Cellular analogs of visual cortical epigenesis. II. Plasticity of binocular integration.
Two differential pairing procedures were applied in the primary visual cortex of anesthetized and paralyzed kittens and cats, to produce changes in ocular dominance and interocular orientation disparity (IOD) during the time of recording of a single neuron. A first experiment was devised to demonstrate plasticity in the balance of monocular responses. The visual activity of the cell was driven iontophoretically to either a "high" or a "low" level, depending on the ocularity of the visual stimulation. Ocular dominance measurements before and after pairing revealed significant long-lasting changes in 33% of cases. Relative ocular preference shifted in most cases (87.5%) in favor of the reinforced eye. Similar proportions of significant changes were observed in kitten and adult cortex. The amplitude of the functional modifications was not significantly related with age, although the largest changes in ocular dominance were recorded at the peak of the critical period. The second experiment more specifically addressed the plasticity of binocular interaction. The activity of a binocular cell was driven iontophoretically to either a "high" or a "low" level, depending on the orientation disparity between two oriented stimuli, presented simultaneously and separately in the receptive field of each eye. Significant long-lasting changes in binocular responses were induced in 40% of cases. The relative IOD preference generally shifted (67%) in favor of the reinforced disparity. In half of the modified cells, functional changes were expressed only in the dichoptic viewing condition used during the pairing procedure. These functional modifications of binocular integration, demonstrated at the cellular level, are analogous to those induced by global manipulations of the visual environment (Hubel and Wiesel, 1970; Shinkman and Bruce, 1977). They are interpreted as evidence for synaptic plasticity. Our results support the hypothesis that covariance levels between pre- and postsynaptic activities determine the sign and the amplitude of changes in synaptic efficacy.


Hypothermia after cardiac arrest should be further evaluated--a systematic review of randomised trials with meta-analysis and trial sequential analysis.
Guidelines recommend mild induced hypothermia (MIH) to reduce mortality and neurological impairment after out-of-hospital cardiac arrest. Our objective was to systematically evaluate the evidence for MIH taking into consideration the risks of systematic and random error and to GRADE the evidence. Systematic review with meta-analysis and trial sequential analysis of randomised trials evaluating MIH after cardiac arrest in adults. We searched CENTRAL, MEDLINE, and EMBASE databases until May 2009. Retrieved trials were evaluated with Cochrane methodology. Meta-analytic estimates were calculated with random- and fixed-effects models and random errors were evaluated with trial sequential analysis (TSA). Five randomised trials (478 patients) were included. All trials had substantial risk of bias. The relative risk (RR) for death was 0.84 (95% confidence interval (CI) 0.70 to 1.01) and for poor neurological outcome 0.78 (95% CI 0.64 to 0.95). For the two trials with least risk of bias the RR for death was 0.92 (95% CI 0.56 to 1.51) and for poor neurological outcome 0.92 (95% confidence interval 0.56 to 1.50). TSA indicated lack of firm evidence for a beneficial effect. The substantial risk of bias and concerns with directness rated down the quality of the evidence to low. Evidence regarding MIH after out-of-hospital cardiac arrest is still inconclusive and associated with non-negligible risks of systematic and random errors. Using GRADE-methodology, we conclude that the quality of evidence is low. Our findings demonstrate that clinical equipoise exists and that large well-designed randomised trials with low risk of bias are needed.


Use of a lipophilic cation for determination of membrane potential in neuroblastoma-glioma hybrid cell suspensions.
Neuroblastoma-glioma hybrid cells (NG108-15) in suspension accumulate the permeant lipophilic cation [(3)H]tetraphenylphosphonium (TPP(+)) against a concentration gradient. The steady-state level of TPP(+) accumulation is about twice as great in physiological media of low K(+) concentration (i.e., 5 mM K(+)/135 mM Na(+)) than in a medium of high K(+) concentration (i.e., 121 mM K(+)/13.5 mM Na(+)). The latter manipulation depolarizes the NG108-15 plasma membrane and indicates that the resting membrane potential (DeltaPsi) is due primarily to a K(+) diffusion gradient (K(in) (+) --> K(out) (+)). TPP(+) accumulation is time and temperature dependent, achieving a steady state in 15-20 min at 37 degrees C, and is a linear function of cell number and TPP(+) concentration (i.e., the concentration gradient is constant). The difference in TPP(+) accumulation in low and high K(+) media under various conditions has been used to calculate mean (+/-SD) DeltaPsi values of -56 +/- 3, -63 +/- 4, and -66 +/- 5 mV at 26, 33, and 37 degrees C, respectively. Importantly, these values are virtually identical to those obtained by direct electrophysiological measurements made under the same conditions. TPP(+) accumulation is abolished by the protonophore carbonylcyanide-m-chlorophenylhydrazone, whereas the neurotoxic alkaloid veratridine diminishes uptake to the same level as that observed in high K(+) media. In addition, the effect of veratridine is dependent upon the presence of external Na(+) and is blocked by tetrodotoxin. The steady-state level of TPP(+) accumulation is enhanced by monensin, indicating that this ionophore induces hyperpolarization under appropriate conditions. Finally, ouabain has essentially no effect on the steady-state level of TPP(+) accumulation in short-term experiments, suggesting that Na(+),K(+)-ATPase activity makes little contribution to the resting potential in these cells. Because many of these observations are corroborated by intracellular recording techniques, it is concluded that TPP(+) distribution measurements can provide a biochemical method for determining membrane potentials in populations of cultured neuronal cells.


CT-guided cryoablation for post-thoracotomy pain syndrome: a retrospective analysis.
PURPOSE Post-thoracotomy pain syndrome is a common condition affecting up to 50% of post-thoracotomy patients. However, percutaneous computed tomography (CT)-guided intercostal nerve cryoablation may provide symptomatic benefit in chronic and/or refractory cases. METHODS A retrospective review of our institution's comprehensive case log from October 2017 to September 2018 for patients who underwent cryoablation was analyzed. Thirteen patients with post-thoracotomy pain syndrome, refractory to medical management, were treated with CT-guided intercostal nerve cryoablation. Most patients had treatment of the intercostal nerve at the level of their thoracotomy scar, two levels above and below. The safety and technical success of this technique and the clinical outcomes of the study population were then retrospectively reviewed. RESULTS Of the patients, 69% experienced significant improvement in their pain symptoms with a median pain improvement score of 3 points (range, -1 to 8 points) over a median follow-up of 11 months (range, 2-18.6 months). Complications included pneumothorax in 8% and pseudohernia in 23% of patients. CONCLUSION CT-guided intercostal nerve cryoablation may be an effective technique in the treatment of post-thoracotomy pain syndrome and requires further study.


The abdominal wall incisional hernia repair in cirrhotic patients.
The incidence of abdominal wall hernia in cirrhotic patients with ascites is between 20 and 40%. Controversies regarding the treatment modality and surgical timing of abdominal wall incisional hernia repair in cirrhotic patients remain. The study proposed wants to analyze the abdominal incisional hernia repair in cirrhotic patients with ascites performed in a single center to determine post-operative morbidity, mortality and complication rate. Cirrhotic patients with abdominal incisional hernia that underwent surgical operation for abdominal wall hernia repair at the "Policlinico Paolo Giaccone" at Palermo University Hospital between January 2015 and December 2016 were identified and the data collected were retrospectively reviewed; patients' medical and surgical records were collected from charts and the surgical and ICU registries. The degree of hepatic dysfunction was classified using Child-Pugh classification. Post-operative mortality was considered up to 30-days after surgery. A follow-up period of 6 months at least was performed to evaluate hernia recurrence and complications. Mortality rate is of 18.5% (p 0.002). Recurrence rate (p 0.004) and seroma formation rate (p 0.001) are most frequent in urgency group. The elevated ASA score and the prediction of a complicated post-operative course is higher in urgency group (p 0.004) as higher is the in-hospital stay (p 0.001) and the ICU stay (p 0.001). Elective surgery for abdominal wall hernia repair in cirrhotic patients seems to be successful and associated with lower mortality/morbidity rate and recurrence rate than urgency.


