of the consensus sequence-based synthetic viruses and their comparison to natural isolates demonstrated that CHIKV LS3 and LS3-GFP are suitable and representative tools to study CHIKV-host interactions, screen for antiviral compounds and unravel their mode of action.


Neoadjuvant Nivolumab for Patients With Resectable Merkel Cell Carcinoma in the CheckMate 358 Trial.
Merkel cell carcinoma (MCC) is a rare, aggressive skin cancer commonly driven by the Merkel cell polyomavirus (MCPyV). The programmed death-1 (PD-1)/programmed death-ligand 1 (PD-L1) immunosuppressive pathway is often upregulated in MCC, and advanced metastatic MCC frequently responds to PD-1 blockade. We report what we believe to be the first trial of anti-PD-1 in the neoadjuvant setting for resectable MCC. In the phase I/II CheckMate 358 study of virus-associated cancer types, patients with resectable MCC received nivolumab 240 mg intravenously on days 1 and 15. Surgery was planned on day 29. Tumor regression was assessed radiographically and microscopically. Tumor MCPyV status, PD-L1 expression, and tumor mutational burden (TMB) were assessed in pretreatment tumor biopsies. Thirty-nine patients with American Joint Committee on Cancer stage IIA-IV resectable MCC received ≥ 1 nivolumab dose. Three patients (7.7%) did not undergo surgery because of tumor progression (n = 1) or adverse events (n = 2). Any-grade treatment-related adverse events occurred in 18 patients (46.2%), and grade 3-4 events in 3 patients (7.7%), with no unexpected toxicities. Among 36 patients who underwent surgery, 17 (47.2%) achieved a pathologic complete response (pCR). Among 33 radiographically evaluable patients who underwent surgery, 18 (54.5%) had tumor reductions ≥ 30%. Responses were observed regardless of tumor MCPyV, PD-L1, or TMB status. At a median follow-up of 20.3 months, median recurrence-free survival (RFS) and overall survival were not reached. RFS significantly correlated with pCR and radiographic response at the time of surgery. No patient with a pCR had tumor relapse during observation. Nivolumab administered approximately 4 weeks before surgery in MCC was generally tolerable and induced pCRs and radiographic tumor regressions in approximately one half of treated patients. These early markers of response significantly predicted improved RFS. Additional investigation of these promising findings is warranted.


The incremental prognostic and clinical value of multiple novel biomarkers in heart failure.
In recent years there has been an increase in the number of biomarkers in heart failure (HF). The clinical role for these novel biomarkers in combination is not clear. The following novel biomarkers were measured from 628 patients recently hospitalized with decompensated HF; mid-regional pro-adrenomedullin (MR-proADM), mid-regional pro-atrial natriuretic peptide (MR-proANP), copeptin, high-sensitivity cardiac troponin T (hs-cTnT), ST2, galectin-3, cystatin C, combined free light chains (cFLC) and high sensitivity C-reactive protein (hsCRP). The incremental prognostic value of these novel biomarkers was evaluated within an extensive model containing established predictors of mortality. During a mean (SD) follow-up of 3.2 (1.5) years, 290 (46%) patients died. Elevated concentrations of all novel biomarkers were associated with an increased unadjusted risk of mortality but only two-thirds were independent predictors following multivariable analysis. Using dichotomized cut-points from receiver operating characteristic analysis, MR-proADM, hs-cTnT, cFLC, hsCRP, and ST2 remained independent predictors of mortality. Further dichotomization into low (0-2 elevated biomarkers) or high (at least three of the five biomarkers elevated) risk groups provided greatest incremental prognostic value (hazard ratio 2.20, 95% confidence interval 1.37-3.54; P = 0.001) and improved the performance of the model (C-statistic 0.730 from 0.721, net reclassification index 32.5%). The novel biomarkers included in this study added little, if any, incremental prognostic value on their own to a model containing established predictors of mortality. However, following dichotomization, five of the novel biomarkers provided incremental prognostic value. There was a clear gradient in the risk of death with increasing numbers of elevated novel biomarkers, with the presence of at least three identifying patients at greatest risk of mortality.


