Exploring influence of subliminal interoception on whole-brain functional network connectivity dynamics.
Recent fMRI studies have highlighted a dynamic relation across large-scale intrinsic connectivity networks (ICNs) of human brain. The origin of such temporal variations in functional connectivity especially during the task-free (resting-state) fMRI is still a matter of debate and ongoing investigation. In this exploratory study, we sought to determine whether subliminal differences in interoception (e.g., distention pressure on the viscera) can influence the dynamics of whole-brain functional network connectivity. A group of healthy right-handed female subjects, close in age (n = 15, mean age ± SD = 30.33 ± 8.7 years) underwent a series of eyes-open resting-state fMRI scans under different interoceptive conditions including catheterization and partial bladder filling. Using a high-dimensional independent component analysis, the functional imaging data were parcellated into 75 components, out of which 33 were identified as non-artifactual ICNs. Changes in dynamic functional network connectivity (dFNC) were evaluated using the sliding-time window approach and k-means clustering algorithm. We used subject medians for each cluster state and compared differences in dFNC correlations using a paired t-test. Following a false discovery rate multiple comparison correction threshold of p<;0.05, no significant differences in dFNC were found. However, different dwell times for each (pseudo-)resting-state were observed. More liberal statistical criteria (uncorrected p<;0.005) also indicated differences in dFNC between ICN pairs especially involving the salience, subcortical, sensorimotor, cerebellar and brainstem networks. Further investigations of the effect of internal (bodily) sensations on the time-varying aspects of functional connectivity can improve our understanding of the nature of temporal fluctuations in interrelations between intrinsic brain networks.


Investigating psychological and physiological responses to the Trier Social Stress Test in young adults with insomnia.
Stress and hyperarousal both contribute to insomnia. Elevated stress-related sleep reactivity is associated with hyperarousal, and might constitute a vulnerability to future insomnia. The present study examined acute stress-induced arousal and its association with nocturnal sleep. Participants were 30 healthy adults (66.7% female, Mage = 26.7 years): 10 with insomnia (INS) and 20 good sleepers with high vulnerability (HV) or low vulnerability (LV) to insomnia. They underwent two consecutive nights of polysomnography. During the evening preceding the second night, the Trier Social Stress Test (TSST) was administered, and psychological and physiological arousal indices were assessed. The TSST elicited an increase in psychological and physiological arousal in all three groups. The INS group showed greater acute cortisol response (p < 0.05) and secretion at bedtime (p < 0.05), and higher pre-sleep cognitive arousal (p < 0.01) than the LV group; HV participants did not significantly differ from those in the INS or the LV group. Increased cortisol response and elevated sympathovagal imbalance (ie, low frequency/high frequency ratio) were each significantly associated with longer nocturnal awakenings (p = 0.048, p = 0.037, respectively). Heightened blood pressure was significantly associated with prolonged sleep onset latency, and reduced total sleep time and sleep efficiency (all ps < 0.05). These findings support the hyperarousal conceptualization of insomnia and indirectly suggest that increased stress reactivity and bedtime hyperarousal might represent a trait-like vulnerability in certain good sleepers. More research is warranted to validate and expand these preliminary findings.


A general setting for halo theory.
We describe a general framework for systematically treating halos that are due to refraction in preferentially oriented ice wedges, and we construct an atlas of such halos. Initially we are constrained neither by the interfacial angles nor the orientations of real ice crystals. Instead we consider "all possible" refraction halos. We therefore make no assumption regarding the wedge angle, and only a weak assumption regarding the allowable wedge orientations. The atlas is thus a very general collection of refraction halos that includes known halos as a small fraction. Each halo in the atlas is characterized by three parameters: the wedge angle, the zenith angle of the spin vector, and the spin vector expressed in the wedge frame. Together with the sun elevation, the three parameter values for a halo not only permit calculation of the halo shape, they also give much information about the halo without extensive calculation, so that often a crude estimate of the halo's appearance is possible merely from inspection of its parameters. As a result, the theory reveals order in what seems initially to be a staggering variety of halo shapes, and in particular it explains why halos look the way they do. Having constructed and studied the atlas, we then see where real or conceivable refraction halos, arising in specific crystal shapes and crystal orientations, fit into the atlas. Although our main goal is to understand halos arising in pyramidal crystals, the results also clarify and unify the classical halos arising in hexagonal prismatic crystals.


Convergent translational evidence of a role for anandamide in amygdala-mediated fear extinction, threat processing and stress-reactivity.
Endocannabinoids are released 'on-demand' on the basis of physiological need, and can be pharmacologically augmented by inhibiting their catabolic degradation. The endocannabinoid anandamide is degraded by the catabolic enzyme fatty acid amide hydrolase (FAAH). Anandamide is implicated in the mediation of fear behaviors, including fear extinction, suggesting that selectively elevating brain anandamide could modulate plastic changes in fear. Here we first tested this hypothesis with preclinical experiments employing a novel, potent and selective FAAH inhibitor, AM3506 (5-(4-hydroxyphenyl)pentanesulfonyl fluoride). Systemic AM3506 administration before extinction decreased fear during a retrieval test in a mouse model of impaired extinction. AM3506 had no effects on fear in the absence of extinction training, or on various non-fear-related measures. Anandamide levels in the basolateral amygdala were increased by extinction training and augmented by systemic AM3506, whereas application of AM3506 to amygdala slices promoted long-term depression of inhibitory transmission, a form of synaptic plasticity linked to extinction. Further supporting the amygdala as effect-locus, the fear-reducing effects of systemic AM3506 were blocked by intra-amygdala infusion of a CB1 receptor antagonist and were fully recapitulated by intra-amygdala infusion of AM3506. On the basis of these preclinical findings, we hypothesized that variation in the human FAAH gene would predict individual differences in amygdala threat-processing and stress-coping traits. Consistent with this, carriers of a low-expressing FAAH variant (385A allele; rs324420) exhibited quicker habituation of amygdala reactivity to threat, and had lower scores on the personality trait of stress-reactivity. Our findings show that augmenting amygdala anandamide enables extinction-driven reductions in fear in mouse and may promote stress-coping in humans.


p73 Is Required for Ovarian Follicle Development and Regulates a Gene Network Involved in Cell-to-Cell Adhesion.
We report that p73 is expressed in ovarian granulosa cells and that loss of p73 leads to attenuated follicle development, ovulation, and corpus luteum formation, resulting in decreased levels of circulating progesterone and defects in mammary gland branching. Ectopic progesterone in p73-deficient mice completely rescued the mammary branching and partially rescued the ovarian follicle development defects. Performing RNA sequencing (RNA-seq) on transcripts from murine wild-type and p73-deficient antral follicles, we discovered differentially expressed genes that regulate biological adhesion programs. Through modulation of p73 expression in murine granulosa cells and transformed cell lines, followed by RNA-seq and chromatin immunoprecipitation sequencing, we discovered p73-dependent regulation of a gene set necessary for cell adhesion and migration and components of the focimatrix (focal intra-epithelial matrix), a basal lamina between granulosa cells that promotes follicle maturation. In summary, p73 is essential for ovarian folliculogenesis and functions as a key regulator of a gene network involved in cell-to-cell adhesion and migration.


Spatial patterns of dengue cases in Brazil.
Dengue infection plays a central role in our society, since it is the most prevalent vector-borne viral disease affecting humans. We statistically investigated patterns concerning the spatial spreading of dengue epidemics in Brazil, as well as their temporal evolution in all Brazilian municipalities for a period of 12 years. We showed that the distributions of cases in municipalities follow power laws persistent in time and that the infection scales linearly with the population of the municipalities. We also found that the average number of dengue cases does not have a clear dependence on the longitudinal position of municipalities. On the other hand, we found that the average distribution of cases varies with the latitudinal position of municipalities, displaying an almost constant growth from high latitudes until reaching the Tropic of Capricorn leveling to a plateau closer to the Equator. We also characterized the spatial correlation of the number of dengue cases between pairs of municipalities, where our results showed that the spatial correlation function decays with the increase of distance between municipalities, following a power-law with an exponential cut-off. This regime leads to a typical dengue traveling distance. Finally, we considered modeling this last behaviour within the framework of a Edwards-Wilkinson equation with a fractional derivative on space.


Targeting Acidity in Pancreatic Adenocarcinoma: Multispectral Optoacoustic Tomography Detects pH-Low Insertion Peptide Probes In Vivo.
pH-low insertion peptides (pHLIP) can serve as a targeting moiety that enables pH-sensitive probes to detect solid tumors. Using these probes in conjunction with multispectral optoacoustic tomography (MSOT) is a promising approach to improve imaging for pancreatic cancer. A pH-sensitive pHLIP (V7) was conjugated to 750 NIR fluorescent dye and evaluated as a targeted probe for pancreatic adenocarcinoma. The pH-insensitive K7 pHLIP served as an untargeted control. Probe binding was assessed in vitro at pH 7.4, 6.8, and 6.6 using human pancreatic cell lines S2VP10 and S2013. Using MSOT, semiquantitative probe accumulation was then assessed in vivo with a murine orthotopic pancreatic adenocarcinoma model. In vitro, the V7-750 probe demonstrated significantly higher fluorescence at pH 6.6 compared with pH 7.4 (S2VP10, P = 0.0119; S2013, P = 0.0160), whereas no difference was observed with the K7-750 control (S2VP10, P = 0.8783; S2013, P = 0.921). In the in vivo S2VP10 model, V7-750 probe resulted in 782.5 MSOT a.u. signal compared with 5.3 MSOT a.u. in K7-750 control in tumor (P = 0.0001). Similarly, V7-750 probe signal was 578.3 MSOT a.u. in the S2013 model compared with K7-750 signal at 5.1 MSOT a.u. (P = 0.0005). There was minimal off-target accumulation of the V7-750 probe within the liver or kidney, and probe distribution was confirmed with ex vivo imaging. Compared with pH-insensitive controls, V7-750 pH-sensitive probe specifically targets pancreatic adenocarcinoma and has minimal off-target accumulation. The noninvasive detection of pH-targeted probes by means of MSOT represents a promising modality to improve the detection and monitoring of pancreatic cancer.