Use of Machine Learning on Contact Lens Sensor-Derived Parameters for the Diagnosis of Primary Open-angle Glaucoma.
To test the hypothesis that contact lens sensor (CLS)-based 24-hour profiles of ocular volume changes contain information complementary to intraocular pressure (IOP) to discriminate between primary open-angle glaucoma (POAG) and healthy (H) eyes. Development and evaluation of a diagnostic test with machine learning. Subjects: From 435 subjects (193 healthy and 242 POAG), 136 POAG and 136 age-matched healthy subjects were selected. Subjects with contraindications for CLS wear were excluded. This is a pooled analysis of data from 24 prospective clinical studies and a registry. All subjects underwent 24-hour CLS recording on 1 eye. Statistical and physiological CLS parameters were derived from the signal recorded. CLS parameters frequently associated with the presence of POAG were identified using a random forest modeling approach. Area under the receiver operating characteristic curve (ROC AUC) for feature sets including CLS parameters and Start IOP, as well as a feature set with CLS parameters and Start IOP combined. The CLS parameters feature set discriminated POAG from H eyes with mean ROC AUCs of 0.611, confidence interval (CI) 0.493-0.722. Larger values of a given CLS parameter were in general associated with a diagnosis of POAG. The Start IOP feature set discriminated between POAG and H eyes with a mean ROC AUC of 0.681, CI 0.603-0.765. The combined feature set was the best indicator of POAG with an ROC AUC of 0.759, CI 0.654-0.855. This ROC AUC was statistically higher than for CLS parameters or Start IOP feature sets alone (both P < .0001). CLS recordings contain information complementary to IOP that enable discrimination between H and POAG. The feature set combining CLS parameters and Start IOP provide a better indication of the presence of POAG than each of the feature sets separately. As such, the CLS may be a new biomarker for POAG.


Single channel blind image deconvolution from radially symmetric blur kernels.
The multichannel exact blind image deconvolution theory tells us that exact recovery of unknown blur kernels is possible from multiple measurements of an identical scene through distinct blur channels. However, in many biological applications, there often exist technical difficulties in obtaining multiple distinct blur measurements, since the image content may vary for various reasons, including specimen drift between snapshots, specimen damage due to prolonged exposure, or physiological changes in live cell imaging. The main contribution of this paper is a new non-iterative single channel blind deconvolution method that eliminates the need of multiple blur measurements, but still guarantees an accurate estimation of the blurring kernel. The basic idea behind this novel method is to exploit the radial symmetry of a certain class of PSFs. This approach simplifies the PSF estimation to a 1-D channel identification problem with multiple excitations, which can be solved using a standard subspace method. Since radially symmetric PSFs are quite often encountered in many practical applications, such as optical imaging systems and electron microscopy, our theory may have great influence on many practical imaging applications. Simulation results as well as real experimental results using optical and electron microscopy confirm our theory.


Type-specific proactive interference in patients with semantic and phonological STM deficits.
Prior neuropsychological evidence suggests that semantic and phonological components of short-term memory (STM) are functionally and neurologically distinct. The current paper examines proactive interference (PI) from semantic and phonological information in two STM-impaired patients, DS (semantic STM deficit) and AK (phonological STM deficit). In Experiment 1 probe recognition tasks with open and closed sets of stimuli were used. Phonological PI was assessed using nonword items, and semantic and phonological PI was assessed using words. In Experiment 2 phonological and semantic PI was elicited by an item recognition probe test with stimuli that bore phonological and semantic relations to the probes. The data suggested heightened phonological PI for the semantic STM patient, and exaggerated effects of semantic PI in the phonological STM case. The findings are consistent with an account of extremely rapid decay of activated type-specific representations in cases of severely impaired phonological and semantic STM.


Functional Impact of BeKm-1, a High-Affinity hERG Blocker, on Cardiomyocytes Derived from Human-Induced Pluripotent Stem Cells.
IKr current, a major component of cardiac repolarization, is mediated by human Ether-à-go-go-Related Gene (hERG, Kv11.1) potassium channels. The blockage of these channels by pharmacological compounds is associated to drug-induced long QT syndrome (LQTS), which is a life-threatening disorder characterized by ventricular arrhythmias and defects in cardiac repolarization that can be illustrated using cardiomyocytes derived from human-induced pluripotent stem cells (hiPS-CMs). This study was meant to assess the modification in hiPS-CMs excitability and contractile properties by BeKm-1, a natural scorpion venom peptide that selectively interacts with the extracellular face of hERG, by opposition to reference compounds that act onto the intracellular face. Using an automated patch-clamp system, we compared the affinity of BeKm-1 for hERG channels with some reference compounds. We fully assessed its effects on the electrophysiological, calcium handling, and beating properties of hiPS-CMs. By delaying cardiomyocyte repolarization, the peptide induces early afterdepolarizations and reduces spontaneous action potentials, calcium transients, and contraction frequencies, therefore recapitulating several of the critical phenotype features associated with arrhythmic risk in drug-induced LQTS. BeKm-1 exemplifies an interesting reference compound in the integrated hiPS-CMs cell model for all drugs that may block the hERG channel from the outer face. Being a peptide that is easily modifiable, it will serve as an ideal molecular platform for the design of new hERG modulators displaying additional functionalities.


Major depression symptoms in primary care and psychiatric care settings: a cross-sectional analysis.
We undertook a study to confirm and extend preliminary findings that participants with major depressive disorder (MDD) in primary care and specialty care settings have with equivalent degrees of depression severity and an indistinguishable constellation of symptoms. Baseline data were collected for a distinct validation cohort of 2,541 participants (42% primary care) from 14 US regional centers comprised of 41 clinic sites (18 primary care, 23 specialty care). Participants met broadly inclusive eligibility criteria requiring a Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, diagnosis of MDD and a minimum depressive symptom score on the 17-item Hamilton Rating Scale for Depression. The main outcome measures were the 30-item Inventory of Depressive Symptomatology--Clinician Rated and the Psychiatric Diagnostic Screening Questionnaire. Primary care and specialty care participants had identical levels of moderately severe depression and identical distributions of depressive severity scores. Both primary care and specialty care participants showed considerable suicide risk, with specialty care participants even more likely to report prior suicide attempts. Core depressive symptoms or concurrent psychiatric disorders were not substantially different between settings. One half of participants in each setting had an anxiety disorder (48.6% primary care vs 51.6% specialty care, P = .143), with social phobia being the most common (25.3% primary care vs 32.1% specialty care, P = .002). For outpatients with nonpsychotic MDD, depressive symptoms and severity vary little between primary care and specialty care settings. In this large, broadly inclusive US sample, the risk factors for chronic and recurrent depressive illness were frequently present, highlighting a clear risk for treatment resistance and the need for aggressive management strategies in both settings.


