Genomic Prediction Enhanced Sparse Testing for Multi-environment Trials.
"Sparse testing" refers to reduced multi-environment breeding trials in which not all genotypes of interest are grown in each environment. Using genomic-enabled prediction and a model embracing genotype × environment interaction (GE), the non-observed genotype-in-environment combinations can be predicted. Consequently, the overall costs can be reduced and the testing capacities can be increased. The accuracy of predicting the unobserved data depends on different factors including (1) how many genotypes overlap between environments, (2) in how many environments each genotype is grown, and (3) which prediction method is used. In this research, we studied the predictive ability obtained when using a fixed number of plots and different sparse testing designs. The considered designs included the extreme cases of (1) no overlap of genotypes between environments, and (2) complete overlap of the genotypes between environments. In the latter case, the prediction set fully consists of genotypes that have not been tested at all. Moreover, we gradually go from one extreme to the other considering (3) intermediates between the two previous cases with varying numbers of different or non-overlapping (NO)/overlapping (O) genotypes. The empirical study is built upon two different maize hybrid data sets consisting of different genotypes crossed to two different testers (T1 and T2) and each data set was analyzed separately. For each set, phenotypic records on yield from three different environments are available. Three different prediction models were implemented, two main effects models (M1 and M2), and a model (M3) including GE. The results showed that the genome-based model including GE (M3) captured more phenotypic variation than the models that did not include this component. Also, M3 provided higher prediction accuracy than models M1 and M2 for the different allocation scenarios. Reducing the size of the calibration sets decreased the prediction accuracy under all allocation designs with M3 being the less affected model; however, using the genome-enabled models (i.e., M2 and M3) the predictive ability is recovered when more genotypes are tested across environments. Our results indicate that a substantial part of the testing resources can be saved when using genome-based models including GE for optimizing sparse testing designs.


Noradrenaline- and melatonin-mediated regulation of pigment aggregation in fish melanophores.
The effects of melatonin and noradrenaline (NA) on bi-directional melanosome transport were analysed in primary cultures of melanophores from the Atlantic cod. Both agents mediated rapid melanosome aggregation, and by using receptor antagonists, melatonin was found to bind to a melatonin receptor whereas NA binds to an alpha2-adrenoceptor. It has previously been stated that melatonin-mediated melanosome aggregation in Xenopus is coupled with tyrosine phosphorylation of a so far unidentified high molecular weight protein and we show that although acting through different receptors and through somewhat different downstream signalling events, tyrosine phosphorylation is of the utmost importance for melanosome aggregation mediated by both NA and melatonin in cod melanophores. Together with cyclic adenosine 3-phosphate-fluctuations, tyrosine phosphorylation functions as a switch signal for melanosome aggregation and dispersion in these cells.


Population pharmacokinetic/pharmacodynamic analysis of the bactericidal activities of sutezolid (PNU-100480) and its major metabolite against intracellular Mycobacterium tuberculosis in ex vivo whole-blood cultures of patients with pulmonary tuberculosis.
Sutezolid (PNU-100480 [U-480]) is an oxazolidinone antimicrobial being developed for the treatment of tuberculosis. An active sulfoxide metabolite (PNU-101603 [U-603]), which reaches concentrations in plasma several times those of the parent, has been reported to drive the killing of extracellular Mycobacterium tuberculosis by sutezolid in hollow-fiber culture. However, the relative contributions of the parent and metabolite against intracellular M. tuberculosis in vivo are not fully understood. The relationships between the plasma concentrations of U-480 and U-603 and intracellular whole-blood bactericidal activity (WBA) in ex vivo cultures were examined using a direct competitive population pharmacokinetic (PK)/pharmacodynamic 4-parameter sigmoid model. The data set included 690 PK determinations and 345 WBA determinations from 50 tuberculosis patients enrolled in a phase 2a sutezolid trial. The model parameters were solved iteratively. The median U-603/U-480 concentration ratio was 7.1 (range, 1 to 28). The apparent 50% inhibitory concentration of U-603 for intracellular M. tuberculosis was 17-fold greater than that of U-480 (90% confidence interval [CI], 9.9- to 53-fold). Model parameters were used to simulate in vivo activity after oral dosing with sutezolid at 600 mg twice a day (BID) and 1,200 mg once a day (QD). Divided dosing resulted in greater cumulative activity (-0.269 log10 per day; 90% CI, -0.237 to -0.293 log10 per day) than single daily dosing (-0.186 log10 per day; 90% CI, -0.160 to -0.208 log10 per day). U-480 accounted for 84% and 78% of the activity for BID and QD dosing, respectively, despite the higher concentrations of U-603. Killing of intracellular M. tuberculosis by orally administered sutezolid is mainly due to the activity of the parent compound. Taken together with the findings of other studies in the hollow-fiber model, these findings suggest that sutezolid and its metabolite act on different mycobacterial subpopulations.


Self-reported awake bruxism and chronotype profile: a multicenter study on Brazilian, Portuguese and Italian dental students.
Objective: To assess the association between self-reported awake bruxism (AB) and the chronotype profile among Brazilian, Portuguese, and Italian dental students.Methods: A cross-sectional study involving 205 dental students was developed. Self-reported AB as well as information on sleep characteristics were collected from a questionnaire. The Morningness-Eveningness Questionnaire (MEQ) was used to measure the chronotype profile. Descriptive statistical analysis, the chi-square test, and univariate analysis were used to evaluate the factors associated with AB.Results: The prevalence of AB in all groups was 33.7%. The intermediate chronotype profile was the most prevalent (60.4%), and only 16.7% of the participants had the morning profile. The univariate analysis showed that older dental students (OR = 2.640, 95% CI 1.388-5.021) and those with the eveningness chronotype profile (OR = 3.370, 95% CI 1.302-8.725) are associated with AB.Discussion: Students over the age of 22 and those with the eveningness chronotype profile were prone to AB.


Induction by transforming growth factor beta of pemphigus vulgaris antigen activity in mouse papilloma cells.
Induction of a marker of epidermal spinous cells, pemphigus antigen activity, was detected by indirect immunofluorescence in murine papilloma cells exposed to human transforming growth factor beta (TGF-beta). Detection of pemphigus antigen activity required exposure of cells to 1.4 mM Ca2+ for 3 h just prior to immunoassay. The brief exposure to Ca2+ may be necessary for translocation of intracellular pemphigus antigen to the cell surface, where it is accessible to antibody. Cells grown in medium containing 0.02-0.04 mM Ca2+ were shown previously to be primarily basal cells characterized by pemphigoid antigen activity. Following treatment with 0.25-25 pg/ml TGF-beta for 44 h under 0.02-0.04 mM Ca2+ conditions, 63 +/- 9% (SD) of cells were pemphigus positive. This percentage was comparable to that of positive control cultures exposed to 1.4 mM Ca2+ for 44 h (70 +/- 10%) and was up to 2-fold that of solvent control cultures. Pemphigus antigen activity was significantly induced by 0.1-25 pg/ml TGF-beta, out of a tested range of 10(-5)-10(3) pg/ml. The total number of papilloma cell colonies was unaffected by treatment with 0.1-25 pg/ml TGF-beta but was reduced greater than 90% by treatment with 10(3)-5 x 10(3) pg/ml TGF-beta. The described immunofluorescence assay for pemphigus antigen activity may be useful for preliminary evaluation of differentiation-inducing agents in anticarcinoma therapy.


Sex differences in dogs' social learning of spatial information.
We used a modified version of the Do as I Do paradigm to investigate dogs' preference and flexibility in the acquisition of different types of spatial information in social learning situations. When required to match the location of the demonstration, dogs (N = 16) preferentially relied on allocentric information, i.e., the relationship between the location of the demonstration and the various objects surrounding it. However, when allocentric cues were inadequate to solve the task, dogs learned to rely on egocentric information, i.e., the direction-left/right-taken by the human demonstrator. The ease of resorting to the non-preferred egocentric strategy was sex-dependent with males acquiring the egocentric strategy in fewer trials than females. This study shows that dogs rely preferentially on allocentric cues when recalling socially acquired spatial information. However, they are impressively flexible in switching to egocentric strategies according to the task requirements. Whether preference for the allocentric strategy in processing spatial information is embedded in the nature of social learning or restricted to our paradigm is an open question. This study also supports the idea that sex differences in cognitive domains are widespread among mammals and calls for further investigations aimed at shedding light on the evolution, function and mechanisms of such differences.


Paclitaxel targets FOXM1 to regulate KIF20A in mitotic catastrophe and breast cancer paclitaxel resistance.
FOXM1 has been implicated in taxane resistance, but the molecular mechanism involved remains elusive. In here, we show that FOXM1 depletion can sensitize breast cancer cells and mouse embryonic fibroblasts into entering paclitaxel-induced senescence, with the loss of clonogenic ability, and the induction of senescence-associated β-galactosidase activity and flat cell morphology. We also demonstrate that FOXM1 regulates the expression of the microtubulin-associated kinesin KIF20A at the transcriptional level directly through a Forkhead response element (FHRE) in its promoter. Similar to FOXM1, KIF20A expression is downregulated by paclitaxel in the sensitive MCF-7 breast cancer cells and deregulated in the paclitaxel-resistant MCF-7Tax(R) cells. KIF20A depletion also renders MCF-7 and MCF-7Tax(R) cells more sensitive to paclitaxel-induced cellular senescence. Crucially, resembling paclitaxel treatment, silencing of FOXM1 and KIF20A similarly promotes abnormal mitotic spindle morphology and chromosome alignment, which have been shown to induce mitotic catastrophe-dependent senescence. The physiological relevance of the regulation of KIF20A by FOXM1 is further highlighted by the strong and significant correlations between FOXM1 and KIF20A expression in breast cancer patient samples. Statistical analysis reveals that both FOXM1 and KIF20A protein and mRNA expression significantly associates with poor survival, consistent with a role of FOXM1 and KIF20A in paclitaxel action and resistance. Collectively, our findings suggest that paclitaxel targets the FOXM1-KIF20A axis to drive abnormal mitotic spindle formation and mitotic catastrophe and that deregulated FOXM1 and KIF20A expression may confer paclitaxel resistance. These findings provide insights into the underlying mechanisms of paclitaxel resistance and have implications for the development of predictive biomarkers and novel chemotherapeutic strategies for paclitaxel resistance.