The dual-effects model of social control revisited: relationship satisfaction as a moderator.
The dual-effects model of social control states that receiving social control leads to better health behavior, but also enhances distress in the control recipient. Associated findings, however, are inconsistent. In this study we investigated the role of relationship satisfaction as a moderator of associations of received spousal control with health behavior and affect. In a study with five waves of assessment spanning two weeks to one year following radical prostatectomy (RP), N=109 married or cohabiting prostate-cancer patients repeatedly reported on their pelvic-floor exercise (PFE) to control postsurgery urinary incontinence and affect as primary outcomes, on received PFE-specific spousal control, relationship satisfaction, and covariates. Findings from two-level hierarchical linear models with repeated assessments nested in individuals suggested significant interactions of received spousal control with relationship satisfaction predicting patients' concurrent PFE and positive affect. Patients who were happy with their relationships seemed to benefit from spousal control regarding regular PFE postsurgery while patients less satisfied with their relationships did not. In addition, the latter reported lower levels of positive affect when receiving much spousal control. Results indicate the utility of the inclusion of relationship satisfaction as a moderator of the dual-effects model of social control.


Somatic symptoms beyond those generally associated with a whiplash injury are increased in self-reported chronic whiplash. A population-based cross sectional study: the Hordaland Health Study (HUSK).
Chronic whiplash leads to considerable patient suffering and substantial societal costs. There are two competing hypothesis on the etiology of chronic whiplash. The traditional organic hypothesis considers chronic whiplash and related symptoms a result of a specific injury. In opposition is the hypothesis that chronic whiplash is a functional somatic syndrome, and related symptoms a result of society-induced expectations and amplification of symptoms. According to both hypotheses, patients reporting chronic whiplash are expected to have more neck pain, headache and symptoms of anxiety and depression than the general population. Increased prevalence of somatic symptoms beyond those directly related to a whiplash neck injury is less investigated. The aim of this study was to test an implication derived from the functional hypothesis: Is the prevalence of somatic symptoms as seen in somatization disorder, beyond symptoms related to a whiplash neck injury, increased in individuals self-reporting chronic whiplash? We further aimed to explore recall bias by comparing the symptom profile displayed by individuals self-reporting chronic whiplash to that among those self-reporting a non-functional injury: fractures of the hand or wrist. We explored symptom load, etiologic origin could not be investigated in this study. Data from the Norwegian population-based "Hordaland Health Study" (HUSK, 1997-99); N = 13,986 was employed. Chronic whiplash was self-reported by 403 individuals and fractures by 1,746. Somatization tendency was measured using a list of 17 somatic symptoms arising from different body parts and organ systems, derived from the research criteria for somatization disorder (ICD-10, F45). Chronic whiplash was associated with an increased level of all 17 somatic symptoms investigated (p<0.05). The association was moderately strong (group difference of 0.60 standard deviation), only partly accounted for by confounding. For self-reported fractures symptoms were only slightly elevated. Recent whiplash was more commonly reported than whiplash-injury a long time ago, and the association of interest weakly increased with time since whiplash (r = 0.016, p = 0.032). The increased prevalence of somatic symptoms beyond symptoms expected according to the organic injury model for chronic whiplash, challenges the standard injury model for whiplash, and is indicative evidence of chronic whiplash being a functional somatic syndrome.


Energy demands of diverse spiking cells from the neocortex, hippocampus, and thalamus.
It has long been known that neurons in the brain are not physiologically homogeneous. In response to current stimulus, they can fire several distinct patterns of action potentials that are associated with different physiological classes ranging from regular-spiking cells, fast-spiking cells, intrinsically bursting cells, and low-threshold cells. In this work we show that the high degree of variability in firing characteristics of action potentials among these cells is accompanied with a significant variability in the energy demands required to restore the concentration gradients after an action potential. The values of the metabolic energy were calculated for a wide range of cell temperatures and stimulus intensities following two different approaches. The first one is based on the amount of Na(+) load crossing the membrane during a single action potential, while the second one focuses on the electrochemical energy functions deduced from the dynamics of the computational neuron models. The results show that the thalamocortical relay neuron is the most energy-efficient cell consuming between 7 and 18 nJ/cm(2) for each spike generated, while both the regular and fast spiking cells from somatosensory cortex and the intrinsically-bursting cell from a cat visual cortex are the least energy-efficient, and can consume up to 100 nJ/cm(2) per spike. The lowest values of these energy demands were achieved at higher temperatures and high external stimuli.


Proton currents constrain structural models of voltage sensor activation.
The Hv1 proton channel is evidently unique among voltage sensor domain proteins in mediating an intrinsic 'aqueous' H+ conductance (GAQ). Mutation of a highly conserved 'gating charge' residue in the S4 helix (R1H) confers a resting-state H+ 'shuttle' conductance (GSH) in VGCs and Ci VSP, and we now report that R1H is sufficient to reconstitute GSH in Hv1 without abrogating GAQ. Second-site mutations in S3 (D185A/H) and S4 (N4R) experimentally separate GSH and GAQ gating, which report thermodynamically distinct initial and final steps, respectively, in the Hv1 activation pathway. The effects of Hv1 mutations on GSH and GAQ are used to constrain the positions of key side chains in resting- and activated-state VS model structures, providing new insights into the structural basis of VS activation and H+ transfer mechanisms in Hv1.


Beta subunit of DNA polymerase III holoenzyme is induced upon ultraviolet irradiation or nalidixic acid treatment of Escherichia coli.
Exposure of Escherichia coli to UV irradiation or nalidixic acid, which induce both the SOS and heat shock responses, led to a 3-4-fold increase in the amount of the beta subunit of DNA polymerase III holoenzyme, as assayed by Western blot analysis using anti-beta antibodies. Such an induction was observed also in a delta rpoH mutant lacking the heat shock-specific sigma 32 subunit of RNA polymerase, but it was not observed in recA13 or lexA3 mutants, in which the SOS response cannot be induced. Mapping of transcription initiation sites of the dnaN gene, encoding the beta subunit, using the S1 nuclease protection assay showed essentially no induction of transcription upon UV irradiation, indicating that induction is regulated primarily at the post-transcriptional level. Analysis of translational gene fusions of the dnaN gene, encoding the beta subunit, to the lacZ reporter gene showed induction of beta-galactosidase activity upon UV irradiation of cells harboring the fusion plasmids. Elimination of a 5' flanking DNA sequence in which the dnaN promoters P1 and P2 were located, did not affect the UV inducibility of the gene fusions. Thus, element(s) present from P3 downstream were sufficient for the UV induction. The induction of the dnaN-lacZ gene fusions was dependent on the recA and lexA gene products, but not on the rpoH gene product, in agreement with the immunoblot analysis. The dependence of dnaN induction on the SOS regulators was not mediated via classical repression by the LexA repressor, since the dnaN promoter does not contain a sequence homologous to the LexA binding site, and dnaN mRNA was not inducible by UV light. This suggests that SOS control may be imposed indirectly, by a post-transcriptional mechanism. The increased amount of the beta subunit is needed, most likely, for increased replication and repair activities in cells which have been exposed to UV radiation.


Perceptual and Ventilatory Responses to Hypercapnia in Athletes and Sedentary Individuals.
Hypercapnic chemosensitivity traditionally captures the ventilatory response to elevated pressures of carbon dioxide in the blood. However, hypercapnia also contributes to subjective breathing perceptions, and previously we demonstrated a closer matching of perception to changes in ventilation in athletes compared to controls. Here we investigated any potential underlying hypercapnic chemosensitivity differences between groups, and explored whether these measures relate to ventilatory and perceptual responses during exercise as well as trait levels of affect. A hypercapnic challenge, incremental maximal exercise test and affective questionnaires were completed by 20 endurance athletes and 20 age-/sex-matched sedentary controls. The hypercapnic challenge involved elevating end-tidal PCO2 by 0.8% (6.1 mmHg) and 1.5% (11.2 mmHg) for 3 min each (randomised), with constant end-tidal oxygen. Ventilatory and perceptual responses to hypercapnia were compared between groups, and within each group the relationships between hypercapnic chemosensitivity (slope analyses) and exercising ventilation and perceptions were calculated using Spearman's non-parametric correlations. While absolute ventilation differences during hypercapnia and exercise were observed, no group differences were found across hypercapnic chemosensitivity (slope) measures. Correlation analyses revealed the anxiety hypercapnic response was related to maximal exercise anxiety, but only in sedentary individuals. Ventilatory and perceptual hypercapnic chemosensitivity do not differ between athletes and sedentary individuals. However, ventilatory and anxiety hypercapnic chemosensitivities were related to ventilatory and anxiety responses during exercise in untrained individuals only. Athletes may employ additional strategies during exercise to reduce the influence of chemosensitivity on ventilatory and perceptual responses.


Roseovarius salinarum sp. nov., isolated from a marine solar saltern.
A Gram-stain-negative, rod-shaped, non-motile and halophilic bacterium, designated N53T, was isolated from a marine solar saltern in Wendeng, China. Cells of strain N53T were 0.3-0.4 µm wide and 2.0-5.5 µm long, catalase-positive and oxidase-positive. The bacterium grew optimally at 33 °C, at pH 7.0-8.0 and in the presence of 6.0 % (w/v) NaCl. Bacteriochlorophyll a was not found. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain N53T formed a phylogenetic lineage with members of the genus Roseovarius. Strain N53T exhibited the highest levels of similarity to Roseovarius pacificus (94.6 %) and Roseovarius confluentis (94.6 %), with a lower level to Roseovarius tolerans was 94.0 %. The percentage of conserved proteins and average nucleotide identity values between N53T and the type strain of the type species, Roseovarius tolerans, were 66.1 and 76.4 %, respectively. The genomic DNA G+C content was 68.1 mol%. The sole respiratory quinone was ubiquinone-10. The predominant cellular fatty acids (>10 %) were C18 : 1ω7c (54.0 %) and C16 : 0 (17.9 %). The polar lipids of strain N53T consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, an unidentified aminolipid, two unidentified phospholipids and two unidentified glycolipids. The differential phenotypic properties, together with the chemotaxonomic and genomic distinctiveness, revealed that strain N53T was separate from other recognized species of the genus Roseovarius. On the basis of the data presented here, strain N53T represents a novel species of the genus Roseovarius, for which the name Roseovariussalinarum sp. nov. is proposed. The type strain is N53T (=MCCC 1H00200T=KCTC 52886T).