Effect of a nutrient mixture on the localization of extracellular matrix proteins in HeLa human cervical cancer xenografts in female nude mice.
Cervical cancer is one of the most commonly diagnosed cancers and a significant cause of mortality in women worldwide. Although cervical cancer is fully treatable in the early stages, once it has metastasized, patient outcome is poor. The objective of the present study was to investigate the effect of dietary supplementation with a nutrient mixture (NM) containing lysine, ascorbic acid, proline, green tea extract and other micronutrients on the expression of extracellular matrix (ECM) proteins in HeLa cell xenografts in nude female mice. After housing for 1 week, female athymic nude mice between 5 and 6 weeks of age (n=12) were inoculated subcutaneously with 3×106 HeLa cells in phosphate-buffered saline and Matrigel and randomly divided into two groups. These were the control group, in which the mice were fed with regular mouse chow, and the NM group, in which the mice were fed with the regular diet supplemented with 0.5% NM (w/w). After 4 weeks, the tumors were excised and processed for histology. Tumor growth was evaluated and the tumors were stained for the ECM proteins collagen I, collagen IV, fibronectin, laminin, periodic acid-Schiff (PAS) and elastin. NM strongly inhibited (by 59%, P=0.001) the growth of HeLa xenografts in nude mice. Tumors from control mice exhibited little to no collagen I expression either internally or in the fibrous capsule, while tumors from the NM group expressed collagen I in the fibrous capsule and within the tumor. Tumors from the control group showed diffuse cytoplasmic and capsular collagen IV with abundant nucleated cells. NM treatment substantially increased collagen IV production and induced a dense fibrous network of collagen IV with chambers that surrounded live nucleated cells and large amounts of necrotic cell debris. Tumors from the mice fed with the NM exhibited a well-defined border of fibronectin in the capsule and intense areas of staining internally whereas control group tumors showed less overall fibronectin with sporadic internal staining and little in the fibrous capsule. Although laminin appeared abundantly in control and NM-treated tumors, the NM group tumors exhibited a chamber-like network of laminin internally. Tumors from the control group exhibited internal areas of intense PAS staining, whereas tumors from the NM-treated group exhibited a more uniform diffuse pattern of PAS staining. In conclusion, NM supplementation of HeLa xenograft-bearing female nude mice demonstrated a potent inhibition of tumor growth and enhancement of ECM proteins, suggesting the therapeutic value of this specific nutrient complex in the treatment of cervical cancer.


Modulation of aortic smooth muscle cell membrane potential by extracellular calcium.
Removal of extracellular calcium may result in depolarization of the resting cell membrane potential. This has been attributed to the stabilizing action of calcium on the ionic permeability of the cell membrane. It is unknown whether this phenomenon is exclusively mediated by extracellular calcium or through associated changes in intracellular calcium. To examine this, we exposed rat aortic smooth muscle cells in culture to different calcium concentrations and studied their effects on the resting membrane potential and intracellular calcium activity. The resting membrane potential was dependent on the extracellular potassium concentration. Exposure to reduced extracellular calcium concentrations (0.25 and 0.5 mM) caused a steep and reversible depolarization of the membrane potential, but intracellular calcium, measured with fura 2-AM, was not reduced below that measured in control conditions (1.8 mM). Atomic absorption spectrophotometric measurements did not indicate a measurable gain in cell sodium after reduction of extracellular calcium levels. We conclude that extracellular calcium controls the resting cell membrane potential of vascular smooth muscle through a mechanism that is independent of cytosolic Ca2+ activity.


Use of a bacterial fluorescence imaging device: wound measurement, bacterial detection and targeted debridement.
Diagnostics which provide objective information to facilitate evidence-based treatment decisions could improve the chance of wound healing. Accurate wound measurements, objective bacterial assessment, and the regular, consistent tracking of these parameters are important aspects of wound care. This study aimed to assess the accuracy, clinical incorporation and documentation capabilities of a handheld bacterial fluorescence imaging device (MolecuLight i:X). Benchtop wound models with known dimensions and clinical wound images were repeatedly measured by trained clinicians to quantify accuracy and intra/inter-user coefficients of variation (COV) of the imaging device measurement software. In a clinical trial of 50 wounds, wound dimensions were digitally measured and fluorescence images were acquired to assess for the presence of bacteria at moderate-to-heavy loads. Finally, fluorescence imaging was implemented into the routine assessment of 22 routine diabetic foot ulcers (DFU) to determine appropriate debridement level and location based on bacterial fluorescence signals. Wound measurement accuracy was >95% (COV <3%). In the clinical trial of 50 wounds, 72% of study wounds demonstrated positive bacterial fluorescence signals. Levine sampling of wounds was found to under-report bacterial loads relative to fluorescence-guided curettage samples. Furthermore, fluorescence documentation of bacterial presence and location(s) resulted in more aggressive, fluorescence-targeted debridement in 17/20 DFUs after standard of care debridement failed to eliminate bacterial fluorescence in 100% of DFU debridements. The bacterial fluorescence imaging device can be readily implemented for objective, evidenced-based wound assessment and documentation at the bedside. Bedside localisation of regions with moderate-to-heavy bacterial loads facilitated improved sampling, debridement targeting and improved wound bed preparation.


The molecular architecture of axonemes revealed by cryoelectron tomography.
Eukaryotic flagella and cilia are built on a 9 + 2 array of microtubules plus >250 accessory proteins, forming a biological machine called the axoneme. Here we describe the three-dimensional structure of rapidly frozen axonemes from Chlamydomonas and sea urchin sperm, using cryoelectron tomography and image processing to focus on the motor enzyme dynein. Our images suggest a model for the way dynein generates force to slide microtubules. They also reveal two dynein linkers that may provide "hard-wiring" to coordinate motor enzyme action, both circumferentially and along the axoneme. Periodic densities were also observed inside doublet microtubules; these may contribute to doublet stability.


When we should worry more: using cognitive bias modification to drive adaptive health behaviour.
A lack of behavioural engagement in health promotion or disease prevention is a problem across many health domains. In these cases where people face a genuine danger, a reduced focus on threat and low levels of anxiety or worry are maladaptive in terms of promoting protection or prevention behaviour. Therefore, it is possible that increasing the processing of threat will increase worry and thereby enhance engagement in adaptive behaviour. Laboratory studies have shown that cognitive bias modification (CBM) can increase or decrease anxiety and worry when increased versus decreased processing of threat is encouraged. In the current study, CBM for interpretation (CBM-I) is used to target engagement in sun protection behaviour. The goal was to investigate whether inducing a negative rather than a positive interpretation bias for physical threat information can enhance worry elicited when viewing a health campaign video (warning against melanoma skin cancer), and consequently lead to more adaptive behaviour (sun protection). Participants were successfully trained to either adopt a positive or negative interpretation bias using physical threat scenarios. However, contrary to expectations results showed that participants in the positive training condition reported higher levels of worry elicited by the melanoma video than participants in the negative training condition. Video elicited worry was, however, positively correlated with a measure of engagement in sun protection behaviour, suggesting that higher levels of worry do promote adaptive behaviour. These findings imply that more research is needed to determine under which conditions increased versus decreased processing of threat can drive adaptive worry. Various potential explanations for the current findings and suggestions for future research are discussed.