[Community participation in the control of intestinal parasitoses at a rural site in Argentina].
To design, implement, and evaluate a complex plan of actions aimed at reducing parasitoses in a rural town in the province of Buenos Aires, Argentina, with the involvement of the community. The project took place in General Mansilla, Buenos Aires province, in three stages. First, the initial epidemiological situation of intestinal parasitoses in the community was assessed by analyzing for parasites a series of stool samples and anal swabs taken from a sample of 522 individuals. Second, interventions were implemented: the pharmacological treatment of the infected individuals, and health education for the public involving local promoters. Third, the pharmacological treatment was evaluated with followup stool testing for those treated; the health education effort was evaluated through a survey and stool testing for parasites of those who had participated solely in the education. The frequency of intestinal parasitoses was 58.2%; of the total, 43.9% were protozoa and 35.2%, helminths. The most common pathogens were Enterobius vermicularis, Blastocystis hominis, and Giardia lamblia. The deworming treatment reduced intestinal parasitoses to 15.1% (P < 0.001), and was more effective in helminths than in protozoa. Intestinal parasitoses declined following the health education intervention, in general (from 58.2% to 47.9%; P = 0.019), as well as for helminths (from 35.2% to 20.3%; P < 0.001), and hygiene habits improved significantly. Treatment of parasitosis and the educational intervention provided by local promoters made it possible to reduce parasitic diseases-particularly those caused by helminths-in the community under study. We recommend extending this experience to other rural communities and broadening the interventions to cutoff additional modes of transmission, such as water and food.


Altered chloride homeostasis in neurological disorders: a new target.
Other than modifying either neurotransmitter release or receptor expression and/or properties, one way to modulate gamma-aminobutyric acid (GABA)A and glycine receptor-mediated signalling is to alter the transmembrane gradient for chloride ions. Because intracellular chloride concentration is low in neurons, and the reversal potential for chloride currents is close to the resting membrane potential, small changes in intracellular chloride concentration can dramatically affect the strength and even polarity of GABA/glycine-mediated transmission. It now appears that chloride homeostasis is actively regulated in the adult brain and affected by endogenous neuromodulators such as brain-derived neurotrophic factor. Modulating chloride gradients even emerges as a mechanism by which microglia can control neuronal excitability. These findings, together with the observation of altered chloride homeostasis in several neurological disorders, point to new targets for therapeutic agents.


A method for event-related phase/amplitude coupling.
Phase/amplitude coupling (PAC) is emerging as an important electrophysiological measure of local and long-distance neuronal communication. Current techniques for calculating PAC provide a numerical index that represents an average value across an arbitrarily long time period. This requires researchers to rely on block design experiments and temporal concatenation at the cost of the sub-second temporal resolution afforded by electrophysiological recordings. Here we present a method for calculating event-related phase/amplitude coupling (ERPAC) designed to capture the temporal evolution of task-related changes in PAC across events or between distant brain regions that is applicable to human or animal electromagnetic recording.


Identification of monocarboxylate transporter 8 as a specific thyroid hormone transporter.
Transport of thyroid hormone across the cell membrane is required for its action and metabolism. Recently, a T-type amino acid transporter was cloned which transports aromatic amino acids but not iodothyronines. This transporter belongs to the monocarboxylate transporter (MCT) family and is most homologous with MCT8 (SLC16A2). Therefore, we cloned rat MCT8 and tested it for thyroid hormone transport in Xenopus laevis oocytes. Oocytes were injected with rat MCT8 cRNA, and after 3 days immunofluorescence microscopy demonstrated expression of the protein at the plasma membrane. MCT8 cRNA induced an approximately 10-fold increase in uptake of 10 nM 125I-labeled thyroxine (T4), 3,3',5-triiodothyronine (T3), 3,3',5'-triiodothyronine (rT3) and 3,3'-diiodothyronine. Because of the rapid uptake of the ligands, transport was only linear with time for <4 min. MCT8 did not transport Leu, Phe, Trp, or Tyr. [125I]T4 transport was strongly inhibited by L-T4, D-T4, L-T3, D-T3, 3,3',5-triiodothyroacetic acid, N-bromoacetyl-T3, and bromosulfophthalein. T3 transport was less affected by these inhibitors. Iodothyronine uptake in uninjected oocytes was reduced by albumin, but the stimulation induced by MCT8 was markedly increased. Saturation analysis provided apparent Km values of 2-5 microM for T4, T3, and rT3. Immunohistochemistry showed high expression in liver, kidney, brain, and heart. In conclusion, we have identified MCT8 as a very active and specific thyroid hormone transporter.


Cost of poor adherence to anti-hypertensive therapy in five European countries.
The financial burden for EU health systems associated with cardiovascular disease (CV) has been estimated to be nearly €110 billion in 2006, corresponding to 10% of total healthcare expenditure across EU or a mean €223 annual cost per capita. The main purpose of this study is to estimate the costs related to hypertension and the economic impact of increasing adherence to anti-hypertensive therapy in five European countries (Italy, Germany, France, Spain and England). A probabilistic prevalence-based decision tree model was developed to estimate the direct costs of CV related to hypertension (CV defined as: stroke, heart attack, heart failure) in five European countries. Our model considered adherence to hypertension treatment as a main driver of blood pressure (BP) control (BP < 140/90 mmHg). Relative risk of CV, based on controlled or uncontrolled BP group, was estimated from the Framingham Heart Study and national review data. Prevalence and cost data were estimated from national literature reviews. A national payer (NP) perspective for 10 years was considered. Probabilistic sensitivity analysis was performed in order to evaluate uncertainty around the results (given as 95% confidence intervals). The model estimated a total of 8.6 million (1.4 in Italy, 3.3 in Germany, 1.2 in Spain, 1.8 in France and 0.9 in England) CV events related to hypertension over the 10-year time horizon. Increasing the adherence rate to anti-hypertensive therapy to 70% (baseline value is different for each country) would lead to 82,235 fewer CV events (24,058 in Italy, 7,870 in Germany, 18,870 in Spain, 24,855 in France and 6,553 in England). From the NP perspective, the direct cost associated with hypertension was estimated to be <euro>51.3 billion (8.1 in Italy, 17.1 in Germany, 12.2 in Spain, 8.8 in France and 5.0 in England). Increasing adherence to anti-hypertensive therapy to 70% would save a total of <euro>332 million (CI 95%: €319-346 million) from the NPs perspective. This study is the first attempt to estimate the economic impact of non-adherence amongst patients with diagnosed hypertension in Europe, using data from five European countries (Italy, France, Germany, Spain and England).


The evidence-based management of bilateral ethmoid osteomas: diagnosis, endoscopic resection and review of the literature.
A 56-year old woman presented with headaches and nasal congestion secondary to bilateral ethmoid osteomas and she made a full recovery following endoscopic resection. Paranasal osteomas are common, benign, encapsulated tumours that are often asymptomatic, but may present similarly to rhinosinusitis and occasionally cause complications depending on their anatomical location. Bilateral ethmoidal osteomas, however are a rare occurrence and have not been previously reported in the English literature as far as the authors are aware. Ethmoidal and fronto-ethmoidal osteomas, like other paranasal osteomas, are easily diagnosed using computed tomography and are increasingly amenable to endoscopic resection. An evidence-based management plan is presented.


MicroRNA expression profiling of human blood monocyte subsets highlights functional differences.
Within human blood there are two subsets of monocytes that can be identified by differential expression of CD16. Although numerous phenotypic and functional differences between the subsets have been described, little is known of the mechanisms underlying the distinctive properties of the two subsets. MicroRNAs (miRNAs) are small non-coding RNAs that can regulate gene expression through promoting mRNA degradation or repressing translation, leading to alterations in cellular processes. Their potential influence on the functions of monocyte subsets has not been investigated. In this study, we employed microarray analysis to define the miRNA expression profile of human monocyte subsets. We identified 66 miRNAs that were differentially expressed (DE) between CD16(+) and CD16(-) monocytes. Gene ontology analysis revealed that the predicted targets of the DE miRNAs were predominantly associated with cell death and cellular movement. We validated the functional impacts of selected DE miRNAs in CD16(-) monocytes, over-expression of miR-432 significantly increases apoptosis, and inhibiting miR-19a significantly reduces cell motility. Furthermore, we found that miR-345, another DE miRNA directly targets the transcription factor RelA in monocytes, which resulted in the differential expression of RelA in monocyte subsets. This implicates miR-345 indirect regulation of many genes downstream of RelA, including important inflammatory mediators. Together, our data show that DE miRNAs could contribute substantially to regulating the functions of human blood monocytes.


Differential regulation by melatonin of cell growth and androgen receptor binding to the androgen response element in prostate cancer cells.
The pineal hormone melatonin inhibits the growth of benign human prostate epithelial cells and the androgen-dependent prostate cancer LNCaP cells. In the androgen-nonresponsive prostate carcinoma PC3 cells melatonin inhibits cell growth only at high but not low cell density. We have recently found that melatonin causes nuclear exclusion of the AR and attenuates it transcriptional activity in LNCaP cells as well as PC3 cells stably transfected with a wild type AR expressing vector (PC3-AR). The aim of this study was to investigate whether melatonin inhibits effects of AR on cell growth in PC3-AR cells and whether inhibition of AR DNA binding is involved. The effects of androgen, melatonin and their combination on the growth of the PC3-AR cells and on AR DNA binding in PC3-AR and LNCaP cells were studied. DHT suppressed cell growth in the PC3-AR cells and enhanced AR binding to the androgen responsive element (ARE). Melatonin had no effect on cell growth in the absence of DHT but counteracted the androgen-induced inhibition at low androgen concentrations. Melatonin did not suppress and even slightly enhanced the capacity of AR binding to the ARE in the PC3-AR as well as in LNCaP cells. Attenuation by melatonin of AR activity in the prostate cancer cells is not due to suppression of AR binding to the ARE, and is presumably caused by its effects on AR protein interaction and intracellular trafficking.


