Identification of genes differentially expressed in benign versus malignant thyroid tumors.
Although fine-needle aspiration biopsy is the most useful diagnostic tool in evaluating a thyroid nodule, preoperative diagnosis of thyroid nodules is frequently imprecise, with up to 30% of fine-needle aspiration biopsy cytology samples reported as "suspicious" or "indeterminate." Therefore, other adjuncts, such as molecular-based diagnostic approaches are needed in the preoperative distinction of these lesions. In an attempt to identify diagnostic markers for the preoperative distinction of these lesions, we chose to study by microarray analysis the eight different thyroid tumor subtypes that can present a diagnostic challenge to the clinician. Our microarray-based analysis of 94 thyroid tumors identified 75 genes that are differentially expressed between benign and malignant tumor subtypes. Of these, 33 were overexpressed and 42 were underexpressed in malignant compared with benign thyroid tumors. Statistical analysis of these genes, using nearest-neighbor classification, showed a 73% sensitivity and 82% specificity in predicting malignancy. Real-time reverse transcription-PCR validation for 12 of these genes was confirmatory. Western blot and immunohistochemical analyses of one of the genes, high mobility group AT-hook 2, further validated the microarray and real-time reverse transcription-PCR data. Our results suggest that these 12 genes could be useful in the development of a panel of markers to differentiate benign from malignant tumors and thus serve as an important first step in solving the clinical problem associated with suspicious thyroid lesions.


Both induction and morphogenesis of cyst nematode feeding cells are mediated by auxin.
Various lines of evidence show that local changes in the auxin concentration are involved in the initiation and directional expansion of syncytia induced by cyst nematodes. Analysis of nematode infections on auxin-insensitive tomato and Arabidopsis mutants revealed various phenotypes ranging from complete inhibition of syncytium development to a decrease in hypertrophy and lateral root formation at the infection site. Specific activation of an auxin-responsive promoter confirmed the role of auxin and pointed at a local accumulation of auxin in developing syncytia Disturbance of auxin gradients by inhibiting polar auxin transport with N-(1-naphthyl)phtalamic acid (NPA) resulted in abnormal feeding cells, which were characterized by extreme galling, massive disordered cell divisions in the cortex, and absence of radial expansion of the syncytium initial toward the vascular bundle. The role of auxin gradients in guiding feeding cell morphogenesis and the cross-talk between auxin and ethylene resulting in a local activation of cell wall degrading enzymes are discussed.


Diagnostic implications of tear protein profiles.
Concentrations of lysozyme, lactoferrin, ceruloplasmin, IgA, and IgG have been measured in tears by the ELISA (enzyme-linked immunosorbent assay) technique. Tears were collected on weighed filter paper discs, after which they were eluted into buffer and transported frozen to a remote laboratory for assay. Patients with sicca, questionably dry eyes and ocular pemphigoid were sampled, as were 54 normal volunteers. Tear protein profiles were established which were unique for each condition and clearly differed from the normal controls. The assay developed is considered suitable for other proteins such as IgE, and could also be used for monitoring the effects of drugs on the lacrimal gland.


Norepinephrine transporter inhibition with desipramine exacerbates L-DOPA-induced dyskinesia: role for synaptic dopamine regulation in denervated nigrostriatal terminals.
Pharmacological dopamine (DA) replacement with Levodopa [L-dihydroxyphenylalanine (L-DOPA)] is the gold standard treatment of Parkinson's disease (PD). However, long-term L-DOPA treatment is complicated by eventual debilitating abnormal involuntary movements termed L-DOPA-induced dyskinesia (LID), a clinically significant obstacle for the majority of patients who rely on L-DOPA to alleviate PD-related motor symptoms. The manifestation of LID may in part be driven by excessive extracellular DA derived from L-DOPA, but potential involvement of DA reuptake in LID severity or expression is unknown. We recently reported that in 6-hydroxydopamine (6-OHDA)-lesioned striatum, norepinephrine transporter (NET) expression increases and may play a significant role in DA transport. Furthermore, L-DOPA preferentially inhibits DA uptake in lesioned striatum. Therefore, we hypothesized that desipramine (DMI), a NET antagonist, could affect the severity of LID in an established LID model. Whereas DMI alone elicited no dyskinetic effects in lesioned rats, DMI + L-DOPA-treated rats gradually expressed more severe dyskinesia compared with L-DOPA alone over time. At the conclusion of the study, we observed reduced NET expression and norepinephrine-mediated inhibition of DA uptake in the DMI + L-DOPA group compared with L-DOPA-alone group in lesioned striatum. LID severity positively correlated with striatal extracellular signal-regulated protein kinase phosphorylation among the three treatment groups, with increased ppERK1/2 in DMI + L-DOPA group compared with the L-DOPA- and DMI-alone groups. Taken together, these results indicate that the combination of chronic L-DOPA and NET-mediated DA reuptake in lesioned nigrostriatal terminals may have a role in LID severity in experimental Parkinsonism.


Father-Inclusive Perinatal Parent Education Programs: A Systematic Review.
Fathers contribute to their children's health starting at the beginning of life. Few parent education programs include fathers. Among those that do, there is little effort to report program effects on father outcomes. In this systematic review, we examined father-inclusive perinatal parent education programs in the United States as they relate to a range of father outcomes. The databases searched were PubMed, Cumulative Index to Nursing and Allied Health Literature, Embase, Ovid Medline, Cochrane Central Register of Controlled Trials, and PsycINFO. Studies were included if they included an evaluation of a parent education program and a report of father outcomes measured within 1 year of the child's birth and were conducted within the United States. Of 1353 total articles, 21 met study criteria. The overall state of the father-inclusive perinatal parent education program literature was poor, with few interventions available to fathers. Available programs were associated with increased father involvement, coparenting relationship, partner relationship quality, father's mental health, and father's supportive behaviors. Program effects on father-infant interaction, parenting knowledge, and attitudes and parenting self-efficacy were inconclusive. Three programs emerged as best evidence-based interventions. Risk of bias was high for many studies. Outcome variability, small sample size, and publication bias contributed to the weak evidence base. There is a need for more evidence-based interventions to support fathers. Clinicians play a key role in engaging fathers in early parent education programs and health care settings. PROSPERO registration number: CRD42017050099.


Synchronized moving aperture radiation therapy (SMART): superimposing tumor motion on IMRT MLC leaf sequences under realistic delivery conditions.
Synchronized moving aperture radiation therapy (SMART) has been proposed to account for tumor motions during radiotherapy in prior work. The basic idea of SMART is to synchronize the moving radiation beam aperture formed by a dynamic multileaf collimator (DMLC) with the tumor motion induced by respiration. In this paper, a two-dimensional (2D) superimposing leaf sequencing method is presented for SMART. A leaf sequence optimization strategy was generated to assure the SMART delivery under realistic delivery conditions. The study of delivery performance using the Varian LINAC and the Millennium DMLC showed that clinical factors such as collimator angle, dose rate, initial phase and machine tolerance affect the delivery accuracy and efficiency. An in-house leaf sequencing software was developed to implement the 2D superimposing leaf sequencing method and optimize the motion-corrected leaf sequence under realistic clinical conditions. The analysis of dynamic log (Dynalog) files showed that optimization of the leaf sequence for various clinical factors can avoid beam hold-offs which break the synchronization of SMART and fail the SMART dose delivery. Through comparison between the simulated delivered fluence map and the planed fluence map, it was shown that the motion-corrected leaf sequence can greatly reduce the dose error.


Design and characterization of modular scaffolds for tubulin assembly.
In cells, microtubule dynamics is regulated by stabilizing and destabilizing factors. Whereas proteins in both categories have been identified, their mechanism of action is rarely understood at the molecular level. This is due in part to the difficulties faced in structural approaches to obtain atomic models when tubulin is involved. Here, we design and characterize new stathmin-like domain (SLD) proteins that sequester tubulins in numbers different from two, the number of tubulins bound by stathmin or by the SLD of RB3, two stathmin family members that have been extensively studied. We established rules for the design of tight tubulin-SLD assemblies and applied them to complexes containing one to four tubulin heterodimers. Biochemical and structural experiments showed that the engineered SLDs behaved as expected. The new SLDs will be tools for structural studies of microtubule regulation. The larger complexes will be useful for cryo-electron microscopy, whereas crystallography or nuclear magnetic resonance will benefit from the 1:1 tubulin-SLD assembly. Finally, our results provide new insight into SLD function, suggesting that a major effect of these phosphorylatable proteins is the programmed release of sequestered tubulin for microtubule assembly at the specific cellular locations of members of the stathmin family.


Promoting healthy teenage behaviour across three European countries through the use of a novel smartphone technology platform, PEGASO fit for future: study protocol of a quasi-experimental, controlled, multi-Centre trial.
Behaviour change interventions targeting physical activity, diet, sleep and sedentary behaviour of teenagers show promise when delivered through smartphones. However, to date there is no evidence of effectiveness of multicomponent smartphone-based interventions. Utilising a user-centred design approach, we developed a theory-based, multi-dimensional system, PEGASO Fit For Future (PEGASO F4F), which exploits sophisticated game mechanics involving smartphone applications, a smartphone game and activity sensors to motivate teenagers to take an active role in adopting and maintaining a healthy lifestyle. This paper describes the study protocol to assess the feasibility, usability and effectiveness (knowledge/awareness and behavioural change in lifestyle) of the PEGASO system. We are conducting a quasi-experimental controlled cluster trial in 4 sites in Spain, Italy, and UK (England, Scotland) over 6 months. We plan to recruit 525, in a 2:1 basis, teenagers aged 13-16 years from secondary schools. The intervention group is provided with the PEGASO system whereas the comparison group continues their usual educational routine. Outcomes include feasibility, acceptance, and usability of the PEGASO system as well as between and within group changes in motivation, self-reported diet, physical activity, sedentary and sleeping behaviour, anthropometric measures and knowledge about a healthy lifestyle. PEGASO F4F will provide evidence into the cross-cultural similarities and differences in the feasibility, acceptability and usability of a multi-dimensional smartphone based behaviour change intervention for teenagers. The study will explore facilitating factors, challenges and barriers of engaging teenagers to adapt and maintain a healthy lifestyle when using smartphone technology. Positive results from this ICT based multi component intervention may have significant implications both at clinical level, improving teenagers health and at public health level since it can present an influential tool against the development of chronic disease during adulthood. https://clinicaltrials.gov Registration number: NCT02930148, registered 4 October 2016.