Individual differences in expert motor coordination associated with white matter microstructure in the cerebellum.
Recent investigations into the neural basis of elite sporting performance have focused on whether cortical activity might characterize individual differences in ability. However, very little is understood about how changes in brain structure might contribute to individual differences in expert motor control. We compared the behavior and brain structure of healthy controls with a group of karate black belts, an expert group who are able to perform rapid, complex movements that require years of training. Using 3D motion tracking, we investigated whether the ability to control ballistic arm movements was associated with differences in white matter microstructure. We found that karate experts are better able than novices to coordinate the timing of inter-segmental joint velocities. Diffusion tensor imaging revealed significant differences between the groups in the microstructure of white matter in the superior cerebellar peduncles (SCPs) and primary motor cortex-brain regions that are critical to the voluntary control of movement. Motor coordination, the amount of experience, and the age at which training began were all associated with individual differences in white matter integrity in the cerebellum within the karate groups. These findings suggest a role for the white matter pathways of the SCPs in motor expertise.


The persistence length of adsorbed dendronized polymers.
The persistence length of cationic dendronized polymers adsorbed onto oppositely charged substrates was studied by atomic force microscopy (AFM) and quantitative image analysis. One can find that a decrease in the ionic strength leads to an increase of the persistence length, but the nature of the substrate and of the generation of the side dendrons influence the persistence length substantially. The strongest effects as the ionic strength is being changed are observed for the fourth generation polymer adsorbed on mica, which is a hydrophilic and highly charged substrate. However, the observed dependence on the ionic strength is much weaker than the one predicted by the Odijk, Skolnik, and Fixman (OSF) theory for semi-flexible chains. Low-generation polymers show a variation with the ionic strength that resembles the one observed for simple and flexible polyelectrolytes in solution. For high-generation polymers, this dependence is weaker. Similar dependencies are found for silica and gold substrates. The observed behavior is probably caused by different extents of screening of the charged groups, which is modified by the polymer generation, and to a lesser extent, the nature of the substrate. For highly ordered pyrolytic graphite (HOPG), which is a hydrophobic and weakly charged substrate, the electrostatic contribution to the persistence length is much smaller. In the latter case, we suspect that specific interactions between the polymer and the substrate also play an important role.


Neural evidence for a distinction between short-term memory and the focus of attention.
It is widely assumed that the short-term retention of information is accomplished via maintenance of an active neural trace. However, we demonstrate that memory can be preserved across a brief delay despite the apparent loss of sustained representations. Delay period activity may, in fact, reflect the focus of attention, rather than STM. We unconfounded attention and memory by causing external and internal shifts of attention away from items that were being actively retained. Multivariate pattern analysis of fMRI indicated that only items within the focus of attention elicited an active neural trace. Activity corresponding to representations of items outside the focus quickly dropped to baseline. Nevertheless, this information was remembered after a brief delay. Our data also show that refocusing attention toward a previously unattended memory item can reactivate its neural signature. The loss of sustained activity has long been thought to indicate a disruption of STM, but our results suggest that, even for small memory loads not exceeding the capacity limits of STM, the active maintenance of a stimulus representation may not be necessary for its short-term retention.


Different chromatin interfaces of the Drosophila dosage compensation complex revealed by high-shear ChIP-seq.
Transcriptional enhancement of X-linked genes to compensate for the sex chromosome monosomy in Drosophila males is brought about by a ribonucleoprotein assembly called Male-Specific-Lethal or Dosage Compensation Complex (MSL-DCC). This machinery is formed in male flies and specifically associates with active genes on the X chromosome. After assembly at dedicated high-affinity "entry" sites (HAS) on the X chromosome, the complex distributes to the nearby active chromatin. High-resolution, genome-wide mapping of the MSL-DCC subunits by chromatin immunoprecipitation (ChIP) on oligonucleotide tiling arrays suggests a rather homogenous spreading of the intact complex onto transcribed chromatin. Coupling ChIP to deep sequencing (ChIP-seq) promises to map the chromosomal interactions of the DCC with improved resolution. We present ChIP-seq binding profiles for all complex subunits, including the first description of the RNA helicase MLE binding pattern. Exploiting the preferential representation of direct chromatin contacts upon high-energy shearing, we report a surprising functional and topological separation of MSL protein contacts at three classes of chromosomal binding sites. Furthermore, precise determination of DNA fragment lengths by paired-end ChIP-seq allows decrypting of the local complex architecture. Primary contacts of MSL-2 and MLE define HAS for the DCC. In contrast, association of the DCC with actively transcribed gene bodies is mediated by MSL-3 binding to nucleosomes. We identify robust MSL-1/MOF binding at a fraction of active promoters genome-wide. Correlation analyses suggest that this association reflects a function outside dosage compensation. Our comprehensive analysis provides a new level of information on different interaction modes of a multiprotein complex at distinct regions within the genome.


[Growth of breastfed and bottle-fed infants up to 2 years of age: CLACYD (Lactation, Alimentation, Growth and Development) study 1993-1995].
Studies done in various countries show important differences in the growth of breastfed and bottle-fed children. In addition, it has been found that breast-fed children grow more slowly beginning at the age of 2 or 3 months in comparison with the reference pattern of the U.S. National Center for Health Statistics (NCHS) and the World Health Organization (WHO). These results cast doubt on whether maximum growth is the same as optimal growth. The objective of this study was to compare the growth in weight and length, from birth to 24 months, for a group of children who were breast-fed with that of a group who were bottle-fed. The study was also intended to describe the growth of the breastfed group in relation to the NCHS/WHO norms and a WHO "12-month breast-fed pooled data set." For this research, data were analyzed from the "Cordoba: lactation, feeding, growth, and development" study (or CLACYD study, for its Spanish-language acronym). That study looked at a representative cohort, stratified by social class, of children born in 1993 in the city of Cordoba, Argentina. The researchers analyzed anthropometric data on 74 children who were breast-fed during the first year of life and on 108 bottle-fed children. The data had been recorded, using standardized techniques, at birth and at 6, 12, and 24 months of age. Both groups were homogenous with respect to the age and schooling of the parents, social stratum, birth order, maternal height, and child's weight and length at birth. The living conditions (housing construction and availability of water and sewer services) were better among the group that was bottle-fed (P = 0.04). The breast-fed children had a lower weight and a shorter length at 6, 12, and 24 months than did the bottle-fed children. The breast-fed children also showed a slowing in growth with respect to the NCHS/WHO guidelines beginning in the second semester. This indicates that the NCHS/WHO norms are not totally adequate for evaluating the growth of breast-fed children in Cordoba, Argentina. In the high and middle social strata, the values for the breast-fed group resembled those for the WHO "pooled data set," both in weight and length. In the low and very low social strata, weight values were satisfactory, but the figures for length were less than those of the "pooled data set." The gap in length found among the low and very low social strata does not reduce the validity of the WHO "pooled data set" reference, but rather indicates the influence of living conditions on linear growth.


Bladder outlet physiology in the context of lower urinary tract dysfunction.
Physiological function of the bladder outlet is complex and symptomatic consequences can result from outlet dysfunction. Within the outlet, smooth, and skeletal muscles constitute the contractile apparatus, but additional cell types include interstitial cells and neuroendocrine cells, and various transmitters are present in the innervation, raising the possibility of unrecognized functional subtleties. Key outlet functions are; maintained closure for urine storage, increased closure (guarding) during exertion, sustained opening for voiding, transient opening for territorial marking in animals and orthograde male ejaculation. These are co-ordinated by several spinal and higher CNS centers, with overlap of the somatic, sympathetic and parasympathetic nervous systems. During voiding, urethral afferents may be important in maintaining detrusor contraction until completion of bladder emptying. Some of the bladder outlet afferents may be shared with the anal sphincter. Dysfunction of the outlet leads to conditions such as retrograde ejaculation, Fowler's syndrome, and detrusor sphincter dyssynergia. Urethral relaxation during urine storage may lead to urinary urgency, which may be misleadingly labeled as overactive bladder. Research priorities are numerous, including; peripheral cellular integrative physiology, interactions with other pelvic organs, interconnectivity of the CNS centers at all levels of the neuraxis, and standardized animal models of outlet functions such as reflex-driven voiding.