Coronary bifurcations as you have never seen them: the Visible Heart® Laboratory bifurcation programme.
The Visible Heart® Laboratory is an original experimental laboratory in which harvested animal hearts are resuscitated and connected to a support machine in order to beat outside the animal body. Resuscitated animal hearts may be exposed to various types of endovascular intervention under full, multimodality inspection. This unique experimental setting allows the performance of percutaneous coronary intervention (PCI) in a setting which resembles a standard catheterisation laboratory set-up, and contemporaneously allows unique multimodality imaging. For these reasons, the performance of PCI on bifurcations in the Visible Heart® Laboratory may improve the knowledge of the dynamic stent deformations and stent-vessel wall interactions associated with the different steps of the various techniques for bifurcation stenting. Furthermore, the collected images may also serve as a novel educative resource for physicians. The performance of bifurcation stenting in the Visible Heart® Laboratory is a promising experimental setting to gain novel information regarding any existing or future PCI technique to treat coronary bifurcations.


Physical mechanism for gating and mechanosensitivity of the human TRAAK K+ channel.
Activation of mechanosensitive ion channels by physical force underlies many physiological processes including the sensation of touch, hearing and pain. TRAAK (also known as KCNK4) ion channels are neuronally expressed members of the two-pore domain K(+) (K2P) channel family and are mechanosensitive. They are involved in controlling mechanical and temperature nociception in mice. Mechanosensitivity of TRAAK is mediated directly through the lipid bilayer--it is a membrane-tension-gated channel. However, the molecular mechanism of TRAAK channel gating and mechanosensitivity is unknown. Here we present crystal structures of TRAAK in conductive and non-conductive conformations defined by the presence of permeant ions along the conduction pathway. In the non-conductive state, a lipid acyl chain accesses the channel cavity through a 5 Å-wide lateral opening in the membrane inner leaflet and physically blocks ion passage. In the conductive state, rotation of a transmembrane helix (TM4) about a central hinge seals the intramembrane opening, preventing lipid block of the cavity and permitting ion entry. Additional rotation of a membrane interacting TM2-TM3 segment, unique to mechanosensitive K2Ps, against TM4 may further stabilize the conductive conformation. Comparison of the structures reveals a biophysical explanation for TRAAK mechanosensitivity--an expansion in cross-sectional area up to 2.7 nm(2) in the conductive state is expected to create a membrane-tension-dependent energy difference between conformations that promotes force activation. Our results show how tension of the lipid bilayer can be harnessed to control gating and mechanosensitivity of a eukaryotic ion channel.


Evolutionary genomics of nucleo-cytoplasmic large DNA viruses.
A previous comparative-genomic study of large nuclear and cytoplasmic DNA viruses (NCLDVs) of eukaryotes revealed the monophyletic origin of four viral families: poxviruses, asfarviruses, iridoviruses, and phycodnaviruses [Iyer, L.M., Aravind, L., Koonin, E.V., 2001. Common origin of four diverse families of large eukaryotic DNA viruses. J. Virol. 75 (23), 11720-11734]. Here we update this analysis by including the recently sequenced giant genome of the mimiviruses and several additional genomes of iridoviruses, phycodnaviruses, and poxviruses. The parsimonious reconstruction of the gene complement of the ancestral NCLDV shows that it was a complex virus with at least 41 genes that encoded the replication machinery, up to four RNA polymerase subunits, at least three transcription factors, capping and polyadenylation enzymes, the DNA packaging apparatus, and structural components of an icosahedral capsid and the viral membrane. The phylogeny of the NCLDVs is reconstructed by cladistic analysis of the viral gene complements, and it is shown that the two principal lineages of NCLDVs are comprised of poxviruses grouped with asfarviruses and iridoviruses grouped with phycodnaviruses-mimiviruses. The phycodna-mimivirus grouping was strongly supported by several derived shared characters, which seemed to rule out the previously suggested basal position of the mimivirus [Raoult, D., Audic, S., Robert, C., Abergel, C., Renesto, P., Ogata, H., La Scola, B., Suzan, M., Claverie, J.M. 2004. The 1.2-megabase genome sequence of Mimivirus. Science 306 (5700), 1344-1350]. These results indicate that the divergence of the major NCLDV families occurred at an early stage of evolution, prior to the divergence of the major eukaryotic lineages. It is shown that subsequent evolution of the NCLDV genomes involved lineage-specific expansion of paralogous gene families and acquisition of numerous genes via horizontal gene transfer from the eukaryotic hosts, other viruses, and bacteria (primarily, endosymbionts and parasites). Amongst the expansions, there are multiple families of predicted virus-specific signaling and regulatory domains. Most NCLDVs have also acquired large arrays of genes related to ubiquitin signaling, and the animal viruses in particular have independently evolved several defenses against apoptosis and immune response, including growth factors and potential inhibitors of cytokine signaling. The mimivirus displays an enormous array of genes of bacterial provenance, including a representative of a new class of predicted papain-like peptidases. It is further demonstrated that a significant number of genes found in NCLDVs also have homologs in bacteriophages, although a vertical relationship between the NCLDVs and a particular bacteriophage group could not be established. On the basis of these observations, two alternative scenarios for the origin of the NCLDVs and other groups of large DNA viruses of eukaryotes are considered. One of these scenarios posits an early assembly of an already large DNA virus precursor from which various large DNA viruses diverged through an ongoing process of displacement of the original genes by xenologous or non-orthologous genes from various sources. The second scenario posits convergent emergence, on multiple occasions, of large DNA viruses from small plasmid-like precursors through independent accretion of similar sets of genes due to strong selective pressures imposed by their life cycles and hosts.


Taxol (paclitaxel): mechanisms of action.
Paclitaxel, an antitumor drug that is demonstrating encouraging activity in human malignancies, is likely to play a major role in cancer chemotherapy. Paclitaxel has an unusual chemical structure--it is a complex diterpene having a taxane ring with a four-membered oxetane ring and an ester side chain at position C-13--and a unique mechanism of action. In vitro, paclitaxel enhances the polymerization of tubulin to stable microtubules and also interacts directly with microtubules, stabilizing them against depolymerization by cold and calcium, which readily depolymerize normal microtubules. The fact that the drug has a specific binding site on the microtubule polymer makes it unique among chemotherapeutic agents, and the ability of paclitaxel to polymerize tubulin in the absence of cofactors like guanosine triphosphate and microtubule-associated proteins is unusual. When paclitaxel and microtubule protein are irradiated with ultraviolet light, the drug preferentially binds covalently to the beta-subunit of tubulin. Paclitaxel binds to cells in a specific and saturable manner with a single set of high-affinity binding sites. The microtubule cytoskeleton is reorganized in the presence of paclitaxel and extensive parallel arrays or stable bundles of microtubules are formed in cells growing in tissue culture. Paclitaxel blocks cells in the G2/M phase of the cell cycle and such cells are unable to form a normal mitotic apparatus.(ABSTRACT TRUNCATED AT 250 WORDS)


BRCA1 and BRCA2 Gene Expression: Diurnal Variability and Influence of Shift Work
BRCA1 and BRCA2 genes are involved in DNA double-strand break repair and related to breast cancer. Shift work is associated with biological clock alterations and with a higher risk of breast cancer. The aim of this study was to investigate the variability of expression of BRCA genes through the day in healthy subjects and to measure BRCA expression levels in shift workers. The study was approached in two ways. First, we examined diurnal variation of BRCA1 and BRCA2 genes in lymphocytes of 15 volunteers over a 24-hour period. Second, we measured the expression of these genes in lymphocytes from a group of shift and daytime workers. The change in 24-hour expression levels of BRCA1 and BRCA2 genes was statistically significant, decreasing from the peak at midday to the lowest level at midnight. Lower levels for both genes were found in shift workers compared to daytime workers. Diurnal variability of BRCA1 and BRCA2 expression suggests a relation of DNA double-strand break repair system with biological clock. Lower levels of BRCA1 and BRCA2 found in shift workers may be one of the potential factors related to the higher risk of breast cancer.


[Diagnosis of attention-deficit hyperactivity disorder in childhood and adolescence].
Attention-deficit hyperactivity disorder (ADHD) has severe psychosocial consequences over the whole life course of patients and their families. During childhood and adolescence ADHD needs to be discriminated from other psychiatric or somatic disorders. Clinical diagnosis is met according to the diagnostic criteria by integrating several anamnestic sources and standardised diagnostic means. Diagnosis of comorbid disorders is obligatory due to their high prevalence rates. In some cases successful treatment of a child suffering from ADHD requires the additional diagnosis and treatment of ADHD in further family members.


Prescription Drug Coverage and Outcomes of Myeloma Therapy Among Medicare Beneficiaries.
Novel parenteral (bortezomib) and oral (lenalidomide) therapies have improved survival in myeloma, but the standard Medicare benefit covers only parenteral drugs. We examined the association between prescription drug coverage, receipt of therapy, and survival among Medicare beneficiaries with myeloma. Using SEER-Medicare data, we identified enrollment in a Medicare Part D plan (PDP) or other creditable prescription drug coverage (OCC) among 9,755 beneficiaries who were diagnosed with myeloma in 2006 to 2011. We examined the receipt of active myeloma therapy and that of classic cytotoxic agents or bortezomib as first-line regimen and overall survival. We report relative risk (RR) for binary outcome comparisons and 3-year restricted mean survival time (RMST) ratios, with 95% CI, adjusting for baseline patient- and disease-related characteristics. Beneficiaries with diffuse large B-cell lymphoma, a cancer that is uniformly treated with parenteral chemotherapy, served as a comparison cohort. Compared with beneficiaries without prescription drug coverage, PDP or OCC enrollees were more likely to receive active myeloma care, and PDP enrollees were less frequently treated with parenteral agents (adjusted RR, 0.86; 95% CI, 0.80 to 0.93) or classic cytotoxic agents in particular (RR, 0.62; 95% CI, 0.51 to 0.76). Overall survival was significantly better for beneficiaries with PDP coverage (adjusted RMST ratio, 1.16; 95% CI, 1.11 to 1.20) or OCC (RMST ratio, 1.16; 95% CI, 1.12 to 1.21). In contrast, we observed no survival differences by prescription drug coverage status in the control cohort with lymphoma. Prescription drug coverage is associated with decreased use of classic cytotoxic chemotherapy and better survival among Medicare beneficiaries with myeloma, which suggests improved access to all existing treatment options. As oral targeted agents increasingly replace parenteral chemotherapy in oncology, adjustments in coverage policy are needed to ensure access to optimal treatment.