Synaptic relationships of serotonin-immunoreactive terminal baskets on GABA neurons in the cat auditory cortex.
Correlative light and electron microscopic immunocytochemical methods were used to analyze the 5-HT innervation of the primary auditory area (AI) of the cat cerebral cortex and to examine the synaptic relationships of 5-HT basket terminations on target neurons in that area. Three morphological types of 5-HT-immunoreactive fibers are present: type I, which is very thin and very finely beaded; type II, which is thin and coarsely beaded; and type III, which has a relatively thick main shaft and very few beads. Type I is the most abundant, type II is relatively less common, and type III is the least abundant type. The 3 types of fibers are present through the thickness of AI and in the subjacent white matter, but the densest plexus is found in layers I-III. One of the most characteristic features of type II fibers is that they commonly form small, dense clusters that resemble baskets apposed to the somata and primary dendrites of unstained neurons. The basket formations are more frequently found in layers I and II, and they vary in complexity. Simultaneous immunostaining for GABA and 5-HT reveals that many 5-HT baskets surround the somata and dendrites of GABA neurons. In 2-microns-thick plastic sections, each basket formation can be seen surrounding 1 or a group of 2 or 3 cells. In the latter case, one cell is much larger and at the electron microscope level is identified as a neuron, while the other cells are neuroglial cells. Reconstructions were made from serial electron micrographs of 135 5-HT-immunoreactive boutons. Of these boutons, 110 belonged to basket formations, 14 to type I axons located in the neuropil, and the remaining 11 to type II fibers located in the white matter. Only 4 of the 135 boutons made conventional synaptic contacts. These were of the asymmetrical type. Most of the boutons made very small, indistinct membrane specializations or none at all. The present results therefore suggest a strong interaction between 5-HT axon terminals and specific GABA neurons, which may be mediated by release sites that are not associated with morphologically distinct synaptic contacts.


Senescent CD14+CD16+ monocytes exhibit proinflammatory and proatherosclerotic activity.
In elderly subjects and in patients with chronic inflammatory diseases, there is an increased subset of monocytes with a CD14(+)CD16(+) phenotype, whose origin and functional relevance has not been well characterized. In this study, we determined whether prolonged survival of human CD14(++)CD16(-) monocytes promotes the emergence of senescent cells, and we analyzed their molecular phenotypic and functional characteristics. We used an in vitro model to prolong the life span of healthy monocytes. We determined cell senescence, intracellular cytokine expression, ability to interact with endothelial cells, and APC activity. CD14(+)CD16(+) monocytes were senescent cells with shortened telomeres (215 ± 37 relative telomere length) versus CD14(++)CD16(-) cells (339 ± 44 relative telomere length; p < 0.05) and increased expression of β-galactosidase (86.4 ± 16.4% versus 10.3 ± 7.5%, respectively; p = 0.002). CD14(+)CD16(+) monocytes exhibited features of activated cells that included expression of CD209, release of cytokines in response to low-intensity stimulus, and increased capacity to sustain lymphocyte proliferation. Finally, compared with CD14(++)CD16(-) cells, CD14(+)CD16(+) monocytes showed elevated expression of chemokine receptors and increased adhesion to endothelial cells (19.6 ± 8.1% versus 5.3 ± 4.1%; p = 0.033). In summary, our data indicated that the senescent CD14(+)CD16(+) monocytes are activated cells, with increased inflammatory activity and ability to interact with endothelial cells. Therefore, accumulation of senescent monocytes may explain, in part, the development of chronic inflammation and atherosclerosis in elderly subjects and in patients with chronic inflammatory diseases.


Genetic variability and evolutionary analysis of parietaria mottle virus: role of selection and genetic exchange.
The genetic variability and evolution of parietaria mottle virus (PMoV) of the genus Ilarvirus was studied by analyzing nucleotide sequences of 2b and CP genes from isolates collected in different countries. Phylogenetic analysis showed that PMoV isolates clustered in different clades: one (clade I) composed of only Italian isolates and three clades (clades II-IV) including the Spanish isolates. The Greek isolate GrT-1 used in this study was in clade IV for the CP phylogenetic tree whereas it formed a separate branch in the 2b phylogenetic tree. The nucleotide sequence diversity of both the 2b and CP genes was low (0.062 ± 0.006 and 0.063 ± 0.006 for 2b and CP, respectively) but higher than those of other ilarviruses. Distribution of synonymous and nonsynonymous substitutions revealed that 2b and CP proteins are under purifying selection, with some positions under diversifying selection. Genetic exchange among Spanish isolates was also detected.


Different Phenotypes of Non-Steroidal Anti-Inflammatory Drug Hypersensitivity during Childhood.
Although non-steroidal anti-inflammatory drug hypersensitivity (NSAID-H) has been widely studied in adults, there is still a lack of data regarding the features and phenotypes of NSAID-H in children. Our aim was to define risk factors and different phenotypes according to clinical patterns. Patients with a history of reaction to any NSAIDs referred between January 2012 and October 2014 were included. After completing a European Network for Drug Allergy (ENDA) questionnaire, initial skin and/or oral provocation tests (OPTs) were performed for the offending drug. Additional OPTs were done with aspirin in case of NSAID-H to determine cross-reactivity. NSAID-hypersensitive patients were defined as being either a selective responder (SR) or cross-intolerant (CI) and further categorized according to either the ENDA/GA2LEN classification or an alternative scheme by Caimmi et al. [Int Arch Allergy Immunol 2012;159:306-312]. Among 121 patients [58.7% male, average age 7.8 years (4.7-10.8)] with 161 NSAID-related reactions, 110 patients with 148 reactions were assessed. NSAID-H was diagnosed in 30 (27%) patients with 37 (25%) reactions. Multivariate regression analysis revealed that an immediate-type reaction and respiratory symptoms during the reaction increased the risk of a reproducible NSAID-related reaction (OR 3.508, 95% CI 1.42-8.7, p = 0.007; OR 3.951, 95% CI 1.33-11.77, p = 0.014, respectively). Additional OPTs revealed 13 SRs and 14 CIs. A family history of allergic disease was more frequent in CIs compared to SRs (57.1 vs. 15.4%, p = 0.031). Reactions belonging to CIs were more frequently characterized by angioedema compared to those of SRs (81.3 vs. 46.2%, p = 0.019). SRs and CIs were further classified as single NSAID-induced urticaria/angioedema and/or anaphylaxis (n = 13), NSAID-induced urticaria/angioedema (n = 7), NSAID-exacerbated cutaneous disease (n = 2) and NSAID-exacerbated respiratory disease (n = 1). Four CIs could not be categorized according to either classification system. One SR could not be categorized according to ENDA/GA2LEN. During childhood, NSAID-H exhibits different phenotypes and the majority of them can be categorized with current classification systems; however, classifications based on adult data may not exactly fit NSAID-H in paediatric patients.


Drosophila Jun kinase regulates expression of decapentaplegic via the ETS-domain protein Aop and the AP-1 transcription factor DJun during dorsal closure.
During Drosophila embryogenesis, ectodermal cells of the lateral epithelium stretch in a coordinated fashion to internalize the amnioserosa cells and close the embryo dorsally. This process, dorsal closure, requires two signaling pathways: the Drosophila Jun-amino-terminal kinase (DJNK) pathway and the Dpp pathway. We have identified mutations in DJun and show that DJNK controls dorsal closure by activating DJun and inactivating the ETS repressor Aop/Yan by phosphorylation. DJun and Aop regulate dpp expression in the most dorsal row of cells. Secreted Dpp then instructs more ventrally located cells to stretch. Our results provide a causal link between the DJNK and Dpp pathways during dorsal closure. Interestingly, in vertebrates, transforming growth factor-beta and c-Jun regulate collagenase gene expression during wound healing, a process that also involves the closing of an epithelial sheath.


Comparison of interreader reproducibility of the prostate imaging reporting and data system and likert scales for evaluation of multiparametric prostate MRI.
The objective of our study was to compare interreader reproducibility of the recently proposed "Prostate Imaging Reporting and Data System," or "PI-RADS," scale incorporating fixed criteria and a standard Likert scale based on overall impression for prostate cancer localization using multiparametric MRI. Fifty-five patients who underwent a 3-T prostate MRI examination using a pelvic phased-array coil and incorporating T2-weighted imaging, diffusion-weighted imaging, and dynamic contrast-enhanced imaging were included in the study. Three radiologists (6, 4, and 1 year of experience) independently scored 18 regions (12 in the peripheral zone [PZ] and six in the transition zone [TZ]) using PI-RADS (range, 3-15) and Likert (range, 1-5) scales, which were based on fixed criteria and overall impression, respectively. Interreader reproducibility was evaluated using the concordance correlation coefficient (CCC), which assesses exact agreement between scores (minimal, < 0.2; poor, 0.2-<0.4; moderate, 0.4-<0.6; strong, 0.6-<0.8; almost perfect, ≥ 0.8). Agreement between experienced readers was strong in the PZ and TZ combined and in the PZ for both the PI-RADS and Likert scales (CCC = 0.608-0.677), moderate in the TZ for the Likert scale (CCC = 0.519), and poor in the TZ for PI-RADS (CCC = 0.376). Agreement between experienced and inexperienced readers was moderate to poor in the PZ and TZ combined for PI-RADS (CCC = 0.340-0.477), moderate in the PZ and TZ combined for the Likert scale (CCC = 0.471-0.497), moderate in the PZ for PI-RADS and Likert scales (CCC = 0.472-0.542), minimal to poor in the TZ for PI-RADS (CCC = 0.094-0.283), and poor in the TZ for the Likert scale (CCC = 0.287-0.400). Interreader reproducibility tended to be higher for relatively experienced readers than for less experienced readers and to be higher in the PZ than in the TZ. For the relatively experienced readers, reproducibility was similar for PI-RADS and Likert scales in the PZ but was somewhat higher for the Likert scale than for PI-RADS in the TZ.