A systematic review of studies measuring health-related quality of life of general injury populations.
It is important to obtain greater insight into health-related quality of life (HRQL) of injury patients in order to document people's pathways to recovery and to quantify the impact of injury on population health over time. We performed a systematic review of studies measuring HRQL in general injury populations with a generic health state measure to summarize existing knowledge. Injury studies (1995-2009) were identified with main inclusion criteria being the use of a generic health status measure and not being restricted to one specific type of injury. Articles were collated by study design, HRQL instrument used, timing of assessment(s), predictive variables and ability to detect change over time. Forty one studies met inclusion criteria, using 24 different generic HRQL and functional status measures (most used were SF-36, FIM, GOS, EQ-5D). The majority of the studies used a longitudinal design, but with different lengths and timings of follow-up (mostly 6, 12, and 24 months). Different generic health measures were able to discriminate between the health status of subgroups and picked up changes in health status between discharge and 12 month follow-up. Most studies reported high prevalences of health problems within the first year after injury. The twelve studies that reported HRQL utility scores showed considerable but incomplete recovery in the first year after discharge. This systematic review demonstrates large variation in use of HRQL instruments, study populations, and assessment time points used in studies measuring HRQL of general injury populations. This variability impedes comparison of HRQL summary scores between studies and prevented formal meta-analyses aiming to quantify and improve precision of the impact of injury on population health over time.


When psychotherapists disclose personal information about themselves to clients.
Psychotherapists sometimes disclose personal information to their clients during therapeutic sessions. We report here our analysis of how these 'therapist self-disclosures' are done. In a sample of 15 sessions involving four therapists, we find that all therapists use them sparingly and some not at all. When they do, they 'match' something in the client's preceding turn. Vehicles for the match can range from comparatively simple agreements to more complicated 'second stories', which use analogies from the therapists' own current life. We find that these 'personal' disclosures are invariably rather ordinary but are made to bear visibly on the therapeutic business at hand, though not always in obvious ways. The ordinariness of therapist's self-disclosures underpins what seems to be one of their main actions-to 'normalize', for a number of disparate local interactional contingencies, the clients' experience. We discuss the practice of using one's own life experiences to bear on one's client's troubles, noting the recurrent features of extreme case formulations and explicit recipient design. We conclude with a brief discussion of the relation between our analyses and those which might be offered by members of the therapeutic community.


Senescent Changes and Topography of the Dark-Adapted Multifocal Electroretinogram.
To investigate the topographic changes of the dark-adapted multifocal electroretinogram (mfERG) across adulthood in the central retina and compare the topography between macular versus extramacular, nasal versus temporal, and inferior versus superior retinal areas. Sixty-five subjects (18-88 years) received a comprehensive dilated eye examination to ensure the health of their retina and were tested with a dark-adapted mfERG protocol using a 61-hexagon pattern. The lens absorption of each subject was also estimated using a heterochromatic flicker photometry (HFP) paradigm. The response amplitude and latency of the dark-adapted mfERG showed a significant change with age, which was best described with a linear model. All the retinal areas examined demonstrated similar aging effects. The extramacular and temporal retina showed higher response amplitude and faster response latency when compared with the macular and nasal retinae, respectively. No difference was found in response amplitude and latency between the inferior and superior retina. The HFP results also showed a significant correlation with age, consistent with senescent increases in short wavelength absorption by the crystalline lens. However, the change in lens absorption did not exceed the magnitude of the change in response amplitude and latency. Our results indicate that there is a decline in dark-adapted retinal activity as measured with the mfERG. These aging processes affect rods and rod-bipolar cells. Their decrease in response can be attributed to both optical and neural factors.


Evolution of cooperation, differentiation, complexity, and diversity in an iterated three-person game.
A nonzero-sum three-person coalition game is presented to study the evolution of complexity and diversity in cooperation, where the population dynamics of players with strategies is given according to their scores in the iterated game and mutations. Two types of differentiation emerge initially: a biased one to classes and a temporal one to change their roles for coalition. Rules to change the hands are self-organized in a society through evolution. The coevolution of diversity and complexity of strategies and interactions (or communications) are found at later stages of the simulation. Relevance of our results to the biological society is briefly discussed.


Methods used to reduce the prevalence of hepatitis C in a dialysis unit.
In the present study, we report on the follow-up of the epidemiology of hepatitis C viremia in our dialysis unit after our previous report, over the period from July 1, 2003 to December 31, 2005. The methods to reduce the prevalence of hepatitis C viremia in our center included: strict adherence to universal infection control precautions, separation of hepatitis C virus (HCV) positive patients from the negative patients and using specially designated machines for them, and from July 2003, periodic testing of all patients for HCV-RNA. Following the application of the above mentioned methods, we have not had, since 31 December 2005, any case of sero-conversion from HCV-negative to HCV-positive in our dialysis unit and the only HCV-positive patients present were those who were already positive at entry. The overall prevalence of HCV-RNA positive patients in our unit has presently come down to 6.5%. Although isolation and use of designated machines for HCV-RNA positive patients is not recommended following the latest guidelines of "Kidney Disease: Improving Global Outcomes" (KDIGO, 2008), the present study supports previous reports that these measures might be beneficial, when there is a high prevalence of HCV-RNA positive patients, and in units where due to understaffing or other causes, break in infection control procedures is likely to occur.


In vitro oncosphere-killing assays to determine immunity to the larvae of Taenia pisiformis, Taenia ovis, Taenia saginata, and Taenia solium.
Taeniid cestodes infect humans and livestock, causing considerable morbidity and mortality, as well as economic loss. Substantial progress has been made toward the production of recombinant vaccines against cysticercosis in livestock animals. Further development of these vaccines would be aided if a reliable in vitro test were available to measure host-protective immune responses in vaccinated animals. Here, we describe in vitro oncosphere-killing assays for the quantification of host-protective serum antibodies against Taenia pisiformis, Taenia ovis, Taenia saginata, and Taenia solium in rabbits, sheep, cattle, and pigs, respectively. Activated oncospheres of T. pisiformis, T. ovis, T. saginata, and T. solium were incubated in vitro in culture medium, test serum, and a source of complement, and oncosphere killing was assessed after 10 days of culture. In vitro oncosphere killing reflected the presence of specific antibody, and the oncosphere-killing assay typically indicated immunity to the homologous parasite that had been determined in vivo. This study describes the first reliable oncosphere-killing assays for T. pisiformis, T. ovis, T. saginata, and T. solium. These assays will be used for further research into the optimization of recombinant vaccines against cysticercosis.