Modeling human interindividual variability in metabolism and risk: the example of 4-aminobiphenyl.
We investigate, through modeling, the impact of interindividual heterogeneity in the metabolism of 4-aminobiphenyl (ABP) and in physiological factors on human cancer risk: A physiological pharmacokinetic model was used to quantify the time course of the formation of the proximate carcinogen, N-hydroxy-4-ABP and the DNA-binding of the active species in the bladder. The metabolic and physiologic model parameters were randomly varied, via Monte Carlo simulations, to reproduce interindividual variability. The sampling means for most parameters were scaled from values developed by Kadlubar et al. (Cancer Res., 51: 4371, 1991) for dogs; variances were obtained primarily from published human data (e.g., measurements of ABP N-oxidation, and arylamine N-acetylation in human liver tissue). In 500 simulations, theoretically representing 500 humans, DNA-adduct levels in the bladder of the most susceptible individuals are ten thousand times higher than for the least susceptible, and the 5th and 95th percentiles differ by a factor of 160. DNA binding for the most susceptible individual (with low urine pH, low N-acetylation and high N-oxidation activities) is theoretically one million-fold higher than for the least susceptible (with high urine pH, high N-acetylation and low N-oxidation activities). The simulations also suggest that the four factors contributing most significantly to interindividual differences in DNA-binding of ABP in human bladder are urine pH, ABP N-oxidation, ABP N-acetylation and urination frequency.


Transport of estrogen-induced oxytocin receptors in the ventromedial hypothalamus.
The modulation of oxytocin (OT) receptors (OTRs) by estrogen was investigated in the ventromedial hypothalamus by in vitro receptor autoradiography. Treatment of ovariectomized and adrenalectomized rats with various doses of estradiol benzoate (EB) increased OTR binding not only in the ventromedial nuclei of the hypothalamus (VMN), but also in the area lateral to the nuclei (IVMN). After a single injection of EB, OTRs first were induced within the ventrolateral parts of the VMN, and only hours later they appeared in the IVMN. This is consistent with the interpretation that OTRs are first induced within the estrogen-sensitive neurons of the ventrolateral VMN and then are transported laterally out of the nuclei. Two additional experiments confirmed this interpretation. First, local infusion of a low dose (10 micrograms) of the neuronal transport inhibitor vinblastine blocked the appearance of OTRs in the IVMN but did not prevent the induction of OTRs by EB within the nuclei. Second, a knife cut placed lateral to the VMN prevented the spread of OTRs out of the nuclei. However, even after treatment with a high dose of EB (2 x 10 micrograms), progesterone (P) was required for a maximal extension of the area covered by OTRs. Thus, the OTR is an estrogen-induced neurotransmitter receptor that is transported to its site of action, the lateral ventromedial hypothalamus, where it is modulated by P and where estrogen-induced OT immunoreactivity is found.


Infective endocarditis--a twelve year surgical outcome series.
To review the clinical course and outcome of patients with infective endocarditis proceeding to surgical treatment in the South Island of New Zealand. A retrospective review of all cases of infective endocarditis requiring cardiac surgery, excepting homograft replacement between 1989 and June 2001 was performed. All patients treated at both cardiothoracic units over this time frame in the South Island of New Zealand were included. A total of 29 patients, ten females and nineteen males, age range 31-79 years (mean 55) underwent surgery. 27 patients had native valve endocarditis, two infection of prosthetic valves. A variety of causative micro-organisms were isolated, and all patients received aggressive intravenous antibiotic therapy. Heart failure was the predominant indication for surgical intervention. Fifteen patients underwent aortic, nine mitral, three combined and two replacement of infected prosthetic valves. There were five peri-operative deaths (17% mortality) and significant morbidity in a further eleven patients (38%). Of the 23 survivors available to follow-up none have recurrent endocarditis, with an average disease free survival of 35 months. Patients who require valve surgery for endocarditis have significant peri-operative morbidity and mortality. Long-term outcome in survivors, however, is extremely good with a prognosis similar to those undergoing elective valve replacement surgery. Mycotic cerebral aneurysms are an emerging important cause of early deaths.


Compulsivity in Alcohol Use Disorder and Obsessive Compulsive Disorder: Implications for Neuromodulation.
Alcohol use Disorder (AUD) is one of the leading causes of morbidity and mortality worldwide. The progression of the disorder is associated with the development of compulsive alcohol use, which in turn contributes to the high relapse rate and poor longer term functioning reported in most patients, even with treatment. While the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) defines AUD by a cluster of symptoms, parsing its heterogeneous phenotype by domains of behavior such as compulsivity may be a critical step to improve outcomes of this condition. Still, neurobiological underpinnings of compulsivity need to be fully elucidated in AUD in order to better design targeted treatment strategies. In this manuscript, we review and discuss findings supporting common mechanisms between AUD and OCD, dissecting the construct of compulsivity and focusing specifically on characteristic disruptions in habit learning and cognitive control in the two disorders. Finally, neuromodulatory interventions are proposed as a probe to test compulsivity as key pathophysiologic feature of AUD, and as a potential therapy for the subgroup of individuals with compulsive alcohol use, i.e., the more resistant stage of the disorder. This transdiagnostic approach may help to destigmatize the disorder, and suggest potential treatment targets across different conditions.


N-Substituted carbazolyloxyacetic acids modulate Alzheimer associated gamma-secretase.
N-Sulfonylated and N-alkylated carbazolyloxyacetic acids were investigated for the inhibition and modulation of the Alzheimer's disease associated gamma-secretase. The introduction of a lipophilic substituent, which may vary from arylsulfone to alkyl, turned 2-carbazolyloxyacetic acids into potent gamma-secretase modulators. This resulted in the selective reduction of Abeta(42) and an increase of the less aggregatory Abeta(38) fragment by several compounds (e.g., 7d and 8c). Introduction of an electron donating group at position 6 and 8 of N-substituted carbazolyloxyacetic acids either decreased the activity or inversed modulation. The most active compounds displayed activity on amyloid precursor protein (APP) overexpressing cell lines in the low micromolar range and little or no effect on the gamma-secretase cleavage at the epsilon-site.


Geographic variation of individual venom profile of Crotalus durissus snakes.
South American rattlesnakes are represented in Brazil by a single species, Crotalus durissus, which has public health importance due to the severity of its envenomation and to its wide geographical distribution. The species is subdivided into several subspecies, but the current classification is controversial. In Brazil, the venoms of C. d. terrificus and C. d. collilineatus are used for hyperimmunization of horses for antivenom production, even though the distinction of these two subspecies are mostly by their geographical distribution. In this context, we described a comparative compositional and functional characterization of individual C. d. collilineatus and C. d. terrificus venoms from three Brazilian states. We compared the compositional patterns of C. d. terrificus and C. d. collilineatus individual venoms by 1-DE and RP-HPLC. For functional analyzes, the enzymatic activities of PLA2, LAAO, and coagulant activity were evaluated. Finally, the immunorecognition of venom toxins by the crotalic antivenom produced at Butantan Institute was evaluated using Western blotting. The protein profile of individual venoms from C. d. collilineatus and C. d. terrificus showed a comparable overall composition, despite some intraspecific variation, especially regarding crotamine and LAAO. Interestingly, HPLC analysis showed a geographic pattern concerning PLA2. In addition, a remarkable intraspecific variation was also observed in PLA2, LAAO and coagulant activities. The immunorecognition pattern of individual venoms from C. d. collilineatus and C. d. terrificus by crotalic antivenom produced at Butantan Institute was similar. The results highlighted the individual variability among the venoms of C. durissus ssp. specimens. Importantly, our data point to a geographical variation of C. durissus ssp. venom profile, regardless of the subspecies, as evidenced by PLA2 isoforms complexity, which may explain the increase in venom neurotoxicity from Northeastern through Southern Brazil reported for the species.


Rodent models of social stress and neuronal plasticity: Relevance to depressive-like disorders.
Exposure to severe or persistent social stress may lead to the development of psychiatric disorders such as anxiety and depression. These mood disorders are associated with structural alterations of neural architecture in limbic brain regions that control emotion, mood and cognition. Structural remodeling may either be a sign of successful adaptation, or of failure to do so. In neuropsychiatric disorders like depression structural remodeling involves apoptosis, reduced neurogenesis, and structural remodeling of neuronal dendrites which most likely reflects the latter. Here we review key findings from animal models of psychosocial stress that have been used to gain insights into the relation between stress-related behavioral disorders like depression and structural plasticity. Specifically, we focus on models having a high face validity like social defeat stress in the resident-intruder paradigm and chronic stress of social subordination in social housing conditions. Moderate to severe social stress appears to stimulate plasticity and neuronal growth in regions of the amygdala, whereas the effects in the hippocampus and prefrontal cortex tend to be opposite. A major focus of the current review is to characterize social stress induced structural changes in these brain regions, aiming to provide insight in pathways and factors that underlie behavioral effects of stress and depression.


Deletion of PIK3C3/Vps34 in sensory neurons causes rapid neurodegeneration by disrupting the endosomal but not the autophagic pathway.
The lipid kinase PIK3C3 (also called Vps34) regulates both the endosomal and autophagic pathways. However, the effect of inactivating PIK3C3 on neuronal endosomal versus autophagic processes in vivo has not been studied. We generated mice in which Pik3c3 was conditionally deleted in differentiated sensory neurons. Within a few days after Pik3c3 deletion, mutant large-diameter myelinated neurons accumulated numerous enlarged vacuoles and ubiquitin-positive aggregates and underwent rapid degeneration. By contrast, Pik3c3-deficient small-diameter unmyelinated neurons accumulated excessive numbers of lysosome-like organelles and degenerated more slowly. These differential degenerative phenotypes are unlikely caused by a disruption in the autophagy pathway, because inhibiting autophagy alone by conditional deletion of Atg7 results in a completely distinct phenotype in all sensory neurons (i.e., formation of very large intracellular inclusion bodies and slow degeneration over a period of several months). More surprisingly, a noncanonical PIK3C3-independent LC3-positive autophagosome formation pathway was activated in Pik3c3-deficient small-diameter neurons. Analyses of Pik3c3/Atg7 double mutant neurons revealed that this unconventional initiation pathway still depends on ATG7. Our studies represent in vivo characterization of PIK3C3 functions in mammals and provide insights into the complexity of neuronal endo-lysosomal and autophagic pathways.