Ecological role of volatiles produced by Epichloë: differences in antifungal toxicity.
Species of Epichloë (Ascomycota, Clavicipitaceae) are endophytic symbionts of pooid grasses. Sexual reproduction of the fungus depends on gamete-transferring Botanophila flies, which in earlier studies were shown to be specifically attracted by the fungal volatiles chokol K and methyl (Z)-3-methyldodec-2-enoate. As several Epichloë volatiles are known to have antimicrobial properties, it was hypothesised that the original function of insect-attracting volatiles is microbial deterrence. However, the origin of volatile compounds and their toxicity within an ecological context has not yet been clarified. We examined the inhibitory effect of chokol K and methyl (Z)-3-methyldodec-2-enoate on mycoparasites, plant pathogenic fungi and on Epichloë itself at ecologically relevant concentrations, and assessed volatile production in pure cultures of Epichloë on complex and defined media supplemented with inorganic sources of carbon and nitrogen. Chokol K reduced the spore germination of all tested fungi, whereas methyl (Z)-3-methyldodec-2-enoate had no inhibitory effect. Moreover, only chokol K was produced in culture, confirming its fungal origin. Our findings are consistent with the proposed scenario that fungal volatile substances have followed an evolutionary pathway from defence to attraction.


The solution structure of a DNA*RNA duplex containing 5-propynyl U and C; comparison with 5-Me modifications.
The addition of the propynyl group at the 5 position of pyrimidine nucleotides is highly stabilising. We have determined the thermodynamic stability of the DNA.RNA hybrid r(GAAGAGAAGC)*d(GC(p)U(p)U(p)C(p)U(p) C(p)U(p)U(p)C) where p is the propynyl group at the 5 position and compared it with that of the unmodified duplex and the effects of methyl substitutions. The incorporation of the propyne group at the 5 position gives rise to a very large stabilisation of the hybrid duplex compared with the analogous 5-Me modification. The duplexes have been characterised by gel electrophoresis and NMR spectroscopy, which indicate that methyl substitutions have a smaller influence on local and global conformation than the propynyl groups. The increased NMR spectral dispersion of the propyne-modified duplex allowed a larger number of experimental restraints to be measured. Restrained molecular dynamics in a fully solvated system showed that the propyne modification leads to substantial conformational rearrangements stabilising a more A-like structure. The propynyl groups occupy a large part of the major groove and make favourable van der Waals interactions with their nearest neighbours and the atoms of the rings. This enhanced overlap may account at least in part for the increased thermodynamic stability. Furthermore, the simulations show a spine of hydration in the major groove as well as in the minor groove involving the RNA hydroxyl groups.


Genetic dissection of a cell-autonomous neurodegenerative disorder: lessons learned from mouse models of Niemann-Pick disease type C.
Understanding neurodegenerative disease progression and its treatment requires the systematic characterization and manipulation of relevant cell types and molecular pathways. The neurodegenerative lysosomal storage disorder Niemann-Pick disease type C (NPC) is highly amenable to genetic approaches that allow exploration of the disease biology at the organismal, cellular and molecular level. Although NPC is a rare disease, genetic analysis of the associated neuropathology promises to provide insight into the logic of disease neural circuitry, selective neuron vulnerability and neural-glial interactions. The ability to control the disorder cell-autonomously and in naturally occurring spontaneous animal models that recapitulate many aspects of the human disease allows for an unparalleled dissection of the disease neurobiology in vivo. Here, we review progress in mouse-model-based studies of NPC disease, specifically focusing on the subtype that is caused by a deficiency in NPC1, a sterol-binding late endosomal membrane protein involved in lipid trafficking. We also discuss recent findings and future directions in NPC disease research that are pertinent to understanding the cellular and molecular mechanisms underlying neurodegeneration in general.


Magnetic microbubble mediated chemo-sonodynamic therapy using a combined magnetic-acoustic device.
Recent pre-clinical studies have demonstrated the potential of combining chemotherapy and sonodynamic therapy for the treatment of pancreatic cancer. Oxygen-loaded magnetic microbubbles have been explored as a targeted delivery vehicle for this application. Despite preliminary positive results, a previous study identified a significant practical challenge regarding the co-alignment of the magnetic and ultrasound fields. The aim of this study was to determine whether this challenge could be addressed through the use of a magnetic-acoustic device (MAD) combining a magnetic array and ultrasound transducer in a single unit, to simultaneously concentrate and activate the microbubbles at the target site. in vitro experiments were performed in tissue phantoms and followed by in vivo treatment of xenograft pancreatic cancer (BxPC-3) tumours in a murine model. In vitro, a 1.4-fold (p < .01) increase in the deposition of a model therapeutic payload within the phantom was achieved using the MAD compared to separate magnetic and ultrasound devices. In vivo, tumours treated with the MAD had a 9% smaller mean volume 8 days after treatment, while tumours treated with separate devices or microbubbles alone were respectively 45% and 112% larger. This substantial and sustained decrease in tumour volume suggests that the proposed drug delivery approach has the potential to be an effective neoadjuvant therapy for pancreatic cancer patients.