Transforming clinical practice amongst community nurses: mentoring for COPD patient self-management.
To report on the process of transforming clinical practice amongst community nurses through a mentoring programme implemented to support self-management amongst community-based sufferers of chronic obstructive pulmonary disease. The increasing incidence of and health burden from, chronic diseases has led to the emergence of more proactive, integrated chronic disease management approaches across the acute and primary care sectors. An important part of these approaches is the direct involvement of patients in their own care. Despite some difficulties with comparing the benefits of chronic disease self-management programmes, many evaluations report some benefit and all highlight the importance of health professionals in supporting self-management behaviours. In the primary care sector, community nurses are ideally situated to support these behaviours, but to do this effectively transformation of nursing practice must occur. Qualitative, longitudinal study informed by action research methods and involving monthly group discussions with community nurse mentors. Community nurses from four community health centres in Tasmania were trained in motivational interviewing techniques to promote self-management amongst chronic obstructive pulmonary disease patients. Nurses' mentoring experiences were monitored during group discussions and subjected to thematic analysis. The paper reports the findings of the first 12 months of the project. In this phase, nurses experienced a transformation in their constructions of chronic obstructive pulmonary disease and their clinical practice. This involved a shift from a fatalistic, prescriptive, biomedical approach to a primary healthcare approach characterised by empathy, consultation, facilitation and a holistic focus. Community nurses face challenges in supporting chronic disease self-management. These challenges can be overcome and a transformation in clinical practice instilled. This study highlights that it is possible to support community nurses to take a lead role in the ongoing management of chronic disease in the community.


Your Metric Matters! Choose Wisely to Assess User Performance With Tourniquets in Simulated First Aid.
Readiness to perform lifesaving interventions during emergencies is based on a person's preparation to proficiently execute the skills required. Graphically plotting the performance of a tourniquet user in simulation has previously aided us in developing our understanding of how the user actually behaves. The purpose of this study was to explore performance assessment and learning curves to better understand how to develop best teaching practices. These were retrospective analyses of a convenience sample of data from a prior manikin study of 200 tourniquet uses among 10 users. We sought to generate hypotheses about performance assessments relevant to developing best teaching practices. The focus was on different metrics of user performance. When one metric was chosen over another, failure counts summed cumulatively over 200 uses differed as much as 12-fold. That difference also indicated that the degree of challenge posed to user performance differed by the metric chosen. When we ranked user performance with one metric and then with another, most (90%; nine of 10) users changed rank: five rose and four fell. Substantial differences in performance outcomes resulted from the difference in metric chosen, which, in turn, changed how the outcome was portrayed and thus interpreted. Hypotheses generated included the following: The usefulness of a specific metric may vary by the user's level of skill from novice to expert; demonstration of the step order in skill performance may suffice for initial training of novices; a mechanical metric of effectiveness, like pulse stoppage, may aid in later training of novices; and training users how to practice on their own and self-assess performance may aid their self-development. The outcome of the performance assessments varied depending on the choice of metric in this study of simulated use of tourniquets.


Scaling of the kinetics of slow aggregation and gel formation for a fluorinated polymer colloid.
The aggregation and gelation kinetics in moderately concentrated (0.004 </= phi(0) </= 0.1) colloidal dispersions of fluorinated polymer particles has been studied. The aggregation was adjusted to proceed slowly enough to allow a convenient characterization of the kinetics through static and dynamic light scattering on quenched and diluted samples. A population balance model based on second-order aggregation rates is developed to compute the time evolution of the cluster mass distribution, from which we calculate the values of the average radii and structure factor measured by light scattering, so as to allow a direct comparison between measured and calculated quantities. The model suggests the introduction of a dimensionless time which allows the scaling of all the aggregation data on unique master curves defined by only two parameters: the exponent of the power-law aggregation kernel, lambda, and the aggregate fractal dimension, d(f). The predicted master curves were observed experimentally, which confirms the validity of the aggregation model and allows the unique determination of the kinetic and structural parameters of the aggregation process. The cluster growth behavior, although significantly slower than DLCA, shows power-law kinetics rather than the exponential one typical of RLCA and the cluster structure is characterized by an unexpectedly small fractal dimension, d(f) = 1.7. The occurrence of gelation has been characterized using small amplitude oscillatory shearing to monitor the time evolution of the elastic modulus. It is found that also these curves, together with the gel time value, scale with the stability ratio of primary particles for a given solid volume fraction. We further use the model to calculate the cumulative occupied volume fraction of the growing aggregates and quantify in this way the increasing space filling, which is solid volume fraction dependent. The experimentally determined dimensionless gel times, which are also solid volume fraction dependent, scale then directly with the dimensionless time to reach a certain degree of the space filling in the model. This finding suggests that, like the aggregation kinetics, the gelation kinetics is governed by a second-order rate process.


Past Decline Versus Current eGFR and Subsequent Mortality Risk.
A single determination of eGFR associates with subsequent mortality risk. Prior decline in eGFR indicates loss of kidney function, but the relationship to mortality risk is uncertain. We conducted an individual-level meta-analysis of the risk of mortality associated with antecedent eGFR slope, adjusting for established risk factors, including last eGFR, among 1.2 million subjects from 12 CKD and 22 other cohorts within the CKD Prognosis Consortium. Over a 3-year antecedent period, 12% of participants in the CKD cohorts and 11% in the other cohorts had an eGFR slope <-5 ml/min per 1.73 m(2) per year, whereas 7% and 4% had a slope >5 ml/min per 1.73 m(2) per year, respectively. Compared with a slope of 0 ml/min per 1.73 m(2) per year, a slope of -6 ml/min per 1.73 m(2) per year associated with adjusted hazard ratios for all-cause mortality of 1.25 (95% confidence interval [95% CI], 1.09 to 1.44) among CKD cohorts and 1.15 (95% CI, 1.01 to 1.31) among other cohorts during a follow-up of 3.2 years. A slope of +6 ml/min per 1.73 m(2) per year also associated with higher all-cause mortality risk, with adjusted hazard ratios of 1.58 (95% CI, 1.29 to 1.95) among CKD cohorts and 1.43 (95% CI, 1.11 to 1.84) among other cohorts. Results were similar for cardiovascular and noncardiovascular causes of death and stronger for longer antecedent periods (3 versus <3 years). We conclude that prior decline or rise in eGFR associates with an increased risk of mortality, independent of current eGFR.


Sorting of Leishmania-bearing dendritic cells reveals subtle parasite-induced modulation of host-cell gene expression.
Once in the mouse skin, Leishmania (L) amazonensis amastigotes are hosted by professional mononuclear phagocytes such as dendritic cells (DCs). When monitored after parasite inoculation, the frequency of amastigote-hosting DCs is very low (<1%) in both the skin and skin-draining lymph nodes. Therefore, we designed and validated an efficient procedure to purify live amastigotes-hosting DCs with the objective to facilitate quantitative and qualitative analysis of such rare cells. To this end, a L. amazonensis transgenic parasite expressing DsRed2 fluorescent protein was generated and added to mouse bone marrow-derived DC cultures. Then, a high speed sorting procedure, performed in BSL-2 containment, was setup to pick out only DCs hosting live amastigotes. This study reveals, for the first time, a unique transcript pattern from sorted live amastigotes-hosting DCs that would have been undetectable in unsorted samples. It was indeed possible to highlight a significant and coordinated up-regulation of L-arginine transporter and arginase2 transcripts in Leishmania-hosting DCs compared to un-parasitized DCs. These results indicate that arginine catabolism for polyamine generation is dominating over L-arginine catabolism for NO generation. In conclusion, this approach provides a powerful method for further characterisation, of amastigote-hosting DCs in the skin and the skin-draining lymph nodes.