The dual impact of ecology and management on social incentives in marine common-pool resource systems.
Understanding how and when cooperative human behaviour forms in common-pool resource systems is critical to illuminating social-ecological systems and designing governance institutions that promote sustainable resource use. Before assessing the full complexity of social dynamics, it is essential to understand, concretely and mechanistically, how resource dynamics and human actions interact to create incentives and pay-offs for social behaviours. Here, we investigated how such incentives for information sharing are affected by spatial dynamics and management in a common-pool resource system. Using interviews with fishermen to inform an agent-based model, we reveal generic mechanisms through which, for a given ecological setting characterized by the spatial dynamics of the resource, the two 'human factors' of information sharing and management may heterogeneously impact various members of a group for whom theory would otherwise predict the same strategy. When users can deplete the resource, these interactions are further affected by the management approach. Finally, we discuss the implications of alternative motivations, such as equity among fishermen and consistency of the fleet's output. Our results indicate that resource spatial dynamics, form of management and level of depletion can interact to alter the sociality of people in common-pool resource systems, providing necessary insight for future study of strategic decision processes.


Sibanye Methods for Prevention Packages Program Project Protocol: Pilot Study of HIV Prevention Interventions for Men Who Have Sex With Men in South Africa.
Human immunodeficiency virus (HIV) prevention intervention programs and related research for men who have sex with men (MSM) in the southern African region remain limited, despite the emergence of a severe epidemic among this group. With a lack of understanding of their social and sexual lives and HIV risks, and with MSM being a hidden and stigmatized group in the region, optimized HIV prevention packages for southern African MSM are an urgent public health and research priority. The objective of the Sibanye Health Project is to develop and evaluate a combination package of biomedical, behavioral, and community-level HIV prevention interventions and services for MSM in South Africa. The project consists of three phases: (1) a comprehensive literature review and summary of current HIV prevention interventions (Phase I), (2) agent-based mathematical modeling of HIV transmission in southern African MSM (Phase II), and (3) formative and stigma-related qualitative research, community engagement, training on providing health care to MSM, and the pilot study (Phase III). The pilot study is a prospective one-year study of 200 men in Cape Town and Port Elizabeth, South Africa. The study will assess a package of HIV prevention services, including condom and condom-compatible lubricant choices, risk-reduction counseling, couples HIV testing and counseling, pre-exposure prophylaxis (PrEP) for eligible men, and non-occupational post-exposure prophylaxis for men with a high risk exposure. The pilot study will begin in October 2014. Preliminary results from all components but the pilot study are available. We developed a literature review database with meta-data extracted from 3800 documents from 67 countries. Modeling results indicate that regular HIV testing and promotion of condom use can significantly impact new HIV infections among South African MSM, even in the context of high coverage of early treatment of HIV-positive men and high coverage of PrEP for at-risk HIV-negative men. Formative qualitative research consisted of 79 in-depth interviews, and six focus group discussions in Cape Town and Port Elizabeth. Analysis of these data has informed pilot study protocol development and has been documented in peer-reviewed manuscripts. Qualitative work regarding stigma faced by South African MSM resulted in finalized scales for use in the pilot study questionnaire. A total of 37 health care providers completed training designed to facilitate clinically and culturally competent care for MSM in the Eastern Cape. The design of a future, larger study of the HIV prevention package will be conducted at the end of the pilot study, powered to detect efficacy of the prevention package. Data from the updated mathematical model, results of the pilot study, acceptability data, and advancements in HIV prevention sciences will be considered in developing the final proposed package and study design. ClinicalTrials.gov NCT02043015; http://clinicaltrials.gov/show/NCT02043015 (Archived by WebCite at http://www.webcitation.org/6THvp7rAj).


TwitterSensing: An Event-Based Approach for Wireless Sensor Networks Optimization Exploiting Social Media in Smart City Applications.
Modern cities are subject to periodic or unexpected critical events, which may bring economic losses or even put people in danger. When some monitoring systems based on wireless sensor networks are deployed, sensing and transmission configurations of sensor nodes may be adjusted exploiting the relevance of the considered events, but efficient detection and classification of events of interest may be hard to achieve. In Smart City environments, several people spontaneously post information in social media about some event that is being observed and such information may be mined and processed for detection and classification of critical events. This article proposes an integrated approach to detect and classify events of interest posted in social media, notably in Twitter, and the assignment of sensing priorities to source nodes. By doing so, wireless sensor networks deployed in Smart City scenarios can be optimized for higher efficiency when monitoring areas under the influence of the detected events.


Qualitative identification of food materials by complex refractive index mapping in the terahertz range.
We investigated the feasibility of qualitative food analysis using complex refractive index mapping of food materials in the terahertz (THz) frequency range. We studied optical properties such as the refractive index and absorption coefficient of food materials, including insects as foreign substances, from 0.2 to 1.3 THz. Although some food materials had a complex composition, their refractive indices were approximated with effective medium values, and therefore, they could be discriminated on the complex refractive index map. To demonstrate food quality inspection with THz imaging, we obtained THz reflective images and time-of-flight imaging of hidden defects in a sugar and milk powder matrix by using time domain THz pulses. Our results indicate that foreign substances can be clearly classified and detected according to the optical parameters of the foods and insects by using THz pulses.


Oxidant-induced hepatocyte injury from menadione is regulated by ERK and AP-1 signaling.
Oxidative stress has been implicated as a mechanism for a variety of forms of liver injury. Although reactive oxygen species (ROS) may damage cellular macromolecules directly, oxidant-induced cell death may result from redox effects on signal transduction pathways. To understand the mechanisms of hepatocyte death from oxidative stress, the functions of the mitogen-activated protein kinases (MAPKs) were determined during oxidant-induced hepatocyte injury from menadione. Low, nontoxic, and high toxic concentrations of the superoxide generator menadione were established in the RALA255-10G rat hepatocyte cell line. Death from menadione was blocked by catalase and ebselen, indicating that death was secondary to oxidant generation and not arylation. Treatment with a nontoxic menadione concentration resulted in a brief activation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). In contrast, treatment with a toxic menadione concentration induced a prolonged activation of both ERK and JNK. Chemical inhibition of ERK function sensitized RALA hepatocytes to death from previously nontoxic menadione concentrations in association with sustained JNK activation. Adenoviral expression of a dominant-negative protein for c-Jun, a downstream substrate for JNK, blocked death from menadione. The pro-apoptotic effect of c-Jun was not mediated through the mitochondrial death pathway. In conclusion, RALA hepatocyte resistance to oxidant-induced death from menadione is dependent on ERK, whereas cell death is mediated by AP-1 activation. These findings identify signaling pathways that may be therapeutic targets in the prevention or treatment of oxidant-induced liver injury.


Surgically-assisted abdominal wall blocks for analgesia after abdominoplasty: A prospective randomized trial.
Abdominoplasty is a common aesthetic procedure. The transversus abdominis plane block (TAPB) and rectus sheath block (RSB) have proven efficacy as analgesic modality for abdominal surgeries. This study demonstrates post-abdominoplasty analgesic duration consequent to the three surgically infiltrated local anesthetic techniques: bilateral TAPB, bilateral RSB, and subcutaneous infiltration (SCI) of 0.25% bupivacaine. In this prospective randomized study, 48 adult patients scheduled for abdominoplasty were randomized into three groups: TAPB group (n = 16), RSB group (n = 16), and SCI group (n = 16) utilizing 40 mL of 0.25% bupivacaine for each block. In both TAPB and RSB groups, the block was performed bilaterally after plication of anterior abdominal wall, while in SCI group, the surgical incisional area was infiltrated before skin closure. Main outcome measures included visual analogue scale (VAS), at rest and during movement; the analgesic duration; and the total required doses of morphine in the first postoperative day. A statistically significant longer analgesia was recorded in the TABP group compared with both the RSB and SCI groups. Statistically significant higher VAS scores in the SCI group 4 hours postoperatively was recorded, both at rest and during movement, compared with both TABP and RSB groups. Significant higher morphine consumption in the SCI group was compared with the other two groups. Among the surgically infiltrated anesthetic techniques for abdominoplasty, bilateral TAPB was associated with longer postoperatively analgesic duration with lower morphine consumption in the first 24 hours compared with RSB and SCI.


Individual and Sexual Network Predictors of HIV Incidence Among Men Who Have Sex With Men in Nigeria.
To date, HIV incidence studies among men who have sex with men (MSM) across sub-Saharan Africa have focused on studying sexual risk practices with less focus on sexual networks. TRUST/RV368 conducted in Abuja and Lagos, Nigeria, recruited MSM using respondent-driven sampling and followed HIV-negative men for incident infection over 4 years. Four-hundred forty-one HIV-uninfected MSM underwent a parallel rapid HIV testing algorithm every 3 months for up to 18 months. HIV incidence per 100 person-years (PY) and 95% confidence intervals (CIs) were estimated using Poisson regression. Individual and network characteristics were examined using multivariable Cox-proportional hazards regression adjusted and unadjusted for respondent-driven sampling weights. Among cohort members with a median age of 23 years [interquartile range (IQR): 20-27], 81 HIV infections occurred over 527 PY (incidence 15.4/100 PY; 95% CI: 12.3 to 19.0). The incidence rate was highest among 16-19 year olds as compared to those 25 years or older (30.9/100 PY; 95% CI: 22.1 to 45.3 vs. 6.9/100 PY; 95% CI: 4.2 to 10.9, respectively). Individual determinants included receptive partnerships, condomless sex, no history of testing for HIV, and rectal gonorrhea. Sexual networks were larger and consisted of an older sexual partner, although there was no clustering by recruitment networks. These HIV incidence data reinforce the unmet HIV prevention needs among young MSM in Nigeria. Even in the context of emerging HIV diagnostic and prevention strategies, structural challenges including stigma and criminalization of same-sex practices highlight the need for novel implementation approaches in the context of MSM-friendly services.


Loss of cardiac myofibrils: mechanism of contractile deficits induced by taurine deficiency.
It is well established that taurine deficiency is associated with myocardial contractile dysfunction; however, the mechanism is unknown. As a follow-up to finding reduced force generation in taurine-depleted rat cardiac trabeculae, using either calcium or strontium activation, this study examined alterations in ventricular fine structure and contractile proteins in animals made taurine deficient by in vivo treatment with a taurine transport antagonist, guanidinoethane sulfonate. Observations of ventricular ultrastructure showed disordered contractile filaments and clear losses of myofibrillar bundles in association with taurine deficiency. Biochemical analyses of ventricular contractile proteins using polyacrylamide gel electrophoresis confirmed losses of the major sarcomeric proteins, myosin and actin. These findings provide a possible mechanism for the contractile deficits and cardiomyopathy described in taurine-deficient animals.


