Evolution of vertebrate forebrain development: how many different mechanisms?
Over the past 50 years and more, many models have been proposed to explain how the nervous system is initially induced and how it becomes subdivided into gross regions such as forebrain, midbrain, hindbrain and spinal cord. Among these models is the 2-signal model of Nieuwkoop & Nigtevecht (1954), who suggested that an initial signal ('activation') from the organiser both neuralises and specifies the forebrain, while later signals ('transformation') from the same region progressively caudalise portions of this initial territory. An opposing idea emerged from the work of Otto Mangold (1933) and other members of the Spemann laboratory: 2 or more distinct organisers, emitting different signals, were proposed to be responsible for inducing the head, trunk and tail regions. Since then, evidence has accumulated that supports one or the other model, but it has been very difficult to distinguish between them. Recently, a considerable body of work from mouse embryos has been interpreted as favouring the latter model, and as suggesting that a 'head organiser', required for the induction of the forebrain, is spatially separate from the classic organiser (Hensen's node). An extraembryonic tissue, the 'anterior visceral endoderm' (AVE), was proposed to be the source of forebrain-inducing signals. It is difficult to find tissues that are directly equivalent embryologically or functionally to the AVE in other vertebrates, which led some (e.g. Kessel, 1998) to propose that mammals have evolved a new way of patterning the head. We will present evidence from the chick embryo showing that the hypoblast is embryologically and functionally equivalent to the mouse AVE. Like the latter, the hypoblast also plays a role in head development. However, it does not act like a true organiser. It induces pre-neural and pre-forebrain markers, but only transiently. Further development of neural and forebrain phenotypes requires additional signals not provided by the hypoblast. In addition, the hypoblast plays a role in directing cell movements in the adjacent epiblast. These movements distance the future forebrain territory from the developing organiser (Hensen's node), and we suggest that this is a mechanism to protect the forebrain from caudalising signals from the node. These mechanisms are consistent with all the findings obtained from the mouse to date. We conclude that the mechanisms responsible for setting up the forebrain and more caudal regions of the nervous system are probably similar among different classes of higher vertebrates. Moreover, while reconciling the two main models, our findings provide stronger support for Nieuwkoop's ideas than for the concept of multiple organisers, each inducing a distinct region of the CNS.


A review for the neuroprotective effects of andrographolide in the central nervous system.
Andrographolide is compound extracted from Andrographis paniculata (A. paniculata), a traditional herb that has been used in ancient China and other parts of eastern Asia to treat an array of disorders, such as cancer, rheumatoid arthritis, diarrhea, upper respiratory tract infection, and laryngitis, for a very long history. The mechanisms of action of andrographolide in disease prevention and/or therapy include anti-inflammation, anti-oxidative stress, anti-apoptosis, and/or pro-apoptosis. Pharmacodynamic studies have shown that andrographolide can cross the blood brain barrier and distribute into different brain regions, and therefore its pharmacological effects in the central nervous system (CNS) have begun to be revealed in recent years. For example, andrographolide has been reported to reduce brain infarct volume in several models of cerebral ischemia. In models of Alzheimer's disease (AD), andrographolide not only reduces Aβ aggregation, but suppresses neuroinflammatory response and synaptic dysfunction, which could be evidenced by the reversal of microglia-mediated production of pro-inflammatory cytokines as well as AD-associated decreases in synaptic proteins, such as postsynaptic membrane dense substance-95. Andrographolide may also inhibit the onset and/or progression of Parkinson's disease, multiple sclerosis, and surgery- or diabetes-induced cognitive impairment. Further, andrographolide has been shown to inhibit chronic stress-induced abnormalities in serum corticosterone levels, mood-associated behavior, and hippocampal neurogenesis, suggesting that andrographolide may have a potential to treat psychiatric disorders, such as anxiety and depression. In this review, we summarize and discuss the pharmacological effects of andrographolide in the CNS in hope of revealing more possibilities of andrographolide in disease prevention and/or therapy.


Quantum spin Hall effect in inverted type-II semiconductors.
The quantum spin Hall (QSH) state is a topologically nontrivial state of quantum matter which preserves time-reversal symmetry; it has an energy gap in the bulk, but topologically robust gapless states at the edge. Recently, this novel effect has been predicted and observed in HgTe quantum wells and in this Letter we predict a similar effect arising in Type-II semiconductor quantum wells made from InAs/GaSb/AlSb. The quantum well exhibits an "inverted" phase similar to HgTe/CdTe quantum wells, which is a QSH state when the Fermi level lies inside the gap. Due to the asymmetric structure of this quantum well, the effects of inversion symmetry breaking are essential. Remarkably, the topological quantum phase transition between the conventional insulating state and the quantum spin Hall state can be continuously tuned by the gate voltage, enabling quantitative investigation of this novel phase transition.


HIV surveillance by testing saliva.
Saliva specimens were tested for HIV antibody (anti-HIV) by an immunoglobulin G (IgG) antibody capture radioimmunoassay (GACRIA) and three sensitive commercial assays. In tests on 460 seronegative subjects and 196 seropositive subjects GACRIA was 99.8% specific and 100% sensitive. The Wellcome HIV monoclonal and Abbott recombinant DNA enzyme-linked immunosorbent assays (ELISAs) were also highly specific (99.8%, 100%) but they were less sensitive (90.9%, 82.0%). The Fujirebio particle agglutination assay was sensitive (97.8%) but its specificity was poor (84.1%). In testing saliva specimens from populations with an anti-HIV prevalence greater than 0.5%, sampling by GACRIA alone could provide a good estimate of the true prevalence. For true prevalences less than 0.5% good estimates could only be obtained if positive GACRIA reactions were confirmed by another independent salivary assay. Salivary testing for anti HIV is a convenient and potentially an accurate epidemiological tool.


Aberrant phosphorylation of human LRH1 at serine 510 is predictable of hepatocellular carcinoma recurrence.
We previously identified the AKT-phosphorylation sites in nuclear receptors and showed that phosphorylation of S379 in mouse retinoic acid γ and S518 in human estrogen receptor α regulate their activity independently of the ligands. Since this site is conserved at S510 in human liver receptor homolog 1 (hLRH1), we developed a monoclonal antibody (mAb) that recognized the phosphorylation form of hLRH1S510 (hLRH1pS510) and verified its clinicopathological significance in hepatocellular carcinoma (HCC). We generated the anti-hLRH1pS510 mAb and assessed its selectivity. We then evaluated the hLRH1pS510 signals in 157 cases of HCC tissues by immunohistochemistry because LRH1 contributes to the pathogenesis of diverse cancers. The developed mAb specifically recognized hLRH1pS510 and worked for immunohistochemistry of formalin-fixed paraffin-embedded tissues. hLRH1pS510 was exclusively localized in the nucleus of HCC cells, but the signal intensity and positive rates varied among the subjects. According to the semi-quantification, 45 cases (34.9%) showed hLRH1pS510-high, and the remaining 112 cases (65.1%) exhibited hLRH1pS510-low. There were significant differences in the recurrence-free survival (RFS) between the two groups, and the 5-year RFS rates in the hLRH1pS510-high and hLRH1pS510-low groups were 26.5% and 46.1%, respectively. In addition, high hLRH1pS510 was significantly correlated with portal vein invasion, hepatic vein invasion, and high levels of serum alpha-fetoprotein (AFP). Furthermore, multivariable analysis revealed that hLRH1pS510-high was an independent biomarker for HCC recurrence. We conclude that aberrant phosphorylation of hLRH1S510 is a predictor of poor prognosis for HCC. The anti-hLRH1pS510 mAb could provide a powerful tool to validate the relevance of hLRH1pS510 in pathological processes such as tumor development and progression.


A comparison of clinical judgment vs the modified Alvarado score in acute appendicitis.
To investigate the value of the modified Alvarado score (MAS) in helping Accident & Emergency (A&E) doctors decide which patients with suspected acute appendicitis need surgical referral. 11,258 patients presented to a University Hospital A&E Department over a two-month period; 82 were triaged as 'abdominal pain' or 'suspected appendicitis'. Ten patients self-discharged prior to seeing a doctor. The remaining case notes (72) were reviewed and MAS's calculated. The Alvarado guidelines suggested an MAS > or = 5 (high) needed admission and an MAS < 5 (low) excluded appendicitis and was appropriate for discharge. Two patients had proven pancreatitis and were excluded. 24/70 patients were admitted for suspected appendicitis; all were referred by the A&E doctor (sensitivity 100%) but only 12 had a high MAS (sensitivity 50%). Twelve patients were therefore admitted despite having a low MAS on retrospective analysis. 46/70 patients were discharged (none re-presented with the same complaint) of which 40/46 were sent home without surgical referral (specificity 87%), but only 44/46 patients discharged had a low MAS (specificity 96%). It is more important to refer every case that needs referral (sensitivity) than to discharge those not needing referral (specificity). We cannot exclude the possibility that morbidity would result were the MAS used in lieu of clinical judgment in deciding whether referral is necessary in cases of suspected acute appendicitis.


Developmental activation of the lysozyme gene in chicken macrophage cells is linked to core histone acetylation at its enhancer elements.
Native chromatin IP assays were used to define changes in core histone acetylation at the lysozyme locus during developmental maturation of chicken macrophages and stimulation to high-level expression by lipo-polysaccharide. In pluripotent precursors the lysozyme gene (Lys) is inactive and there is no acetylation of core histones at the gene, its promoter or at the upstream cis-control elements. In myeloblasts, where there is a very low level of Lys expression, H4 acetylation appears at the cis-control elements but not at the Lys gene or its promoter: neither H3 nor H2B become significantly acetylated in myeloblasts. In mature macrophages, Lys expression increases 5-fold: H4, H2B and H2A.Z are all acetylated at the cis-control elements but H3 remains unacetylated except at the -2.4 S silencer. Stimulation with LPS increases Lys expression a further 10-fold: this is accompanied by a rise in H3 acetylation throughout the cis-control elements; H4 and H2B acetylation remain substantial but acetylation at the Lys gene and its promoter remains low. Acetylation is thus concentrated at the cis-control elements, not at the Lys gene or its immediate promoter. H4 acetylation precedes H3 acetylation during development and H3 acetylation is most directly linked to high-level Lys expression.