Evaluation of prices, availability and affordability of essential medicines in Lahore Division, Pakistan: A cross-sectional survey using WHO/HAI methodology.
Inadequate access to medicines affected by un-controlled prices is a major concern in developing countries, including Pakistan, which lacks comprehensive data on medicine prices. Thus, the objective of this study was to evaluate the prices, availability and affordability of essential medicines in Lahore division, Pakistan. The survey was undertaken from November, 2016 till March, 2017 by including 50 medicines, 14 from the WHO/HAI core list and 36 supplementary medicines from national essential medicine list (NEML) at public (n = 16) and private (n = 16) health facilities. The prices, availability and affordability of selected medicines were measured using a variant of the WHO/HAI standard methodology available on HAI website and WHO/HAI manual. A questionnaire was used for data collection from Lahore division. The prices were compared to International reference prices (IRPs) and the daily wage of a lowest paid unskilled government worker was used to calculate medicine affordability. Data suggested poor availability of originator brands (OB) in public and private sector facilities, i.e., 6.8% and 55.0%, respectively. Similarly, low availability was observed for lowest price generics (LPGs), both in public (35.3%) and private sector (20.3%) facilities-far below the WHO global action plan targets of 80% availability of essential medicines by 2025. In private sector, 53% OB and 38% LPG medicines were found excessively priced. The cost of standard treatment with OBs was unaffordable, i.e., above a single daily wage (1.4 day's wages) was demanded to purchase the standard treatment for the selected diseases in case of OBs medicines. Whereas, the cost of LPGs medicine required to purchase the standard treatment of the selected diseases was 0.6 day's wage (median), below a single daily wage. In conclusion, access to essential medicines, especially at public sector facilities was affected by low availability, particularly of OBs in comparison to LPGs. Thus, the better availability of LPGs might be a rational basis of transition into a generic system of prescribing that may improve the availability and accessibility of essential medicines in Lahore division. Medicine prices in Lahore division were found higher in comparison to IRPs. Thus, the efforts must be made to formulate patient's pocket friendly drug pricing policy that favors price cuts and improves affordability.


Trial of an attenuated bovine rotavirus vaccine (RIT 4237) in Gambian infants.
A randomised, controlled trial of bovine rotavirus vaccine was undertaken in Gambian infants. Three doses were administered, from the age of ten weeks, concurrently with oral or killed polio vaccine. Prevaccination rotavirus neutralising antibody levels were high. 84/185 infants (45%) showed an increase in neutralising antibody titre after receiving rotavirus vaccine, compared with 20/91 (22%) unvaccinated infants. Clinical rotavirus infection was detected in 24/78 (31%) children in the rotavirus/oral polio group, 34/83 (41%) children in the placebo/oral polio group, and 23/92 (25%) children in the rotavirus/killed polio group, giving an overall vaccine efficacy of 33% (95% CI 4-53%). RIT 4237 did not appear to reduce the severity of clinical infections. Most cases (92%) were caused by rotaviruses with short RNA electropherotypes. Serological responses to rotavirus vaccination appeared unaffected by the concurrent administration of oral polio vaccine. Lower types 1 and 3 polio antibody levels were found in children who received oral polio and rotavirus vaccines but the differences were not statistically significant.


A bioprosthetic ovary created using 3D printed microporous scaffolds restores ovarian function in sterilized mice.
Emerging additive manufacturing techniques enable investigation of the effects of pore geometry on cell behavior and function. Here, we 3D print microporous hydrogel scaffolds to test how varying pore geometry, accomplished by manipulating the advancing angle between printed layers, affects the survival of ovarian follicles. 30° and 60° scaffolds provide corners that surround follicles on multiple sides while 90° scaffolds have an open porosity that limits follicle-scaffold interaction. As the amount of scaffold interaction increases, follicle spreading is limited and survival increases. Follicle-seeded scaffolds become highly vascularized and ovarian function is fully restored when implanted in surgically sterilized mice. Moreover, pups are born through natural mating and thrive through maternal lactation. These findings present an in vivo functional ovarian implant designed with 3D printing, and indicate that scaffold pore architecture is a critical variable in additively manufactured scaffold design for functional tissue engineering.


Honey reduces the metastatic characteristics of prostate cancer cell lines by promoting a loss of adhesion.
Honey has been shown to have a range of therapeutic effects in humans, with anti-inflammatory and anti-bacterial effects among those previously characterised. Here, we examine the possibility of New Zealand thyme, manuka and honeydew honeys, and their major sugar and phenolic components, reducing the development of metastatic cancer. Their activity was examined in vitro, in PC3 and DU145 prostate cancer cell lines, through measuring the compounds' effects on the metastatic characteristics of migration, invasion and adhesion. First, the phenolic compounds gallic acid, caffeic acid, quercetin, kaempferol and chrysin were quantified in the honeys using high performance liquid chromatography, and found in nanomolar concentrations. In a Boyden chamber-based migration assay, non-toxic concentrations of thyme and honeydew honeys reduced cell migration by 20%, and all phenolic compounds except caffeic acid also lowered migration, although a mixture of only the sugars found in honey had no effect. All of the honeys, phenolics and the sugar-only mixture reduced invasive movement of cells through extracellular matrix by up to 75%. Most notably, each of the three honeys and the sugar-only mixture reduced cell adhesion to collagen I by 90%. With the exception of quercetin, phenolic compounds did not reduce adhesion. Therefore, honey and its sugar and phenolic components can lower the metastatic properties of cancer cells, and may do this by preventing effective cell adhesion to the extracellular matrix. The sugars and phenol compounds of honey are much more effective in combination than individually.


Dendritic cells and pluripotency: unlikely allies in the pursuit of immunotherapy.
As the fulcrum on which the balance between the opposing forces of tolerance and immunity has been shown to pivot, dendritic cells (DC) hold significant promise for immune intervention in a variety of disease states. Here we discuss how the directed differentiation of human pluripotent stem cells may address many of the current obstacles to the use of monocyte-derived DC in immunotherapy, providing a novel source of previously inaccessible DC subsets and opportunities for their scale-up, quality control and genetic modification. Indeed, given that it is the immunological legacy DC leave behind that is of therapeutic value, rather than their persistence per se, we propose that immunotherapy should serve as an early target for the clinical application of pluripotent stem cells.


Quality assurance of the gene ontology using abstraction networks.
The gene ontology (GO) is used extensively in the field of genomics. Like other large and complex ontologies, quality assurance (QA) efforts for GO's content can be laborious and time consuming. Abstraction networks (AbNs) are summarization networks that reveal and highlight high-level structural and hierarchical aggregation patterns in an ontology. They have been shown to successfully support QA work in the context of various ontologies. Two kinds of AbNs, called the area taxonomy and the partial-area taxonomy, are developed for GO hierarchies and derived specifically for the biological process (BP) hierarchy. Within this framework, several QA heuristics, based on the identification of groups of anomalous terms which exhibit certain taxonomy-defined characteristics, are introduced. Such groups are expected to have higher error rates when compared to other terms. Thus, by focusing QA efforts on anomalous terms one would expect to find relatively more erroneous content. By automatically identifying these potential problem areas within an ontology, time and effort will be saved during manual reviews of GO's content. BP is used as a testbed, with samples of three kinds of anomalous BP terms chosen for a taxonomy-based QA review. Additional heuristics for QA are demonstrated. From the results of this QA effort, it is observed that different kinds of inconsistencies in the modeling of GO can be exposed with the use of the proposed heuristics. For comparison, the results of QA work on a sample of terms chosen from GO's general population are presented.


Structural insights into E1 recognition and the ubiquitin-conjugating activity of the E2 enzyme Cdc34.
Ubiquitin (Ub) signaling requires the sequential interactions and activities of three enzymes, E1, E2, and E3. Cdc34 is an E2 that plays a key role in regulating cell cycle progression and requires unique structural elements to function. The molecular basis by which Cdc34 engages its E1 and the structural mechanisms by which its unique C-terminal extension functions in Cdc34 activity are unknown. Here, we present crystal structures of Cdc34 alone and in complex with E1, and a Cdc34~Ub thioester mimetic that represents the product of Uba1-Cdc34 Ub transthiolation. These structures reveal conformational changes in Uba1 and Cdc34 and a unique binding mode that are required for transthiolation. The Cdc34~Ub structure reveals contacts between the Cdc34 C-terminal extension and Ub that stabilize Cdc34~Ub in a closed conformation and are critical for Ub discharge. Altogether, our structural, biochemical, and cell-based studies provide insights into the molecular mechanisms by which Cdc34 function in cells.


Full-length transcriptome sequences of ephemeral plant Arabidopsis pumila provides insight into gene expression dynamics during continuous salt stress.
Arabidopsis pumila is native to the desert region of northwest China and it is extraordinarily well adapted to the local semi-desert saline soil, thus providing a candidate plant system for environmental adaptation and salt-tolerance gene mining. However, understanding of the salt-adaptation mechanism of this species is limited because of genomic sequences scarcity. In the present study, the transcriptome profiles of A. pumila leaf tissues treated with 250 mM NaCl for 0, 0.5, 3, 6, 12, 24 and 48 h were analyzed using a combination of second-generation sequencing (SGS) and third-generation single-molecule real-time (SMRT) sequencing. Correction of SMRT long reads by SGS short reads resulted in 59,328 transcripts. We found 8075 differentially expressed genes (DEGs) between salt-stressed tissues and controls, of which 483 were transcription factors and 1157 were transport proteins. Most DEGs were activated within 6 h of salt stress and their expression stabilized after 48 h; the number of DEGs was greatest within 12 h of salt stress. Gene annotation and functional analyses revealed that expression of genes associated with the osmotic and ionic phases rapidly and coordinately changed during the continuous salt stress in this species, and salt stress-related categories were highly enriched among these DEGs, including oxidation-reduction, transmembrane transport, transcription factor activity and ion channel activity. Orphan, MYB, HB, bHLH, C3H, PHD, bZIP, ARF and NAC TFs were most enriched in DEGs; ABCB1, CLC-A, CPK30, KEA2, KUP9, NHX1, SOS1, VHA-A and VP1 TPs were extensively up-regulated in salt-stressed samples, suggesting that they play important roles in slat tolerance. Importantly, further experimental studies identified a mitogen-activated protein kinase (MAPK) gene MAPKKK18 as continuously up-regulated throughout salt stress, suggesting its crucial role in salt tolerance. The expression patterns of the salt-responsive 24 genes resulted from quantitative real-time PCR were basically consistent with their transcript abundance changes identified by RNA-Seq. The full-length transcripts generated in this study provide a more accurate depiction of gene transcription of A. pumila. We identified potential genes involved in salt tolerance of A. pumila. These data present a genetic resource and facilitate better understanding of salt-adaptation mechanism for ephemeral plants.