Strategies for Developing Students' Empathy and Awareness for the Needs of People with Disabilities: Contributions to Design Education.
The design of products for people with disabilities requires the understanding of a wide range of factors related to users' health, functional abilities, needs, expectations and preferences. Such multifactorial perspective is often perceived as beyond the reach by the students of both graphic and product design, as it comprises knowledge from different areas such are not usually part of design curriculum as health, rehabilitation, computer science and biomedical engineering. Here, we report on strategies for developing design students' empathy and awareness for the needs and expectations of people with disabilities. By means of a combination of theoretical and practical approaches, a course on Inclusive Design was developed as part of the regular curriculum of the Bachelor Programme in Design at Sao Paulo State University (UNESP, Bauru campus, Brazil), with the collaborative participation of members of SORRI BAURU Rehabilitation Center. The final projects developed by the students were based on the demands presented by SORRI BAURU's rehabilitation team, and results reveal that the theoretical-practical approach based on interdisciplinarity was shown to provide the design students a learning experience that, ultimately, supports the quality decision-making in the design process. This paper describes the pedagogical approach, theoretical contents and practical activities developed during the Inclusive Design course. The challenges, benefits, results, and contributions of this experience from the perspective of the design education are also discussed.


Incidence of intraoperative seizures during motor evoked potential monitoring in a large cohort of patients undergoing different surgical procedures.
OBJECTIVE The purpose of this study was to investigate the incidence of seizures during the intraoperative monitoring of motor evoked potentials (MEPs) elicited by electrical brain stimulation in a wide spectrum of surgeries such as those of the orthopedic spine, spinal cord, and peripheral nerves, interventional radiology procedures, and craniotomies for supra- and infratentorial tumors and vascular lesions. METHODS The authors retrospectively analyzed data from 4179 consecutive patients who underwent surgery or an interventional radiology procedure with MEP monitoring. RESULTS Of 4179 patients, only 32 (0.8%) had 1 or more intraoperative seizures. The incidence of seizures in cranial procedures, including craniotomies and interventional neuroradiology, was 1.8%. In craniotomies in which transcranial electrical stimulation (TES) was applied to elicit MEPs, the incidence of seizures was 0.7% (6/850). When direct cortical stimulation was additionally applied, the incidence of seizures increased to 5.4% (23/422). Patients undergoing craniotomies for the excision of extraaxial brain tumors, particularly meningiomas (15 patients), exhibited the highest risk of developing an intraoperative seizure (16 patients). The incidence of seizures in orthopedic spine surgeries was 0.2% (3/1664). None of the patients who underwent surgery for conditions of the spinal cord, neck, or peripheral nerves or who underwent cranial or noncranial interventional radiology procedures had intraoperative seizures elicited by TES during MEP monitoring. CONCLUSIONS In this largest such study to date, the authors report the incidence of intraoperative seizures in patients who underwent MEP monitoring during a wide spectrum of surgeries such as those of the orthopedic spine, spinal cord, and peripheral nerves, interventional radiology procedures, and craniotomies for supra- and infratentorial tumors and vascular lesions. The low incidence of seizures induced by electrical brain stimulation, particularly short-train TES, demonstrates that MEP monitoring is a safe technique that should not be avoided due to the risk of inducing seizures.


The contributions of visual and central attention to visual working memory.
We investigated the role of two kinds of attention-visual and central attention-for the maintenance of visual representations in working memory (WM). In Experiment 1 we directed attention to individual items in WM by presenting cues during the retention interval of a continuous delayed-estimation task, and instructing participants to think of the cued items. Attending to items improved recall commensurate with the frequency with which items were attended (0, 1, or 2 times). Experiments 1 and 3 further tested which kind of attention-visual or central-was involved in WM maintenance. We assessed the dual-task costs of two types of distractor tasks, one tapping sustained visual attention and one tapping central attention. Only the central attention task yielded substantial dual-task costs, implying that central attention substantially contributes to maintenance of visual information in WM. Experiment 2 confirmed that the visual-attention distractor task was demanding enough to disrupt performance in a task relying on visual attention. We combined the visual-attention and the central-attention distractor tasks with a multiple object tracking (MOT) task. Distracting visual attention, but not central attention, impaired MOT performance. Jointly, the three experiments provide a double dissociation between visual and central attention, and between visual WM and visual object tracking: Whereas tracking multiple targets across the visual filed depends on visual attention, visual WM depends mostly on central attention.


Periprosthetic humeral fractures during shoulder arthroplasty.
Currently, there is little information available on the treatment and outcome of intraoperative periprosthetic humeral fractures that occur during shoulder arthroplasty. The purpose of this study was to report on the incidence, treatment, and outcome of, as well as the risk factors for, intraoperative periprosthetic humeral fractures. Between 1980 and 2002, forty-five intraoperative periprosthetic humeral fractures occurred during shoulder arthroplasty at our institution. Twenty-eight fractures occurred during primary total shoulder arthroplasty, three occurred during primary hemiarthroplasty, and fourteen occurred during revision arthroplasty. Nineteen fractures involved the greater tuberosity, sixteen involved the humeral shaft, six involved the metaphysis, three involved the greater tuberosity and the humeral shaft, and one involved both the greater and lesser tuberosities. All patients were followed for a minimum of two years. At the time of the latest follow-up, outcomes were assessed, radiographs were examined, and relative risks were calculated. Over the twenty-two-year study period, the rate of intraoperative humeral fractures at our institution was 1.5%. All fractures healed at a mean of seventeen weeks. In the primary arthroplasty group (thirty-one patients), range of motion and pain scores improved significantly (p < 0.05) at the time of follow-up. In the revision arthroplasty group (fourteen patients), range of motion remained unchanged whereas pain scores improved significantly (p < 0.005). Transient nerve injuries occurred in six patients. Four fractures displaced postoperatively and were then treated nonoperatively; all four healed. Significant relative risks for intraoperative fracture were female sex, revision surgery, and press-fit implants (p < 0.05). The data from the present study suggest that although intraoperative humeral fractures are associated with a high rate of healing, there was a substantial rate of associated complications, including transient nerve injuries and fracture displacement. Significant risk factors for intraoperative fractures include female sex, revision surgery, and press-fit humeral implants.


Adipocyte telomere length associates negatively with adipocyte size, whereas adipose tissue telomere length associates negatively with the extent of fibrosis in severely obese women.
Telomere length can be considered as a biological marker for cell proliferation and aging. Obesity is associated with adipocyte hypertrophy and proliferation as well as with shorter telomeres in adipose tissue. As adipose tissue is a mixture of different cell types and the cellular composition of adipose tissue changes with obesity, it is unclear what determines telomere length of whole adipose tissue. We aimed to investigate telomere length in whole adipose tissue and isolated adipocytes in relation to adiposity, adipocyte hypertrophy and adipose tissue inflammation and fibrosis. Telomere length was measured by real-time PCR in visceral adipose tissue, and isolated adipocytes of 21 obese women with a waist ranging from 110 to 147 cm and age from 31 to 61 years. Telomere length in adipocytes was shorter than in whole adipose tissue. Telomere length of adipocytes but not whole adipose tissue correlated negatively with waist and adipocyte size, which was still significant after correction for age. Telomere length of whole adipose tissue associated negatively with fibrosis as determined by collagen content. Thus, in extremely obese individuals, adipocyte telomere length is a marker of adiposity, whereas whole adipose tissue telomere length reflects the extent of fibrosis and may indicate adipose tissue dysfunction.


Population Pharmacokinetics of Fluconazole in Premature Infants with Birth Weights Less than 750 Grams.
Fluconazole is an effective agent for prophylaxis of invasive candidiasis in premature infants. The objective of this study was to characterize the population pharmacokinetics (PK) and dosing requirements of fluconazole in infants with birth weights of <750 g. As part of a randomized clinical trial, infants born at <750 g birth weight received intravenous (i.v.) or oral fluconazole at 6 mg/kg of body weight twice weekly. Fluconazole plasma concentrations from samples obtained by either scheduled or scavenged sampling were measured using a liquid chromatography-tandem mass spectrometry assay. Population PK analysis was conducted using NONMEM 7.2. Population PK parameters were allometrically scaled by body weight. Covariates were evaluated by univariable screening followed by multivariable assessment. Fluconazole exposures were simulated in premature infants using the final PK model. A population PK model was developed from 141 infants using 604 plasma samples. Plasma fluconazole PK were best described by a one-compartment model with first-order elimination. Only serum creatinine was an independent predictor for clearance in the final model. The typical population parameter estimate for oral bioavailability in the final model was 99.5%. Scavenged samples did not bias the parameter estimates and were as informative as scheduled samples. Simulations indicated that the study dose maintained fluconazole troughs of >2,000 ng/ml in 80% of simulated infants at week 1 and 59% at week 4 of treatment. Developmental changes in fluconazole clearance are best predicted by serum creatinine in this population. A twice-weekly dose of 6 mg/kg achieves appropriate levels for prevention of invasive candidiasis in extremely premature infants.


The effectiveness and safety of computed tomographic peritoneography and video-assisted thoracic surgery for hydrothorax in peritoneal dialysis patients: A retrospective cohort study in Japan.
Albeit uncommon, hydrothorax is an important complication of peritoneal dialysis (PD). Due to paucity of evidence for optimal treatment, this study aimed to evaluate the effectiveness and safety of computed tomographic (CT) peritoneography and surgical intervention involving video-assisted thoracic surgery (VATS) for hydrothorax in a retrospective cohort of patients who underwent PD in Japan. Of the 982 patients who underwent PD from six centers in Japan between 2007 and 2019, 25 (2.5%) with diagnosed hydrothorax were enrolled in this study. PD withdrawal rates were compared between patients who underwent VATS for diaphragm repair (surgical group) and those who did not (non-surgical group) using the Kaplan-Meier method and log-rank test. The surgical and non-surgical groups comprised a total of 11 (44%) and 14 (56%) patients, respectively. Following hydrothorax diagnosis by thoracentesis and detection of penetrated sites on the diaphragm using CT peritoneography, VATS was performed at a median time of 31 days (interquartile range [IQR], 20-96 days). During follow-up (median, 26 months; IQR, 10-51 months), 9 (64.3%) and 2 (18.2%) patients in the non-surgical and surgical groups, respectively, withdrew from PD (P = 0.021). There were no surgery-related complications or hydrothorax relapse in the surgical group. This study demonstrated the effectiveness and safety of CT peritoneography and VATS for hydrothorax. This approach may be useful in hydrothorax cases to avoid early drop out of PD and continue PD in the long term. Further studies are warranted to confirm these results.