Biomechanical approaches to understanding the potentially injurious demands of gymnastic-style impact landings.
Gymnasts are exposed to a high incidence of impact landings due to the execution of repeated dismount performances. Biomechanical research can help inform recent discussions surrounding a proposed rule change in potentially injurious gymnastic dismounting. The review examines existing understanding of the mechanisms influencing the impact loads incurred in gymnastic-style landings achieved using biomechanical approaches. Laboratory-based and theoretical modelling research of inherent and regulatory mechanisms is appraised. The integration of the existing insights into injury prevention interventions studies is further considered in the appraisals. While laboratory-based studies have traditionally been favoured, the difficulty in controlling and isolating mechanisms of interest has partially restricted the understanding gained. An increase in the use of theoretical approaches has been evident over the past two decades, which has successfully enhanced insight into less readily modified mechanisms. For example, the important contribution of mass compositions and 'tuned' mass coupling responses to impact loading has been evidenced. While theoretical studies have advanced knowledge in impact landing mechanics, restrictions in the availability of laboratory-based input data have suppressed the benefits gained. The advantages of integrating laboratory-based and theoretical approaches in furthering scientific understanding of loading mechanisms have been recognised in the literature. Since a multi-mechanism contribution to impact loading has been evident, a deviation away from studies examining isolated mechanisms may be supported for the future. A further scientific understanding of the use of regulatory mechanisms in alleviating a performer's inherent injury predisposition may subsequently be gained and used to inform potential rule changes in gymnastics. While the use of controlled studies for providing scientific evidence for the effectiveness of gymnastics injury counter measures has been advocated over the past decade, a lack of information based on randomised controlled studies or actual evaluation of counter measures in the field setting has been highlighted. The subsequent integration of insight into biomechanical risk factors of landing with clinical practice interventions has been recently advocated.


Dynamic Nuclear Polarization enhanced NMR at 187 GHz/284 MHz using an Extended Interaction Klystron amplifier.
A Dynamic Nuclear Polarisation (DNP) enhanced solid-state Magic Angle Spinning (MAS) NMR spectrometer which uses a 187 GHz (corresponding to (1)H NMR frequency of 284 MHz) Extended Interaction Klystron (EIK) amplifier as the microwave source is briefly described. Its performance is demonstrated for a biomolecule (bacteriorhodopsin), a pharmaceutical, and surface functionalised silica. The EIK is very compact and easily incorporated into an existing spectrometer. The bandwidth of the amplifier is sufficient that it obviates the need for a sweepable magnetic field, once set, for all commonly used radicals. The variable power (CW or pulsed) output from the EIK is transmitted to the DNP-NMR probe using a quasi-optic system with a high power isolator and a corrugated waveguide which feeds the microwaves into the DNP-NMR probe. Curved mirrors inside the probe project the microwaves down the axis of the MAS rotor, giving a very efficient system such that maximum DNP enhancement is achieved with less than 3 W output from the microwave source. The DNP-NMR probe operates with a sample temperature down to 90K whilst spinning at 8 kHz. Significant enhancements, in excess of 100 for bacteriorhodopsin in purple membrane (bR in PM), are shown along with spectra which are enhanced by ≈25 with respect to room temperature, for both the pharmaceutical furosemide and surface functionalised silica. These enhancements allow hitherto prohibitively time consuming experiments to be undertaken. The power at which the DNP enhancement in bR in PM saturates does not change significantly between 90K and 170 K even though the enhancement drops by a factor of ≈11. As the DNP build up time decreases by a factor 3 over this temperature range, the reduction in T1n is presumably a significant contribution to the drop in enhancement.


Microsatellite instability in inflammatory bowel disease-associated neoplastic lesions is associated with hypermethylation and diminished expression of the DNA mismatch repair gene, hMLH1.
Twelve to 15% of sporadic colorectal cancers display defective DNA mismatch repair (MMR), manifested as microsatellite instability (MSI). In this group of cancers, promoter hypermethylation of the MMR gene hMLH1 is strongly associated with, and believed to be the cause of, MSI. A subset of colorectal neoplastic lesions arising in inflammatory bowel disease (IBD) is also characterized by MSI. We wished to determine whether hMLH1 hypermethylation was associated with diminished hMLH1 protein expression and MSI in IBD neoplasms. We studied 148 patients with IBD neoplasms, defined as carcinoma or dysplasia occurring in patients with ulcerative colitis or Crohn's disease. MSI was evaluated using multiplex fluorescent PCR to amplify loci D2S123, BAT-25, BAT-26, D5S346, and D17S250 in all cases. Lesions were characterized as high-frequency MSI (MSI-H) if they manifested instability at two or more loci, low-frequency MSI (MSI-L) if unstable at only one locus, or MS-stable (MSS) if showing no instability at any loci. Methylation-specific PCR was performed to determine the methylation status of the hMLH1 promoter region. hMLH1 protein expression was also evaluated by immunohistochemistry. Thirteen (9%) of 148 neoplasms arising in IBD were MSI-H, comprising 11 carcinomas and 2 dysplastic lesions. Sixteen additional lesions (11%) were MSI-L, comprising 11 carcinomas and 5 dysplastic lesions. The remaining 118 neoplasms (80%) were MSS. Six (46%) of 13 MSI-H, 1 (6%) of 16 MSI-L, and 4 (15%) of 27 MSS lesions showed hMLH1 hypermethylation (P = 0.013). Diminished hMLH1 protein expression in neoplastic cell nuclei relative to surrounding normal cell nuclei was demonstrated immunohistochemically in four of four (100%) hypermethylated lesions tested. In IBD neoplasia, hMLH1 promoter hypermethylation occurs frequently in the setting of MSI, particularly MSI-H. Furthermore, hMLH1 hypermethylation and MSI are strongly associated with diminished hMLH1 protein expression in IBD neoplasms. These findings suggest that hMLH1 hypermethylation causes defective DNA MMR in at least a subset of IBD neoplasms.


Q-switched mode locking with acousto-optic modulator in a diode-pumped Nd:YVO4 laser.
The Q-Switched Mode Locking (QML) regime provides the generation of relatively high peak power picosecond pulses train with energies of a few muJ each in a simple resonator. The fully modulated efficient QML regime was demonstrated in the diode-pumped Nd:YVO4 laser. The acousto-optic cell playing a double role of Q-switch and Mode Locker was located near flat output coupler. The two folding mirrors were mounted on the translation stages for matching the resonance frequency of the cavity to the radio frequency of acousto-optic modulator. The QML pulses with envelope durations of 100-150 ns and near 100% modulation depth were observed for wide range of pump powers and repetition rates. Up to 3 W of output average power, 100 muJ of the envelope energy, having approximately 5-8 mode-locked pulses were achieved.


GB virus C/hepatitis G virus replicates in human haematopoietic cells and vascular endothelial cells.
A novel flavivirus, GB virus C (GBV-C)/hepatitis G virus (HGV), has been detected in chronic liver disease patients. It is known that the viral RNA can be detected in approximately 5% of American blood donors. However, the implications for liver disease and the sites of virus replication remain unknown. Possible sites of virus replication were studied by using cell lines and/or primary cells derived from human lymphoid cells, myeloid cells, hepatocytes and endothelial cells. RNA was detected by virus strand-specific RT-PCR and GBV-C/HGV antigen was detected with a rabbit polyclonal anti-E2 (envelope 2) antibody by Western blot analysis. Negative-strand RNA, representative of replicating virus, was detected in lymphoid and megakaryocytoid cell lines and primary vascular endothelial cells. In addition, an increase in virus titre over time was demonstrated and viral antigen was detected, and virus could be passaged to infect fresh cells. However, viral RNA or antigen could not be detected in any of the hepatocyte lines tested. These results indicate that the replication site of GBV-C/HGV is not primarily in hepatocytes and that detection of replicating virus in hepatic tissue may reflect virus replication in haematopoietic cells and/or vascular endothelial cells present in the liver.


High-performance ion-exclusion/cation-exchange chromatography of anions and cations in acid rain waters on a weakly acidic cation-exchange resin.
A new method for the simultaneous determination of anions (sulfate, nitrate, and chloride) and cations (sodium, ammonium, potassium, magnesium, and calcium) in acid rain waters was investigated using high-performance ion-exclusion/cation-exchange chromatography with conductimetric detection on a separation column packed with a polymethacrylate-based weakly acidic cation-exchange resin in the hydrogen-form and an eluent comprising 1.5 mM sulfosalicylic acid-6 mM 18-crown-6 at pH 2.6, operated at 1.5 ml/min. Effective separation and highly sensitive conductimetric detection for the anions and the cations was achieved in about 14 min. Since the ionic balance (equivalents of anions/equivalents of cations) of acid rain waters of different pH (4.40-4.67) ranged from 0.97 to 0.94, evaluation of the water quality of acid rain was possible. This method was successfully applied to the simultaneous determination of the anions and the cations in acid rain transported from mainland China and North Korea to central Japan monitored by a meteorological satellite data analyzer.


Residual amplitude modulation in interferometric gravitational wave detectors.
The effects of residual amplitude modulation (RAM) in laser interferometers using heterodyne sensing can be substantial and difficult to mitigate. In this work, we analyze the effects of RAM on a complex laser interferometer used for gravitational wave detection. The RAM introduces unwanted offsets in the cavity length signals and thereby shifts the operating point of the optical cavities from the nominal point via feedback control. This shift causes variations in the sensing matrix, and leads to degradation in the performance of the precision noise subtraction scheme of the multiple-degree-of-freedom control system. In addition, such detuned optical cavities produce an optomechanical spring, which also perturbs the sensing matrix. We use our simulations to derive requirements on RAM for the Advanced LIGO (aLIGO) detectors, and show that the RAM expected in aLIGO will not limit its sensitivity.


Differential Expression of Vascular Endothelial Growth Factor-A165 Isoforms Between Intracranial Atherosclerosis and Moyamoya Disease.
Vascular endothelial growth factor-A165 (VEGF-A165) has been identified as a combination of 2 alternative splice variants: proangiogenic VEGF-A165a and antiangiogenic VEGF-A165b. Intracranial atherosclerotic disease (ICAD) and moyamoya disease (MMD) are 2 main types of intracranial arterial steno-occlusive disorders with distinct capacities for collateral formation. Recent studies indicate that VEGF-A165 regulates collateral growth in ischemia. Therefore, we investigated if there is a distinctive composition of VEGF-A165 isoforms in ICAD and MMD. Sixty-six ICAD patients, 6 MMD patients, and 5 controls were enrolled in this prospective study. ICAD and MMD patients received intensive medical management upon enrollment. Surgery was offered to 9 ICAD patients who had recurrent ischemic events, 6 MMD patients, and 5 surgical controls without ICAD. VEGF-A165a and VEGF-A165b plasma levels were measured at baseline, within 1 week after patients having surgery, and at 1, 3, and 6 months after treatment. A significantly higher baseline VEGF-A165a/b ratio was observed in MMD compared to ICAD (P = .016). The VEGF-A165a/b ratio increased significantly and rapidly after surgical treatment in ICAD (P = .026) more so than in MMD and surgical controls. In patients with ICAD receiving intensive medical management, there was also an elevation of the VEGF-A165a/b ratio, but at a slower rate, reaching the peak at 3 months after initiation of treatment (baseline versus 3 months VEGF-A165a/b ratio, P = .028). Our study shows an increased VEGF-A165a/b ratio in MMD compared to ICAD, and suggests that both intensive medical management and surgical revascularization elevate the VEGF-A165a/b ratio in ICAD patients.