Hearing Loss and Age-Induced Changes in the Central Auditory System Measured by the P3 Response to Small Changes in Frequency.
Recent behavioral studies have suggested that individuals with sloping audiograms exhibit localized improvements in frequency discrimination in the frequency region near the drop in hearing. Auditory-evoked potentials may provide evidence of such cortical plasticity and reorganization of frequency maps. The objective of this study was to evaluate electrophysiological evidence of cortical plasticity related to cortical frequency representation and discrimination abilities in older individuals with high-frequency sensorineural hearing loss (SNHL). It was hypothesized that the P3 response in this group would show evidence of physiological reorganization of frequency maps and enhanced neural representation at the edge of their high-frequency loss due to their restricted SNHL. The P3 auditory event-related potential in response to small frequency changes was recorded in a repeated measures design using an oddball paradigm that presented upward and downward frequency changes of 2%, 5%, and 20% to three groups of listeners. P3 recordings from a group of seven older individuals with a restricted sloping hearing loss >1000 or 2000 Hz was compared to two control groups of younger (n = 7) and older (n = 7) individuals with normal hearing/borderline normal hearing through 4000 Hz. The auditory P3 was recorded using an oddball paradigm (80%/20%) with the standard tone at the highest frequency of normal hearing in the hearing-impaired participants, also known as the edge frequency (EF). EFs were either 1000 or 2000 Hz for all participants. The target tones represented upward and downward frequency changes of 2%, 5%, and 20% from the standard tones of either 1000 or 2000 Hz. Waveforms were recorded using a two-channel clinical-evoked potential system. Latency and amplitude of the P300 peak were analyzed across groups for the three frequency conditions using repeated measures analysis of variance. The results of this study suggest that the P3 response can be elicited by frequency changes as small as 2-5%. P3 responses at the EF of hearing loss were present and larger in amplitude for more participants with a sloping hearing loss compared to age-matched normal-hearing peers tested at the same frequencies. As a result, the older participants with sloping hearing losses had P3 responses more similar to the younger normal-hearing participants than their age-matched peers with normal hearing. These preliminary results partially support the idea of enhanced cortical representation of frequency at the EF of localized SNHL in older adults that is not purely due to age.


Artemia as a possible vector for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus transmission (XSV) to Macrobrachium rosenbergii post-larvae.
Five developmental stages of Artemia were exposed to Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) by immersion and oral routes in order to investigate the possibility of Artemia acting as a reservoir or carrier of these viruses. The second objective was to determine if virus-exposed Artemia were capable of transmitting the disease to post-larvae (PL) of M. rosenbergii. There was no significant difference in percent mortality between Artemia control groups and groups challenged with these viruses. On the other hand, all the developmental stages of Artemia were positive for both viruses by nested RT-PCR, regardless of the challenge route. In horizontal transmission experiments, 100% mortality was observed in M. rosenbergii PL fed with Artemia nauplii exposed to MrNV and XSV by either challenge route. However, no mortality was observed in PL fed with virus-free Artemia. RT-PCR analysis of the M. rosenbergii PL confirmed the presence of MrNV and XSV in the challenge group and absence in the control group.


Expanding the treatment window with mechanical thrombectomy in acute ischemic stroke.
Acute ischemic stroke is a common disease associated with high mortality and significant long-term disability. Treatment options for acute ischemic stroke continue to evolve and include pharmaceutical and mechanical therapies. With the recent US Food and Drug Administration approval of a new device for mechanical thrombectomy, the options available for treatment of acute ischemic stroke have been expanded. Thrombolytic therapy is generally given intravenously in the first 3 h and up to 6 h via the intraarterial route for pharmacological clot disruption. The maximum time-frame for mechanical thrombectomy devices has yet to be determined. A 78-year-old female presented to the emergency room with a dense right hemiparesis, leftward gaze preference and dense global aphasia. Eight hours after symptom onset, left carotid angiography confirmed a left internal carotid artery terminus occlusion. A single pass was made through the clot with an X6 Merci Retriever device. After a single pass, the vessel was reopened and normal flow in the left internal carotid artery was demonstrated. At the time of discharge, her neurological deficits had improved significantly. Furthermore, the final infarct area, as demonstrated on magnetic resonance imaging, was probably much smaller than it would have been if the vessel had not been recanalized. We report the use of a new mechanical thrombectomy device 8 h after onset of ischemic symptoms, with substantial subsequent improvement in neurological outcome. In selected cases, use of the Merci Retriever can result in improved outcomes beyond the traditional 6-h window used for intraarterial pharmacological thrombolysis.


Impact of Ohmic Resistance on Measured Electrode Potentials and Maximum Power Production in Microbial Fuel Cells.
Low solution conductivity is known to adversely impact power generation in microbial fuel cells (MFCs), but its impact on measured electrode potentials has often been neglected in the reporting of electrode potentials. While errors in the working electrode (typically the anode) are usually small, larger errors can result in reported counter electrode potentials (typically the cathode) due to large distances between the reference and working electrodes or the use of whole cell voltages to calculate counter electrode potentials. As shown here, inaccurate electrode potentials impact conclusions concerning factors limiting power production in MFCs at higher current densities. To demonstrate how the electrochemical measurements should be adjusted using the solution conductivity, electrode potentials were estimated in MFCs with brush anodes placed close to the cathode (1 cm) or with flat felt anodes placed further from the cathode (3 cm) to avoid oxygen crossover to the anodes. The errors in the cathode potential for MFCs with brush anodes reached 94 mV using acetate in a 50 mM phosphate buffer solution. With a felt anode and acetate, cathode potential errors increased to 394 mV. While brush anode MFCs produced much higher power densities than flat anode MFCs under these conditions, this better performance was shown primarily to result from electrode spacing following correction of electrode potentials. Brush anode potentials corrected for solution conductivity were the same for brushes set 1 or 3 cm from the cathode, although the range of current produced was different due to ohmic losses with the larger distance. These results demonstrate the critical importance of using corrected electrode potentials to understand factors limiting power production in MFCs.


Evidence-based decision-making on water quality in domestic water supply in Malawi, Ecuador, and Brazil.
Scholars and practitioners advocate evidence-based decision-making (EBDM) because facts lead to beneficial outcomes. At the same time, EBDM to reduce risks in water quality is under-studied. We examine types of decision-making by water utilities and government agencies, and the nature and origin of evidence available to them in their work on delivering safe drinking water to households. Using qualitative content analysis, we comparatively analyze water utilities and government agencies in Malawi, Ecuador, and Brazil. The results show that the water utilities perform combinations of decision-making types on water quality such as implementation, intelligence-gathering, and evaluation and choice, while government agencies perform more intelligence gathering. Sources and types of information are mainly water consumers, guidelines for water quality standards, and self-monitoring from water utilities. The analysis is useful in establishing a foundation for developing evidence-based management within water supply services, and potentially other water resources management activities.


Predicting venous thrombosis in women using a combination of genetic markers and clinical risk factors.
Family history of venous thromboembolism (VTE) has been suggested to be more useful in risk assessment than thrombophilia testing. We investigated established genetic susceptibility variants for association with VTE and evaluated a genetic risk score in isolation and combined with known trigger factors, including family history of VTE. A total of 18 single nucleotide polymorphisms (SNPs) selected from the literature were genotyped in 2835 women participating in a Swedish nationwide case-control study (the ThromboEmbolism Hormone Study [TEHS]). Association with VTE was assessed by odds ratios (ORs) with 95% confidence interval (CI) using logistic regression. Clinical and genetic predictors that contributed significantly to the fit of the logistic regression model were included in the prediction models. SNP-SNP interactions were investigated and incorporated into the models if found significant. Risk scores were evaluated by calculating the area under the receiver-operating characteristics curve (AUC). Seven SNPs (F5 rs6025, F2 rs1799963, ABO rs514659, FGG rs2066865, F11 rs2289252, PROC rs1799810 and KNG1 rs710446) with four SNP-SNP interactions contributed to the genetic risk score for VTE, with an AUC of 0.66 (95% CI, 0.64-0.68). After adding clinical risk factors, which included family history of VTE, the AUC reached 0.84 (95% CI, 0.82-0.85). The goodness of fit of the genetic and combined scores improved when significant SNP-SNP interaction terms were included. Prediction of VTE in high-risk individuals was more accurate when a combination of clinical and genetic predictors with SNP-SNP interactions was included in a risk score.


Knowledge-based 3D segmentation and reconstruction of coronary arteries using CT images.
An approach for the 3D segmentation and reconstruction of human left coronary arteries using angio-CT images is presented in This work. Each voxel in the 3D dataset is assumed to belong to one of the three homogeneous regions: blood, myocardium, and lung. A priori knowledge of the regions is introduced via Bayes' rule. Posterior probabilities obtained using Bayes' rule are anisotropically smoothed, and the 3D segmentation is obtained via MAP classifications of the smoothed posteriors. An active contour model is then applied to extract the coronary arteries from the rest of the volumetric data with subvoxel accuracy. The geometric model of the left coronary arteries obtained in this work may be used to provide accurate boundary conditions for hemodynamic simulations, or to provide objective measurements of clinically relevant parameters such as lumen sizes in a 3D sense.


Discovery of a ternary pseudobrookite phase in the earth-abundant Ti-Zn-O system.
We combine theory with experiment in searching for "missing", stable materials within the Zn-Ti-O chemical system, leading to the discovery of a new pseudobrookite phase, ZnxTi3-xO5-δ. This ternary system was chosen for (1) technological relevance, (2) earth abundance, and (3) the fact that many compounds in this system are predicted from enthalpies of formation to be borderline stable, suggesting an important role of entropic contributions in their stabilization and making this chemical system a perfect test bed for exploring the limits of theoretical predictions. The initial set of exploratory experimental syntheses, via sintering in evacuated ampoules and quenching, resulted in a single phase ZnxTi3-xO5-δ composition with x ≈ 0.6 and an almost stoichiometric oxygen content, as evaluated by X-ray fluorescence, energy dispersive spectroscopy, thermogravimetric analysis, and X-ray photoelectron spectroscopy. The theoretically calculated lowest energy crystal structure for the closest stoichiometric ZnTi5O10 composition matched that measured experimentally by synchrotron X-ray diffraction (allowing for differences attributable to cation disorder). The measured broad optical absorption, n-type electrical conductivity, and stability in acidic media are comparable to those of other ternary pseudobrookites and Ti-O Magnéli phases, suggesting comparable applicability as a robust electrode or catalyst support in electrochemical devices or water remediation. However, the new phase decomposes upon heating in air as it oxidizes. The success of the present approach to identify a "missing material" in an earth-abundant and applications-rich system suggests that future efforts to experimentally realize and theoretically confirm missing materials in this and similar systems are warranted, both scientifically and technologically.