Efficacy of irradiation and external hyperthermia in locally advanced, hormone-refractory or radiation recurrent prostate cancer: a preliminary report.
To present a preliminary report on the feasibility, efficacy, and toxicity of irradiation (RT) and hyperthermia (HT) in patients with locally advanced, hormone-refractory prostate cancer (LAHRPC) who may or may not have received prior RT. Between 1997 and 2002, 13 consecutive patients with LAHRPC or RT-recurrent prostate cancer were treated with RT and HT on a Phase I-II protocol. Eight patients had RT-recurrent LAHRPC (Group A) and 5 had LAHRPC without prior RT (Group B). All patients had large and clinically symptomatic tumors. The median RT dose was 39.6 Gy and 66.6 Gy in Groups A and B, respectively. External deep HT was delivered using a BSD-2000 Sigma-60 applicator. The median number of HT treatments was 8 in group A and 10 in group B. The median follow-up was 14 and 13 months for Groups A and B, respectively. All patients achieved a complete or partial response (CR/PR) and complete palliation of symptoms. Eleven patients had follow-up CT scans that demonstrated a CR in six and a PR in five. Two patients, who died of metastasis, did not have CT scans and had a PR on digital rectal examination. Two patients demonstrated a biochemical CR. The median duration of the CR/PR among Group A patients was 12 months after therapy. Three patients in Group A developed tumor recurrence at 9, 17, and 27 months after repeat RT to doses of 39.6, 36, and 50 Gy, respectively. At last follow-up, no Group B patient developed local recurrence. Grade 1-2 rectal bleeding was noted in 3 patients. RT and HT were generally well tolerated by all patients who had not previously undergone RT. Of the 8 patients who had, 6 (75%) tolerated retreatment well with minimal or no complications. Two patients in the repeat RT group had severe complications. One patient with lymphoma and factor XI deficiency developed Grade 4 hemorrhagic cystitis. Another previously irradiated patient developed a rectovesical fistula 4 months after retreatment, after disappearance of a large, invasive, and necrotic tumor. This preliminary report demonstrates the feasibility and efficacy of RT and HT in patients with LAHRPC, who may or may not have received prior RT. Presently, such patients who have undergone previous RT have no effective treatment options. RT and HT were generally well tolerated by patients who were not previously undergone RT. Of those who had been, most (6 of 8) tolerated retreatment well with minimal or no complications. The high-risk factors for treatment- and tumor regression-related side effects include the presence of large necrotic tumors, previous RT with a large dose/fraction, and the presence of bleeding disorders. Despite the size of these large tumors, RT and HT resulted in significant tumor shrinkage, rapid serum prostate-specific antigen decline, durable treatment responses, and durable palliation of symptoms. Additional clinical studies are warranted.


A cohort study of the relationship of diabetic retinopathy to blood pressure.
To determine whether blood pressure at baseline was associated with incidence or progression of diabetic retinopathy during a 10-year interval in a population-based cohort. A probability sample of all persons receiving primary care for diabetes in an 11-county area of southern Wisconsin were invited to participate in the study examinations in 1980 to 1982, 1984 to 1986, and 1990 to 1992. Blood pressure measurements, height, weight, ocular photographs, and glycosylated hemoglobin measurements were obtained and ocular examinations and a medical history interview were performed at each evaluation. Fundus photographs of seven standard photographic fields were obtained and graded according to the modified Airlie House Classification scheme. End points were incidence, any progression, or progression to proliferative diabetic retinopathy. At the baseline examination, 996 subjects were determined to have younger-onset diabetes and 1370 subjects had older-onset diabetes compared with 891 and 987, respectively, at the 4-year follow-up examination and 765 and 533, respectively, at the 10-year follow-up examination. In discrete linear logistic analyses, a 10-mm Hg increase in systolic blood pressure was significantly associated with incidence of retinopathy in subjects with younger-onset diabetes 10 years after the baseline examination (odds ratio, 1.27; 95% confidence interval, 1.03 to 1.57). No consistent association of blood pressure and retinopathy in subjects with older-onset diabetes was observed. Mortality did not affect the relationship of blood pressure and retinopathy. Increased systolic blood pressure at baseline was associated with a modest subsequent increased risk of incidence of diabetic retinopathy in subjects with younger-onset diabetes.


Survival and cure of acute myeloid leukaemia in England, 1971-2006: a population-based study.
The 5-year relative survival of adults diagnosed with acute myeloid leukaemia (AML) was less than 10% during the 1970s and 1980s in England. This population-based study estimated the 5-year relative survival and 'cure' for 48 380 adult patients diagnosed with AML in England during 1971-2006. Relative survival and cure mixture models were used to produce estimates of 5-year relative survival and the percentage 'cured'. 'Cure' was defined as the proportion of a group of survivors for whom there is no excess mortality compared with the general population. The 5-year relative survival and the percentage 'cured' increased for patients aged under 70 years at diagnosis during 1971-2006, but advancing age was associated with poorer outcome. During the study period a dramatic increase in 5-year relative survival occurred in those aged 15-24 years, from 7% to 53%. The percentage 'cured' was less than 10% for all ages in 1975, but increased to 45% for those aged 15-24 years in 2000. Cure could not be estimated for patients over 70 years, because survival was consistently low (<5%). The long-term outcome of patients with AML has improved substantially, particularly in younger patients. The potential exists for further increasing levels of 'cure'.


The Effect of Helicobacter pylori Eradication on the Gastrointestinal Microbiota in Patients with Duodenal Ulcer.
Recent reports have indicated that Helicobacter pylori (H. pylori) might have an effect on gastrointestinal flora; moreover, gastric commensual bacteria have been observed in the development of duodenal ulcer (DU). In our study, we aimed to evaluate the effect of H. pylori eradication on gastrointestinal flora in DU patients. A case-control study was performed at Jiangsu Shengze Hospital between December, 2013 and April, 2014. The patients received antibiotic eradication therapy if H. pylori testing was positive. At least four weeks after cessation of the eradication therapy, a repeat gastroscopy was performed to collect biopsies again in the same position. Gastric mucosa samples and feces specimens were collected to extract bacteria DNA and then to quantify by real-time polymerase chain reaction (PCR). After the eradication of H. pylori, an increase of Lactobacillus group, Clostridium leptum subgroup, Enterobacteria and a decrease of Clostridium coccoides subgroup were found in the antrum. In the corpus, the number of bacteria in the Lactobacillus group was increased and the expression of Clostridium coccoides subgroup was significantly down-regulated. In the feces samples, only the number in the Lactobacillus group was increased. Moreover, the distribution was significantly different between female and the male patients. The presence of H. pylori in the stomach suppressed the colonization with Lactobacillus group, Clostridium leptum subgroup and Enterobacteria. Gender might affect the distribution and/or recolonization of the bacteria in DU patients.


Survival after liver transplantation for hepatocellular carcinoma.
Selection criteria for patients with hepatocellular carcinoma (HCC) suitable for liver transplantation (LT) include tumor size and number and vascular invasion. There has been a recent trend to expand the transplant criteria for HCC. We reviewed our experience to determine survival following LT based on tumor characteristics. A retrospective analysis was performed on 72 patients with HCC who underwent LT between 1985 and July 2002. The Milan criteria were applied for LT candidacy for HCCs that were deemed unresectable from anatomical considerations and/or the severity of underlying cirrhosis. Patients were divided into four groups: group 1: patients with known HCC who satisfied the selection criteria (n = 22); group 2: patients with known HCC that exceeded the criteria (n = 17); group 3: patients with incidental HCC found at pathological examination of the explant (n = 33); group 4: contemporary LT recipients without HCC (n = 935). In the known HCC group, the interval between listing as status 2 and transplantation was 72.2 +/- 133.6 days (median 23 days). Three-year patient survival was 80.2% in group 1, 35.8% in group 2, 63.2% in group 3, and 81.5% in group 4. In group 2 patients, the tumors were significantly larger, had more nodules, and were more often bilobar. In group 3, five (15%) exceeded the criteria mainly because of tumor size and four patients died within 3 years post-LT (three from tumor recurrence). Liver transplantation for HCC yields acceptable survival in early-stage tumors, particularly if transplanted soon after listing. Long-term survival was inferior in patients with multiple tumors and tumors that were greater than 5 cm in diameter.


Stumbling toward equity: the role of government in kidney transplantation.
In Mortal Peril, Professor Epstein is critical of the current, regulated system for organ donation and suggests that a market for organ tissue would better meet the needs of patients. In this response to Professor Epstein, Professor Laura Dooley and Dr. Robert Gaston pair their skills to attack Professor Epstein's analysis. As they have done on several other occasions, Professors Dooley and Gaston argue that the kidney donation and transplantation arena is fraught with racial inequity, and that Professor Epstein's proposal for a market in kidneys will exacerbate this inequity. The authors maintain that to prevent the poor from being excluded from transplants, the government plays a critical (if imperfect) role in the allocation of these scarce resources. Furthermore, government intervention is acceptable to correct past discrimination because there is scientific evidence that the disproportionate incidence of kidney failure in African Americans is related to the evolutionary pressures of slave trading and slavery. Professors Dooley and Gaston also defend their previous efforts to change the government system of allocation and characterize the government's willingness to adopt their recommendations as an appropriate response to scientific research rather than a governmental susceptibility to lobbying from special interest groups. Finally, the authors criticize Professor Epstein's argument that dialysis is a viable alternative to transplantation because there are significant differences in "quality of life, morbidity and survival." Professors Dooley and Gaston conclude that government intervention is necessary for maintaining the equity in kidney transplantation that a market system would not.