Human neural progenitor cell engraftment increases neurogenesis and microglial recruitment in the brain of rats with stroke.
Stem cell transplantation is to date one of the most promising therapies for chronic ischemic stroke. The human conditionally immortalised neural stem cell line, CTX0E03, has demonstrable efficacy in a rodent model of stroke and is currently in clinical trials. Nonetheless, the mechanisms by which it promotes brain repair are not fully characterised. This study investigated the cellular events occurring after CTX0E03 transplantation in the brains of rats that underwent ischemic stroke. We focused on the endogenous proliferative activity of the host brain in response to cell transplantation and determined the identity of the proliferating cells using markers for young neurons (doublecortin, Dcx) and microglia (CD11b). So as to determine the chronology of events occurring post-transplantation, we analysed the engrafted brains one week and four weeks post-transplantation. We observed a significantly greater endogenous proliferation in the striatum of ischemic brains receiving a CTX0E03 graft compared to vehicle-treated ischemic brains. A significant proportion of these proliferative cells were found to be Dcx+ striatal neuroblasts. Further, we describe an enhanced immune response after CTX0E03 engraftment, as shown by a significant increase of proliferating CD11b+ microglial cells. Our study demonstrates that few Dcx+ neuroblasts are proliferative in normal conditions, and that this population of proliferative neuroblasts is increased in response to stroke. We further show that CTX0E03 transplantation after stroke leads to the maintenance of this proliferative activity. Interestingly, the preservation of neuronal proliferative activity upon CTX0E03 transplantation is preceded and accompanied by a high rate of proliferating microglia. Our study suggests that microglia might mediate in part the effect of CTX0E03 transplantation on neuronal proliferation in ischemic stroke conditions.


The Methods Behind 2015 Informatics Capacity and Needs Assessment Study.
The 2015 Informatics Needs and Capacity of Local Health Departments (LHDs) survey is the most recent comprehensive source of quantitative data on LHD informatics. Conducted by the National Association of County & City Health Officials (NACCHO), this is the third nationally representative quantitative study of LHD informatics since 2009. The previous 2 comprehensive quantitative assessments were conducted by NACCHO in 2009-2010 and 2011. Given that public health informatics is rapidly evolving, the 2015 Informatics survey is a much-needed country-wide assessment of the current informatics needs and capacities of LHDs. This article outlines detailed methodology used in the 2015 Informatics survey, including instrument development, pretesting, sampling design and sample size, survey administration, and sampling weights. A 9-member advisory committee representing federal, state, and local health agency representatives guided the design and implementation of this study. The survey instrument was organized into 6 topic areas: demographics, physical infrastructure, skills and capacity available, public health workforce development needs, electronic health records, and health information exchange. The instrument was pretested with a sample of 20 LHDs and subsequently pilot-tested with 30 LHDs. The survey was administered via the Qualtrics survey software to the sample of 650 LHDs, selected using stratified random sampling. The survey was fielded for approximately 8 weeks and 324 usable responses were received, constituting a response rate of 50%. Statistical weights were developed to account for 3 factors: (a) disproportionate response rate by population size (using 7 population strata), (b) oversampling of LHDs with larger population sizes, and