The Open System Interconnection as a building block in a health sciences information network.
The interconnection of integrated health sciences library systems with other health sciences computer systems to achieve information networks will require either custom linkages among specific devices or the adoption of standards that all systems support. The most appropriate standards appear to be those being developed under the Open System Interconnection (OSI) reference model, which specifies a set of rules and functions that computers must follow to exchange information. The protocols have been modularized into seven different layers. The lowest three layers are generally available as off-the-shelf interfacing products. The higher layers require special development for particular applications. This paper describes the OSI, its application in health sciences networks, and specific tasks that remain to be undertaken.


Generation, Characterization, and Application of Hierarchically Structured Self-Assembly Induced by the Combined Effect of Self-Emulsification and Phase Separation.
Hierarchically structured magnetic single-hole hollow spheres (MSHS) have been successfully obtained via a facile self-assembly strategy. This methodology allows the double emulsions generated via the combined effect of self-emulsification and phase separation to provide confinement for directing the self-assembly of magnetic nanoparticles (MNPs). The resulting MSHS fully capitalize on both the multifunctional properties of MNPs and container features of single-hole hollow spheres. Moreover, the magnetic properties showed obvious improvement and can be tuned by modulating the assembled structure. Thus, MSHS can be used as a smart platform with multiple functionalities including image contrast enhancement, selective encapsulation for biomacromolecules, on-demand release, and magnetically guided transport. This strategy is very promising in the design of hierarchically structured assemblies for desired applications in biomedicine and other fields.


Parental inheritance and psychological disability in fragile X females.
Studies of adult female carriers of the fragile X chromosome indicate that certain psychological problems occur with a greater frequency and severity than expected. This study examines the association of parental origin of the fragile X chromosome and of fragility detected in the karyotype with measures of social, educational, and psychological functioning in a group of adult fragile X females of normal intelligence. The results show that, as a group, women who inherit the fragile X chromosome from their mother and who demonstrate positive fragility in the karyotype (MI+ group = [maternal inheritance with positive fragility]) manifest significantly more impairment of social, educational, and psychological functioning when compared with fragile X females with paternal inheritance or negative fragility or with a matched control group comprising non-fragile X women. In particular, MI+ women show lower levels of both educational achievement and socioeconomic status and a greater degree of disturbance in communication, socialization, affect, and thought processes. These clinical findings are consistent with the recently advanced hypothesis which proposes that a two-stage process leading to chromosome imprinting in a preoogonial cell causes the fragile X syndrome.


Structure-function-folding relationships and native energy landscape of dynein light chain protein: nuclear magnetic resonance insights.
The detailed characterization of the structure, dynamics and folding process of a protein is crucial for understanding the biological functions it performs. Modern biophysical and nuclear magnetic resonance (NMR) techniques have provided a way to obtain accurate structural and thermodynamic information on various species populated on the energy landscape of a given protein. In this context, we review here the structure-function-folding relationship of an important protein, namely, dynein light chain protein (DLC8). DLC8, the smallest subunit of the dynein motor complex, acts as a cargo adaptor. The protein exists as a dimer under physiological conditions and dissociates into a pure monomer below pH 4. Cargo binding occurs at the dimer interface. Dimer stability and relay of perturbations through the dimer interface are anticipated to be playing crucial roles in the variety of functions the protein performs. NMR investigations have provided great insights into these aspects of DLC8 in recent years.


A widespread bacterial phenazine forms S-conjugates with biogenic thiols and crosslinks proteins.
Phenazines are redox-active compounds produced by a range of bacteria, including many pathogens. Endowed with various biological activities, these ubiquitous N-heterocycles are well known for their ability to generate reactive oxygen species by redox cycling. Phenazines may lead to an irreversible depletion of glutathione, but a detailed mechanism has remained elusive. Furthermore, it is not understood why phenazines have so many protein targets and cause protein misfolding as well as their aggregation. Here we report the discovery of unprecedented conjugates (panphenazines A, B) of panthetheine and phenazine-1-carboxylic (PCA) acid from a Kitasatospora sp., which prompted us to investigate their biogenesis. We found that PCA reacts with diverse biogenic thiols under radical-forming conditions, which provides a plausible model for irreversible glutathione depletion. To evaluate the scope of the reaction in cells we designed biotin and rhodamine conjugates for protein labelling and examined their covalent fusion with model proteins (ketosynthase, carbonic anhydrase III, albumin). Our results reveal important, yet overlooked biological roles of phenazines and show for the first time their function in protein conjugation and crosslinking.


Infective endocarditis: improving the diagnostic yield.
Isolating aetiological agents in patients with infective endocarditis (IE) remains problematical. We postulated that the high local incidence of culture-negative IE resulted from antibiotic exposure prior to blood cultures and that a structured delay in therapy in the subacute presentation would improve the diagnostic yield. We aimed to prospectively observe the diagnostic approach and give an overview of supplementary laboratory tests. Patients with suspected IE were enrolled into this analytical observational study and followed up for six months (n = 92). We compared the diagnostic yield and outcome in cases where antibiotics were withheld for 72 hours, with those cases who received early antimicrobials, despite being deemed safe for delayed therapy. Definitive diagnoses (definite or excluded IE) were made in 92.8% of patients where antibiotics were delayed, compared to 60% of patients who received empirical treatment (p = 0.08). The mortality rates were 18.4% and 30.0% (p = 0.18). Twenty-three of 26 patients with definite culture-negative IE received antibiotics during the 48 hours preceding cultures, compared to eight of 21 culture-positive patients (P < 0.001). Screening for atypical bacteria did not improve the yield. C-reactive protein (CRP) had a sensitivity of 97.0% (negative predictive value 87.5%), whereas a positive rheumatoid factor (RF) had a specificity of 93.8% (positive predictive value 91.7%). We observed tendencies towards a greater diagnostic yield and lower mortality where antibiotics were initially withheld. Antibiotic prior to blood cultures were an important cause of culture-negative IE. A normal CRP proved useful in excluding IE; a positive RF strongly favoured IE.


Randomized, placebo-controlled trial of the anti-tumor necrosis factor antibody fragment afelimomab in hyperinflammatory response during severe sepsis: The RAMSES Study.
This study investigated whether treatment with the anti-tumor necrosis factor-alpha monoclonal antibody afelimomab would improve survival in septic patients with serum interleukin (IL)-6 concentrations of >1000 pg/mL. Multicenter, double-blind, randomized, placebo-controlled study. Eighty-four intensive care units in academic medical centers in Europe and Israel. A total of 944 septic patients were screened and stratified by the results of a rapid qualitative immunostrip test for serum IL-6 concentrations. Patients with a positive test kit result indicating IL-6 concentrations of >1000 pg/mL were randomized to receive either afelimomab (n = 224) or placebo (n = 222). Patients with a negative IL-6 test (n = 498) were not randomized and were followed up for 28 days. Treatment consisted of 15-min infusions of 1 mg/kg afelimomab or matching placebo every 8 hrs for 3 days. Standard surgical and intensive care therapy was otherwise delivered. The study was terminated prematurely after an interim analysis estimated that the primary efficacy end points would not be met. The 28-day mortality rate in the nonrandomized patients (39.6%, 197 of 498) was significantly lower (p <.001) than that found in the randomized patients (55.8%, 249 of 446). The mortality rates in the IL-6 test kit positive patients randomized to afelimomab and placebo were similar, 54.0% (121 of 224) vs. 57.7% (128 of 222), respectively. Treatment with afelimomab was not associated with any particular adverse events. The IL-6 immunostrip test identified two distinct sepsis populations with significantly different mortality rates. A small (3.7%) absolute reduction in mortality rate was found in the afelimomab-treated patients. The treatment difference did not reach statistical significance.


The Epidemiology and Management of Exertional Heat Illnesses in High School Sports During the 2012/2013–2016/2017 Academic Years.
Context: Recent data on exertional heat illness (EHI) in high school sports are limited yet warranted to identify specific settings with the highest risk of EHI. Objective: To describe the epidemiology of EHI in high school sports during the 2012/2013-2016/2017 academic years. Design: Descriptive epidemiology study. Setting: Aggregate injury and exposure data collected from athletic trainers working in high school sports in the United States. Patients or Other Participants: High school athletes during the 2012/2013-2016/2017 academic years. Intervention: High School Reporting Information Online surveillance system data from the 2012/2013-2016/2017 academic years were analyzed. Main Outcome Measures: EHI counts, rates per 10,000 athlete exposures (AEs), and distributions were examined by sport, event type, and US census region. EHI management strategies provided by athletic trainers were analyzed. Injury rate ratios with 95% confidence intervals (CIs) compared EHI rates. Results: Overall, 300 EHIs were reported for an overall rate of 0.13/10,000 AE (95% CI, 0.11 to 0.14). Of these, 44.3% occurred in American football preseason practices; 20.7% occurred in American football preseason practices with a registered air temperature ≥90°F and ≥1 hour into practice. The EHI rate was higher in American football than all other sports (0.52 vs 0.04/10,000 AE; injury rate ratio = 11.87; 95% CI, 9.22 to 15.27). However, girls' cross-country had the highest competition EHI rate (1.18/10,000 AE). The EHI rate was higher in the South US census region than all other US census regions (0.23 vs 0.08/10,000 AE; injury rate ratio = 2.96; 95% CI, 2.35 to 3.74). Common EHI management strategies included having medical staff on-site at the onset of EHI (92.7%), removing athlete from play (85.0%), and giving athlete fluids via the mouth (77.7%). Conclusions: American football continues to have the highest overall EHI rate although the high competition EHI rate in girls' cross-country merits additional examination. Regional differences in EHI incidence, coupled with sport-specific variations in management, may highlight the need for region- and sport-specific EHI prevention guidelines.