Modeling personal particle-bound polycyclic aromatic hydrocarbon (pb-pah) exposure in human subjects in Southern California.
Exposure to polycyclic aromatic hydrocarbon (PAH) has been linked to various adverse health outcomes. Personal PAH exposures are usually measured by personal monitoring or biomarkers, which are costly and impractical for a large population. Modeling is a cost-effective alternative to characterize personal PAH exposure although challenges exist because the PAH exposure can be highly variable between locations and individuals in non-occupational settings. In this study we developed models to estimate personal inhalation exposures to particle-bound PAH (PB-PAH) using data from global positioning system (GPS) time-activity tracking data, traffic activity, and questionnaire information. We conducted real-time (1-min interval) personal PB-PAH exposure sampling coupled with GPS tracking in 28 non-smoking women for one to three sessions and one to nine days each session from August 2009 to November 2010 in Los Angeles and Orange Counties, California. Each subject filled out a baseline questionnaire and environmental and behavior questionnaires on their typical activities in the previous three months. A validated model was used to classify major time-activity patterns (indoor, in-vehicle, and other) based on the raw GPS data. Multiple-linear regression and mixed effect models were developed to estimate averaged daily and subject-level PB-PAH exposures. The covariates we examined included day of week and time of day, GPS-based time-activity and GPS speed, traffic- and roadway-related parameters, meteorological variables (i.e. temperature, wind speed, relative humidity), and socio-demographic variables and occupational exposures from the questionnaire. We measured personal PB-PAH exposures for 180 days with more than 6 h of valid data on each day. The adjusted R2 of the model was 0.58 for personal daily exposures, 0.61 for subject-level personal exposures, and 0.75 for subject-level micro-environmental exposures. The amount of time in vehicle (averaging 4.5% of total sampling time) explained 48% of the variance in daily personal PB-PAH exposure and 39% of the variance in subject-level exposure. The other major predictors of PB-PAH exposures included length-weighted traffic count, work-related exposures, and percent of weekday time. We successfully developed regression models to estimate PB-PAH exposures based on GPS-tracking data, traffic data, and simple questionnaire information. Time in vehicle was the most important determinant of personal PB-PAH exposure in this population. We demonstrated the importance of coupling real-time exposure measures with GPS time-activity tracking in personal air pollution exposure assessment.


Debunking Cannabidiol as a Treatment for COVID-19: Time for the FDA to Adopt a Focused Deterrence Model?
Many cannabidiol (CBD) retailers make unsupported medical claims about their product. In recent years, the U.S. Food and Drug Administration (FDA) has sent warning letters to CBD retailers who promoted CBD to treat Alzheimer's disease, cancer, diabetes, and other serious conditions for which there is no evidence of its efficacy as a treatment or preventive. Compliance with these warning letters has been low. During the novel coronavirus disease 2019 (COVID-19) pandemic, the FDA has begun sending more strongly worded warning letters that appear to have better compliance in that most of these companies have removed COVID-19-related claims. However, many continue to present other unsupported medical claims on other serious medical conditions like cancer, depression, addiction, and bone fractures, among many others. We argue that adopting a strategy of focused deterrence where the FDA prioritizes enforcement related to COVID-19 claims - but when COVID-19-related claims are found, pursues all other violations by that company - would present an opportunity to efficiently cut down on harmful claims overstating CBD's benefits.


Pediatric issues in disaster management, part 2: evacuation centers and family separation/reunification.
Although children and infants are likely to be victims in a disaster and are more vulnerable in a disaster than adults, disaster planning and management has often overlooked the specific needs of pediatric patients. We discuss key components of disaster planning and management for pediatric patients including emergency medical services, hospital/facility issues, evacuation centers, family separation/reunification, children with special healthcare needs, mental health issues, and overcrowding/surge capacity. Specific policy recommendations and an appendix with detailed practical information and algorithms are included. The first part of this three part series on pediatric issues in disaster management addresses the emergency medical system from the field to the hospital and surge capacity including the impact of crowding. The second part addresses the appropriate set up and functioning of evacuation centers and family separation and reunification. The third part deals with special patient populations: the special healthcare needs patient and mental health issues.


Complexity in transcription control at the activation domain-mediator interface.
Transcript elongation by polymerase II paused at the Egr1 promoter is activated by mitogen-activated protein kinase phosphorylation of the ternary complex factor (TCF) ELK1 bound at multiple upstream sites and subsequent phospho-ELK1 interaction with mediator through the MED23 subunit. Consequently, Med23 knockout (KO) nearly eliminates Egr1 (early growth response factor 1) transcription in embryonic stem (ES) cells, leaving a paused polymerase at the promoter. Med23 KO did not, however, eliminate Egr1 transcription in fibroblasts. Chromatin immunoprecipitation analysis and direct visualization of fluorescently labeled TCF derivatives and mediator subunits revealed that three closely related TCFs bound to the same control regions. The relative amounts of these TCFs, which responded differently to the loss of MED23, differed in ES cells and fibroblasts. Transcriptome analysis suggests that most genes expressed in both cell types, such as Egr1, are regulated by alternative transcription factors in the two cell types that respond differently to the same signal transduction pathways.


Off-Label and Unlicensed Drug Use in Neonatology: Reality in a Portuguese University Hospital.
This study aims to characterize the drugs used in a Portuguese Neonatal Intensive Unit Care, assess the rate of off-label or unlicensed drugs use according to the information available in the Summary of Product Characteristics and compare results between preterm and full-term neonates. A 6-month period retrospective cross-sectional study was conducted in our Neonatal Intensive Unit Care in 2013. Prescribed drugs data were recorded and compared with the paediatric information contained in drugs Summary of Product Characteristics. We analyzed 1011 prescriptions of 84 active substances, made in 218 admissions. In 42.9% of the cases, medicines were used according to Summary of Product Characteristics information; 27.9% of drugs were approved for neonatal period but used in an off-label manner; off-label drugs for neonates were used in 10.1%, whereas those with undetermined approval state and contraindicated were used 6.0% and 8.7% of the cases, respectively. Unlicensed prescriptions accounted for 4.4% of total. Preterm received a higher rate of drugs used according to Summary of Product Characteristics (p < 0.0001), whereas full-term received more off-label drugs for dose/frequency (p < 0.0001) and contra-indicated for neonates (p < 0.012). Preterm neonates received a higher median number of drugs, since they stayed longer in the unit. The main reason for off-label prescribing was the use of doses/frequencies of administration different from those stated in the Summary of Product Characteristics, suggesting that updating these documents is necessary. Manipulation of medicines is one of the causes for unlicensed drugs use, emphasizing the lack of appropriate formulations for neonatal age. Progresses have been made to reduce the risks of off-label/unlicensed prescriptions, but competent authorities must continue their efforts to develop safer and more effective drugs for neonatal period.


Unc-51-like kinase 1/2-mediated endocytic processes regulate filopodia extension and branching of sensory axons.
The molecular mechanism and significance of endocytic processes involved in directional axon elongation are not well understood. The Unc-51 family of serine/threonine kinases was shown to be important for axon growth and was also linked to endocytosis, providing an entry point to study this problem. We found that mouse Unc-51-like kinase 1/2 (Ulk1/2) proteins are localized to vesicular structures in growth cones of mouse spinal sensory neurons. RNAi-mediated knockdown of Ulk1 and/or Ulk2 resulted in impaired endocytosis of nerve growth factor (NGF), excessive axon arborization, and severely stunted axon elongation. The evidence also indicates that Ulk1/2 mediates a non-clathrin-coated endocytosis in sensory growth cones. Interestingly, NGF can induce the interaction of Ulk1 with TrkA receptor complexes through promoting K63-polyubiquitination of Ulk1 and binding of Ulk1 to the scaffolding protein p62. These results and additional studies suggest that Ulk1/2 proteins regulate filopodia extension and neurite branching during sensory axon outgrowth, probably through regulating TrkA receptor trafficking and signaling.


Optimisation of AAV-NDI1 Significantly Enhances Its Therapeutic Value for Correcting Retinal Mitochondrial Dysfunction.
AAV gene therapy for ocular disease has become a reality with the market authorisation of LuxturnaTM for RPE65-linked inherited retinal degenerations and many AAV gene therapies currently undergoing phase III clinical trials. Many ocular disorders have a mitochondrial involvement from primary mitochondrial disorders such as Leber hereditary optic neuropathy (LHON), predominantly due to mutations in genes encoding subunits of complex I, to Mendelian and multifactorial ocular conditions such as dominant optic atrophy, glaucoma and age-related macular degeneration. In this study, we have optimised the nuclear yeast gene, NADH-quinone oxidoreductase (NDI1), which encodes a single subunit complex I equivalent, creating a candidate gene therapy to improve mitochondrial function, independent of the genetic mutation driving disease. Optimisation of NDI1 (ophNdi1) substantially increased expression in vivo, protected RGCs and increased visual function, as assessed by optokinetic and photonegative response, in a rotenone-induced murine model. In addition, ophNdi1 increased cellular oxidative phosphorylation and ATP production and protected cells from rotenone insult to a significantly greater extent than wild type NDI1. Significantly, ophNdi1 treatment of complex I deficient patient-derived fibroblasts increased oxygen consumption and ATP production rates, demonstrating the potential of ophNdi1 as a candidate therapy for ocular disorders where mitochondrial deficits comprise an important feature.


A map of direct TF-DNA interactions in the human genome.
Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is the most popular assay to identify genomic regions, called ChIP-seq peaks, that are bound in vivo by transcription factors (TFs). These regions are derived from direct TF-DNA interactions, indirect binding of the TF to the DNA (through a co-binding partner), nonspecific binding to the DNA, and noise/bias/artifacts. Delineating the bona fide direct TF-DNA interactions within the ChIP-seq peaks remains challenging. We developed a dedicated software, ChIP-eat, that combines computational TF binding models and ChIP-seq peaks to automatically predict direct TF-DNA interactions. Our work culminated with predicted interactions covering >4% of the human genome, obtained by uniformly processing 1983 ChIP-seq peak data sets from the ReMap database for 232 unique TFs. The predictions were a posteriori assessed using protein binding microarray and ChIP-exo data, and were predominantly found in high quality ChIP-seq peaks. The set of predicted direct TF-DNA interactions suggested that high-occupancy target regions are likely not derived from direct binding of the TFs to the DNA. Our predictions derived co-binding TFs supported by protein-protein interaction data and defined cis-regulatory modules enriched for disease- and trait-associated SNPs. We provide this collection of direct TF-DNA interactions and cis-regulatory modules through the UniBind web-interface (http://unibind.uio.no).