Changes in bioturbation of iron biogeochemistry and in molecular response of the clam Ruditapes decussates upon Perkinsus olseni infection.
A series of artificial microcosms was used to test the effect of clam density on benthic iron biogeochemistry and, subsequently, if the response of clam Ruditapes decussatus to infection with Perkinsus olseni, a common opportunistic parasite known to be iron dependent, was correlated with the dynamics of iron sediment pore waters within the chambers. Three series of benthic microcosms were used in the experiment, comparing similar densities of clams (none, one, two, three, or four individuals/chamber) between a control set (no deliberate infection) and two parallel sets of clams that were deliberately infected with the parasite after 10 days of incubation. Fifteen chambers were used simultaneously and the experiment was conducted for 35 days. In order to avoid spurious effects of differential organic loading and clam feeding efficiency on the oxidative state of the sediment, the iron balance was tentatively shifted during incubation toward decreased dissolved iron in pore water. This was done by applying a constant flow of air to all chambers and refraining from supplying extra organic matter during the experimental run, which led to the reduction of benthic oxygen demand as the experiment progressed. Results showed that microcosms bearing both higher clam densities and lower infection levels were able to exert a quantitative influence in iron biogeochemistry through bioturbation activity. This effect was significantly depressed in chambers hosting clams with high infection levels. In addition, analysis of molecular markers responsive to iron and parasite stress revealed an upper regulation of HSP70 and ferritin in infected clams, thus suggesting a role of those molecules on both host protection and response to parasite presence by limiting iron availability. Together, these findings suggest a correlation between the expression of clam molecular iron/stress markers and iron bioavailability, which can be modified by the presence or absence of Perkinsus infection. In turn, we propose that clam lethargy in response to parasite invasion might help to combat infection by reducing iron mobilization in the surrounding sediment through a decrease in bioturbation activity, thus reducing its availability to the parasite.


Inflammatory Biomarker Levels After Propofol or Sevoflurane Anesthesia: A Meta-analysis.
The perioperative inflammatory response may be implicated in adverse outcomes including neurocognitive dysfunction and cancer recurrence after oncological surgery. The immunomodulatory role of anesthetic agents has been demonstrated in vitro; however, its clinical relevance is unclear. The purpose of this meta-analysis was to compare propofol and sevoflurane with respect to biomarkers of perioperative inflammation. The secondary aim was to correlate markers of inflammation with clinical measures of perioperative cognition. Databases were searched for randomized controlled trials examining perioperative inflammation after general anesthesia using propofol compared to sevoflurane. Inflammatory biomarkers investigated were interleukin (IL)-6, IL-10, tissue necrosis factor alpha (TNF-α), and C-reactive protein (CRP). The secondary outcome was incidence of perioperative neurocognitive disorders. Meta-analysis with metaregression was performed to determine the difference between propofol and sevoflurane. Twenty-three studies were included with 1611 participants. Studies varied by surgery type, duration, and participant age. There was an increase in the mean inflammatory biomarker levels following surgery, with meta-analysis revealing no difference in effect between propofol and sevoflurane. Heterogeneity between studies was high, with surgery type, duration, and patient age contributing to the variance across studies. Only 5 studies examined postoperative cognitive outcomes; thus, a meta-analysis could not be performed. Nonetheless, of these 5 studies, 4 reported a reduced incidence of cognitive decline associated with propofol use. Surgery induces an inflammatory response; however, the inflammatory response did not differ as a function of anesthetic technique. This absence of an effect suggests that patient and surgical variables may have a far more significant impact on the postoperative inflammatory responses than anesthetic technique. The majority of studies assessing perioperative cognition in older patients reported a benefit associated with the use of propofol; however, larger trials using homogenous outcomes are needed to demonstrate such an effect.


[Proton therapy].
This review summarises the potential usage of proton therapy in Denmark. About one third of Danes are diagnosed with cancer, and half of these need radiotherapy in the course of treatment. Radiation dose cannot be adequately increased without giving rise to unacceptable, high risk of toxicity, but proton therapy is encouraging due to a unique depth dose distribution. In some cases, the benefit of proton therapy is obvious, but in most cases the gain is less obvious, and patients should only receive treatment within clinical trials. Clinical studies on proton therapy with focus on reduction of radiation-induced side effects and improvement of quality of life should be conducted.


Adipocytokines in obesity and metabolic disease.
The current global obesity pandemic is the leading cause for the soaring rates of metabolic diseases, especially diabetes, cardiovascular disease, hypertension, and non-alcoholic hepatosteatosis. Efforts devoted to find cures for obesity and associated disorders in the past two decades have prompted intensive interest in adipocyte biology, and have led to major advances in the mechanistic understanding of adipose tissue as an essential endocrine organ. Adipose tissue secretes an array of hormones (adipokines) that signal key organs to maintain metabolic homeostasis, and their dysfunction has been causally linked to a wide range of metabolic diseases. In addition, obesity induces production of inflammatory cytokines (often referred to together with adipokines as adipocytokines) and infiltration of immune cells into adipose tissue, which creates a state of chronic low-grade inflammation. Metabolic inflammation has been increasingly recognized as a unifying mechanism linking obesity to a broad spectrum of pathological conditions. This review focuses on classic examples of adipocytokines that have helped to form the basis of the endocrine and inflammatory roles of adipose tissue, and it also details a few newly characterized adipocytokines that provide fresh insights into adipose biology. Studies of adipocytokines in clinical settings and their therapeutic potential are also discussed.