Imaging mass spectrometry of proteins and peptides: 3D volume reconstruction.
As large genomic and proteomic datasets are generated from homogenates of various tissues, the need for information on the spatial localization of their encoded products has become more pressing. Matrix-assisted laser desorption-ionization (MALDI) imaging mass spectrometry (IMS) offers investigators the means with which to unambiguously study peptides and proteins with molecular specificity, and to determine their distribution in two and three dimensions. In the past few years, several parameters have been optimized for IMS, including sample preparation, matrix application and instrumental acquisition parameters (Box 1). These developments have resulted in a high degree of reproducibility in mass accuracy and peak intensities (Supplementary Fig. 1 online). Recently, we have optimized our protocol to be able to increase the number of molecular species analyzed by collecting two sets of sections, covering one set of sections with sinapinic acid for optimal detection of proteins and adjacent sections with 2,5-dihydroxybenzoic acid (DHB) matrix for the optimal detection of low-mass species, including peptides. Approximately 1,000 peaks can be observed in each dataset (Fig. 1). Furthermore, the sections are collected at an equal distance, 200 mum instead of 400-500 mum used previously, thus enabling the use of virtual z-stacks and three-dimensional (3D) volume renderings to investigate differential localization patterns in much smaller brain structures such as the substantia nigra and the interpeduncular nucleus. Here we present our optimized step-by-step procedure based on previous work in our laboratory, describing how to make 3D volume reconstructions of MALDI IMS data, as applied to the rat brain.


Cooking and Meal Planning as Predictors of Fruit and Vegetable Intake and BMI in First-Year College Students.
The objective was to determine if cooking skills and meal planning behaviors are associated with greater fruit and vegetable intake and lower body mass index (BMI) in first-year college students who are at risk for excessive weight gain. A cross-sectional analysis was conducted using baseline data from a multi-state research project aimed at preventing weight gain in first-year college students. Cooking type, frequency and confidence, self-instruction for healthful mealtime behavior intention, self-regulation of healthful mealtime behavior, and cup equivalents of fruits and vegetables (FV) were measured using validated surveys. BMI was calculated from measured height and weight. First-year students (n = 1108) considered at risk for weight gain from eight universities completed baseline assessments within the first month of entering college. Multiple linear regression was used to determine associations among independent variables of cooking patterns, meal planning behaviors, and dependent variables of fruit and vegetable intake and BMI, after controlling for the influence of sex. Cooking more frequently, cooking with greater skills, and practicing meal planning behaviors are associated with greater fruit and vegetable intake and lower BMI in first-year college students. Interventions aimed at improving health in college students may be enhanced by incorporating cooking and meal planning components.


Relation between temperature and mortality in thirteen Spanish cities.
In this study we examined the shape of the association between temperature and mortality in 13 Spanish cities representing a wide range of climatic and socio-demographic conditions. The temperature value linked with minimum mortality (MMT) and the slopes before and after the turning point (MMT) were calculated. Most cities showed a V-shaped temperature-mortality relationship. MMTs were generally higher in cities with warmer climates. Cold and heat effects also depended on climate: effects were greater in hotter cities but lesser in cities with higher variability. The effect of heat was greater than the effect of cold. The effect of cold and MMT was, in general, greater for cardio-respiratory mortality than for total mortality, while the effect of heat was, in general, greater among the elderly.


Core filaments of the nuclear matrix.
The nuclear matrix is concealed by a much larger mass of chromatin, which can be removed selectively by digesting nuclei with DNase I followed by elution of chromatin with 0.25 M ammonium sulfate. This mild procedure removes chromatin almost completely and preserves nuclear matrix morphology. The complete nuclear matrix consists of a nuclear lamina with an interior matrix composed of thick, polymorphic fibers and large masses that resemble remnant nucleoli. Further extraction of the nuclear matrices of HeLa or MCF-7 cells with 2 M sodium chloride uncovered a network of core filaments. A few dark masses remained enmeshed in the filament network and may be remnants of the nuclear matrix thick fibers and nucleoli. The highly branched core filaments had diameters of 9 and 13 nm measured relative to the intermediate filaments. They may serve as the core structure around which the matrix is constructed. The core filaments retained 70% of nuclear RNA. This RNA consisted both of ribosomal RNA precursors and of very high molecular weight hnRNA with a modal size of 20 kb. Treatment with RNase A removed the core filaments. When 2 M sodium chloride was used directly to remove chromatin after DNase I digestion without a preceding 0.25 M ammonium sulfate extraction, the core filaments were not revealed. Instead, the nuclear interior was filled with amorphous masses that may cover the filaments. This reflected a requirement for a stepwise increase in ionic strength because gradual addition of sodium chloride to a final concentration of 2 M without an 0.25 M ammonium sulfate extraction uncovered core filaments.


DNA methylation patterns are associated with n-3 fatty acid intake in Yup'ik people.
A large body of evidence links a high dietary intake of n-3 (ω-3) polyunsaturated fatty acids (PUFAs) with improved cardiometabolic outcomes. Recent studies suggested that the biologic processes underlying the observed associations may involve epigenetic changes, specifically DNA methylation. To evaluate changes in methylation associated with n-3 PUFA intake, we conducted an epigenome-wide methylation association study of long-chain n-3 PUFA intake and tested associations between the diabetes- and cardiovascular disease-related traits. We assessed DNA methylation at ∼470,000 cytosine-phosphate-guanine (CpG) sites in a cross-sectional study of 185 Yup'ik Alaska Native individuals representing the top and bottom deciles of PUFA intake. Linear regression models were used to test for the associations of interest, adjusting for age, sex, and community group. We identified 27 differentially methylated CpG sites at biologically relevant regions that reached epigenome-wide significance (P < 1 × 10⁻⁷). Specifically, regions on chromosomes 3 (helicase-like transcription factor), 10 (actin α 2 smooth muscle/Fas cell surface death receptor), and 16 (protease serine 36/C16 open reading frame 67) each harbored 2 significant correlates of n-3 PUFA intake. In conclusion, we present promising evidence of association between several biologically relevant epigenetic markers and long-term intake of marine-derived n-3 PUFAs.


Methylated lysine 79 of histone H3 targets 53BP1 to DNA double-strand breaks.
The mechanisms by which eukaryotic cells sense DNA double-strand breaks (DSBs) in order to initiate checkpoint responses are poorly understood. 53BP1 is a conserved checkpoint protein with properties of a DNA DSB sensor. Here, we solved the structure of the domain of 53BP1 that recruits it to sites of DSBs. This domain consists of two tandem tudor folds with a deep pocket at their interface formed by residues conserved in the budding yeast Rad9 and fission yeast Rhp9/Crb2 orthologues. In vitro, the 53BP1 tandem tudor domain bound histone H3 methylated on Lys 79 using residues that form the walls of the pocket; these residues were also required for recruitment of 53BP1 to DSBs. Suppression of DOT1L, the enzyme that methylates Lys 79 of histone H3, also inhibited recruitment of 53BP1 to DSBs. Because methylation of histone H3 Lys 79 was unaltered in response to DNA damage, we propose that 53BP1 senses DSBs indirectly through changes in higher-order chromatin structure that expose the 53BP1 binding site.


Chinese red yeast rice (Monascus purpureus) for primary hyperlipidemia: a meta-analysis of randomized controlled trials.
Extracts of Chinese red yeast rice (RYR, a traditional dietary seasoning of Monascus purpureus) contains several active ingredients including lovastatin, and several trials of its possible lipid-lowering effects have been conducted. This meta-analysis assesses the effectiveness and safety of RYR preparations on lipid modification in primary hyperlipidemia. We included randomized controlled trials testing RYR preparation, compared with placebo, no treatment, statins, or other active lipid-lowering agents in people with hyperlipidemia through searching PubMed, CBMdisk, TCMLARS, the Cochrane Library, and AMED up to December 2004. Ninety-three randomized trials (9625 participants) were included and three RYR preparations (Cholestin, Xuezhikang and Zhibituo) were tested. The methodological quality of trial reports was generally low in terms of generation of the allocation sequence, allocation concealment, blinding, and intention-to-treat. The combined results showed significant reduction of serum total cholesterol levels (weighted mean difference -0.91 mmol/L, 95% confidence interval -1.12 to -0.71), triglycerides levels (-0.41 mmol/L, -0.6 to -0.22), and LDL-cholesterol levels (-0.73 mmol/L, -1.02 to -0.043), and increase of HDL-cholesterol levels (0.15 mmol/L, 0.09 to 0.22) by RYR treatment compared with placebo. The lipid modification effects appeared to be similar to pravastatin, simvastatin, lovastatin, atorvastatin, or fluvastatin. Compared with non-statin lipid lowering agents, RYR preparations appeared superior to nicotinate and fish oils, but equal to or less effective than fenofibrate and gemfibrozil. No significant difference in lipid profile was found between Xuezhikang and Zhibituo. RYR preparations were associated with non-serious adverse effects such as dizziness and gastrointestinal discomfort. Current evidence shows short-term beneficial effects of RYR preparations on lipid modification. More rigorous trials are needed, and long-term effects and safety should be investigated if RYR preparations are to be recommended as one of the alternative treatments for primary hyperlipidemia.


Predicting suicide risk in 137,112 people with severe mental illness in Finland: external validation of the Oxford Mental Illness and Suicide tool (OxMIS).
Oxford Mental Illness and Suicide tool (OxMIS) is a standardised, scalable, and transparent instrument for suicide risk assessment in people with severe mental illness (SMI) based on 17 sociodemographic, criminal history, familial, and clinical risk factors. However, alongside most prediction models in psychiatry, external validations are currently lacking. We utilised a Finnish population sample of all persons diagnosed by mental health services with SMI (schizophrenia-spectrum and bipolar disorders) between 1996 and 2017 (n = 137,112). To evaluate the performance of OxMIS, we initially calculated the predicted 12-month suicide risk for each individual by weighting risk factors by effect sizes reported in the original OxMIS prediction model and converted to a probability. This probability was then used to assess the discrimination and calibration of the OxMIS model in this external sample. Within a year of assessment, 1.1% of people with SMI (n = 1475) had died by suicide. The overall discrimination of the tool was good, with an area under the curve of 0.70 (95% confidence interval: 0.69-0.71). The model initially overestimated suicide risks in those with elevated predicted risks of >5% over 12 months (Harrell's Emax = 0.114), which applied to 1.3% (n = 1780) of the cohort. However, when we used a 5% maximum predicted suicide risk threshold as is recommended clinically, the calibration was excellent (ICI = 0.002; Emax = 0.005). Validating clinical prediction tools using routinely collected data can address research gaps in prediction psychiatry and is a necessary step to translating such models into clinical practice.