Role of cryoanalgesia in the control of pain after thoracotomy.
Thoracotomy causes severe postoperative pain, which is difficult to manage since the use of systemic analgesics often causes respiratory depression. Cryoanalgesia of the intercostal nerves has been advocated as an effective means of local analgesia without serious side effects. A prospective randomised blind trial to investigate the efficacy of the technique was carried out. A total of 53 patients undergoing thoracotomy were allocated to either the trial or a control group. At thoracotomy the surgeon was informed of the patient's trial allocation. The trial group received one minute of direct cryotherapy to at least five intercostal nerves related to the incision. All patients received methadone via the lumbar epidural route in a dose calculated according to their weight. A linear analogue assessment of postoperative pain was made by the patients as soon as they were sufficiently awake. An independent record of all postoperative analgesia was kept. After discharge from hospital further assessments were made at least six weeks after operation. Statistical analysis of the scores of postoperative pain and analgesic consumption showed that there was no significant difference between the trial and the control group. There was, however, a suggestion of an increase in the long term morbidity, although these figures were not amenable to statistical analysis. Thus is has not been possible to demonstrate a role for cryoanalgesia in the control of post thoracotomy pain.


Tick-borne encephalitis among U.S. travelers to Europe and Asia - 2000-2009.
Tick-borne encephalitis virus (TBEV) is the most common arbovirus transmitted by ticks in Europe. Approximately 10,000 cases of tick-borne encephalitis (TBE) are reported annually in Europe and Russia. Although TBE is endemic in parts of China, information regarding its incidence is limited. TBEV is closely related to Powassan virus (POWV), another tick-borne flavivirus that is a rare cause of encephalitis in North America and Russia; TBEV and POWV can cross-react in serologic tests. Before 2000, two cases of TBE in North American travelers to Europe were reported. State health officials or clinicians send specimens from patients with unexplained encephalitis to CDC as part of routine surveillance and diagnostic testing. CDC recently reviewed all 2000-2009 laboratory records to identify cases of TBE among U.S. travelers; the five cases identified are summarized in this report. All five cases had TBEV or POWV immunoglobulin M (IgM) antibodies in serum and were confirmed as acute TBE cases by plaque-reduction neutralization tests against both viruses. All four patients who had traveled to Europe or Russia had biphasic illnesses (a common feature of TBE) and made nearly complete recoveries. The fifth patient, the first reported case of TBE in a U.S. traveler to China, had a monophasic illness with severe encephalitis and neurologic sequelae. Health-care providers should be aware of TBE, should counsel travelers about measures to reduce exposure to tick bites, and should consider the diagnosis of TBE in travelers returning from TBE-endemic countries with meningitis or encephalitis.


Investigation of single and synergic effects of NLRC5 and PD-L1 variants on the risk of colorectal cancer.
Constitutive activation of interferon signaling pathways has been reported in colorectal cancer (CRC), leading to a strong CD8+ T cell response through stimulation of NLRC5 expression. Primed CD8+ T cell expansion, however, may be negatively regulated by PD-L1 expression. Additionally, aberrant PD-L1 expression enables cancer cells to escape the immune attack. Our study aimed to select potential regulatory variants in the NLRC5 and PD-L1 genes by using several online in silico tools, such as UCSC browser, HaploReg, Regulome DB, Gtex Portal, microRNA and transcription factor binding site prediction tools and to investigate their influence on CRC risk in a Czech cohort of 1424 CRC patients and 1114 healthy controls. Logistic regression analysis adjusted for age and gender reported a moderate association between rectal cancer risk and two NLRC5 SNPs, rs1684575 T>G (OR: 1.60, 95% CI: 1.13-2.27, recessive model) and rs3751710 (OR: 0.70, 95% CI: 0.51-0.96, dominant model). Given that a combination of genetic variants, rather than a single polymorphism, may explain better the genetic etiology of CRC, we studied the interplay between the variants within NLRC5, PD-L1 and the previously genotyped IFNGR1 and IFNGR2 variants, to evaluate their involvement in the risk of CRC development. Overall we obtained 18 pair-wise interactions within and between the NLRC5 ad PD-L1 genes and 6 more when IFNGR variants were added. Thirteen out of the 24 interactions were below the threshold for the FDR calculated and controlled at an arbitrary level q*<0.10. Furthermore, the interaction IFNGR2 rs1059293 C>T-NLRC5 rs289747 G>A (P<0.0001) remained statistically significant even after Bonferroni correction. Our data suggest that not only a single genetic variant but also an interaction between two or more variants within genes involved in immune regulation may play important roles in the onset of CRC, providing therefore novel biological information, which could eventually improve CRC risk management but also PD-1-based immunotherapy in CRC.


Studies on transmitted beam modulation effect from laser induced damage on fused silica optics.
UV laser induced damage (LID) on exit surface of fused silica could cause modulation effect to transmitted beam and further influence downstream propagation properties. This paper presents our experimental and analytical studies on this topic. In experiment, a series of measurement instruments are applied, including beam profiler, interferometer, microscope, and optical coherent tomography (OCT). Creating and characterizing of LID on fused silica sample have been implemented. Morphological features are studied based on their particular modulation effects on transmitted beam. In theoretical investigation, analytical modeling and numerical simulation are performed. Modulation effects from amplitude, phase, and size factors are analyzed respectively. Furthermore, we have novelly designed a simplified polygon model to simulate actual damage site with multiform modulation features, and the simulation results demonstrate that the modeling is usable and representative.


Impact of different HIV resistance interpretation by distinct systems on clinical utility of resistance testing.
Genotypic assays are widely used tools for determining HIV-1 drug resistance and for guiding treatment. Several systems have been developed to interpret the complex influence of amino acid substitutions in HIV reverse transcriptase or protease on the phenotypic susceptibility or clinical response to the 18 available antiretroviral agents. In this review we analyse both studies comparing interpretations by different systems and studies showing correlation of interpretations with clinical outcome, in order to identify discordance and how this may affect prediction of subsequent therapy outcomes. During the last year, several studies analysing interpretation systems, individually or comparatively, have shown substantial variability of the predicted drug activities and therapeutic outcomes. Discrepant interpretation was detected mostly for nucleoside reverse transcriptase inhibitors and rarely for non-nucleoside reverse transcriptase inhibitors. Better correlation with treatment outcome was found with most recently updated systems, while a weaker prediction was found with systems interpreting activity of nucleoside reverse transcriptase inhibitors solely on the basis of phenotypic susceptibility. Virological, patient-related and treatment-related factors can all affect the results of systems' clinical validations. Refinement of resistance interpretation is possible by introducing rules derived from genotype-outcomes correlation or, at least for protease inhibitors, genotype-phenotype correlation. Papers showing clinical validation of the available interpretation systems are presented with a critical view to help the readers' evaluation of their possible use. There is a need for developing a consensus towards common interpretations. Large clinical and virological databases with quality data will be useful for future improvements.


Full-field 3D deformation measurement: comparison between speckle phase and displacement evaluation.
The objective of this paper is to describe a full-field deformation measurement method based on 3D speckle displacements. The deformation is evaluated from the slope of the speckle displacement function that connects the different reconstruction planes. For our experiment, a symmetrical arrangement with four illuminations parallel to the planes (x,z) and (y,z) was used. Four sets of speckle patterns were sequentially recorded by illuminating an object from the four directions, respectively. A single camera is used to record the holograms before and after deformations. Digital speckle photography is then used to calculate relative speckle displacements in each direction between two numerically propagated planes. The 3D speckle displacements vector is calculated as a combination of the speckle displacements from the holograms recorded in each illumination direction. Using the speckle displacements, problems associated with rigid body movements and phase wrapping are avoided. In our experiment, the procedure is shown to give the theoretical accuracy of 0.17 pixels yielding the accuracy of 2×10-3 in the measurement of deformation gradients.