Do a robot's social skills and its objection discourage interactants from switching the robot off?
Building on the notion that people respond to media as if they were real, switching off a robot which exhibits lifelike behavior implies an interesting situation. In an experimental lab study with a 2x2 between-subjects-design (N = 85), people were given the choice to switch off a robot with which they had just interacted. The style of the interaction was either social (mimicking human behavior) or functional (displaying machinelike behavior). Additionally, the robot either voiced an objection against being switched off or it remained silent. Results show that participants rather let the robot stay switched on when the robot objected. After the functional interaction, people evaluated the robot as less likeable, which in turn led to a reduced stress experience after the switching off situation. Furthermore, individuals hesitated longest when they had experienced a functional interaction in combination with an objecting robot. This unexpected result might be due to the fact that the impression people had formed based on the task-focused behavior of the robot conflicted with the emotional nature of the objection.


A Framework for Applying Natural Language Processing in Digital Health Interventions.
Digital health interventions (DHIs) are poised to reduce target symptoms in a scalable, affordable, and empirically supported way. DHIs that involve coaching or clinical support often collect text data from 2 sources: (1) open correspondence between users and the trained practitioners supporting them through a messaging system and (2) text data recorded during the intervention by users, such as diary entries. Natural language processing (NLP) offers methods for analyzing text, augmenting the understanding of intervention effects, and informing therapeutic decision making. This study aimed to present a technical framework that supports the automated analysis of both types of text data often present in DHIs. This framework generates text features and helps to build statistical models to predict target variables, including user engagement, symptom change, and therapeutic outcomes. We first discussed various NLP techniques and demonstrated how they are implemented in the presented framework. We then applied the framework in a case study of the Healthy Body Image Program, a Web-based intervention trial for eating disorders (EDs). A total of 372 participants who screened positive for an ED received a DHI aimed at reducing ED psychopathology (including binge eating and purging behaviors) and improving body image. These users generated 37,228 intervention text snippets and exchanged 4285 user-coach messages, which were analyzed using the proposed model. We applied the framework to predict binge eating behavior, resulting in an area under the curve between 0.57 (when applied to new users) and 0.72 (when applied to new symptom reports of known users). In addition, initial evidence indicated that specific text features predicted the therapeutic outcome of reducing ED symptoms. The case study demonstrates the usefulness of a structured approach to text data analytics. NLP techniques improve the prediction of symptom changes in DHIs. We present a technical framework that can be easily applied in other clinical trials and clinical presentations and encourage other groups to apply the framework in similar contexts.


Genuair® Usability Test: Results of a National Public Survey of the Elderly.
Proper use of inhaler devices may be problematic in elderly patients due to age-related difficulties. A survey was administered to elderly patients to investigate the usability of the Genuair® device and patients' subjective viewpoint on the device. A representative sample of the Italian population aged ≥ 65 years was completed with a pre-defined sample of 89 patients with hand arthritis/arthrosis. Of 526 respondents, 88 were not self-sufficient. Only the replies of the 438 self-sufficient respondents were analyzed. A total of 107 participants (24%) reported having respiratory diseases, and 81 of these (76%) were users of inhaler devices. After the first test, the device was considered "practical/handy" by 90% of patients and "easy to use" by 89%. After the second test, in which patients received a demonstration of the correct inhalation maneuver, the percentage of patients scoring ≥ 7 increased to 93% for the first characteristic and was confirmed for the second, with no differences between the groups in terms of age, educational level, use of devices, and presence of arthritis/arthrosis. The mean time to explain the inhaler technique and to perform a correct inhalation was 1'38"± 1'37", and 70% of the respondents required less than 2 minutes, with no differences between the groups in terms of age, education level, use of devices, and presence of arthritis/arthrosis. In conclusion, Genuair® was well accepted and easy to use in a representative sample of the Italian population aged ≥ 65 years. These characteristics make it a valid choice in the elderly, thus enabling patients to better cope with the problems and difficulties that are common to this age group.