Predictors of outcome in routine care for Cryptococcal meningitis in Western Kenya: lessons for HIV outpatient care in resource-limited settings.
Cryptococcal meningitis is a leading cause of mortality among HIV-infected individuals in sub-Saharan Africa but little is known about its treatment and outcomes in decentralised HIV outpatient settings. We assessed adherence to treatment guidelines and determined predictors of survival. A computerised laboratory database identified HIV-infected adults with cryptococcal meningitis at Family AIDS Care and Education Services in Nyanza Province, Kenya, between 2005-2009. Medical records were reviewed. Kaplan-Meier survival curves were generated. Bivariate and multivariate Cox proportional hazards models were used to determine associations between key clinical characteristics and survival. Medical records were located for 79% (71/90). Mortality was 38% (27/71) over a median follow-up period of 201 days (IQR: 10-705 days). Adherence to local guidelines for treatment of cryptococcal meningitis was 48% (34/71). Higher body mass index was associated with improved survival (HR: 0.82, 95% CI (0.68 to 0.99)) even after controlling for factors such as age, CD4 cell count, receipt of highly active anti-retroviral therapy, and treatment with any anti-fungal therapy. Cryptococcal meningitis diagnosed in routine HIV outpatient settings is largely treated as an outpatient and adherence to treatment guidelines is poor. Body mass index is a critical independent predictor of outcome. Additional research to determine the most effective strategies to reduce premature mortality is urgently needed.


Pain in Paget's disease: a retrospective study of treatment efficacy.
The authors addressed the role and the management of pain in Paget's disease by a retrospective study. The objectives were: to assess the presence of pain in Paget's disease; to look for a relationship between pain and the levels of total alkaline phosphatase (total ALP); to verify if the most commonly used drugs in Paget's disease, calcitonin and bisphosphonates, were able to reduce the pain and the levels of total ALP. The study analyzed 107 Italian patients with Paget's disease who were hospitalized at the same Institute between 1970 and 2010; all patients affected by severe arthritis were excluded. From the analysis of the clinical records it emerged that as many as 85% of patients had pain and that total ALP was also increased in most of the patients with pain in comparison with patients without pain. The clinical and metabolic effects of different therapies were then assessed: many patients had not received any specific therapy (58%), others had been treated with calcitonin (25%) and others with bisphosphonates (17%). In fact, the patients treated with bisphosphonates had significantly lower levels both of pain and total ALP. The authors hypothesize that the pain in Paget's disease has a primary origin and is correlated to the degree of bone metabolic hyperactivity. Finally, treatment with bisphosphonates appeared to be the most appropriate treatment, having been able to control both the pain and the metabolic hyperactivity.


Antibodies Against Immune Checkpoint Molecules Restore Functions of Tumor-Infiltrating T Cells in Hepatocellular Carcinomas.
Ligand binding to inhibitory receptors on immune cells, such as programmed cell death 1 (PD-1) and cytotoxic T-lymphocyte associated protein 4 (CTLA4), down-regulates the T-cell-mediated immune response (called immune checkpoints). Antibodies that block these receptors increase antitumor immunity in patients with melanoma, non-small-cell lung cancer, and renal cell cancer. Tumor-infiltrating CD4+ and CD8+ T cells in patients with hepatocellular carcinoma (HCC) have been found to be functionally compromised. We analyzed HCC samples from patients to determine if these inhibitory pathways prevent T-cell responses in HCCs and to find ways to restore their antitumor functions. We collected HCC samples from 59 patients who underwent surgical resection from November 2013 through May 2017, along with tumor-free liver tissues (control tissues) and peripheral blood samples. We isolated tumor-infiltrating lymphocytes (TIL) and intra-hepatic lymphocytes. We used flow cytometry to quantify expression of the inhibitory receptors PD-1, hepatitis A virus cellular receptor 2 (TIM3), lymphocyte activating 3 (LAG3), and CTLA4 on CD8+ and CD4+ T cells from tumor, control tissue, and blood; we studied the effects of antibodies that block these pathways in T-cell activation assays. Expression of PD-1, TIM3, LAG3, and CTLA4 was significantly higher on CD8+ and CD4+ T cells isolated from HCC tissue than control tissue or blood. Dendritic cells, monocytes, and B cells in HCC tumors expressed ligands for these receptors. Expression of PD-1, TIM3, and LAG3 was higher on tumor-associated antigen (TAA)-specific CD8+ TIL, compared with other CD8+ TIL. Compared with TIL that did not express these inhibitory receptors, CD8+ and CD4+ TIL that did express these receptors had higher levels of markers of activation, but similar or decreased levels of granzyme B and effector cytokines. Antibodies against CD274 (PD-ligand1 [PD-L1]), TIM3, or LAG3 increased proliferation of CD8+ and CD4+ TIL and cytokine production in response to stimulation with polyclonal antigens or TAA. Importantly, combining antibody against PD-L1 with antibodies against TIM3, LAG3, or CTLA4 further increased TIL functions. The immune checkpoint inhibitory molecules PD-1, TIM3, and LAG3 are up-regulated on TAA-specific T cells isolated from human HCC tissues, compared with T cells from tumor-free liver tissues or blood. Antibodies against PD-L1, TIM3, or LAG3 restore responses of HCC-derived T cells to tumor antigens, and combinations of the antibodies have additive effects. Strategies to block PD-L1, TIM3, and LAG3 might be developed for treatment of primary liver cancer.


The NYCKidSeq project: study protocol for a randomized controlled trial incorporating genomics into the clinical care of diverse New York City children.
Increasingly, genomics is informing clinical practice, but challenges remain for medical professionals lacking genetics expertise, and in access to and clinical utility of genomic testing for minority and underrepresented populations. The latter is a particularly pernicious problem due to the historical lack of inclusion of racially and ethnically diverse populations in genomic research and genomic medicine. A further challenge is the rapidly changing landscape of genetic tests and considerations of cost, interpretation, and diagnostic yield for emerging modalities like whole-genome sequencing. The NYCKidSeq project is a randomized controlled trial recruiting 1130 children and young adults predominantly from Harlem and the Bronx with suspected genetic disorders in three disease categories: neurologic, cardiovascular, and immunologic. Two clinical genetic tests will be performed for each participant, either proband, duo, or trio whole-genome sequencing (depending on sample availability) and proband targeted gene panels. Clinical utility, cost, and diagnostic yield of both testing modalities will be assessed. This study will evaluate the use of a novel, digital platform (GUÍA) to digitize the return of genomic results experience and improve participant understanding for English- and Spanish-speaking families. Surveys will collect data at three study visits: baseline (0 months), result disclosure visit (ROR1, + 3 months), and follow-up visit (ROR2, + 9 months). Outcomes will assess parental understanding of and attitudes toward receiving genomic results for their child and behavioral, psychological, and social impact of results. We will also conduct a pilot study to assess a digital tool called GenomeDiver designed to enhance communication between clinicians and genetic testing labs. We will evaluate GenomeDiver's ability to increase the diagnostic yield compared to standard practices, improve clinician's ability to perform targeted reverse phenotyping, and increase the efficiency of genetic testing lab personnel. The NYCKidSeq project will contribute to the innovations and best practices in communicating genomic test results to diverse populations. This work will inform strategies for implementing genomic medicine in health systems serving diverse populations using methods that are clinically useful, technologically savvy, culturally sensitive, and ethically sound. ClinicalTrials.gov NCT03738098 . Registered on November 13, 2018 Trial Sponsor: Icahn School of Medicine at Mount Sinai Contact Name: Eimear Kenny, PhD (Principal Investigator) Address: Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Pl., Box 1003, New York, NY 10029 Email: firstname.lastname@example.com.


Using Think Aloud Protocols to Assess E-Prescribing in Community Pharmacies.
Think aloud protocol has rarely been used as a method of data collection in community pharmacies. The aim of the report is to describe how think aloud protocols were used to identify issues that arise when using e-prescribing technology in pharmacies. In this paper, we report on the benefits and challenges of using think aloud protocols in pharmacies to examine the use of e-prescribing systems. Sixteen pharmacists and pharmacy technicians were recruited from seven community pharmacies in Wisconsin. Data were collected using direct observation alongside think aloud protocol. Direct observations and think aloud protocols took place between January-February, 2011. Participants were asked to verbalize their thoughts as they process electronic prescriptions. Participants identify weaknesses in e-prescribing that they had previously not conceived. This created heightened awareness for vigilance when processing e-prescriptions. The main challenge with using think aloud protocols were interruptions in the pharmacy. Some participants found it difficult to remember to continue verbalizing during think aloud sessions. The use of think aloud protocols as method of data collection is a new way for understanding the issues related to technology use in community pharmacy practice. Think aloud protocol was beneficial in providing objective information on e-prescribing not based on pharmacist's or technician's opinion of the technology. This method provided detailed information and also a wide variety of real time challenges with e-prescribing technology in community pharmacies. Using this data collection method can help identify potential patient safety issues when using e-prescribing and suggestions for redesign.


Proteomic analysis defines kinase taxonomies specific for subtypes of breast cancer.
Multiplexed small molecule inhibitors covalently bound to Sepharose beads (MIBs) were used to capture functional kinases in luminal, HER2-enriched and triple negative (basal-like and claudin-low) breast cancer cell lines and tumors. Kinase MIB-binding profiles at baseline without perturbation proteomically distinguished the four breast cancer subtypes. Understudied kinases, whose disease associations and pharmacology are generally unexplored, were highly represented in MIB-binding taxonomies and are integrated into signaling subnetworks with kinases that have been previously well characterized in breast cancer. Computationally it was possible to define subtypes using profiles of less than 50 of the more than 300 kinases bound to MIBs that included understudied as well as metabolic and lipid kinases. Furthermore, analysis of MIB-binding profiles established potential functional annotations for these understudied kinases. Thus, comprehensive MIBs-based capture of kinases provides a unique proteomics-based method for integration of poorly characterized kinases of the understudied kinome into functional subnetworks in breast cancer cells and tumors that is not possible using genomic strategies. The MIB-binding profiles readily defined subtype-selective differential adaptive kinome reprogramming in response to targeted kinase inhibition, demonstrating how MIB profiles can be used in determining dynamic kinome changes that result in subtype selective phenotypic state changes.