Augmented Reality in Pediatric Septic Shock Simulation: Randomized Controlled Feasibility Trial.
Septic shock is a low-frequency but high-stakes condition in children requiring prompt resuscitation, which makes it an important target for simulation-based education. In this study, we aimed to design and implement an augmented reality app (PediSepsisAR) for septic shock simulation, test the feasibility of measuring the timing and volume of fluid administration during septic shock simulation with and without PediSepsisAR, and describe PediSepsisAR as an educational tool. We hypothesized that we could feasibly measure our desired data during the simulation in 90% of the participants in each group. With regard to using PediSepsisAR as an educational tool, we hypothesized that the PediSepsisAR group would report that it enhanced their awareness of simulated patient blood flow and would more rapidly verbalize recognition of abnormal patient status and desired management steps. We performed a randomized controlled feasibility trial with a convenience sample of pediatric care providers at a large tertiary care pediatric center. Participants completed a prestudy questionnaire and were randomized to either the PediSepsisAR or control (traditional simulation) arms. We measured the participants' time to administer 20, 40, and 60 cc/kg of intravenous fluids during a septic shock simulation using each modality. In addition, facilitators timed how long participants took to verbalize they had recognized tachycardia, hypotension, or septic shock and desired to initiate the sepsis pathway and administer antibiotics. Participants in the PediSepsisAR arm completed a poststudy questionnaire. We analyzed data using descriptive statistics and a Wilcoxon rank-sum test to compare the median time with event variables between groups. We enrolled 50 participants (n=25 in each arm). The timing and volume of fluid administration were captured in all the participants in each group. There was no statistically significant difference regarding time to administration of intravenous fluids between the two groups. Similarly, there was no statistically significant difference between the groups regarding time to verbalized recognition of patient status or desired management steps. Most participants in the PediSepsisAR group reported that PediSepsisAR enhanced their awareness of the patient's perfusion. We developed an augmented reality app for use in pediatric septic shock simulations and demonstrated the feasibility of measuring the volume and timing of fluid administration during simulation using this modality. In addition, our findings suggest that PediSepsisAR may enhance participants' awareness of abnormal perfusion.


Investigation of a mpox outbreak in Central African Republic, 2021-2022.
Human monkeypox virus is spreading globally, and more information is required about its epidemiological and clinical disease characteristics in endemic countries. We report the investigation of an outbreak in November 2021 in Central African Republic (CAR). The primary case, a hunter, fell ill after contact with a non-human primate at the frontier between forest and savannah. The ensuing investigation in a small nearby town concerned two families and four waves of inter-human transmission, with 14 confirmed cases, 11 suspected cases and 17 non-infected contacts, and a secondary attack rate of 59.5% (25/42). Complications were observed in 12 of the 19 (63.2%) confirmed and suspected cases with available clinical follow-up data: eight cases of bronchopneumonia, two of severe dehydration, one corneal ulcer, one abscess, two cutaneous superinfections, and six cutaneous sequelae (cheloid scars, or depigmentation). There was one death, giving a case fatality ratio of 1/25 (4.0%) for confirmed and suspected cases. This outbreak, with the largest number of confirmed cases ever described in CAR, confirms the potential severity of the disease associated with clade I monkeypox viruses, and highlights the need for rapid control over virus circulation to prevent the further national and international spread of infection.


Structural and evolutionary inference from molecular variation in Neisseria porins.
The porin proteins of the pathogenic Neisseria species, Neisseria gonorrhoeae and Neisseria meningitidis, are important as serotyping antigens, putative vaccine components, and for their proposed role in the intracellular colonization of humans. A three-dimensional structural homology model for Neisseria porins was generated from Escherichia coli porin structures and N. meningitidis PorA and PorB sequences. The Neisseria sequences were readily assembled into the 16-strand beta-barrel fold characteristic of porins, despite relatively low sequence identity with the Escherichia proteins. The model provided information on the spatial relationships of variable regions of peptide sequences in the PorA and PorB trimers and insights relevant to the use of these proteins in vaccines. The nucleotide sequences of the porin genes from a number of other Neisseria species were obtained by PCR direct sequencing and from GenBank. Alignment and analysis of all available Neisseria porin sequences by use of the structurally conserved regions derived from the PorA and PorB structural models resulted in the recovery of an improved phylogenetic signal. Phylogenetic analyses were consistent with an important role for horizontal genetic exchange in the emergence of different porin classes and confirmed the close evolutionary relationships of the porins from N. meningitidis, N. gonorrhoeae, Neisseria lactamica, and Neisseria polysaccharea. Only members of this group contained three conserved lysine residues which form a potential GTP binding site implicated in pathogenesis. The model placed these residues on the inside of the pore, in close proximity, consistent with their role in regulating pore function when inserted into host cells.


Impact of pharmacist involvement in the transitional care of high-risk patients through medication reconciliation, medication education, and postdischarge call-backs (IPITCH Study).
Previous data suggest that direct pharmacist interaction with patients through medication reconciliation, discharge counseling, and postdischarge phone calls decreases the number of adverse drug events (ADEs) and plays an overall positive role in transitional care. Previous studies have evaluated pharmacist involvement in improving transitional care, but these studies did not include multiple postdischarge follow-up phone calls. The objectives of this study were to assess the impact of pharmacist involvement in transitions of care as measured by decreased medication errors (MEs) and ADEs, patients' knowledge related to communication about their medications as measured by improvement in the Hospital Consumer Assessment of Healthcare Providers and Systems (HCAHPS) scores, and 30-day all-cause inpatient readmissions and emergency department (ED) visits. This was a prospective, randomized, single-period longitudinal study that occurred from November 2012 through June 2013 at an urban, tertiary, academic medical center. Patients admitted to 2 designated internal medicine units on high-risk medications or with greater than 3 prescription medications upon discharge were included for randomization. The control group received the usual hospital standard of care. The study group received face-to-face medication reconciliation, a patient-specific pharmaceutical care plan, discharge counseling, and postdischarge phone calls on days 3, 14, and 30 to provide education and assess study endpoints. A total of 278 patients were included in the final analysis, with 141 in the control group and 137 in the study group. Fifty-five patients (39%) in the control arm experienced an inpatient readmission or ED visit within 30-days postdischarge compared to 34 patients (24.8%) in the study arm (P = 0.01). Eighteen patients (12.8%) in the control group experienced an ADEs or MEs compared to 11 patients (8%) in the study group (P > 0.05). The HCAHPS scores during the study period showed a 9% improvement for the assessed questionnaire domain (P > 0.05). This study demonstrated that pharmacist involvement in hospital discharge transitions of care had a positive impact on decreasing composite inpatient readmissions and ED visits. Statistically significant difference in medication-related events and HCAHPS scores were not observed. Patients with moderately complex medication regimens benefited from a continuity of care involving a pharmacy team during transitions in care.


Effect of the antipsychotic drug haloperidol on arrhythmias during acute myocardial infarction in a porcine model.
Patients receiving psychiatric medication, like the antipsychotic drug haloperidol, are at an increased risk of sudden cardiac death (SCD). Haloperidol blocks the cardiac rapidly-activating delayed rectifier potassium current, thereby increasing electrical dispersion of repolarization which can potentially lead to arrhythmias. Whether these patients are also at a higher risk to develop SCD during an acute myocardial infarction (AMI) is unknown. AMI locally shortens action potential duration, which might further increase repolarization dispersion and increase the risk of arrhythmia in the presence of haloperidol compared to without. Our aim was to test whether treatment with haloperidol implies an increased risk of SCD when eventually experiencing AMI. Twenty-eight female Danish Landrace pigs were randomized into three groups: low dose haloperidol (0.1 mg/kg), high dose (1.0 mg/kg) or vehicle-control group. One hour after haloperidol/vehicle infusion, AMI was induced by balloon-occlusion of the mid-left anterior descending coronary artery and maintained for 120 min, followed by 60 min of reperfusion. VF occurred during occlusion in 7/11 pigs in the control group, 3/11 in the low dose (p = 0.198) and 2/6 in the high dose group (p = 0.335). High dose haloperidol significantly prolonged QT, and reduced heart rate, vascular resistance and blood pressure before and during AMI. Premature ventricular contractions in phase 1b during AMI were reduced with high dose haloperidol. AMI-induced arrhythmia was not aggravated in pigs with haloperidol treatment. Our results do not suggest that AMI is contributing to the excess mortality in patients treated with antipsychotic drugs seen in epidemiological studies.


Regulation of in vitro capillary tube formation by anti-integrin antibodies.
Human endothelial cells are induced to form an anastomosing network of capillary tubes on a gel of collagen I in the presence of PMA. We show here that the addition of mAbs, AK7, or RMAC11 directed to the alpha chain of the major collagen receptor on endothelial cells, the integrin alpha 2 beta 1, enhance the number, length, and width of capillary tubes formed by endothelial cells derived from umbilical vein or neonatal foreskins. The anti-alpha 2 beta 1 antibodies maintained the endothelial cells in a rounded morphology and inhibited both their attachment to and proliferation on collagen but not on fibronectin, laminin, or gelatin matrices. Furthermore, RMAC11 promoted tube formation in collagen gels of increased density which in the absence of RMAC11 did not allow tube formation. Neither RMAC11 or AK7 enhanced capillary formation in the absence of PMA. Lumen structure and size were also altered by antibody RMAC11. In the absence of antibody the majority of lumina were formed intracellularly from single cells, but in the presence of RMAC11, multiple cells were involved and the lumen size was correspondingly increased. Endothelial cells were also induced to undergo capillary formation in fibrin gels after PMA stimulation. The addition of anti-alpha v beta 3 antibodies promoted tube formation in fibrin gels and inhibited EC adhesion to and proliferation on a fibrinogen matrix. The enhancement of capillary formation by the anti-integrin antibodies was matrix specific; that is, anti-alpha v beta 3 antibodies only enhanced tube formation on fibrin gels and not on collagen gels while anti-alpha v beta 1 antibodies only enhanced tubes on collagen and not on fibrin gels. Thus we postulate that changes in the adhesive nature of endothelial cells for their extracellular matrix can profoundly effect their function. Anti-integrin antibodies which inhibit cell-matrix interactions convert endothelial cells from a proliferative phenotype towards differentiation which results in enhanced capillary tube formation.