Enhanced resting-state oscillations in schizophrenia are associated with decreased synchronization during inattentional blindness.
Patients suffering from schizophrenia have been characterized by an apparent lack of theta (around 6 Hz) and gamma (>40 Hz) brain oscillatory activity during task execution. The neurocognitive reasons for these abnormal synchronization patterns, however, remain elusive. Recording the electroencephalogramm (EEG) during a selective visual attention task, the current study investigates whether abnormal brain oscillatory resting-state activity in the theta band might account for a lack of task-related brain oscillatory activity in schizophrenia. EEGs were recorded from 26 patients with schizophrenia and 26 healthy matched controls during rest and during the execution of a selective visual attention task, in which an unexpected object (monkey) appeared on the screen. On a behavioral level, patients were less likely to report perceiving the unexpected event than controls. Controls showed a stronger increase in task-related theta power than patients in prefrontal, parietal, and occipital brain regions. Task-related theta power change differed between patients who perceived, and patients who did not perceive the unexpected event. Moreover, patients showed higher levels of theta power during rest than controls, whereas the absolute theta power values during the selective attention task did not differ between groups. These results suggest that the failure to increase oscillatory activity during a cognitive task can be accounted for by abnormally high oscillatory activity in a resting state. This finding has important implications for future studies examining abnormal brain oscillatory activity in schizophrenia, which usually treat resting-state activity as a baseline for task-related activity.


The RNA binding and transport proteins staufen and fragile X mental retardation protein are expressed by rat primary afferent neurons and localize to peripheral and central axons.
Neuronal proteins have been traditionally viewed as being derived solely from the soma; however, accumulating evidence indicates that dendritic and axonal sites are capable of a more autonomous role in terms of new protein synthesis. Such extra-somal translation allows for more rapid, on-demand regulation of neuronal structure and function than would otherwise be possible. While mechanisms of dendritic RNA transport have been elucidated, it remains unclear how RNA is trafficked into the axon for this purpose. Primary afferent neurons of the dorsal root (DRG) and trigeminal (TG) ganglia have among the longest axons in the neuraxis and such axonal protein synthesis would be advantageous, given the greater time involved for protein trafficking to occur via axonal transport. Therefore, we hypothesized that these primary sensory neurons might express proteins involved in RNA transport. Rat DRG and TG neurons expressed staufen (stau) 1 and 2 (detected at the mRNA level) and stau2 and fragile x mental retardation protein (FMRP; detected at the protein level). Stau2 mRNA was also detected in human TG neurons. Stau2 and FMRP protein were localized to the sciatic nerve and dorsal roots by immunohistochemistry and to dorsal roots by Western blot. Stau2 and FMRP immunoreactivities colocalized with transient receptor potential channel type 1 immunoreactivity in sensory axons of the sciatic nerve and dorsal root, suggesting that these proteins are being transported into the peripheral and central terminals of nociceptive sensory axons. Based on these findings, we propose that stau2 and FMRP proteins are attractive candidates to subserve RNA transport in sensory neurons, linking somal transcriptional events to axonal translation.


Inhaled nitric oxide improves outcomes after successful cardiopulmonary resuscitation in mice.
Sudden cardiac arrest (CA) is a leading cause of death worldwide. Breathing nitric oxide (NO) reduces ischemia/reperfusion injury in animal models and in patients. The objective of this study was to learn whether inhaled NO improves outcomes after CA and cardiopulmonary resuscitation (CPR). Adult male mice were subjected to potassium-induced CA for 7.5 minutes whereupon CPR was performed with chest compression and mechanical ventilation. One hour after CPR, mice were extubated and breathed air alone or air supplemented with 40 ppm NO for 23 hours. Mice that were subjected to CA/CPR and breathed air exhibited a poor 10-day survival rate (4 of 13), depressed neurological and left ventricular function, and increased caspase-3 activation and inflammatory cytokine induction in the brain. Magnetic resonance imaging revealed brain regions with marked water diffusion abnormality 24 hours after CA/CPR in mice that breathed air. Breathing air supplemented with NO for 23 hours starting 1 hour after CPR attenuated neurological and left ventricular dysfunction 4 days after CA/CPR and markedly improved 10-day survival rate (11 of 13; P=0.003 versus mice breathing air). The protective effects of inhaled NO on the outcome after CA/CPR were associated with reduced water diffusion abnormality, caspase-3 activation, and cytokine induction in the brain and increased serum nitrate/nitrite levels. Deficiency of the α1 subunit of soluble guanylate cyclase, a primary target of NO, abrogated the ability of inhaled NO to improve outcomes after CA/CPR. These results suggest that NO inhalation after CA and successful CPR improves outcome via soluble guanylate cyclase-dependent mechanisms.


Osteopontin enhances endogenous repair after neonatal hypoxic-ischemic brain injury.
Hypoxic-ischemic (HI) brain injury is a frequent cause of perinatal morbidity and mortality with limited therapeutic options. To identify molecules important for cerebral damage and repair, we investigated the growth factor-related gene expression profile after neonatal cerebral HI. We identified osteopontin (OPN) as the most highly upregulated factor early after HI. We therefore explored the role of endogenous OPN in brain damage and repair. Nine-day-old wild-type mice were exposed to cerebral HI; growth factor-related gene expression profiles were analyzed 1 to 7 days later by reverse transcriptase-polymerase chain reaction arrays. To determine the contribution of OPN to brain damage, we used p9 OPN(-/-) and wild-type mice. HI brain damage, sensorimotor function, and cell proliferation and differentiation were compared. Gene expression profiling of 150 genes related to growth factors and neurotrophins showed that expression of 52 genes changed during the first 7 days after HI. OPN was the gene with the strongest increase expression at all time points measured. We show here for the first time that in response to neonatal HI, OPN-deficient mice developed increased gray and white matter loss and more pronounced sensorimotor deficits as compared with wild-type littermates. Furthermore, OPN deficiency decreases HI-induced cell proliferation/survival and oligodendrogenesis without affecting neuronal differentiation. OPN plays an important role in repairing brain injury after neonatal HI by regulating cerebral cell proliferation/survival and oligodendrocyte differentiation after injury. The observed promyelinative effect of OPN may offer novel possibilities for a therapy targeting white matter injury.


What is happening to the health of the Croatian population?
To describe the problems in the interpretation of Croatian mortality data and explore possible reasons for the recorded increase in mortality in the 1990-1999 period, particularly related to different methods of collection and estimation of data on deaths and population. Numbers of recorded deaths and population estimates were first obtained from the Croatian Institute for Public Health and examined in detail. The Institute used population estimates supplied by the Croatian Statistics Bureau, which included de jure population data (including all Croatian citizens wherever they live) until 1996 and de facto population data (including only population living in Croatia at least for a year, irrespective of citizenship) since 1996. A different set of population estimates based on de facto estimates since 1992 was obtained from the Croatian Bureau of Statistics. We examined trends in age- and sex-specific death rates from major causes in 1990-1999 period, using the mortality data from the Croatian Institute for Public Health and both sets of population estimates. Lung cancer as a cause of death was examined in more detail, since it is relatively stable over short periods of time. Interviews were undertaken with key informants to identify the reasons for any discrepancies. In Croatia, relatively stable death rates from lung cancer in men ranged from 84/100,000 in 1990 to 79/ 100,000 in 1995. In 1996, a marked discontinuity appeared in the Croatian data, with a 14% increase compared to 1995 (from 79/100,000 to 91/100,000) and a further increase in 1999 (94/100,000), which is not credible on the basis of the natural history of lung cancer. Analysis of mortality rates with de facto population estimates showed more gradual increase from 1992-1996. Methods used to estimate population and mortality during the 1990s were inconsistent and misleading. At present, it is impossible to be certain about the true level of mortality in Croatia during 1990s, as the numerator (deaths) and denominator (population) were incompatible until 1998. Major problems in data collection would have been identified if the investigation of unexpected mortality trends in Croatia in the 1990s had been done. Systematic analysis of health patterns should be done as soon as data from the 2001 census become available. Capacities in public health should be strengthened to make this possible. This issue has received little recognition from the international donor organizations, particularly those that use health data.


Influences of inbreeding and genetics on telomere length in mice.
We measured telomere lengths of blood leukocytes in several inbred and outbred mammalian species, using a telomere-specific fluorescent probe and flow cytometry. Humans, non-human primates, and three outbred populations of Peromyscus mice ( Peromyscus leucopus, Peromyscus maniculatus, and Peromyscus polionotus) have short telomeres. Two common strains of laboratory mice, C57BL/6J and DBA/2J, have telomeres several times longer than most other mammals surveyed. Moreover, the two inbred laboratory mouse strains display significantly different telomere lengths, suggesting the existence of strain-specific genetic determinants. To further examine the effects of inbreeding, we studied three Peromyscus leucopus inbred lines (GS109, GS16A1, and GS16B), all derived from the outbred P. leucopus stock. Telomeres of all three inbred lines are significantly lengthened relative to outbred P. leucopus, and the three lines display strain-specific significantly different telomere lengths, much like the C57BL/6J and DBA/2J strains of M. musculus. To further characterize the genetic inheritance of telomere length, we carried out several crosses to obtain hybrid F(1) mice between parental strains displaying the phenotype of long and short telomeres. In all F(1) mice assayed, peripheral blood leukocyte telomere length was intermediate to that of the parents. Additionally, we generated F(2) mice from a cross of the ( P. leucopus outbred x GS16B)F(1). Based on the distribution of telomere length in the F(2) population, we determined that more than five loci contribute to telomere length regulation in Peromyscus. We concluded that inbreeding, through unknown mechanisms, results in the elongation of telomeres, and that telomere length for a given species and/or sub-strain is genetically determined by multiple segregating loci.


Characterizing herpes simplex virus type 1 and type 2 seroprevalence declines and epidemiological association in the United States.
Assessing the epidemiological association between herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) infections in the United States, and characterizing the trends in the standardized HSV-1 and HSV-2 antibody prevalences (seroprevalences), 1999-2016. Source of data was the cross-sectional and nationally-representative biennial surveys of the National Health and Nutrition Examination Survey (NHANES). All nine NHANES rounds for 1999-2016 were included in analysis. Datasets of these rounds were combined and analyzed accounting for survey design and applying weighting procedures. Logistic regressions were used to identify associations with seropositivity. Sensitivity analyses were conducted. Odds of HSV-1 infection declined by 2.84% (95% CI: 1.70%-4.00%) annually among men, and by 2.22% (95% CI: 1.23%-3.21%) among women. Declines were highest at younger ages. Odds of HSV-2 infection declined by 2.23% (95% CI: 0.71%-3.82%) annually among men, and by 2.89% (95% CI: 1.57%-4.28%) among women. Odds ratio of the association between HSV-2 and HSV-1 seropositivity was 0.71 (95% CI: 0.60-0.84) for men and 0.81 (95% CI: 0.72-0.91) for women, after adjustment for age, ethnicity, and year. HSV-1 and HSV-2 seroprevalences showed a strong declining trend for at least two decades, for both sexes and for the different ethnicities, possibly reflecting improvements in hygiene and living conditions (for HSV-1), and safer sexual behavior (for HSV-2). HSV-1 seroprevalence declines are most pronounced among young individuals. There is evidence for cross protection between the two infections, suggestive of HSV-1 seropositivity being partially protective against HSV-2 infection.