Human papillomavirus and cancer: the epidemiological evidence.
Summary of the studies carried out by the IARC on HPV and cervical cancer is presented. the first one was the international prevalence survey of HPV types in invasive cervical cancer (ICC) conducted in up to 22 countries. Overall, 99.7% of 1000 cases with histologically confirmed ICC were also shown to be HPV DNA positive using the GP5+/GP6+ or E7 primers, indicating that HPV is a necessary cause of cervical cancer. The most prevalent HPV types were HPV 16 (53%), HPV 18 (15%), HPV 45 (9%), HPV 31 (6%) and HPV 33 (3%). HPV 16 was the most common type in all geographical regions, followed by HPV 18 that was particularly, common in South-East Asia. The second set of studies included case-control studies carried out in 13 countries. They included about 2000 cases and 2000 controls. Positivity, for any HPV DNA yielded a pooled odds ratio (OR) of 70. The association was equally strong for both squamous cell (OR=74) and adenocarcinoma (OR=50) and for HPV 16 and 18 as well as for the less common HPV types. Our results indicate that in addition to HPV 16 and 18, HPV 31, 33, 35, 45, 51, 52, 58 and 59 now can be considered as carcinogenic. The third group of studies is aimed to determine the HPV DNA prevalence in random, age-stratified (by 5 years, 15-19 to 65+) subsamples (1100 women) of the general population. Two age-peaks (<25 and >59 years), have been found in some countries (Costa-Rica, Mexico, Colombia) but not in all (Argentina). Whether the second peak is due to viral reactivation, variations in screening or represents a birth-cohort effect remains to be determined. The distribution of the most prevalent HPV types in the general population (HPV 16, 18, 45, 31, 58, 33, 35) resembles that for cervical cancer cases. our studies provide the most solid epidemiological evidence, to conclude that HPV is not only the central cause of cervical cancer worldwide but also a necessary cause.


Public health consequences on vulnerable populations from acute chemical releases.
Data from a large, multi-state surveillance system on acute chemical releases were analyzed to describe the type of events that are potentially affecting vulnerable populations (children, elderly and hospitalized patients) in order to better prevent and plan for these types of incidents in the future. During 2003-2005, there were 231 events where vulnerable populations were within ¼ mile of the event and the area of impact was greater than 200 feet from the facility/point of release. Most events occurred on a weekday during times when day care centers or schools were likely to be in session. Equipment failure and human error caused a majority of the releases. Agencies involved in preparing for and responding to chemical emergencies should work with hospitals, nursing homes, day care centers, and schools to develop policies and procedures for initiating appropriate protective measures and managing the medical needs of patients. Chemical emergency response drills should involve the entire community to protect those that may be more susceptible to harm.


Educational attainment, perceived control and the quality of women's diets.
Data from the Southampton Women's Survey have established that women of lower educational attainment have poorer quality diets than those of higher educational attainment. This relationship is strong and graded such that for every increase in level of educational qualification, there is an increase in the likelihood that a woman will have a better quality diet. It is not wholly explained by socio-economic status. Qualitative research carried out in Southampton suggests that women of lower educational attainment may have a poorer diet because they feel they lack control over the food choices they make for themselves and their families. We set out to investigate the relationship between educational attainment, perceived control and quality of diet in a sample of women from Southampton. Cross-sectional study using structured interviews in which women's diet, educational attainment and perceived control were assessed. 19 Children's Centres and baby clinics in Southampton, UK. 372 women, median age 28 years. Quality of diet assessed by prudent diet score produced from principal components analysis of 20-item food frequency questionnaire, and perceived control assessed by a validated questionnaire. Women of lower educational attainment tended to have lower prudent diet scores and lower perceived control scores than women of higher educational attainment. Having a lower prudent diet score was associated with consuming fewer vegetables and vegetable dishes, less wholemeal bread and vegetarian food, and more chips and roast potatoes, meat pies, Yorkshire puddings and pancakes, crisps and snacks, white bread and added sugar. In a regression model both lower educational attainment and lower perceived control were associated with lower prudent diet scores, independent of the effects of confounding factors. However there was an interaction effect such that lower perceived control was only related to prudent diet score in the group of women of lower educational attainment. Women of lower educational attainment perceive themselves to have less control over their lives than women of higher educational attainment, and this perceived lack of control is reflected in their diets being of poorer quality. Our findings suggest that level of perceived control over life is a more important predictor of quality of diet in women of lower educational attainment than in those of higher educational attainment. It may be that psychological and social difficulties disproportionately affect the diets of women of lower educational attainment. We are currently exploring variations in quality of diet among women of lower educational attainment in relation to a range of psychological and social factors.


Use of artificial intelligence to shorten the behavioral diagnosis of autism.
The Autism Diagnostic Interview-Revised (ADI-R) is one of the most commonly used instruments for assisting in the behavioral diagnosis of autism. The exam consists of 93 questions that must be answered by a care provider within a focused session that often spans 2.5 hours. We used machine learning techniques to study the complete sets of answers to the ADI-R available at the Autism Genetic Research Exchange (AGRE) for 891 individuals diagnosed with autism and 75 individuals who did not meet the criteria for an autism diagnosis. Our analysis showed that 7 of the 93 items contained in the ADI-R were sufficient to classify autism with 99.9% statistical accuracy. We further tested the accuracy of this 7-question classifier against complete sets of answers from two independent sources, a collection of 1654 individuals with autism from the Simons Foundation and a collection of 322 individuals with autism from the Boston Autism Consortium. In both cases, our classifier performed with nearly 100% statistical accuracy, properly categorizing all but one of the individuals from these two resources who previously had been diagnosed with autism through the standard ADI-R. Our ability to measure specificity was limited by the small numbers of non-spectrum cases in the research data used, however, both real and simulated data demonstrated a range in specificity from 99% to 93.8%. With incidence rates rising, the capacity to diagnose autism quickly and effectively requires careful design of behavioral assessment methods. Ours is an initial attempt to retrospectively analyze large data repositories to derive an accurate, but significantly abbreviated approach that may be used for rapid detection and clinical prioritization of individuals likely to have an autism spectrum disorder. Such a tool could assist in streamlining the clinical diagnostic process overall, leading to faster screening and earlier treatment of individuals with autism.


Oxidative damage increases with reproductive energy expenditure and is reduced by food-supplementation.
A central principle in life-history theory is that reproductive effort negatively affects survival. Costs of reproduction are thought to be physiologically based, but the underlying mechanisms remain poorly understood. Using female North American red squirrels (Tamiasciurus hudsonicus), we test the hypothesis that energetic investment in reproduction overwhelms investment in antioxidant protection, leading to oxidative damage. In support of this hypothesis we found that the highest levels of plasma protein oxidative damage in squirrels occurred during the energetically demanding period of lactation. Moreover, plasma protein oxidative damage was also elevated in squirrels that expended the most energy and had the lowest antioxidant protection. Finally, we found that squirrels that were food-supplemented during lactation and winter had increased antioxidant protection and reduced plasma protein oxidative damage providing the first experimental evidence in the wild that access to abundant resources can reduce this physiological cost.


Effects of bacterial lipopolysaccharide injection on white blood cell counts, hematological variables, and serum glucose, insulin, and cortisol concentrations in ewes fed low- or high-protein diets.
Bacterial lipopolysaccharide endotoxins (LPS) elicit inflammatory responses reflective of acute bacterial infection. We determined if feeding ewes high-CP (15.5%) or low-CP (8.5%) diets for 10 d altered inflammatory responses to an intravenous bolus of 0 (control), 0.75 (L75), or 1.50 (L150) μg of LPS/kg of BW in a 2 × 3 factorial arrangement of treatments (n = 5/treatment). Rectal temperatures, heart and respiratory rates, blood leukocyte concentrations, and serum cortisol, insulin, and glucose concentrations were measured for 24 h after an LPS bolus (bolus = 0 h). In general, rectal temperatures were greater (P ≤ 0.05) in control ewes fed high CP, but LPS increased (P ≤ 0.05) rectal temperatures in a dose-dependent manner at most times between 2 and 24 h after the bolus. Peak rectal temperatures in L75 and L150 occurred 4 h after the bolus. A monophasic, dose-independent increase (P ≤ 0.023) in serum cortisol occurred from 0.5 to 24 h after the bolus, with peak cortisol at 4 h. Serum insulin was increased (P ≤ 0.016) by LPS in a dose-dependent manner from 4 to 24 h after the bolus. Insulin did not differ between control ewes fed high- and low-CP diets but was greater (P < 0.001) in L75 ewes fed low CP compared with high CP and in L150 ewes fed high CP compared with low CP. Increased insulin was not preceded by increased serum glucose. Total white blood cell concentrations were not affected (P ≥ 0.135) by LPS, but the neutrophil and monocyte fractions of white blood cells were increased (P ≤ 0.047) by LPS at 12 and 24 h and at 24 h after the bolus, respectively, and the lymphocyte fraction was increased (P = 0.037) at 2 h and decreased (P ≤ 0.006) at 12 and 24 h after the bolus. Red blood cell and hemoglobin concentrations and hematocrit (%) were increased (P ≤ 0.022) by LPS at 2 and 4 h after the bolus. Rectal temperatures and serum glucose were greater (P ≤ 0.033) in ewes fed a high-CP diet before LPS injection, but these effects were lost at and within 2.5 h of the bolus, respectively. Feeding high-CP diets for 10 d did not reduce inflammation in ewes during the first 24 h after LPS exposure but may benefit livestock by preventing acute insulin resistance when endotoxin exposure is mild.


Dual-responsive core-crosslinked polyphosphoester-based nanoparticles for pH/redox-triggered anticancer drug delivery.
"Intelligent" crosslinked nanoparticles (NPs) provide great advantages in enhancing drug bioavailability and reducing side effects in anticancer therapeutics. In this study, a novel biodegradable polyphosphoester-based functional copolymer prodrug PTX-(PBYP-g-MPA)-b-PEEP was prepared to construct pH/redox dual-responsive core-crosslinked nanoparticles (DOX/CCL NPs), in which paclitaxel (PTX) was conjugated to the polyphosphoester to form an amphiphilic prodrug and doxorubicin (DOX) was encapsulated inside the prodrug NPs. At first, PTX was used as an initiator to polymerize 2-(but-3-yn-1-yloxy)-2-oxo-1,3,2-dioxaphospholane (BYP) and 2-ethoxy-2-oxo-1,3,2-dioxaphospholane (EOP) by one-pot sequential ring-opening polymerization, yielding a biodegradable polymeric prodrug PTX-PBYP-b-PEEP. Subsequently, a radical-mediated thiol-yne "click" reaction was performed between the alkynyl groups on the PBYP segment and the thiol group of 3-mercaptopropionic acid (MPA) to form a functional carboxyl group at the side chain. The potential positively charged DOX·HCl can be physically encapsulated via electrostatic interaction with the carboxyl group and hydrophobic interaction. Afterwards, the DOX/CCL NPs with cleavable disulfide (S-S) linkages can be formed by partial crosslinking through amidation between the pendant carboxyl groups and cystamine. These NPs possess multifunctional characteristics used for in vitro drug release. Notably, a redox-responsive crosslinker, cystamine dihydrochloride, and synergetic non-covalent interactions not only stabilize the nanoparticles, achieve high DOX-loading capacity of drug loading content (DLC, 14.6%) and drug loading efficiency (DLE, 73.1%), but also endow the DOX/CCL NPs with controlled drug release capacity, which is due to the cleavage of S-S bonds in the presence of 10 mM glutathione (GSH) and weakened electrostatic interaction caused by the protonation of carboxyl groups at a lower pH (5.0). Moreover, these pH/redox dual-responsive DOX/CCL NPs can be steadily internalized by HeLa cells, exhibiting high-efficiency cellular proliferation inhibition. This study presents a promising strategy for controlled intracellular drug release in cancer therapy.