Treatment at the front end of the criminal justice continuum: the association between arrest and admission into specialty substance abuse treatment.
To reduce criminal recidivism and drug use, it has been proposed that the substance abuse treatment delivery system cut across different components of the criminal justice continuum. Arrest, at the front end of this continuum, may represent a critical moment to motivate people with substance use disorders (SUD) to seek treatment but is often over looked as an intervention point. We used data from the 2002 National Survey on Drug Use and Health (NSDUH) to compare treatment need and recent treatment admission for participants with no criminal justice (CJ) involvement in the past year, past-year arrest, and CJ supervision (i.e., probation or parole status). Of those arrested, 44.8% met criteria for an SUD. However, only 14% of those arrested with an SUD received treatment in the year of their arrest. In multivariate modelling, arrest was an independent predictor of treatment admission (odds ratio (OR) = 8.74) similar in magnitude to meeting criteria for an SUD (OR = 8.22). Those further along the continuum--under supervision--were most likely to receive treatment (OR = 22.62). Arrest involves the largest number of individuals entering the criminal justice system. The NSDUH suggests that nearly 6 million individuals in the US experience an arrest annually and that nearly half meet criteria for an SUD. Although arrest involves the largest number of individuals entering the criminal justice system, it is also the most fleeting point as individuals can move in and out rather quickly. Minimally, arrest imposes contact between the individual and a law enforcement person and can be an opportunity for early intervention strategies such as pre-arraignment diversion into treatment or brief intervention strategies. Using brief intervention at this early point in the continuum may motivate a greater number of individuals to seek treatment or decrease drug and alcohol use. Training and procedural shifts at this point of contact could have important policy implications in reducing the number of subsequent arrests or preventing individuals moving further along the criminal justice continuum, as well as decreasing the fiscal and resource burdens associated with criminal justice processing and confinement.


Characterisation of shops selling veterinary medicines in a tsetse-infested area of Kenya.
In a tsetse-infested area of Kenya, we characterised veterinary-drug outlets based on personnel and business characteristics to assess their capacities to provide clinical veterinary services. Structured questionnaires were administered to the retailers and sought information on the characteristics of the owners, salespersons and businesses. A total of 41 retail outlets (20 agro-veterinary, 11 pharmacy, and 10 general shops) were identified. There was poor response to questions on owner characteristics. Proprietors, who had no more than secondary education owned 15 out of 28 shops. Few shops (4/29) were owned by proprietors, who had professional qualifications (in animal health). Most salespersons had only secondary education but no qualifications. Animal-health assistants (AHAs), veterinarians and manufacturer's package inserts (drug leaflets) were the preferred information sources for the retailers. We concluded that drug retailers were poorly equipped with the technical knowledge necessary for drug dispensation and advice.


Skin mini-erosion technique for monitoring metabolites in interstitial fluid: its feasibility demonstrated by OGTT results in diabetic and non-diabetic subjects.
The aim of this study was to evaluate a new interstitial fluid (IF) sampling technique and its application in diabetology. IF and venous whole blood were sampled serially during an oral glucose tolerance test (OGTT) on days 1 and 3 after formation of a mini-erosion in eight non-diabetic controls and eight Type 2 diabetic subjects. Glucose, lactate, glycerol, 3-hydroxybutyrate and insulin were assayed in IF and plasma. With solitary exceptions, the IF sample volumes were in excess of those required for measurement of all five substances. However, mean IF sampling rates differed significantly both between the non-diabetic and diabetic groups and between days 1 and 3 (p < 0.001 in all cases). In both groups, the OGTT curves of glucose, glycerol and 3-hydroxybutyrate were similar to the respective curves for plasma, whereas for lactate concentrations they were markedly greater in IF than in plasma (40% and 93% in the control group and 57% and 100% greater in the diabetic group on days 1 and 3, respectively). The reverse was true of insulin concentrations in the non-diabetic group, which were 57% and 74% lower in IF than in plasma on days 1 and 3, respectively. In the non-diabetic group, the baseline (pre-OGTT) insulin level in IF increased from 49 +/- 22% (SD) of that in plasma on day 1 to 74 +/- 19% of that in plasma on day 3 (p=0.005). Sampling site re-epithelialization was rapid. In conclusion, the feasibility of transdermal sampling of IF via a skin mini-erosion has been demonstrated in both diabetic and non-diabetic subjects.


Does a delay in performing an activated clotting (ACT) test really matter? A study in nonheparinized blood and a single ACT machine.
Activating clotting time (ACT) is a point-of-care, blood clotting test used to monitor anticoagulation. Recently, institutional requirements have required that ACT testing be completed outside the operating room with trained, certified personnel other than anesthesia staff. For this reason, in this study, we looked at whether a delay in processing an ACT makes a significant difference to the ACT results. Twenty patients between 18 and 65 years of age consented to the study, each undergoing non-cardiac surgery, with no intraoperative administration of heparin. The study was approved by our Institutional Review Board. A blood sample was taken from the patient's arterial line in the operating room. Immediately afterward, 1 mL was placed into each of two ACT cartridges and the measurement was done in a Medtronic ACT2 machine. The first ACT value was 126.9 +/- 14.5 seconds. The ACT value at approximately 30 minutes was 108.3 +/- 20.3 seconds (p < .0001). The time between the first and last measurements was 29.4 +/- 3.0 minutes. The results suggest that the ACT values decrease over time between sampling all measurements. At approximately 30 minutes, the ACT values average 15% less than the control measurements. Therefore, it would seem prudent to determine ACT values immediately in the operating room without any delay, using point-of-care testing.


Model of End-Stage Liver Disease-eXcluding International Normalized Ratio (MELD-XI) Scoring System to Predict Outcomes in Patients Who Undergo Left Ventricular Assist Device Implantation.
The use of continuous-flow left ventricular assist devices (CF-LVADs) to treat advanced heart failure is increasing. Although risk scores, such as Model for End-Stage Liver Disease and the HeartMate II Risk Score, require the use of the international normalized ratio, many patients are on anticoagulation before CF-LVAD implantation. This study evaluated the ability of the Model of End-Stage Liver Disease-eXcluding International Normalized Ratio (MELD-XI) scoring system to predict clinical outcomes in patients with advanced heart failure who undergo CF-LVAD implantation. A single-center retrospective review was performed of 524 patients who were implanted with the HeartMate II LVAD (Thoratec Corporation, Pleasanton, CA) or the HeartWare HVAD (HeartWare International Inc, Framingham, MA) between 2004 and 2016. Patients were stratified into two cohorts: those with a MELD-XI score of less than 14 (n = 301) and 14 or higher (n = 223). Patients with the higher-risk MELD-XI score of 14 or higher demonstrated lower survival rates at 1, 3, 6, 12, and 24 months (p < 0.001 for all) and increased risk of early right heart failure and infections compared with patients with MELD-XI score of less than 14. MELD-XI was not significantly inferior at predicting 90-day mortality compared with the HeartMate II Risk Score (p = 0.92). Patients with elevated MELD-XI scores at follow-up demonstrated higher rates of mortality. These findings suggest that a MELD-XI score of 14 or higher was associated with a higher postoperative mortality rate than that seen in patients with a lower MELD-XI score. The MELD-XI scoring system can be used to predict outcomes in patients with advanced heart failure who undergo CF-LVAD implantation.


GMCSF-armed vaccinia virus induces an antitumor immune response.
Oncolytic Western Reserve strain vaccinia virus selective for epidermal growth factor receptor pathway mutations and tumor-associated hypermetabolism was armed with human granulocyte-macrophage colony-stimulating factor (GMCSF) and a tdTomato fluorophore. As the assessment of immunological responses to human transgenes is challenging in the most commonly used animal models, we used immunocompetent Syrian golden hamsters, known to be sensitive to human GMCSF and semipermissive to vaccinia virus. Efficacy was initially tested in vitro on various human and hamster cell lines and oncolytic potency of transgene-carrying viruses was similar to unarmed virus. The hGMCSF-encoding virus was able to completely eradicate subcutaneous pancreatic tumors in hamsters, and to fully protect the animals from subsequent rechallenge with the same tumor. Induction of specific antitumor immunity was also shown by ex vivo co-culture experiments with hamster splenocytes. In addition, histological examination revealed increased infiltration of neutrophils and macrophages in GMCSF-virus-treated tumors. These findings help clarify the mechanism of action of GMCSF-armed vaccinia viruses undergoing clinical trials.


Dual-Action Heteromultivalent Glycopolymers Stringently Block and Arrest Influenza A Virus Infection In Vitro and Ex Vivo.
Here, we demonstrate the concerted inhibition of different influenza A virus (IAV) strains using a low-molecular-weight dual-action linear polymer. The 6'-sialyllactose and zanamivir conjugates of linear polyglycerol are optimized for simultaneous targeting of hemagglutinin and neuraminidase on the IAV surface. Independent of IAV subtypes, hemagglutination inhibition data suggest better adsorption of the heteromultivalent polymer than homomultivalent analogs onto the virus surface. Cryo-TEM images imply heteromultivalent compound-mediated virus aggregation. The optimized polymeric nanomaterial inhibits >99.9% propagation of various IAV strains 24 h postinfection in vitro at low nM concentrations and is up to 10000× more effective than the commercial zanamivir drug. In a human lung ex vivo multicyclic infection setup, the heteromultivalent polymer outperforms the commercial drug zanamivir and homomultivalent analogs or their physical mixtures. This study authenticates the translational potential of the dual-action targeting approach using small polymers for broad and high antiviral efficacy.