Lanthanide complexes as chiral probes exploiting circularly polarized luminescence.
Optical probes that provide information about local chirality have been developed based on changes to the circular polarisation of emitted light. Highly emissive complexes of lanthanide ions are ideally suited for CPL spectroscopy and the design criteria for developing such probes are defined, based on a rigorous stereochemical analysis. The perturbation of a dynamically racemic complex may occur either by a change in complex constitution or by non-covalent association. With complexes of enantiopure ligands, perturbation may involve either dynamic helicity inversion or a reversible change in the lanthanide coordination environment.


Mortality in patients with TIMI 3 flow after PCI in relation to time delay to reperfusion.
Percutaneous coronary intervention (PCI) performed within 12 h from symptom onset enables complete blood flow restoration in infarct-related artery in 90% of patients. Nevertheless, even with complete restoration of epicardial blood flow in culprit vessel (postprocedural Thrombolysis in Myocardial Infarction (TIMI) flow grade 3), myocardial perfusion at tissue level may be insufficient. We hypothesized that the outcome of patients with STEMI/bundle branch block (BBB)-myocardial infarction and post-PCI TIMI 3 flow is related to the time to reperfusion. Observational study based on a retrospective analysis of population of 635 consecutive patients with STEMI/BBB-MI and post-PCI TIMI 3 flow from January 2009 to December 2011 (mean age 63 years, 69.6% males). Mortality of patients was evaluated in relation to the time from symptom onset to reperfusion. A total of 83 patients (13.07%) with postprocedural TIMI 3 flow after PCI had died at 1-year follow-up. Median TD in patients who survived was 3.92 h (iqr 5.43), in patients who died 6.0 h (iqr 11.42), P = 0.004. Multiple logistic regression analysis identified time delay ≥ 9 h as significantly related to 1-year mortality of patients with STEMI/BBB-MI and post-PCI TIMI 3 flow (OR 1.958, P = 0.026). Other significant variables associated with mortality in multivariate regression analysis were: left ventricle ejection fraction < 30% (P = 0.006), age > 65 years (P < 0.001), Killip class >2 (P <0.001), female gender (P = 0.019), and creatinine clearance < 30 mL/min (P < 0.001). Time delay to reperfusion is significantly related to 1-year mortality of patients with STEMI/BBB-MI and complete restoration of epicardial blood flow in culprit vessel after PCI.


Identification of amino acids critical for the DNA binding and dimerization properties of the human retinoic acid receptor alpha. Importance of lysine 360, lysine 365, and valine 361.
Retinoic acid receptors (RARs) and retinoid X receptors (RXRs) activate target genes by binding to retinoic acid response elements (RAREs) as heterodimeric, asymmetrical complexes, and display a high degree of cooperativity in binding to RAREs. We have examined here the effect of lysine, cysteine, arginine, histidine, and tyrosine side chain chemical modification on the DNA binding, homo- and heterodimerization properties of the full-length human retinoic acid receptor alpha (hRARalpha). Lysines are the only residues to be engaged in the dimerization with human retinoid X receptor alpha (hRXRalpha) in the absence of DNA, whereas histidines are selectively involved in the homodimerization of hRARalpha in the presence of a RARE. Arginine modification affected the DNA binding activity of each type of dimer, whereas cysteines and tyrosines were primarily involved in the homo- or heterodimerization process in the presence of the same RARE. Modified lysines, interfering with the dimerization with hRXRalpha, were identified by receptor labeling and peptide mapping. They are located in the hormone binding domain eighth heptad repeat, at positions 360 and 365. In keeping with these results, mutation of Lys360, Val361, and Lys365 diminished strongly the DNA binding activity of hRARalpha as a homodimer or a heterodimer. Our results thus provide direct evidence for the differential involvement of basic, polar, or aromatic amino acids in the DNA binding, homodimerization, and heterodimerization properties of hRARalpha. Furthermore, they demonstrate the use of distinct dimerization interfaces and identify the type of amino acids involved in these protein-protein interactions.