African genetic diversity: implications for human demographic history, modern human origins, and complex disease mapping.
Comparative studies of ethnically diverse human populations, particularly in Africa, are important for reconstructing human evolutionary history and for understanding the genetic basis of phenotypic adaptation and complex disease. African populations are characterized by greater levels of genetic diversity, extensive population substructure, and less linkage disequilibrium (LD) among loci compared to non-African populations. Africans also possess a number of genetic adaptations that have evolved in response to diverse climates and diets, as well as exposure to infectious disease. This review summarizes patterns and the evolutionary origins of genetic diversity present in African populations, as well as their implications for the mapping of complex traits, including disease susceptibility.


Programmatically selected multidrug-resistant strains drive the emergence of extensively drug-resistant tuberculosis in South Africa.
South Africa shows one of the highest global burdens of multidrug-resistant (MDR) and extensively drug-resistant (XDR) tuberculosis (TB). Since 2002, MDR-TB in South Africa has been treated by a standardized combination therapy, which until 2010 included ofloxacin, kanamycin, ethionamide, ethambutol and pyrazinamide. Since 2010, ethambutol has been replaced by cycloserine or terizidone. The effect of standardized treatment on the acquisition of XDR-TB is not currently known. We genetically characterized a random sample of 4,667 patient isolates of drug-sensitive, MDR and XDR-TB cases collected from three South African provinces, namely, the Western Cape, Eastern Cape and KwaZulu-Natal. Drug resistance patterns of a subset of isolates were analyzed for the presence of commonly observed resistance mutations. Our analyses revealed a strong association between distinct strain genotypes and the emergence of XDR-TB in three neighbouring provinces of South Africa. Strains predominant in XDR-TB increased in proportion by more than 20-fold from drug-sensitive to XDR-TB and accounted for up to 95% of the XDR-TB cases. A high degree of clustering for drug resistance mutation patterns was detected. For example, the largest cluster of XDR-TB associated strains in the Eastern Cape, affecting more than 40% of all MDR patients in this province, harboured identical mutations concurrently conferring resistance to isoniazid, rifampicin, pyrazinamide, ethambutol, streptomycin, ethionamide, kanamycin, amikacin and capreomycin. XDR-TB associated genotypes in South Africa probably were programmatically selected as a result of the standard treatment regimen being ineffective in preventing their transmission. Our findings call for an immediate adaptation of standard treatment regimens for M/XDR-TB in South Africa.


A combined metabolomic and proteomic investigation of the effects of a failure to express dystrophin in the mouse heart.
Muscle degeneration in the heart of 1-9 month-old mdx mice (a model for Duchenne muscular dystrophy) has been monitored using metabolomic and proteomic approaches. In both data sets, a pronounced aging trend was detected in control and mdx mice, and this trend was separate from the disease process. In addition, the characteristic increase in taurine associated with dystrophic tissue is correlated with proteins associated with oxidative phosphorylation and mitochondrial metabolism.


Storage and retrieval of highly repetitive sequence collections.
A repetitive sequence collection is a set of sequences which are small variations of each other. A prominent example are genome sequences of individuals of the same or close species, where the differences can be expressed by short lists of basic edit operations. Flexible and efficient data analysis on such a typically huge collection is plausible using suffix trees. However, the suffix tree occupies much space, which very soon inhibits in-memory analyses. Recent advances in full-text indexing reduce the space of the suffix tree to, essentially, that of the compressed sequences, while retaining its functionality with only a polylogarithmic slowdown. However, the underlying compression model considers only the predictability of the next sequence symbol given the k previous ones, where k is a small integer. This is unable to capture longer-term repetitiveness. For example, r identical copies of an incompressible sequence will be incompressible under this model. We develop new static and dynamic full-text indexes that are able of capturing the fact that a collection is highly repetitive, and require space basically proportional to the length of one typical sequence plus the total number of edit operations. The new indexes can be plugged into a recent dynamic fully-compressed suffix tree, achieving full functionality for sequence analysis, while retaining the reduced space and the polylogarithmic slowdown. Our experimental results confirm the practicality of our proposal.


Hypocretin (orexin) biology and the pathophysiology of narcolepsy with cataplexy.
The discovery of hypocretins (orexins) and their causal implication in narcolepsy is the most important advance in sleep research and sleep medicine since the discovery of rapid eye movement sleep. Narcolepsy with cataplexy is caused by hypocretin deficiency owing to destruction of most of the hypocretin-producing neurons in the hypothalamus. Ablation of hypocretin or hypocretin receptors also leads to narcolepsy phenotypes in animal models. Although the exact mechanism of hypocretin deficiency is unknown, evidence from the past 20 years strongly favours an immune-mediated or autoimmune attack, targeting specifically hypocretin neurons in genetically predisposed individuals. These neurons form an extensive network of projections throughout the brain and show activity linked to motivational behaviours. The hypothesis that a targeted immune-mediated or autoimmune attack causes the specific degeneration of hypocretin neurons arose mainly through the discovery of genetic associations, first with the HLA-DQB1*06:02 allele and then with the T-cell receptor α locus. Guided by these genetic findings and now awaiting experimental testing are models of the possible immune mechanisms by which a specific and localised brain cell population could become targeted by T-cell subsets. Great hopes for the identification of new targets for therapeutic intervention in narcolepsy also reside in the development of patient-derived induced pluripotent stem cell systems.


A review of elasmobranch research in the Red Sea.
Given the global concern about the status of elasmobranch fishes, the paucity of information on elasmobranchs in the Red Sea is worrisome. Management of elasmobranchs in areas other than the Red Sea has been helped by research on population ecology, reproductive biology and resource partitioning, subjects that are virtually absent in the Red Sea elasmobranch literature. This review provides the first comprehensive summary of elasmobranch biology in the Red Sea with the aim of facilitating research in a region that remains remarkably under-studied.


The role of genetic factors for hearing deterioration across 20 years: a twin study.
Hearing deterioration at advanced ages is associated with environmental exposures (eg, to noise and solvents) and genetic influences may also be important. Little is known about the role of genetic influences on hearing when evaluated longitudinally. We sought to investigate longitudinal hearing loss in a cohort of adult male twins to evaluate the importance of genetic and environmental factors for hearing deterioration over time. Hearing using conventional clinical audiometry was assessed in 583 male twins (128 monozygotic twin pairs and 111 dizygotic twin pairs) aged 34-79 at baseline and again two decades later. The hearing thresholds at two time points were compared at each frequency and in two different frequency regions. Genetic analyses were based on structural equation models. Bivariate Cholesky decomposition was used for longitudinal analysis. The prevalence of hearing loss increased over time in better and worse ear. The hearing threshold shift was more pronounced in the high-frequency region, especially at 8000 Hz. Genetic influences were moderate (heritability: 53%-65%) for pure-tone averages at both lower and higher frequencies, and were of equal magnitude at baseline and follow-up. In contrast, environmental influences were of substantial importance (55%-88%) for rate of change of the hearing threshold over the 18-year period. Genetic factors are of considerable importance for level of hearing acuity, but environmental factors are more important for rate of change over an 18-year period.


Designing an LED luminaire with balance between uniformity of luminance and illuminance for non-Lambertian road surfaces.
We present a method of designing a light-emitting diode (LED) luminaire for non-Lambertian road surfaces, in which we optimized the overlap between the lighting areas of a luminaire and its adjacent luminaires to attain balance between uniformity of luminance and illuminance. The goal of the balance is to approach highly uniform luminance and raise illuminance uniformity as high as possible. The optimal illuminance rendered by a single luminaire and the corresponding far-field intensity were derived from the optimal overlap condition, and, thus, the optical model was established. The performance of the model is as follows: overall uniformity of luminance (UO) is 0.91, longitudinal uniformity of Luminance (UL) is 0.92, threshold increment (TI) is 1.64%, uniformity of horizontal illuminance (UHE) is 0.66, and surround ratio (SR) is 0.52. Finally, a compact freeform lens capping an LED was constructed to realize the optical model. The performance of the luminaire designed with arrayed units of an LED capped by the freeform lens is as follows: UO is 0.80, UL is 0.85, TI is 3.57%, UHE is 0.56, and SR is 0.51. In addition, the luminaire provides 19,600 lumens of luminous flux, of which 68% and 54% are projected on the target area and roadway alone, respectively, while 27% is absorbed by the surrounding materials.


Whole-genome validation of high-information-content fingerprinting.
Fluorescent-based high-information-content fingerprinting (HICF) techniques have recently been developed for physical mapping. These techniques make use of automated capillary DNA sequencing instruments to enable both high-resolution and high-throughput fingerprinting. In this article, we report the construction of a whole-genome HICF FPC map for maize (Zea mays subsp. mays cv B73), using a variant of HICF in which a type IIS restriction enzyme is used to generate the fluorescently labeled fragments. The HICF maize map was constructed from the same three maize bacterial artificial chromosome libraries as previously used for the whole-genome agarose FPC map, providing a unique opportunity for direct comparison of the agarose and HICF methods; as a result, it was found that HICF has substantially greater sensitivity in forming contigs. An improved assembly procedure is also described that uses automatic end-merging of contigs to reduce the effects of contamination and repetitive bands. Several new features in FPC v7.2 are presented, including shared-memory multiprocessing, which allows dramatically faster assemblies, and automatic end-merging, which permits more accurate assemblies. It is further shown that sequenced clones may be digested in silico and located accurately on the HICF assembly, despite size deviations that prevent the precise prediction of experimental fingerprints. Finally, repetitive bands are isolated, and their effect on the assembly is studied.