Malaria PCR detection in Cambodian low-transmission settings: dried blood spots versus venous blood samples.
In the context of malaria elimination, novel strategies for detecting very low malaria parasite densities in asymptomatic individuals are needed. One of the major limitations of the malaria parasite detection methods is the volume of blood samples being analyzed. The objective of the study was to compare the diagnostic accuracy of a malaria polymerase chain reaction assay, from dried blood spots (DBS, 5 μL) and different volumes of venous blood (50 μL, 200 μL, and 1 mL). The limit of detection of the polymerase chain reaction assay, using calibrated Plasmodium falciparum blood dilutions, showed that venous blood samples (50 μL, 200 μL, 1 mL) combined with Qiagen extraction methods gave a similar threshold of 100 parasites/mL, ∼100-fold lower than 5 μL DBS/Instagene method. On a set of 521 field samples, collected in two different transmission areas in northern Cambodia, no significant difference in the proportion of parasite carriers, regardless of the methods used was found. The 5 μL DBS method missed 27% of the samples detected by the 1 mL venous blood method, but most of the missed parasites carriers were infected by Plasmodium vivax (84%). The remaining missed P. falciparum parasite carriers (N = 3) were only detected in high-transmission areas.


Involvement of the HtrA family of proteases in the protection of the cyanobacterium Synechocystis PCC 6803 from light stress and in the repair of photosystem II.
Photosystem II (PSII) is prone to irreversible light-induced damage, with the D1 polypeptide a major target. Repair processes operate in the cell to replace a damaged D1 subunit within the complex with a newly synthesized copy. As yet, the molecular details of PSII repair are relatively obscure despite the critical importance of this process for maintaining PSII activity and cell viability. We are using the cyanobacterium Synechocystis sp. PCC 6803 to identify the various proteases and chaperones involved in D1 turnover in vivo. Two families of proteases are being studied: the FtsH family (four members) of Zn(2+)-activated nucleotide-dependent proteases; and the HtrA (or DegP) family (three members) of serine-type proteases. In this paper, we report the results of our studies on a triple mutant in which all three copies of the htrA gene family have been inactivated. Growth of the mutant on agar plates was inhibited at high light intensities, especially in the presence of glucose. Oxygen evolution measurements indicated that, under conditions of high light, the rate of synthesis of functional PSII was less in the mutant than in the wild-type. Immunoblotting experiments conducted on cells blocked in protein synthesis further indicated that degradation of D1 was slowed in the mutant. Overall, our observations indicate that the HtrA family of proteases are involved in the resistance of Synechocystis 6803 to light stress and play a part, either directly or indirectly, in the repair of PSII in vivo.


Plasmon nanomechanical coupling for nanoscale transduction.
We demonstrate plasmon-mechanical coupling in a metalized nanomechanical oscillator. A coupled surface plasmon is excited in the 25 nm wide gap between two metalized silicon nitride beams. The strong plasmonic dispersion allows the nanomechanical beams' thermal motion at a frequency of 4.4 MHz to be efficiently transduced to the optical transmission, with a measured displacement spectral density of 1.11 × 10(-13) m/Hz(1/2). When exciting the second-order plasmonic mode at λ = 780 nm we observe optical-power-induced frequency shifts of the mechanical oscillator. Our results show that novel functionality of plasmonic nanostructures can be achieved through coupling to engineered nanoscale mechanical oscillators.


Dynamic increase in extracellular ATP accelerates photoreceptor cell apoptosis via ligation of P2RX7 in subretinal hemorrhage.
Photoreceptor degeneration is the most critical cause of visual impairment in age-related macular degeneration (AMD). In neovascular form of AMD, severe photoreceptor loss develops with subretinal hemorrhage due to choroidal neovascularization (CNV), growth of abnormal blood vessels from choroidal circulation. However, the detailed mechanisms of this process remain elusive. Here we demonstrate that neovascular AMD with subretinal hemorrhage accompanies a significant increase in extracellular ATP, and that extracellular ATP initiates neurodegenerative processes through specific ligation of Purinergic receptor P2X, ligand-gated ion channel, 7 (P2RX7; P2X7 receptor). Increased extracellular ATP levels were found in the vitreous samples of AMD patients with subretinal hemorrhage compared to control vitreous samples. Extravascular blood induced a massive release of ATP and photoreceptor cell apoptosis in co-culture with primary retinal cells. Photoreceptor cell apoptosis accompanied mitochondrial apoptotic pathways, namely activation of caspase-9 and translocation of apoptosis-inducing factor (AIF) from mitochondria to nuclei, as well as TUNEL-detectable DNA fragmentation. These hallmarks of photoreceptor cell apoptosis were prevented by brilliant blue G (BBG), a selective P2RX7 antagonist, which is an approved adjuvant in ocular surgery. Finally, in a mouse model of subretinal hemorrhage, photoreceptor cells degenerated through BBG-inhibitable apoptosis, suggesting that ligation of P2RX7 by extracellular ATP may accelerate photoreceptor cell apoptosis in AMD with subretinal hemorrhage. Our results indicate a novel mechanism that could involve neuronal cell death not only in AMD but also in hemorrhagic disorders in the CNS and encourage the potential application of BBG as a neuroprotective therapy.


Molecular characterization of the conoid complex in Toxoplasma reveals its conservation in all apicomplexans, including Plasmodium species.
The apical complex is the instrument of invasion used by apicomplexan parasites, and the conoid is a conspicuous feature of this apparatus found throughout this phylum. The conoid, however, is believed to be heavily reduced or missing from Plasmodium species and other members of the class Aconoidasida. Relatively few conoid proteins have previously been identified, making it difficult to address how conserved this feature is throughout the phylum, and whether it is genuinely missing from some major groups. Moreover, parasites such as Plasmodium species cycle through 3 invasive forms, and there is the possibility of differential presence of the conoid between these stages. We have applied spatial proteomics and high-resolution microscopy to develop a more complete molecular inventory and understanding of the organisation of conoid-associated proteins in the model apicomplexan Toxoplasma gondii. These data revealed molecular conservation of all conoid substructures throughout Apicomplexa, including Plasmodium, and even in allied Myzozoa such as Chromera and dinoflagellates. We reporter-tagged and observed the expression and location of several conoid complex proteins in the malaria model P. berghei and revealed equivalent structures in all of its zoite forms, as well as evidence of molecular differentiation between blood-stage merozoites and the ookinetes and sporozoites of the mosquito vector. Collectively, we show that the conoid is a conserved apicomplexan element at the heart of the invasion mechanisms of these highly successful and often devastating parasites.


Proteomic analysis of human prostate cancer.
Proteomics is a promising approach in the identification of proteins and biochemical pathways involved in tumorigenesis. In an effort to discover such proteins and pathways that are deregulated in prostate tumorigenesis, cellular proteomes of matched normal prostate epithelial cells and high-grade prostate cancer cells were analyzed by tissue microdissection, two-dimensional electrophoresis, and mass spectrometry. Forty protein alterations were detected in the tumors; however, the majority of these changes were not shared among the 12 neoplasms. In contrast, parallel cDNA microarray analysis identified a number of common gene expression changes. The marked heterogeneity of the observed protein alterations may have significance with regard to tumor biology and research strategies for molecular profiling analyses of human prostate cancer.


Sixteen-kinase gene expression identifies luminal breast cancers with poor prognosis.
Breast cancer is a heterogeneous disease made of various molecular subtypes with different prognosis. However, evolution remains difficult to predict within some subtypes, such as luminal A, and treatment is not as adapted as it should be. Refinement of prognostic classification and identification of new therapeutic targets are needed. Using oligonucleotide microarrays, we profiled 227 breast cancers. We focused our analysis on two major breast cancer subtypes with opposite prognosis, luminal A (n = 80) and basal (n = 58), and on genes encoding protein kinases. Whole-kinome expression separated luminal A and basal tumors. The expression (measured by a kinase score) of 16 genes encoding serine/threonine kinases involved in mitosis distinguished two subgroups of luminal A tumors: Aa, of good prognosis and Ab, of poor prognosis. This classification and its prognostic effect were validated in 276 luminal A cases from three independent series profiled across different microarray platforms. The classification outperformed the current prognostic factors in univariate and multivariate analyses in both training and validation sets. The luminal Ab subgroup, characterized by high mitotic activity compared with luminal Aa tumors, displayed clinical characteristics and a kinase score intermediate between the luminal Aa subgroup and the luminal B subtype, suggesting a continuum in luminal tumors. Some of the mitotic kinases of the signature represent therapeutic targets under investigation. The identification of luminal A cases of poor prognosis should help select appropriate treatment, whereas the identification of a relevant kinase set provides potential targets.


A novel mutation in the mitochondrial tRNA(Ser(UCN)) gene in a family with non-syndromic sensorineural hearing impairment.
We describe a family with non-syndromic sensorineural hearing impairment inherited in a manner consistent with maternal transmission. Affected members were found to have a novel heteroplasmic mtDNA mutation, T7510C, in the tRNA(Ser(UCN)) gene. This mutation was not found in 661 controls, is well conserved between species, and disrupts base pairing in the acceptor stem of the tRNA, making it the probable cause of hearing impairment in this family. Sequencing of the other mitochondrial tRNA genes did not show any other pathogenic mutations. Four other mutations causing hearing impairment have been reported in the tRNA(Ser(UCN)) gene, two having been shown to affect tRNA(Ser(UCN)) levels. With increasing numbers of reports of mtDNA mutations causing hearing impairment, screening for such mutations should be considered in all cases unless mitochondrial inheritance can be excluded for certain.