Effective glomerular filtration pressure and single nephron filtration rate during hydropenia, elevated ureteral pressure, and acute volume expansion with isotonic saline.
Free-flow and stop-flow intratubular pressures were measured in rats with an improved Gertz technique using Landis micropipets or a Kulite microtransducer. In hydropenia, average single nephron glomerular filtration rate was 29.3 nl/min, glomerular hydrostatic pressure (stop-flow pressure + plasma colloid osmotic pressure) was 70 cm H(2)O and mean glomerular effective filtration pressure was 12.7-14.3 cm H(2)O, approaching zero at the efferent end of the glomerulus. Thus, the glomerulus is extremely permeable, having a filtration coefficient four to five times greater than previously estimated. Mean effective filtration pressure and single nephron glomerular filtartion rate fell with elevated ureteral pressure and rose with volume expansion, more or less proportionately. Changes in effective filtration pressure were due primarily to increased intratubular pressure in ureteral obstruction and to reduced plasma colloid osmotic pressure in volume expansion; glomerular hydrostatic pressure remained constant in both conditions and thus played no role in regulation of filtration rate.


Redox regulation of mitochondrial biogenesis.
The cell renews, adapts, or expands its mitochondrial population during episodes of cell damage or periods of intensified energy demand by the induction of mitochondrial biogenesis. This bigenomic program is modulated by redox-sensitive signals that respond to physiological nitric oxide (NO), carbon monoxide (CO), and mitochondrial reactive oxygen species production. This review summarizes our current ideas about the pathways involved in the activation of mitochondrial biogenesis by the physiological gases leading to changes in the redox milieu of the cell, with an emphasis on the responses to oxidative stress and inflammation. The cell's energy supply is protected from conditions that damage mitochondria by an inducible transcriptional program of mitochondrial biogenesis that operates in large part through redox signals involving the nitric oxide synthase and the heme oxygenase-1/CO systems. These redox events stimulate the coordinated activities of several multifunctional transcription factors and coactivators also involved in the elimination of defective mitochondria and the expression of counterinflammatory and antioxidant genes, such as IL10 and SOD2, as part of a unified damage-control network. The redox-regulated mechanisms of mitochondrial biogenesis schematically outlined in the graphical abstract link mitochondrial quality control to an enhanced capacity to support the cell's metabolic needs while improving its resistance to metabolic failure and avoidance of cell death during periods of oxidative stress.


Blowing big bubbles.
Although street artists have the know-how to blow bubbles over one meter in length, the bubble width is typically determined by the size of the hoop, or wand they use. In this article we explore a regime in which, by blowing gently downwards, we generate bubbles with radii up to ten times larger than the wand. We observe the big bubbles at lowest air speeds, analogous to the dripping mode observed in droplet formation. We also explore the impact of the surfactant chosen to stabilize the bubbles. We are able to create bubbles of comparable size using either Fairy liquid, a commercially available detergent often used by street artists, or sodium dodecyl sulfate (SDS) solutions. The bubbles obtained from Fairy liquid detach from the wand and are stable for several seconds, however those from SDS tend to burst just before detachment.


Association between environmental dust exposure and lung cancer in dogs.
The objective of this study was to investigate the relationship between the accumulation of black dust matter in lungs (anthracosis) and primary lung cancer in dogs. A retrospective study was carried out on material from 35 dogs with primary lung cancer and 160 controls. The amount, histological appearance and birefringence of anthracosis were assessed in pulmonary specimens by light microscopy, and the odds ratio (OR) calculated for dogs with primary lung cancer. The same factors were analysed to identify an association between tumour histotype, histological grade, and clinical stage. Papillary adenocarcinoma was most commonly diagnosed (45.7%). The majority of tumours were of histological grade II, and the lung cancer was more often localised (clinical stage I). An increased risk of lung cancer was observed in dogs with higher amounts of anthracosis (OR: 2.11, CI 95%: 1.20-3.70; P < 0.01), which suggests an association between anthracosis due to inhalation of polluted air and lung cancer in dogs.


The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas.
Lymphocytes of melanoma patients can be restimulated in vitro with autologous tumor cells to generate antitumor cytolytic T lymphocytes (CTL). Previous reports have indicated that, when such CTL are obtained from HLA-A2 melanoma patients, they often display broad reactivity on A2 melanoma cell lines. Such antitumor CTL clones, which appeared to recognize the same antigen, were isolated from two patients. We report here the cloning of a cDNA that directs the expression of the antigen recognized by these CTL. This cDNA corresponds to the transcript of the tyrosinase gene. The gene was found to be active in all tested melanoma samples and in most melanoma cell lines. Among normal cells, only melanocytes appear to express the gene. The tyrosinase antigen presented by HLA-A2 may therefore constitute a useful target for specific immunotherapy of melanoma. But possible adverse effects of antityrosinase immunization, such as the destruction of normal melanocytes and its consequences, will have to be examined before clinical pilot studies can be undertaken.