The association of pipe and cigar use with cotinine levels, lung function, and airflow obstruction: a cross-sectional study.
Cigarette smoking is the major cause of chronic obstructive pulmonary disease, but studies on the contribution of other smoking techniques are sparse. To determine whether pipe and cigar smoking was associated with elevated cotinine levels, decrements in lung function, and increased odds of airflow obstruction. Cross-sectional study. Population-based sample from 6 U.S. communities. Men and women aged 48 to 90 years without clinical cardiovascular disease at enrollment who were part of MESA (Multi-Ethnic Study of Atherosclerosis). The MESA Lung Study measured spirometry according to American Thoracic Society guidelines and urine cotinine levels by immunoassay on a subsample of MESA. Pipe-years and cigar-years were calculated as years from self-reported age of starting to age of quitting (or to current age in current users) multiplied by pipe-bowls or cigars per day. Of 3528 participants, 9% reported pipe smoking (median, 15 pipe-years), 11% reported cigar smoking (median, 6 cigar-years), and 52% reported cigarette smoking (median, 18 pack-years). Self-reported current pipe and cigar smokers had elevated urine cotinine levels compared with never-smokers. Pipe-years were associated with decrements in FEV(1), and cigar-years were associated with decrements in the FEV(1)-FVC ratio. Participants who smoked pipes or cigars had increased odds of airflow obstruction whether they had also smoked cigarettes (odds ratio, 3.43 [95% CI, 1.75 to 6.71]; P < 0.001) or not (odds ratio, 2.31 [CI, 1.04 to 5.11]; P = 0.039) compared with participants with no smoking history. Cross-sectional design. Pipe and cigar smoking increased urine cotinine levels and was associated with decreased lung function and increased odds of airflow obstruction, even in participants who had never smoked cigarettes. National Heart, Lung, and Blood Institute, National Institutes of Health.


Polyphenols journey through blood-brain barrier towards neuronal protection.
Age-related complications such as neurodegenerative disorders are increasing and remain cureless. The possibility of altering the progression or the development of these multifactorial diseases through diet is an emerging and attractive approach with increasing experimental support. We examined the potential of known bioavailable phenolic sulfates, arising from colonic metabolism of berries, to influence hallmarks of neurodegenerative processes. In silico predictions and in vitro transport studies across blood-brain barrier (BBB) endothelial cells, at circulating concentrations, provided evidence for differential transport, likely related to chemical structure. Moreover, endothelial metabolism of these phenolic sulfates produced a plethora of novel chemical entities with further potential bioactivies. Pre-conditioning with phenolic sulfates improved cellular responses to oxidative, excitotoxicity and inflammatory injuries and this attenuation of neuroinflammation was achieved via modulation of NF-κB pathway. Our results support the hypothesis that these small molecules, derived from dietary (poly)phenols may cross the BBB, reach brain cells, modulate microglia-mediated inflammation and exert neuroprotective effects, with potential for alleviation of neurodegenerative diseases.


Antibody escape of SARS-CoV-2 Omicron BA.4 and BA.5 from vaccine and BA.1 serum.
The Omicron lineage of SARS-CoV-2, which was first described in November 2021, spread rapidly to become globally dominant and has split into a number of sublineages. BA.1 dominated the initial wave but has been replaced by BA.2 in many countries. Recent sequencing from South Africa's Gauteng region uncovered two new sublineages, BA.4 and BA.5, which are taking over locally, driving a new wave. BA.4 and BA.5 contain identical spike sequences, and although closely related to BA.2, they contain further mutations in the receptor-binding domain of their spikes. Here, we study the neutralization of BA.4/5 using a range of vaccine and naturally immune serum and panels of monoclonal antibodies. BA.4/5 shows reduced neutralization by the serum from individuals vaccinated with triple doses of AstraZeneca or Pfizer vaccine compared with BA.1 and BA.2. Furthermore, using the serum from BA.1 vaccine breakthrough infections, there are, likewise, significant reductions in the neutralization of BA.4/5, raising the possibility of repeat Omicron infections.


Adverse effects of nonsteroidal antiinflammatory drugs: an update of gastrointestinal, cardiovascular and renal complications.
Non-steroidal anti-inflammatory drugs (NSAIDs) are used chronically to reduce pain and inflammation in patients with arthritic conditions, and also acutely as analgesics by many patients. Both therapeutic and adverse effects of NSAIDs are due to inhibition of cyclooxygenase (COX) enzyme. NSAIDs are classified as non-selective and COX-2-selective inhibitors (COXIBS) based on their extent of selectivity for COX inhibition. However, regardless of their COX selectivity, reports are still appearing on the GI side effect of NSAIDs particularly on the lower gastrointestinal (GI) tract and the harmful role of their controlled release formulations. In addition, previously unpublished data stored in the sponsor's files, question the GI sparing properties of rofecoxib, a COXIB that has been withdrawn due to cardiovascular (CV) side effects. Presently, the major side effects of NSAIDs are the GI complications, renal disturbances and CV events. There is a tendency to believe that all NSAIDs are associated with renal and CV side effects, a belief that is not supported by solid evidence. Indeed, lower but still therapeutics doses of some NSAIDs may be cardioprotective. In this review, we briefly discuss the GI toxicity of the NSAIDs and assess their renal and CV adverse effects in more detail.


A standardized triglyceride and carbohydrate challenge: the oral triglyceride tolerance test.
A standardized method of assessing postprandial triglyceride changes is not available. We evaluated an oral triglyceride tolerance test (OTTT) designed for routine clinical and research use. A 200-ml strawberry-flavored test drink (50 g fat, 50 g carbohydrate) was administered twice to 30 diabetic and 20 nondiabetic subjects. Venous plasma triglyceride and glucose levels were measured when fasting and every 2 h for 8 h after the drink. Fingerprick plasma triglyceride levels were measured when fasting and at 6 and 8 h after the drink. The drink was consumed within 3 min and well tolerated by all subjects. The median triglyceride rise at 6 h was similar in diabetic and nondiabetic subjects (0.23 vs. 0.42 mmol/l, NS) and correlated with glucose increase at 2 h (r = 0.429, P = 0.018 and r = 0.509, P = 0.026; respectively). Diabetic subjects had higher 6-h geometric mean (1 SD range) triglyceride levels (1.82 [1.87 to 3.23] vs. 1.11 [0.66 to 1.11 mmol/l], P < 0.003) but a similar coefficient of variation (17.5 vs. 17.0%, NS) and a similar median (interquartile range) time to achieve maximal concentration (T(max)) (6.0 [4.0 to 6.0] vs. 5.0 [4.0 to 6.0] h, NS). Capillary triglyceride values were equivalent to simultaneous venous samples but consistently 10% greater. The OTTT permits simple evaluation of postchallenge triglyceride levels, is acceptable to subjects, and can be performed with capillary sampling. It could be used to monitor triglyceride-lowering therapies and to provide additional information concerning cardiovascular disease risk, particularly in diabetic subjects.


Control of timing of embryonic M-phase entry and exit is differentially sensitive to CDK1 and PP2A balance.
Harmonious embryo development requires precise coordination between the timing of the cell cycle and the developmental program. Cyclin accumulation determines the timing of the cell cycle M-phase entry and its degradation determines the timing of the M-phase exit. It is well known that CDK1 and PP2A also govern M-phase entry. However, it is unknown how this kinase and phosphatase regulate the precise timing of events at the beginning of the M-phase and how they cooperate with cyclin metabolism. Here we use Xenopus laevis one-cell embryo cell-free extract experiments to answer this question critical for understanding the regulation of embryo development. Using, separately, low concentrations of the chemical inhibitor of CDK1, RO3306 (RO), or the inhibitor of phosphatases, okadaic acid (OA), we show that moderately diminished CDK1 or PP2A activities results in a delay and an acceleration respectively, of M-phase entry. Simultaneous diminution of CDK1 and PP2A activities results in an intermediate timing of M-phase entry, prolongs the duration of M-phase and diminishes the dynamics of cyclin B2 degradation. We thus show, for the first time, that equilibrium between CDK1 and PP2A specifies the timing of M-phase entry and exit and regulates the dynamics of cyclin B degradation upon M-phase exit in Xenopus laevis first embryonic mitosis.


Preparation of pre-confluent retinal cells increases graft viability in vitro and in vivo: a mouse model.
Graft failure remains an obstacle to experimental subretinal cell transplantation. A key step is preparing a viable graft, as high levels of necrosis and apoptosis increase the risk of graft failure. Retinal grafts are commonly harvested from cell cultures. We termed the graft preparation procedure "transplant conditions" (TC). We hypothesized that culture conditions influenced graft viability, and investigated whether viability decreased following TC using a mouse retinal pigment epithelial (RPE) cell line, DH01. Cell viability was assessed by trypan blue exclusion. Levels of apoptosis and necrosis in vitro were determined by flow cytometry for annexin V and propidium iodide and Western blot analysis for the pro- and cleaved forms of caspases 3 and 7. Graft viability in vivo was established by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and cleaved caspase 3 immunolabeling of subretinal allografts. Pre-confluent cultures had significantly less nonviable cells than post-confluent cultures (6.6%±0.8% vs. 13.1%±0.9%, p<0.01). Cell viability in either group was not altered significantly following TC. Caspases 3 and 7 were not altered by levels of confluence or following TC. Pre-confluent cultures had low levels of apoptosis/necrosis (5.6%±1.1%) that did not increase following TC (4.8%±0.5%). However, culturing beyond confluence led to progressively increasing levels of apoptosis and necrosis (up to 16.5%±0.9%). Allografts prepared from post-confluent cultures had significantly more TUNEL-positive cells 3 hours post-operatively than grafts of pre-confluent cells (12.7%±3.1% vs. 4.5%±1.4%, p<0.001). Subretinal grafts of post-confluent cells also had significantly higher rates of cleaved caspase 3 than pre-confluent grafts (20.2%±4.3% vs. 7.8%±1.8%, p<0.001). Pre-confluent cells should be used to maximize graft cell viability.