Cellular actions of somatostatin on rat periaqueductal grey neurons in vitro.
Functional studies indicate that the midbrain periaqueductal grey (PAG) is involved in the analgesic actions of somatostatin; however, the cellular actions of somatostatin in this brain region are unknown. In the present study, whole-cell patch clamp recordings were made from rat PAG neurons in vitro. In 93% of acutely isolated neurons, somatostatin inhibited Ca(2+)-channel currents. This effect was mimicked by the sst-2 selective agonist BIM-23027, but not by the sst-1 and sst-5 selective agonists CH-275 and L-362855. In brain slices, 81% of neurons responded to somatostatin (300 nm) with an increase in K(+) conductance that reversed polarity at -114 mV. A greater proportion of somatostatin-sensitive neurons (93%) than somatostatin-insensitive neurons (53%) responded to the opioid agonist met-enkephalin (10 microm). Somatostatin also reduced the amplitude of evoked GABA(A)-mediated inhibitory postsynaptic currents (IPSCs). The actions of somatostatin in brain slices were mimicked by BIM-23027, but not by CH-275. Somatostatin had a variable effect on the rate of spontaneous miniature IPSCs in normal external potassium solutions. In high external potassium solutions, somatostatin reduced the rate of miniature IPSCs in all neurons, and this inhibition was abolished by addition of Cd(2+) (30 microm). Somatostatin had no effect on the amplitude of miniature IPSCs. These results indicate that somatostatin acts via sst-2 receptors to directly inhibit a subpopulation of PAG neurons by activating a potassium conductance and inhibits GABA release within PAG via a presynaptic Ca(2+)-dependent mechanism. Thus, like opioids, somatostatin has the potential to exert pre- and postsynaptic disinhibitory effects within the PAG.


Neural disruption to theory of mind predicts daily social functioning in individuals at familial high-risk for schizophrenia.
Theory-of-mind (ToM) ability is foundational for successful social relationships, and dependent on a neurocognitive system, which includes temporoparietal junction and medial prefrontal cortex. Schizophrenia is associated with ToM impairments, and initial studies demonstrate similar, though more subtle deficits, in unaffected first-degree relatives, indicating that ToM deficits are a potential biomarker for the disorder. Importantly, the social consequences of ToM deficits could create an additional vulnerability factor for individuals at familial high risk (FHR). However, behavioral studies of ToM are inconsistent and virtually nothing is known about the neural basis of ToM in FHR or the relationship between ToM and social functioning. Here, FHR and non-FHR control participants underwent functional MRI scanning while reasoning about a story character's thoughts, emotions or physical appearance. Afterwards, participants completed a 28-day online 'daily-diary' questionnaire in which they reported daily social interactions and degree of ToM reasoning. FHR participants demonstrated less neural activity in bilateral temporoparietal junction when reasoning about thoughts and emotions. Moreover, across all participants, the degree of neural activity during ToM reasoning predicted several aspects of daily social behavior. Results suggest that vulnerability for schizophrenia is associated with neurocognitive deficits in ToM and the degree of deficit is related to day-to-day social functioning.


Heteronuclear NMR investigations of dynamic regions of intact Escherichia coli ribosomes.
15N-(1)H NMR spectroscopy has been used to probe the dynamic properties of uniformly (15)N labeled Escherichia coli ribosomes. Despite the high molecular weight of the complex ( approximately 2.3 MDa), [(1)H-(15)N] heteronuclear single-quantum correlation spectra contain approximately 100 well resolved resonances, the majority of which arise from two of the four C-terminal domains of the stalk proteins, L7/L12. Heteronuclear pulse-field gradient NMR experiments show that the resonances arise from species with a translational diffusion constant consistent with that of the intact ribosome. Longitudinal relaxation time (T(1)) and T(1 rho) (15)N-spin relaxation measurements show that the observable domains tumble anisotropically, with an apparent rotational correlation time significantly longer than that expected for a free L7/L12 domain but much shorter than expected for a protein rigidly incorporated within the ribosomal particle. The relaxation data allow the ribosomally bound C-terminal domains to be oriented relative to the rotational diffusion tensor. Binding of elongation factor G to the ribosome results in the disappearance of the resonances of the L7/L12 domains, indicating a dramatic reduction in their mobility. This result is in agreement with cryoelectron microscopy studies showing that the ribosomal stalk assumes a single rigid orientation upon elongation factor G binding. As well as providing information about the dynamical properties of L7/L12, these results demonstrate the utility of heteronuclear NMR in the study of mobile regions of large biological complexes and form the basis for further NMR studies of functional ribosomal complexes in the context of protein synthesis.


Conceptualizing performance of nursing care as a prerequisite for better measurement: a systematic and interpretive review.
Despite the critical role of nursing care in determining high-performing healthcare delivery, performance science in this area is still at an early stage of development and nursing's contribution most often remains invisible to policy-makers and managers. The objectives of this study were: 1) to develop a theoretically based framework to conceptualize nursing care performance; 2) to analyze how the different components of the framework have been operationalized in the literature; and 3) to develop a pool of indicators sensitive to various aspects of nursing care that can be used as a basis for designing a performance measurement system. We carried out a systematic review of published literature across three databases (MEDLINE, EMBASE and CINAHL), focusing on literature between 1990 and 2008. Screening of 2,103 papers resulted in final selection of 101 papers. A detailed template was used to extract the data. For the analysis, we used the method of interpretive synthesis, focusing first on 31 papers with theoretical or conceptual frameworks; the remaining 70 articles were used to strengthen and consolidate the findings. Current conceptualizations of nursing care performance mostly reflect a system perspective that builds on system theory, Donabedian's earlier works on healthcare organization, and Parsons' theory of social action. Drawing on these foundational works and the evidence collated, the Nursing Care Performance Framework (NCPF) we developed conceptualizes nursing care performance as resulting from three nursing subsystems that operate together to achieve three key functions: (1) acquiring, deploying and maintaining nursing resources, (2) transforming nursing resources into nursing services, and (3) producing changes in patients' conditions. Based on the literature review, these three functions are operationalized through 14 dimensions that cover 51 variables. The NCPF not only specifies core aspects of nursing performance, it also provides decision-makers with a conceptual tool to serve as a common ground from which to define performance, devise a common and balanced set of performance indicators for a given sector of nursing care, and derive benchmarks for this sector. The NCPF provides a comprehensive, integrated and theoretically based model that allows performance evaluation of both the overall nursing system and its subsystems. Such an approach widens the view of nursing performance to embrace a multidimensional perspective that encompasses the diverse aspects of nursing care.


Mechanical and in vitro biological performance of graphene nanoplatelets reinforced calcium silicate composite.
Calcium silicate (CaSiO3, CS) ceramic composites reinforced with graphene nanoplatelets (GNP) were prepared using hot isostatic pressing (HIP) at 1150°C. Quantitative microstructural analysis suggests that GNP play a role in grain size and is responsible for the improved densification. Raman spectroscopy and scanning electron microscopy showed that GNP survived the harsh processing conditions of the selected HIP processing parameters. The uniform distribution of 1 wt.% GNP in the CS matrix, high densification and fine CS grain size help to improve the fracture toughness by ∼130%, hardness by ∼30% and brittleness index by ∼40% as compared to the CS matrix without GNP. The toughening mechanisms, such as crack bridging, pull-out, branching and deflection induced by GNP are observed and discussed. The GNP/CS composites exhibit good apatite-forming ability in the simulated body fluid (SBF). Our results indicate that the addition of GNP decreased pH value in SBF. Effect of addition of GNP on early adhesion and proliferation of human osteoblast cells (hFOB) was measured in vitro. The GNP/CS composites showed good biocompatibility and promoted cell viability and cell proliferation. The results indicated that the cell viability and proliferation are affected by time and concentration of GNP in the CS matrix.


Graft loss due to recurrent focal segmental glomerulosclerosis in renal transplant recipients in the United States.
Rates of and risk factors for graft loss and graft loss resulting from recurrent focal segmental glomerulosclerosis (FSGS) have not been studied in a national population. A retrospective analysis was performed on a national registry (1999 United States Renal Data System) of 101,808 renal transplant recipients (October 1, 1987, to December 31, 1996). Of these, 3,861 recipients of solitary renal transplants who had end-stage renal disease resulting from FSGS met inclusion criteria. Outcomes were graft loss and graft loss resulting from recurrent FSGS. As a percentage of all graft loss, recurrent FSGS accounted for 18.7% in living donor recipients and 7.8% in cadaveric recipients. In white recipients, the corresponding figures were 27% and 13%. In multivariate analysis, factors associated with graft loss resulting from recurrent FSGS were white recipient, donor African-American kidney in white recipient, younger recipient age, and treatment for rejection. African-American recipients had higher rates of graft loss overall. A living donor was associated with superior overall graft survival. Among renal transplant recipients with FSGS, white recipients had a higher risk of graft loss resulting from recurrent FSGS, disproportionately seen in recipients of African-American kidneys. The role of donor/recipient race pairing on graft loss resulting from recurrent FSGS should be validated. Living donor had no association with graft loss from recurrent FSGS after correction for other factors. African-American recipients with FSGS may have the most to gain from a living donor, given their improved graft survival and decreased risk of graft loss resulting from recurrent FSGS. This is a US government work. There are no restrictions on its use.


Cross-talk between stem cells and the dysfunctional brain is facilitated by manipulating the niche: evidence from an adhesion molecule.
In the injured brain, the behavior of neural stem/progenitor cells (NSCs) is regulated by multiple converging factors encountered in the niche, which is composed of several neural and non-neural cell types. Signals emanating from the host influence the migration, survival, distribution, and fate of transplanted NSCs, which in turn can create host microenvironments that favor a return to homeostasis. We tested the hypothesis that overexpression of key facilitatory molecules that define the injury niche might enhance this bidirectional stem cell-host interaction to therapeutic advantage. As proof of concept, we investigated whether conditioning the niche with the neural cell adhesion molecule L1 might enhance recovery in a prototypical neurodegenerative milieu-the MPTP-induced model of Parkinson's disease in aged mice-where cross-talk between NSCs and imperiled host dopaminergic neurons is known to be pivotal in rescuing the function and connectivity of the latter. In lesioned mice (and in unlesioned controls), we overexpressed L1 in the NSCs to be transplanted into the ventral mesencephalon. Several pairwise experimental combinations were tested based on variations of engrafting L1 overexpressing versus nonoverexpressing NSCs into wild-type (WT) versus L1-overexpressing transgenic mice (specifically L1 transcribed from the GFAP promoter and, hence, overexpressed in host astrocytes). Enrichment for L1-particularly when expressed simultaneously in both donor NSCs and host brain-led to rapid and extensive distribution of exogenous NSCs, which in turn rescued (with an efficacy greater than in nonengineered controls) dysfunctional host dopaminergic nigral neurons, even when grafting was delayed by a month. L1 overexpression by NSCs also enhanced their own differentiation into tyrosine hydroxylase-expressing neurons in both WT and transgenic hosts. Graft-host interactions were thus favored by progressively increasing levels of L1. More broadly, this study supports the view that manipulating components of the niche (such as an adhesion molecule) that facilitate cross-talk between stem cells and the dysfunctional brain may offer new strategies for more efficacious neurotransplantation, particularly when treatment is delayed as in chronic lesions or advanced stages of a neurodegenerative disease.