A donor cornea with metastatic cells from a cutaneous malignant melanoma.
To describe the case of a donor cornea that showed hematogenous metastatic spread of cutaneous melanoma to the sclerocorneal limbus. Corneal tissue obtained from a donor with cutaneous malignant melanoma was evaluated for endothelial cell density, corneal transparency, and epithelial morphology. Subsequently, hematoxylin and eosin staining and immunohistochemical characterization using S100, HMB45, Melan-A, and CD34 antibodies were performed on the corneal sections. The corneal tissue was transparent with high endothelial cell density; it was graded as being suitable for transplantation according to the current eye bank criteria. However, the aggressiveness of the donor's cancer and the diffuse melanosis of the sclera led to the suspicion of malignant melanoma metastasis to the cornea. Histochemical analysis of the corneoscleral rim showed small aggregates rich in pigmented cells that were localized in cleft-like structures in the sclera, at the sclerocorneal interface and in the peripheral avascular portion of the cornea. The aggregates were positive for the melanocytic tumor markers S100, HMB45, and Melan-A; the rims of the clefts expressed the panvascular CD34 antigen, which was suggestive of neovascularization. Corneal tissue from a donor with malignant cutaneous melanoma displayed neoplastic lesions of melanocytic origin that had spread from a primitive melanoma through hematogenous routes to the sclerocorneal limbus. On the basis of this finding, we believe that having a metastatic cutaneous malignant melanoma could in some cases be reviewed as an exclusionary criterion for undergoing cornea transplantation.


On the influence of water on the electronic structure of firefly oxyluciferin anions from absorption spectroscopy of bare and monohydrated ions in vacuo.
A complete understanding of the physics underlying the varied colors of firefly bioluminescence remains elusive because it is difficult to disentangle different enzyme-lumophore interactions. Experiments on isolated ions are useful to establish a proper reference when there are no microenvironmental perturbations. Here, we use action spectroscopy to compare the absorption by the firefly oxyluciferin lumophore isolated in vacuo and complexed with a single water molecule. While the process relevant to bioluminescence within the luciferase cavity is light emission, the absorption data presented here provide a unique insight into how the electronic states of oxyluciferin are altered by microenvironmental perturbations. For the bare ion we observe broad absorption with a maximum at 548 ± 10 nm, and addition of a water molecule is found to blue-shift the absorption by approximately 50 nm (0.23 eV). Test calculations at various levels of theory uniformly predict a blue-shift in absorption caused by a single water molecule, but are only qualitatively in agreement with experiment highlighting limitations in what can be expected from methods commonly used in studies on oxyluciferin. Combined molecular dynamics simulations and time-dependent density functional theory calculations closely reproduce the broad experimental peaks and also indicate that the preferred binding site for the water molecule is the phenolate oxygen of the anion. Predicting the effects of microenvironmental interactions on the electronic structure of the oxyluciferin anion with high accuracy is a nontrivial task for theory, and our experimental results therefore serve as important benchmarks for future calculations.


The C. elegans glycosyltransferase BUS-8 has two distinct and essential roles in epidermal morphogenesis.
Ventral enclosure in Caenorhabditis elegans involves migration of epidermal cells over a neuroblast substrate and subsequent adhesion at the ventral midline. Organisation of the neuroblast layer by ephrins and their receptors is essential for this migration. We show that bus-8, which encodes a predicted glycosyltransferase, is essential for embryonic enclosure and acts in or with ephrin signalling to mediate neuroblast organisation and to permit epidermal migration. BUS-8 acts non-cell-autonomously in this process, and likely modifies an extracellular regulator of ephrin signalling and cell organisation. Weak and cold-sensitive alleles of bus-8 show that the gene has a separate and distinct post-embryonic role, being essential for epidermal integrity and production of the cuticle surface. This disorganisation of the epidermis and cuticle layers causes increased drug sensitivity, which could aid the growing use of C. elegans in drug screening and chemical genomics. The viable mutants are also resistant to infection by the pathogen Microbacterium nematophilum, due to failure of the bacterium to bind to the host surface. The two separate essential roles of BUS-8 in epidermal morphogenesis add to our growing understanding of the widespread importance of glycobiology in development.