Thinking the unthinkable: Alzheimer's, Creutzfeldt-Jakob and Mad Cow disease: the age-related reemergence of virulent, foodborne, bovine tuberculosis or losing your mind for the sake of a shake or burger.
The possibility of the age-related reemergence of foodborne Mycobacterium bovis (bovine tuberculosis) as a vector for Creutzfeldt-Jakob Disease (CJD or human Mad Cow Disease) and Mad Cow disease itself is real. The CDC reported last May of an outbreak of CJD linked to the consumption of meat contaminated "with the agent causing" bovine spongiform encephalopathy (BSE) in a New Jersey racetrack between the time frame 1995-2004. In the opinion of experts, ample justification exists for considering a similar pathogenesis for Alzheimer's, Creutzfeldt-Jakob and the other spongiform encephalopathies such as Mad Cow disease. In fact, Creutzfeldt-Jakob and Alzheimer's often coexist and at this point are thought to differ merely by time-dependent physical changes. A recent study links up to 13% of all "Alzheimer's" victims as really having Creutzfeldt-Jakob disease. Bovine tuberculosis, which includes Mycobacterium bovis and M. avium-intracellulare or paratuberculosis, is and has always been the most prevalent threat to the cattle industry, and the USDA reports that between 20% and 40% of US dairy herds are infected with paratuberculosis alone. The health risk for milk tainted with M. bovis has been known for decades and there was a time not so long ago when "tuberculin-tested" was printed on every milk container. Schliesser stated that meat from tuberculous animals may also constitute a significant risk of infection. At the turn of the 20th century 25% of the many US deaths from TB in adults were caused by M. bovis. Dairy products aside, when past and present meat consumption are factored in, there is three times the risk of developing Alzheimer's in meat eaters as opposed to vegetarians. The investigation into the causal trail for Creutzfeldt-Jakob, indistinguishable from Alzheimer's except for its shorter, lethal course might have grown cold where it not for Roel's and others who linked mad cow in cattle with M. bovis and related paratuberculosis on clinical, pathologic and epidemiological grounds. The southwest of the UK, the very cradle of British BSE and CJD outbreaks, saw an exponential increase in bovine tuberculosis just prior to it's spongiform outbreaks. All of this brings up the unthinkable: that Alzheimer's, Cruetzfeldt-Jackob, and Mad Cow Disease might just be caused by eating the meat or dairy in consumer products or feed. It is only appropriate therefore to explore the role of bovine TB and the atypical mycobacteria in Alzheimer's, JCD and Mad Cow disease and develop better serological surveillance for these pathogens.


SU-E-J-191: A Multivariate Framework for N-Tissue Classification in Treatment Assessment of Glioblastomas.
Glioblastoma is the most common primary brain tumor in adults and is rapidly fatal. Treatment monitoring of these patients has increased awareness that many patients have new areas of contrast enhancement without progressive clinical signs and symptoms. Although the enhancing areas mimic tumor progression, the lesions result from treatment effects and subsequently stabilize or improve without further treatment and are not correlated with poorer outcomes. This phenomenon has been termed pseudoprogression and is hypothesized to occur secondarily to edema and vessel permeability in the tumor area as a result of the combined effects of radiation and chemotherapy. Since the new enhancing lesions of pseudoprogression are indistinguishable from true disease progression, there is a need for a predictive model to distinguish the two phenomena. We developed a classification algorithm that combines perfusion and diffusion MRI imaging to effectively partition the cases as one exhibiting true or pseudo progression based on a vector of features containing T1, rCBV and ADC imaging. The multi-sequence classification algorithm uses an expectation maximization (EM) algorithm that learns from training cases with known clinical outcome to assigns each voxel to a type of tissue. A training set of 20 where the clinical outcome is known from biopsy or from long-term follow-up was used by EM algorithm to model typical imaging values within tissue of pseudo, tumor, edema, necrosis, vessels or brain anatomy to construct a database of expected values for each tissue type. When presented with a new case, the algorithm automatically classifies voxels by their geographical proximities and Mahalanobis distance to the pre-sampled values. Usage of advanced classification techniques allows automated labeling of voxels into normal, pseudoprogression or tumoral tissue types. The technique allows for early detection of pseudo progression to spare patients from unnecessary surgery or toxic chemotherapy.


A Surgeon's guide to advances in the pharmacological management of acute Charcot neuroarthropathy.
Acute Charcot neuroarthropathy is a devastating condition and, its incidence is increasing. Currently, treatment consists of immobilisation and off-loading of the involved extremity. Outcomes are frequently poor and novel treatments are being sought urgently. This review aims to outline advances in the pharmacological treatment of this, condition. PubMed and the Cochrane Database of systematic reviews were searched. Relevant papers were cross referenced. Eleven original studies were identified. The limited data available suggest pamidronate, alendronate and calcitonin provide some clinical and biochemical improvements while zoledronic acid is deleterious and, increases off-loading times. However, the data is not robust enough to convincingly demonstrate clinically meaningful effects. The studies were predominantly low quality and heterogeneous. They differed markedly in study type, pharmacological agent used, dosing regimen, disease, aetiology/stage/location, concurrent off-loading regimen, outcomes and, follow-up. Few were rigorous in controlling for associated confounding variables and none investigated long term outcomes. The routine use of pharmacological treatment modalities for this condition is not recommended in the United States by the Food and Drug Administration or in the United Kingdom by the National Institute for Health and Clinical Excellence. Given the evidence available this is justified and further higher quality research is required.


Chemical composition, antioxidant properties and hepatoprotective effects of chamomile (Matricaria recutita L.) decoction extract against alcohol-induced oxidative stress in rat.
The present study assessed the chemical composition, antioxidant properties, and hepatoprotective effects of subacute pre-treatment with chamomile (Matricaria recutita L.) decoction extract (CDE) against ethanol (EtOH)-induced oxidative stress in rats. The colorimetric analysis demonstrated that the CDE is rich in total polyphenols, total flavonoids and condensed tannins, and exhibited an important in vitro antioxidant activity. The use of LC/MS technique allowed us to identify 10 phenolic compounds in CDE. We found that CDE pretreatment, in vivo, protected against EtOH-induced liver injury evident by plasma transaminases activity and preservation of the hepatic tissue structure. The CDE counteracted EtOH-induced liver lipoperoxidation, preserved thiol -SH groups and prevented the depletion of antioxidant enzyme activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). We also showed that acute alcohol administration increased tissue and plasma hydrogen peroxide (H(2)O(2)), calcium and free iron levels. More importantly, CDE pre-treatment reversed all EtOH-induced disturbances in intracellular mediators. In conclusion, our data suggest that CDE exerted a potential hepatoprotective effect against EtOH-induced oxidative stress in rat, at least in part, by negatively regulating Fenton reaction components such as H(2)O(2) and free iron, which are known to lead to cytotoxicity mediated by intracellular calcium deregulation.


Molecules of the mind: integrating synaptic biochemistry to understand brain function.
The focused meeting entitled 'Molecular Determinants of Synaptic Function: Molecules and Models' brought together several molecules and experimental models that are furthering our understanding of the biochemical basis of integrative brain function. Invited speakers and short communications from more junior scientists highlighted how individual molecules or protein networks underlie defined subcellular functions (e.g. transmitter release, receptor activation and transmitter uptake) can be used to unravel integrative function at cellular, systems and behavioural levels.


Homeostatic levels of p62 control cytoplasmic inclusion body formation in autophagy-deficient mice.
Inactivation of constitutive autophagy results in formation of cytoplasmic protein inclusions and leads to liver injury and neurodegeneration, but the details of abnormalities related to impaired autophagy are largely unknown. Here we used mouse genetic analyses to define the roles of autophagy in the aforementioned events. We report that the ubiquitin- and LC3-binding protein "p62" regulates the formation of protein aggregates and is removed by autophagy. Thus, genetic ablation of p62 suppressed the appearance of ubiquitin-positive protein aggregates in hepatocytes and neurons, indicating that p62 plays an important role in inclusion body formation. Moreover, loss of p62 markedly attenuated liver injury caused by autophagy deficiency, whereas it had little effect on neuronal degeneration. Our findings highlight the unexpected role of homeostatic level of p62, which is regulated by autophagy, in controlling intracellular inclusion body formation, and indicate that the pathologic process associated with autophagic deficiency is cell-type specific.


Autologous blood transfusions. Council on Scientific Affairs.
Blood collected from a patient for retransfusion at a later time into that same individual is called "autologous blood." When the guidelines established by the American Association of Blood Banks are followed, autologous blood is the safest type of blood for transfusion. It also decreases the demand for banked blood and eliminates the risk of infection and alloimmunization from a transfusion. Autologous transfusions are becoming widely available; since 1974 the number of institutions providing autologous transfusion programs has increased more than fourfold. The Council on Scientific Affairs endorses the use of autologous blood transfusions.


Basic fibroblast growth factor reduces functional and structural damage in chronic kidney disease.
Chronic kidney disease (CKD) is characterized by loss of renal function. The pathological processes involved in the progression of this condition are already known, but the molecular mechanisms have not been completely explained. Recent reports have shown the intrinsic capacity of the kidney to undergo repair after acute injury through the reexpression of repairing proteins (Villanueva S, Cespedes C, Vio CP. Am J Physiol Regul Integr Comp Physiol 290: R861-R870, 2006). Stimulation with basic fibroblast growth factor (bFGF) could accelerate this process. However, it is not known whether bFGF can induce this phenomenon in kidney cells affected by CKD. Our aim was to study the evolution of renal damage in animals with CKD treated with bFGF and to relate the amount of repairing proteins with renal damage progression. Male Sprague-Dawley rats were subjected to 5/6 nephrectomy (NPX) and treated with bFGF (30 μg/kg, NPX+bFGF); a control NPX group was treated with saline (NPX+S). Animals were euthanized 35 days after bFGF administration. Functional effects were assessed based on serum creatinine levels; morphological damage was assessed by the presence of macrophages (ED-1), interstitial α-smooth muscle actin (α-SMA), and interstitial collagen through Sirius red staining. The angiogenic factors VEGF and Tie-2 and the epithelial/tubular factors Ncam, bFGF, Pax-2, bone morphogenic protein-7, Noggin, Lim-1, Wnt-4, and Smads were analyzed. Renal stem cells were evaluated by Oct-4. We observed a significant reduction in serum creatinine levels, ED-1, α-SMA, and Sirius red as well as an important induction of Oct-4, angiogenic factors, and repairing proteins in NPX+bFGF animals compared with NPX+S animals. These results open new perspectives toward reducing damage progression in CKD.