Treatment of life-threatening hemorrhage due to acquired factor VIII inhibitor.
An otherwise healthy elderly man developed massive, life-threatening, sublingual bleeding associated with an idiopathic factor VIII inhibitor. The patient was treated wtih cyclophosphamide, steroids, factor VIII concentrates, and repeated plasmapheresis (including three times with NCI-IBM blood-cell separator). Rapid clinical and laboratory improvement occurred, with complete disappearance of the inhibitor. The patient has remained well, without evidence of an inhibitor, for 8 mo. The possible role of each of the therapeutic measures in the disappearance of the inhibitor and the possible pathogenetic mechanism of this disorder are discussed. A high mortality rate and a striking incidence of sublingual hematoma have been observed in cases in the literature.


Additional sauropod dinosaur material from the Callovian Oxford Clay Formation, Peterborough, UK: evidence for higher sauropod diversity.
Four isolated sauropod axial elements from the Oxford Clay Formation (Callovian, Middle Jurassic) of Peterborough, UK, are described. Two associated posterior dorsal vertebrae show a dorsoventrally elongated centrum and short neural arch, and nutrient or pneumatic foramina, most likely belonging to a non-neosauropod eusauropod, but showing ambiguous non-neosauropod eusauropod and neosauropod affinities. An isolated anterior caudal vertebra displays a ventral keel, a 'shoulder' indicating a wing-like transverse process, along with a possible prespinal lamina. This, together with an overall high complexity of the anterior caudal transverse process (ACTP) complex, indicates that this caudal could have belonged to a neosauropod. A second isolated middle-posterior caudal vertebra also shows some diagnostic features, despite the neural spine and neural arch not being preserved and the neurocentral sutures being unfused. The positioning of the neurocentral sutures on the anterior one third of the centrum indicates a middle caudal position, and the presence of faint ventrolateral crests, as well as a rhomboid anterior articulation surface, suggest neosauropod affinities. The presence of possible nutrient foramina are only tentative evidence of a neosauropod origin, as they are also found in Late Jurassic non-neosauropod eusauropods. As the caudals from the two other known sauropods from the Peterborough Oxford Clay, Cetiosauriscus stewarti and an indeterminate non-neosauropod eusauropod, do not show the features seen on either of the new elements described, both isolated caudals indicate a higher sauropod species diversity in the faunal assemblage than previously recognised. An exploratory phylogenetic analysis using characters from all four isolated elements supports a basal neosauropod placement for the anterior caudal, and a diplodocid origin for the middle caudal. The dorsal vertebrae are an unstable OTU, and therefore remain part of an indeterminate eusauropod of uncertain affinities. Together with Cetiosauriscus, and other material assigned to different sauropod groups, this study indicates the presence of a higher sauropod biodiversity in the Oxford Clay Formation than previously recognised. This study shows that it is still beneficial to examine isolated elements, as these may be indicators for higher species richness in deposits that are otherwise poor in terrestrial fauna.


Microenvironmental variables must influence intrinsic phenotypic parameters of cancer stem cells to affect tumourigenicity.
Since the discovery of tumour initiating cells (TICs) in solid tumours, studies focussing on their role in cancer initiation and progression have abounded. The biological interrogation of these cells continues to yield volumes of information on their pro-tumourigenic behaviour, but actionable generalised conclusions have been scarce. Further, new information suggesting a dependence of tumour composition and growth on the microenvironment has yet to be studied theoretically. To address this point, we created a hybrid, discrete/continuous computational cellular automaton model of a generalised stem-cell driven tissue with a simple microenvironment. Using the model we explored the phenotypic traits inherent to the tumour initiating cells and the effect of the microenvironment on tissue growth. We identify the regions in phenotype parameter space where TICs are able to cause a disruption in homeostasis, leading to tissue overgrowth and tumour maintenance. As our parameters and model are non-specific, they could apply to any tissue TIC and do not assume specific genetic mutations. Targeting these phenotypic traits could represent a generalizable therapeutic strategy across cancer types. Further, we find that the microenvironmental variable does not strongly affect the outcomes, suggesting a need for direct feedback from the microenvironment onto stem-cell behaviour in future modelling endeavours.


Shifts in the elemental composition of plants during a very severe drought.
Diverse plant functions (e.g. growth, storage, defense and anti-stress mechanisms) use elements disproportionally. We hypothesized that plants growing under different abiotic and biotic conditions would shift their elemental compositions in response to a very severe drought. We tested this hypothesis by investigating the changes in foliar stoichiometry and species composition from a very severe drought. We also tested the effects of previous droughts (acclimation) on this response. Different species growing in the same community responded more similarly to a very severe drought than did individual species growing in different communities. The stoichiometric shifts were thus more community-dependent than species-dependent. The results also suggested that plants grown in monoculture were less stoichiometrically plastic during the drought than plants grown in a more diverse community. Previous exposure to long-term drought treatments in the same communities did not significantly affect the stoichiometric shifts during the new drought. Differential use of resources may have been responsible for these responses. Monocultured plants, which used the same resources in similar proportions, had more difficulty avoiding direct competition when the resources became scarcer. Moreover, each species tested had a particular elemental composition in all communities and climatic treatments. The differences in foliar elemental compositions were largest between plant functional groups (shrubs and grasses) and smallest among species within the same functional group. Global principal components analyses (PCAs) identified a general tendency for all species, independently of the community in which they grew, toward lower concentrations of K, N, P, Mg and S, and to higher concentrations of C and Fe as the drought advanced. This study has demonstrated the utility of analyses of differences and shifts in plant elemental composition for understanding the processes underlying the responses of plants to changes in biotic and abiotic environmental conditions.


Osteosarcoma, seasonality, and environmental factors in Wisconsin, 1979-1989.
Proxy exposure measures and readily available data from the Wisconsin Cancer Reporting System were used to contrast 167 osteosarcoma cases with 989 frequency-matched cancer referents reported during 1979-1989. Differences in potential exposure to water-borne radiation and fluoridated drinking water, population size for the listed place of residence, and seasonality were assessed. An association was found between osteosarcoma and residence in a population of less than 9,000 (odds ratio = 1.6, 95% confidence interval = 1.1-2.4). In addition, an association between month of birth (May through July versus other months of birth) and osteosarcoma among individuals who were less than 25 y of age (odds ratio = 1.9, 95% confidence interval = 1.1-3.4). Overall, no association was found between potential exposure to fluoridated drinking water and osteosarcoma (odds ratio = 1.0, 95% confidence interval = 0.6-1.5). The association between osteosarcoma and water-borne radiation was weak and was not significant statistically (odds ratio = 1.5, 95% confidence interval = 0.8-2.8).


Studies of the biogenic amine transporters. VI. Characterization of a novel cocaine binding site, identified with [125I]RTI-55, in membranes prepared from whole rat brain minus caudate.
Previous studies showed that the cocaine analog [125I]RTI-55 labels dopamine and serotonergic (5-HT) biogenic amine transporters (BATs) with high affinity. Here we characterized [125I]RTI-55 binding to membranes prepared from whole rat brain minus the caudate nuclei. Paroxetine (50 nM) was used to block [125I]RTI-55 binding to 5-HT transporter sites. Initial experiments identified drugs that displaced [125I]RTI-55 binding with moderately low slope factors. Binding surface analysis of the interaction of 3 beta-(4-chlorophenyl)tropan-2 beta-carboxylic acid phenyl ester hydrochloride (RTI-113) and 3 beta-(4-iodophenyl)tropan-2 beta-carboxylic acid phenyl ester hydrochloride (RTI-122) with [125I]RTI-55 binding sites readily resolved two binding sites for [125I]RTI-55 with Kd values of 0.44 nM and 17 nM and Bmax values of 31 and 245 fmol/mg protein. Potent 5-HT and noradrenergic uptake inhibitors had low affinity for both sites. Whereas cocaine, CFT and WIN35,065-2 were 6.0-, 25- and 14-fold selective for the first site, benztropine, PCP and the novel pyrrole, (+-)-(2RS,3aSR,8bRS)-1,2,3,3a,4,8b-hexahydro- 2-benzyl-1-methylindeno-[1,2-b]pyrrole resorcylate [(+-)-HBMP, formerly called (+-)-RTI-4793-14], were moderately selective for the second site. A single binding site with the characteristics of site 1 was resolved using COS cells transiently expressing the cloned rat dopamine transporter. Lesion studies with 6-hydroxydopamine and 5,7-dihydroxytryptamine were conducted to test the hypothesis that site 1 and site 2 are physically distinct. The data showed that these neurotoxins differentially decreased [125I]RTI-55 binding to sites 1 and 2. The differential distribution of sites 1 and 2 in rat brain provides further support for this hypothesis. Viewed collectively, these data show that [125I]RTI-55 labels a novel binding site in rat brain membranes, termed DATsite2, which is not associated with the classic dopamine, serotonin or norepinephrine transporters.


Cancer-induced hypercalcemia.
Cancer-induced hypercalcemia (CIH) occurs in 5% to 30% of patients with cancer during the course of their disease, depending on the type of tumor. This review provides information on the pathophysiology and treatment of CIH. Enhanced bone resorption is the primary cause of CIH and the release of tumor-derived mediators induces this increase in osteoclast-mediated resorption. The interactions between osteoclasts and cancer cells are mainly mediated by parathyroid hormone-related protein (PTHrP), that activates osteoblasts to produce receptor activator of nuclear factor-kappa ligand (RANKL) and osteoclast precursors, with subsequent bone osteolysis. Low parathyroid hormone serum levels together with high calcium levels in a cancer patient may suggest a CIH. There are two different therapeutic approaches for treating CIH, to increase the urinary excretion of calcium, or to inhibit osteoclastic bone resorption, RANKL or the action of PTHrP. In patients with CIH the first step of therapy is usually to restore renal function which is often impaired due to dehydration. Bisphosphonates administration is at present the main-stay of treatment, while calcitonin, gallium nitrate and mithramycin have limited activity and several side-effects. Anti-RANKL therapy (denosumab) and antibodies against PTHrP are promising therapies, but their clinical use should be further explored to more clearly document the effects.