Faecal calprotectin testing for differentiating amongst inflammatory and non-inflammatory bowel diseases: systematic review and economic evaluation.
Irritable bowel syndrome (IBS) is common, and causes pain, bloating and diarrhoea and/or constipation. It is a troublesome condition that reduces the quality of life but causes no permanent damage. Inflammatory bowel disease (IBD) comprises mainly ulcerative colitis (UC) and Crohn's disease (CD). Both cause serious complications and may lead to sections of the bowel having to be removed, although this is more common with CD. The presenting symptoms of IBS and IBD can be similar. Distinguishing them on clinical signs and symptoms can be difficult. Until recently, colonoscopy was often required to rule out IBD. In younger people, > 60% of colonoscopies showed no abnormality. Faecal calprotectin (FC) is a protein released by the white blood cells, neutrophils, found in inflamed areas of the bowel in IBD. Determining the level of FC in stool samples may help distinguish IBS from IBD. To review the value of FC for distinguishing between IBD and non-IBD. Sources included MEDLINE, EMBASE, The Cochrane Library, Web of Science, websites of journals and the European Crohn's and Colitis Organisation (conference abstracts 2012 and 2013), and contact with experts. Systematic review and economic modelling. Review Manager (RevMan) version 5.2 (The Cochrane Collaboration, The Nordic Cochrane Centre, Copenhagen, Denmark) was used for most analysis, with statistical analyses done in Stata version 12 (StataCorp LP, College Station, TX, USA). Forest plots and receiver operating characteristic curves were produced. Quality Assessment of Diagnostic Accuracy Studies was used for quality assessment. Economic modelling was done in Microsoft Excel 2010 (Microsoft Corporation, Redmond, WA, USA). Studies were often small, most used only one calprotectin cut-off level, and nearly all came from secondary care populations. Twenty-eight studies provided data for 2 × 2 tables and were included in meta-analyses, with seven in the most important comparison in adults (IBS vs. IBD) and eight in the key comparison in paediatrics (IBD vs. non-IBD). Most studies used laboratory enzyme-linked immunosorbent assay (ELISA) tests. For distinguishing between IBD and IBS in adults, these gave pooled sensitivity of 93% and specificity of 94% at FC cut-off level of 50 µg/g. Sensitivities at that cut-off ranged from 83% to 100%, and specificities from 60% to 100%. For distinguishing between IBD and non-IBD in paediatric populations with ELISA tests, sensitivities ranged from 95% to 100% at cut-off of 50 µg/g and specificities of 44-93%. Few studies used point-of-care testing but that seemed as reliable as ELISA, though perhaps less specific. The evidence did not provide any grounds for preferring one test over others on clinical effectiveness grounds. FC testing in primary care could reduce the need for referral and colonoscopies. Any quality-adjusted life-year gains are likely to be small because of the low prevalence of IBD and the high sensitivities of all of the tests, resulting in few false negatives with IBD. However, considerable savings could accrue. Areas of uncertainty include the optimum management of people with borderline results (50-150 µg/g), most of whom do not have IBD. Repeat testing may be appropriate before referral. Faecal calprotectin can be a highly sensitive way of detecting IBD, although there are inevitably trade-offs between sensitivity and specificity, with some false positives (IBS with positive calprotectin) if a low calprotectin cut-off is used. In most cases, a negative calprotectin rules out IBD, thereby sparing most people with IBS from having to have invasive investigations, such as colonoscopy. PROSPERO CRD 42012003287. The National Institute for Health Research Health Technology Assessment programme.


Giving and taking: representational building blocks of active resource-transfer events in human infants.
Active resource transfer is a pervasive and distinctive feature of human sociality. We hypothesized that humans possess an action schema of giving specific for representing social interactions based on material exchange, and specified the set of necessary assumptions about giving events that this action schema should be equipped with. We tested this proposal by investigating how 12-month-old infants interpret abstract resource-transfer events. Across eight looking-time studies using a violation-of-expectation paradigm we found that infants were able to distinguish between kinematically identical giving and taking actions. Despite the surface similarity between these two actions, only giving was represented as an object-mediated social interaction. While we found no evidence that infants expected the target of a giving or taking action to reciprocate, the present results suggest that infants interpret giving as an inherently social action, which they can possibly use to map social relations via observing resource-transfer episodes.


Oxygen treatment of sleep hypoxaemia in Duchenne muscular dystrophy.
Patients with Duchenne muscular dystrophy develop progressive ventilatory muscle weakness and often die of respiratory complications. Recurrent, often profound, hypoxaemia has been shown in a previous study by this group to occur during rapid eye movement (REM) sleep in these patients before they develop sleep symptoms. In this study the efficacy and physiological effects of nocturnal oxygen in such patients have been assessed. Seven patients with Duchenne muscular dystrophy (age range 16-22 years; mean vital capacity 1.37 litres) with normal arterial blood gas tensions when awake were investigated by standard overnight polysomnography on an acclimatization night followed by two successive nights on which they received room air and nasal oxygen (2 litres/min) respectively in random order. Total sleep time, proportion of REM and non-REM sleep, and frequency and duration of arousals were similar on the two nights. When breathing air six of the seven subjects developed oxygen desaturation of more than 5% during REM sleep. With oxygen only one subject showed any oxygen desaturation exceeding 2.5%. Oxygen desaturation was associated with periods of hypopnoea or cessation of respiratory effort. The mean duration of episodes of hypopnoea and apnoea was prolonged during oxygen breathing by 19% and the mean duration of episodes during REM sleep by 33% (the proportion of REM sleep associated with hypopnoea and apnoea increased in all subjects). Heart rate in non-REM sleep fell by 9.3%; heart rate variation in REM and non-REM sleep was unchanged. These acute studies show that oxygen reduces the sleep hypoxaemia associated with respiratory muscle weakness; whether long term treatment will be possible or desirable is not clear as oxygen potentiates the underlying ventilatory disturbance.


Recovery from Spatial Neglect with Intra- and Transhemispheric Functional Connectivity Changes in Vestibular and Visual Cortex Areas-A Case Study.
Vestibular signals are involved in higher cortical functions like spatial orientation and its disorders. Vestibular dysfunction contributes, for example, to spatial neglect which can be transiently improved by caloric stimulation. The exact roles and mechanisms of the vestibular and visual systems for the recovery of neglect are not yet known. Resting-state functional connectivity (fc) magnetic resonance imaging was recorded in a patient with hemispatial neglect during the acute phase and after recovery 6 months later following a right middle cerebral artery infarction before and after caloric vestibular stimulation. Seeds in the vestibular [parietal operculum (OP2)], the parietal [posterior parietal cortex (PPC); 7A, hIP3], and the visual cortex (VC) were used for the analysis. During the acute stage after caloric stimulation the fc of the right OP2 to the left OP2, the anterior cingulum, and the para/hippocampus was increased bilaterally (i.e., the vestibular network), while the interhemispheric fc was reduced between homologous regions in the VC. After 6 months, similar fc increases in the vestibular network were found without stimulation. In addition, fc increases of the OP2 to the PPC and the VC were seen; interhemispherically this was true for both PPCs and for the right PPC to both VCs. Improvement of neglect after caloric stimulation in the acute phase was associated with increased fc of vestibular cortex areas in both hemispheres to the para-hippocampus and the dorsal anterior cingulum, but simultaneously with reduced interhemispheric VC connectivity. This disclosed a, to some extent, similar but also distinct short-term mechanism (vestibular stimulation) of an improvement of spatial orientation compared to the long-term recovery of neglect.


rAAV6-microdystrophin rescues aberrant Golgi complex organization in mdx skeletal muscles.
Muscular dystrophies are a diverse group of severe degenerative muscle diseases. Recent interest in the role of the Golgi complex (GC) in muscle disease has been piqued by findings that several dystrophies result from mutations in putative Golgi-resident glycosyltransferases. Given this new role of the Golgi in sarcolemmal stability, we hypothesized that abnormal Golgi distribution, regulation and/or function may constitute part of the pathology of other dystrophies, where the primary defect is independent of Golgi function. Thus, we investigated GC organization in the dystrophin-deficient muscles of mdx mice, a mouse model for Duchenne muscular dystrophy. We report aberrant organization of the synaptic and extrasynaptic GC in skeletal muscles of mdx mice. The GC is mislocalized and improperly concentrated at the surface and core of mdx myofibers. Golgi complex localization is disrupted after the onset of necrosis and normal redistribution is impaired during regeneration of mdx muscle fibers. Disruption of the microtubule cytoskeleton may account in part for aberrant GC localization in mdx myofibers. Golgi complex distribution is restored to wild type and microtubule cytoskeleton organization is significantly improved by recombinant adeno-associated virus 6-mediated expression of DeltaR4-R23/DeltaCT microdystrophin showing a novel mode of microdystrophin functionality. In summary, GC distribution abnormalities are a novel component of mdx skeletal muscle pathology rescued by microdystrophin expression.


Using ecological momentary assessment in testing the effectiveness of an alcohol intervention: a two-arm parallel group randomized controlled trial.
Alcohol consumption of college students has a fluctuating nature, which might impact the measurement of intervention effects. By using 25 follow-up time-points, this study tested whether intervention effects are robust or might vary over time. Data were used from a two-arm parallel group randomized controlled trial applying ecological momentary assessment (EMA) with 30 data time-points in total. Students between 18 and 24 years old who reported heavy drinking in the past six months and who were ready to change their alcohol consumption were randomly assigned to the experimental (n = 456: web-based brief alcohol intervention) and control condition (n = 451: no intervention). Outcome measures were weekly alcohol consumption, frequency of binge drinking, and heavy drinking status. According to the intention-to-treat principle, regression analyses revealed that intervention effects on alcohol consumption varied when exploring multiple follow-up time-points. Intervention effects were found for a) weekly alcohol consumption at 1, 2, 3, 4, and 7 weeks follow-up, b) frequency of binge drinking at 1, 2, 7, and 12 weeks follow-up, and c) heavy drinking status at 1, 2, 7, and 16 weeks follow-up. This research showed that the commonly used one and six month follow-up time-points are relatively arbitrary and not using EMA might bring forth erroneous conclusions on the effectiveness of interventions. Therefore, future trials in alcohol prevention research and beyond are encouraged to apply EMA when assessing outcome measures and intervention effectiveness. Netherlands Trial Register NTR2665.


Stable isotope and fatty acid analyses reveal significant differences in trophic niches of smooth hammerhead Sphyrna zygaena (Carcharhiniformes) among three nursery areas in northern Humboldt Current System.
Fishery pressure on nursery areas of smooth hammerhead in northern Peruvian coast have become a serious threat to sustainability of this resource. Even though, some management actions focused on conservation of the smooth hammerhead populations were proposed in recent years, their scientific foundations are often limited, and biomass of smooth hammerhead in Peruvian waters continues to decrease. To inform management and conservation, this study aims to evaluate the trophic niche of smooth hammerhead juveniles from three nursery areas in the northern Peruvian coast using stable isotope and fatty acid analyses. First, we compared the environmental characteristics of each nursery area (i.e., sea surface temperature and chlorophyll-a concentration) and concluded that nursery areas differed significantly and consistently in sea surface temperature. Subsequently, we evaluated isotopic composition of carbon and nitrogen and fatty acid profiles of muscle and liver tissues collected from juvenile smooth hammerhead from each nursery area. We found that juvenile smooth hammerhead captured in San José were enriched in heavier 13C and 15N isotopes compared to those captured in Máncora and Salaverry. Furthermore, the broadest isotopic niches were observed in juveniles from Máncora, whereas isotopic niches of juveniles from Salaverry and San José were narrower. This difference is primarily driven by the Humboldt Current System and associated upwelling of cold and nutrient rich water that drives increased primary production in San José and, to a less extent, in Salaverry. Compared to smooth hammerhead juveniles from Máncora, those from San José and Salaverry were characterised by higher essential fatty acid concentrations related to pelagic and migratory prey. We conclude that smooth hammerhead juveniles from three nursery areas in the northern Peruvian coast differ significantly in their trophic niches. Thus, management and conservation efforts should consider each nursery area as a unique juvenile stock associated with a unique ecosystem and recognize the dependence of smooth hammerhead recruitment in San José and Salaverry on the productivity driven by the Humboldt Current System.


Essential role of UCP1 modulating the central effects of thyroid hormones on energy balance.
Classically, metabolic effects of thyroid hormones (THs) have been considered to be peripherally mediated, i.e. different tissues in the body respond directly to thyroid hormones with an increased metabolism. An alternative view is that the metabolic effects are centrally regulated. We have examined here the degree to which prolonged, centrally infused triiodothyronine (T3) could in itself induce total body metabolic effects and the degree to which brown adipose tissue (BAT) thermogenesis was essential for such effects, by examining uncoupling protein 1 (UCP1) KO mice. Wildtype and UPC1 KO mice were centrally-treated with T3 by using minipumps. Metabolic measurements were analyzed by indirect calorimetry and expression analysis by RT-PCR or western blot. BAT morphology and histology were studied by immunohistochemistry. We found that central T3-treatment led to reduced levels of hypothalamic AMP-activated protein kinase (AMPK) and elevated body temperature (0.7 °C). UCP1 was essential for the T3-induced increased rate of energy expenditure, which was only observable at thermoneutrality and notably only during the active phase, for the increased body weight loss, for the increased hypothalamic levels of neuropeptide Y (NPY) and agouti-related peptide (AgRP) and for the increased food intake induced by central T3-treatment. Prolonged central T3-treatment also led to recruitment of BAT and britening/beiging ("browning") of inguinal white adipose tissue (iWAT). We conclude that UCP1 is essential for mediation of the central effects of thyroid hormones on energy balance, and we suggest that similar UCP1-dependent effects may underlie central energy balance effects of other agents.


Conformational changes in Sindbis virus envelope proteins accompanying exposure to low pH.
The attachment of high multiplicities of Sindbis virus to tissue-cultured cells followed by brief treatment at low pH has been shown to produce cell fusion (fusion from without). In this report, experiments to determine the effects of low pH on the physical and biological properties of Sindbis virus are described. Exposure of purified Sindbis virions to mildly acidic conditions resulted in a rapid and irreversible alteration in particle density and sedimentation characteristics, followed by a slower loss of infectivity. Infectivity was not restored by a return to neutral pH; rather, the loss of virus infectivity seemed to be initiated by exposure to low pH but continued at neutral pH. The formation of a virus-cell complex in which virions were attached to the cell surface protected the particles from low-pH inactivation, although low pH could still expose virus functions responsible for cell fusion. Low pH was found to induce a conformational change in the E2 polypeptide of the intact virion. These results are discussed with respect to the process of Sindbis virus infection of tissue-cultured cells.


Structured Population-based Prostate-specific Antigen Screening for Prostate Cancer: The European Association of Urology Position in 2019.
Prostate cancer (PCa) is one of the first three causes of cancer mortality in Europe. Screening in asymptomatic men (aged 55-69yr) using prostate-specific antigen (PSA) is associated with a migration toward lower staged disease and a reduction in cancer-specific mortality. By 20yr after testing, around 100 men need to be screened to prevent one PCa death. While this ratio is smaller than for breast and colon cancer, the long natural history of PCa means many men die from other causes. As such, the nonselective use of PSA testing and radical treatments can lead to overdiagnosis and overtreatment. The European Association of Urology (EAU) supports measures to encourage appropriate PCa detection through PSA testing, while reducing overdiagnosis and overtreatment. These goals may be achieved using personalized risk-stratified approaches. For diagnosis, the greatest benefit from early detection is likely to come in men assessed using baseline PSA levels at the age of 45yr to individualize screening intervals. Multiparametric magnetic resonance imaging as well as risk calculators based on family history, ethnicity, digital rectal examination, and prostate volume should be considered to triage the need for biopsy, thus reducing the risk of overdiagnosis. For treatment, the EAU advocates balancing patient's life expectancy and cancer's mortality risk when deciding an approach. Active surveillance is encouraged in well-informed patients with low-risk and some intermediate-risk cancers, as it decreases the risks of overtreatment without compromising oncological outcomes. Conversely, the EAU advocates radical treatment in suitable men with more aggressive PCa. Multimodal treatment should be considered in locally advanced or high-grade cancers. PATIENT SUMMARY: Implementation of prostate-specific antigen (PSA)-based screening should be considered at a population level. Men at risk of prostate cancer should have a baseline PSA blood test (eg, at 45yr). The level of this test, combined with family history, ethnicity, and other factors, can be used to determine subsequent follow-up. Magnetic resonance imaging scans and novel biomarkers should be used to determine which men need biopsy and how any cancers should be treated.


How microorganisms avoid phagocyte attraction.
Microorganisms have developed several mechanisms to modulate the host immune system to increase their survival and propagation in the host. Their presence in the host is not only revealed by self-produced peptides but also through host-derived chemokines and active complement fragments. These so-called chemoattractants are recognized by G protein-coupled receptors (GPCRs) expressed on leukocyte cell membranes. Activation of GPCRs triggers leukocyte activation and guides their recruitment to the site of infection. Therefore, GPCRs play a central role in leukocyte trafficking leading to microbial clearance. It is therefore not surprising that microorganisms are able to sabotage this arm of the immune response. Different microorganisms have evolved a variety of tactics to modulate GPCR activation. Here, we review the mechanisms and proteins used by major human pathogens and less virulent microorganisms that affect GPCR signaling. While viruses generally produce receptor and chemoattractant mimics, parasites and bacteria such as Staphylococcus aureus, Streptococcus pyogenes, Porphyromonas gingivalis, and Bordetella pertussis secrete proteins that affect receptor signaling, directly antagonize receptors, cleave stimuli, and even prevent stimulus generation. As the large arsenal of GPCR modulators aids prolonged microbial persistence in the host, their study provides us a better understanding of microbial pathogenesis.


The clinical significance of focal active colitis.
Focal crypt injury by neutrophils (cryptitis/crypt abscesses), or focal active colitis (FAC), is a common isolated finding in endoscopic colorectal biopsies. Focal active colitis is often thought of as a feature of Crohn's disease, but may also be seen in ischemia, infections, partially treated ulcerative colitis, and as an isolated finding in patients undergoing endoscopy to exclude neoplasia. Clinical, endoscopic, and pathological data were retrospectively reviewed from 49 patients with focal active colitis, who had no other diagnostic findings on colorectal biopsy and no history of chronic inflammatory bowel disease. The histological findings were correlated with clinical diagnoses. Follow-up information was available for 42 of 49 focal active colitis patients. None developed inflammatory bowel disease; however, 19 patients had an acute self-limited colitis-like diarrheal illness, 11 had incidental focal active colitis (patients without diarrhea that were endoscoped to exclude colonic neoplasia and found to have asymptomatic FAC), 6 had irritable bowel syndrome, 4 had antibiotic-associated colitis, and 2 had ischemic colitis. Twenty patients were immunosuppressed, and 19 were taking nonsteroidal anti-inflammatory drugs. No histological features predicted final diagnoses. FAC did not predict the development of chronic colitis, even when mild crypt distortion or slight basal plasmacytosis was present. The preponderance of acute self-limited colitis and antibiotic-associated colitis among the FAC patients, along with the high number of immunosuppressed patients, support the conclusion that most FAC cases are infectious. The incidental detection of FAC in patients undergoing endoscopy to exclude colonic neoplasia was not clinically significant. The role of nonsteroidal anti-inflammatory drugs in FAC deserves further study.


Off-target-isocentric approach in non-coplanar Volumetric Modulated Arc Therapy (VMAT) planning for lung SBRT treatments.
Volumetric Modulated Arc Therapy (VMAT) has emerged as an efficient alternative to traditional three-dimensional (3D) non-coplanar conformal (3D-NC-C) beams for lung cancer stereotactic body radiotherapy (SBRT) because of its superior dosimetric properties and native ease in planning and treatment delivery. However, patient immobilization in lung SBRT often presents challenging geometrical clearance issues in the execution of large (in excess of 180°) non-coplanar arcs. In this study, we present an off-target-isocentric, non-coplanar VMAT (OTI-NC-VMAT) technique that appears to be simple, dosimetrically robust and allows for ample patient/couch-gantry clearance. We compared this technique to a target-isocentric, non-coplanar VMAT (TI-NC-VMAT) technique and the 3D-NC-C beams for dosimetric evaluations. Nineteen lung cancer patients previously treated with 3D-NC-C SBRT technique at our institution were selected. For each patient, an OTI-NC-VMAT plan and TI-NC-VMAT plan were created and compared to the original 3D-NC-C treatment plan. All of the plans were created for the same prescription dose of 54 Gy total in 3 fractions, covering 95% of the planning target volume (PTV). Nine to ten non-coplanar beams were used for the 3D technique and three non-coplanar arcs were used in both the TI-NC-VMAT and OTI-NC-VMAT plans, with the couch set at ± 20° and 0°, with each arc rotation in excess of 180°. Progressive Resolution Optimizer (PRO) in Varian Eclipse version 11 was used for all of the treatment planning. Conformity Index (CI), conformity number (CN), gradient index (GI), maximum dose at 2 cm away from the PTV (D2cm), mean lung dose (MLD), V20, V5 and mean target dose (MTD) were analyzed for all of the plans. We also performed statistical analysis to examine differences in the dosimetric indices between 3D and VMAT techniques. Dosimetric indices CI, CN, GI, V20 and MTD values were similar, within 5%, for all three plans: 3D-NC-C, TI-NC-VMAT and OTI-NC-VMAT. However, both types of VMAT plans were dosimetrically superior to 3D conformal plans in organ-at-risk (OAR) sparing; D2cm, MLD, and V5 values were significantly lower at 6-8%, 9-12% and 26-30% in VMAT plans, respectively. The OTI-NC-VMAT plans showed equivalent plan quality to the TI-NC-VMAT plans and exhibited robust freedom from limiting arc rotation due to potential patient/couch-gantry collision. The OTI-NC-VMAT plans appear dosimetrically equivalent to TI-NC-VMAT plans for lung SBRT, while permitting large angle arc selection, free from obstructional limitations. Both OTI-NC-VMAT and TI-NC-VMAT plans were dosimetrically superior to 3D-NC-C plans in terms of organ-at-risk (OAR) sparing.


Nanobubble formation on a warmer substrate.
The solvent exchange procedure is an often-used protocol to produce surface nanobubbles. In this procedure, the substrate is exposed to a good solvent for gas which is then mixed and rinsed with a poor solvent for gas and the nanobubbles form on the solid-liquid interface. Here we study the effects of temperatures of the substrate and the first solvent on nanobubble formation. Atomic force microscopy with temperature control was used to examine the formation of nanobubbles at temperatures between 37 °C and 54 °C. It was found that the probability of nanobubble formation was larger on substrates at higher temperatures. Moreover, on warmer substrates we found nanobubbles with lateral extensions up to 8 μm. A morphologic analysis shows that all nanobubbles, including giant nanobubbles, have a similar aspect ratio, independent of the substrate temperature, and that this aspect ratio corresponds to a contact angle between 13° and 22° (on the gas side), much smaller than the macroscopic counterparts. We finally discuss the implications of our results for various theories on nanobubble stability.


Genome-wide association study of lifetime cannabis use based on a large meta-analytic sample of 32 330 subjects from the International Cannabis Consortium.
Cannabis is the most widely produced and consumed illicit psychoactive substance worldwide. Occasional cannabis use can progress to frequent use, abuse and dependence with all known adverse physical, psychological and social consequences. Individual differences in cannabis initiation are heritable (40-48%). The International Cannabis Consortium was established with the aim to identify genetic risk variants of cannabis use. We conducted a meta-analysis of genome-wide association data of 13 cohorts (N=32 330) and four replication samples (N=5627). In addition, we performed a gene-based test of association, estimated single-nucleotide polymorphism (SNP)-based heritability and explored the genetic correlation between lifetime cannabis use and cigarette use using LD score regression. No individual SNPs reached genome-wide significance. Nonetheless, gene-based tests identified four genes significantly associated with lifetime cannabis use: NCAM1, CADM2, SCOC and KCNT2. Previous studies reported associations of NCAM1 with cigarette smoking and other substance use, and those of CADM2 with body mass index, processing speed and autism disorders, which are phenotypes previously reported to be associated with cannabis use. Furthermore, we showed that, combined across the genome, all common SNPs explained 13-20% (P<0.001) of the liability of lifetime cannabis use. Finally, there was a strong genetic correlation (rg=0.83; P=1.85 × 10(-8)) between lifetime cannabis use and lifetime cigarette smoking implying that the SNP effect sizes of the two traits are highly correlated. This is the largest meta-analysis of cannabis GWA studies to date, revealing important new insights into the genetic pathways of lifetime cannabis use. Future functional studies should explore the impact of the identified genes on the biological mechanisms of cannabis use.


RNA and DNA binding of inert oligonuclear ruthenium(II) complexes in live eukaryotic cells.
Confocal microscopy was used to study the intracellular localisation of a series of inert polypyridylruthenium(II) complexes with three eukaryotic cells lines - baby hamster kidney (BHK), human embryonic kidney (HEK-293) and liver carcinoma (Hep-G2). Co-staining experiments with the DNA-selective dye DAPI demonstrated that the di-, tri- and tetra-nuclear polypyridylruthenium(II) complexes that are linked by the bis[4(4'-methyl-2,2'-bipyridyl)]-1,12-dodecane bridging ligand ("bb12") showed a high degree of selectivity for the nucleus of the eukaryotic cells. Additional co-localisation experiments with the general nucleic acid stain SYTO 9 indicated that the ruthenium complexes showed a considerable preference for the RNA-rich nucleolus, rather than chromosomal DNA. No significant differences were observed in the intracellular localisation between the ΔΔ and ΛΛ enantiomers of the dinuclear complex. Cytotoxicity assays carried out over 72 hours indicated that the ruthenium complexes, particularly the tri- and tetra-nuclear species, were significantly toxic to the eukaryotic cells. However, when the activity of the least cytotoxic compound (the ΔΔ enantiomer of the dinuclear species) was determined over a 24 hour period, the results indicated that the ruthenium complex was approximately a 100-fold less toxic to liver and kidney cells than to Gram positive bacteria. Circular dichroism (CD) spectroscopy was used to examine the effect of the ΔΔ and ΛΛ enantiomers of the dinuclear complex on the solution conformations of RNA and DNA. The CD experiments indicated that the RNA maintained the A-type conformation, and the DNA the B-type structure, upon binding by the ruthenium complexes.


International multicenter tool to predict the risk of nonsentinel node metastases in breast cancer.
Axillary treatment of breast cancer patients is undergoing a paradigm shift, as completion axillary lymph node dissections (ALNDs) are being questioned in the treatment of patients with tumor-positive sentinel nodes. This study aims to develop a novel multi-institutional predictive tool to calculate patient-specific risk of residual axillary disease after tumor-positive sentinel node biopsy. Breast cancer patients with a tumor-positive sentinel node and a completion ALND from five European centers formed the original patient series (N = 1000). Statistically significant variables predicting nonsentinel node involvement were identified in logistic regression analysis. A multivariable predictive model was developed and validated by area under the receiver operating characteristics curve (AUC), first internally in 500 additional patients and then externally in 1068 patients from other centers. All statistical tests were two-sided. Nine tumor- and sentinel node-specific variables were identified as statistically significant factors predicting nonsentinel node involvement in logistic regression analysis. A resulting predictive model applied to the internal validation series resulted in an AUC of 0.714 (95% confidence interval [CI] = 0.665 to 0.763). For the external validation series, the AUC was 0.719 (95% CI = 0.689 to 0.750). The model was well calibrated in the external validation series. We present a novel, international, multicenter, predictive tool to assess the risk of additional axillary metastases after tumor-positive sentinel node biopsy in breast cancer. The predictive model performed well in internal and external validation but needs to be further studied in each center before application to clinical use.


Ambiguity in the processing of Mandarin Chinese relative clauses: One factor cannot explain it all.
This study addresses the question of whether native Mandarin Chinese speakers process and comprehend subject-extracted relative clauses (SRC) more readily than object-extracted relative clauses (ORC) in Mandarin Chinese. Presently, this has been a hotly debated issue, with various studies producing contrasting results. Using two eye-tracking experiments with ambiguous and unambiguous RCs, this study shows that both ORCs and SRCs have different processing requirements depending on the locus and time course during reading. The results reveal that ORC reading was possibly facilitated by linear/temporal integration and canonicity. On the other hand, similarity-based interference made ORCs more difficult, and expectation-based processing was more prominent for unambiguous ORCs. Overall, RC processing in Mandarin should not be broken down to a single ORC (dis)advantage, but understood as multiple interdependent factors influencing whether ORCs are either more difficult or easier to parse depending on the task and context at hand.


Coherency-Based Curve Compression for High-Order Finite Element Model Visualization.
Finite element (FE) models are frequently used in engineering and life sciences within time-consuming simulations. In contrast with the regular grid structure facilitated by volumetric data sets, as used in medicine or geosciences, FE models are defined over a non-uniform grid. Elements can have curved faces and their interior can be defined through high-order basis functions, which pose additional challenges when visualizing these models. During ray-casting, the uniformly distributed sample points along each viewing ray must be transformed into the material space defined within each element. The computational complexity of this transformation makes a straightforward approach inadequate for interactive data exploration. In this paper, we introduce a novel coherency-based method which supports the interactive exploration of FE models by decoupling the expensive world-to-material space transformation from the rendering stage, thereby allowing it to be performed within a precomputation stage. Therefore, our approach computes view-independent proxy rays in material space, which are clustered to facilitate data reduction. During rendering, these proxy rays are accessed, and it becomes possible to visually analyze high-order FE models at interactive frame rates, even when they are time-varying or consist of multiple modalities. Within this paper, we provide the necessary background about the FE data, describe our decoupling method, and introduce our interactive rendering algorithm. Furthermore, we provide visual results and analyze the error introduced by the presented approach.


Explosive eruption, flank collapse and megatsunami at Tenerife ca. 170 ka.
Giant mass failures of oceanic shield volcanoes that generate tsunamis potentially represent a high-magnitude but low-frequency hazard, and it is actually difficult to infer the mechanisms and dynamics controlling them. Here we document tsunami deposits at high elevation (up to 132 m) on the north-western slopes of Tenerife, Canary Islands, as a new evidence of megatsunami generated by volcano flank failure. Analyses of the tsunami deposits demonstrate that two main tsunamis impacted the coasts of Tenerife 170 kyr ago. The first tsunami was generated during the submarine stage of a retrogressive failure of the northern flank of the island, whereas the second one followed the debris avalanche of the subaerial edifice and incorporated pumices from an on-going ignimbrite-forming eruption. Coupling between a massive retrogressive flank failure and a large explosive eruption represents a new type of volcano-tectonic event on oceanic shield volcanoes and a new hazard scenario.


Correlation of corrosion and biomechanics in the retrieval of a single modular neck total hip arthroplasty design: modular neck total hip arthroplasty system.
Increased modularity of total hip arthroplasty components has occurred, with theoretical advantages and disadvantages. Recent literature indicates the potential for elevated revision rates of modular neck systems and the potential for local pseudotumor and metallosis formation at the modular neck/stem site. Retrieval analysis of one modular neck implant design including SEM (SCANNING ELECTRON MICROSCOPY) assessment was done and correlated with FEA (finite element analysis) as well as clinical features of patient demographics, implant and laboratory analysis. Correlation of the consistent corrosion locations to FEA indicates that the material and design features of this system may result in a biomechanical reason for failure. The stem aspect of the modular neck/stem junction may be at particular risk.