Morphological sperm alternations in different types of prostatitis.
The possible negative effect of the chronic pelvic pain syndrome (CPPS) on semen parameters and especially sperm morphology is still a controversial matter. The aim of this study was therefore to investigate if different types of prostatitis can have a negative effect on sperm morphology and to compare our results with that of a literature survey. Semen analyses were performed on 34 males with confirmed CPPS (NIH III A), 18 males with CPPS (NIH III B) and 17 males as controls. When sperm morphology was evaluated according to WHO criteria, no differences were found between the mean percentages of morphologically normal spermatozoa for the three groups. An extended sperm morphology evaluation according to strict criteria showed that the NIH III A group had a tendency for a lower percentage of morphologically normal spermatozoa (5.3 +/- 3.1%) and acrosome index (8.7 +/- 4.8%) compared with the control group with values of 7.3 +/- 5.6% and 12.7 +/- 7.3%, respectively. There was a statistically significant higher (P = 0.0186) mean percentage (17.5 +/- 15.7%) of elongated spermatozoa in the NIH III A group compared with the control group (7.2 +/- 9.5%) while the NIH III B group had values between those of the control and NIH III A group. Our results indicate that CPPS NIH III A can have a significant negative effect on sperm morphology parameters, as evaluated by strict criteria, and to a lesser extent in cases of CPPS NIH III B compared with a control group.


Insights in the use of health care services in chronic benign pain in childhood and adolescence.
The utilization of health care services in children and adolescents with chronic benign pain was studied in a Dutch population sample of 254 chronic pain sufferers aged 0-18 years. Children and adolescents who had reported chronic pain (continuous or recurrent pain >3 months) in our previous prevalence study were asked to keep a 3-week diary on their pain and to fill out questionnaires on background factors, health care use and the impact of pain. Parent ratings were used for children aged 0-11 years, self-report was used in adolescents (12-18 years). In a 3-month period, in 53.4% of the cases medication was used for pain, and general practitioners and specialists were consulted for pain in 31.1% and 13.9% of subjects, respectively. Physiotherapists, psychologists and alternative health providers were visited by 11.5, 2.8, and 4.0%, respectively. In the preceding year, 6.4% had been hospitalized due to pain. The most important factors linked to utilizing medical services were gender, various pain characteristics, school absenteeism and disability. Although consulters reported to be less physically fit and less satisfied with health, their parents were better adapted to the pain, by talking and sharing, mutual support, normalization of the child and heightened self-esteem, than non-consulters. Prospective studies are needed to test causality of coping on care-seeking behavior.


Energy decay in a tapped granular column: Can a one-dimensional toy model provide insight into fully three-dimensional systems?
The decay of energy within particulate media subjected to an impulse is an issue of significant scientific interest, but also one with numerous important practical applications. In this paper, we study the dynamics of a granular system exposed to energetic impulses in the form of discrete taps from a solid surface. By considering a one-dimensional toy system, we develop a simple theory, which successfully describes the energy decay within the system following exposure to an impulse. We then extend this theory so as to make it applicable also to more realistic, three-dimensional granular systems, assessing the validity of the model through direct comparison with discrete particle method simulations. The theoretical form presented possesses several notable consequences; in particular, it is demonstrated that for suitably large systems, effects due to the bounding walls may be entirely neglected. We also establish the existence of a threshold system size above which a granular bed may be considered fully three dimensional.


A Fresh Look at Resonances and Complex Absorbing Potentials: Density Matrix-Based Approach.
A new strategy of using complex absorbing potentials (CAPs) within electronic structure calculations of metastable electronic states, which are ubiquitous in chemistry and physics, is presented. The stumbling block in numerical applications of CAPs is the necessity to optimize the CAP strength for each system, state, and one-electron basis set, while there is no clear metric to assess the quality of the results and no simple algorithm of achieving numerical convergence. By analyzing the behavior of resonance wave functions, we found that robust results can be obtained when considering fully stabilized resonance states characterized by constant density at large η (parameter determining the CAP strength). Then the perturbation due to the finite-strength CAP can be removed by a simple energy correction derived from energy decomposition analysis and response theory. The utility of this approach is illustrated by CAP-augmented calculations of several shape resonances using EOM-EA-CCSD with standard Gaussian basis sets.


Structural covariance networks of the dorsal anterior insula predict females' individual differences in empathic responding.
Previous functional imaging studies have shown key roles of the dorsal anterior insula (dAI) and anterior midcingulate cortex (aMCC) in empathy for the suffering of others. The current study mapped structural covariance networks of these regions and assessed the relationship between networks and individual differences in empathic responding in 94 females. Individual differences in empathy were assessed through average state measures in response to a video task showing others' suffering, and through questionnaire-based trait measures of empathic concern. Overall, covariance patterns indicated that dAI and aMCC are principal hubs within prefrontal, temporolimbic, and midline structural covariance networks. Importantly, participants with high empathy state ratings showed increased covariance of dAI, but not aMCC, to prefrontal and limbic brain regions. This relationship was specific for empathy and could not be explained by individual differences in negative affect ratings. Regarding questionnaire-based empathic trait measures, we observed a similar, albeit weaker modulation of dAI covariance, confirming the robustness of our findings. Our analysis, thus, provides novel evidence for a specific contribution of frontolimbic structural covariance networks to individual differences in social emotions beyond negative affect.


A transposon insertion in the Escherichia coli uvrC gene; UvrC protein is absolutely required for the incision step in excision repair.
The formation of single-stranded breaks in DNA following UV irradiation is assessed in uvrC34 mutants. By altering the SOS DNA-repair system, either by additional mutations or by using drugs affecting transcription or translation, it is shown that such single-stranded breaks require one or more DNA-damage-inducible functions. A UV-sensitive strain is characterized as carrying a Tn10 insertion into the uvrC gene. The absence of post-irradiation incision in this strain demonstrates that uvrC function is absolutely required in vivo for the incision stage of excision repair, and suggests that other uvrC mutants are 'leaky'.


The large subunit of HIV-1 reverse transcriptase interacts with beta-actin.
HIV-1 reverse transcriptase is a dimeric enzyme mainly involved in the replication of the viral genome. A filamentous phage cDNA expression library from human lymphocytes was used to select cellular proteins interacting with HIV-1 reverse transcriptase Affinity selections using the bacterially expressed monomeric large subunit of reverse transcriptase (p66) yielded host beta-actin. This clone was expressed as glutathione-S-transferase fusion protein which was identified by using a specific antibody against beta-actin. Furthermore we show that also the eukaryotic beta-actin binds to either the large subunit of reverse transcriptase or to the Pol precursor polyprotein in vitro. The reverse transcriptase/beta-actin interaction might be important for the secretion of HIV-1 virions.


[Sexual abuse of children and adolescents: characteristics of sexual victimization in family relations].
This study analyzes the characteristics of sexual abuse committed within the family against age groups classified according to the Brazilian Statute for Children and Adolescents (the prevailing legislation on matters pertaining to minors) and treated at the Reference Center for Children and Adolescents and the Guardianship Councils in Ribeirão Preto, São Paulo State, Brazil, from 1995 to 2000. Some 234 abuses were identified, committed by 217 aggressors, against 210 families and a total of 226 victims. A total of 131 children (48.7%) and 95 adolescents (41.2%), predominantly females, were victimized. Children ranging from 10 to 12 years were the most frequently abused (19.5%), as well as adolescents from 12 to 14 years old (17.3%). The majority of the victims live in families with 3 (19.9%) or 4 children (177%), and the firstborn are the most frequently abused (33.6%). The majority of aggressors who acted alone victimized only one individual (86.7%). Fathers (34.2%) and stepfathers (30.3%) were the most frequent aggressors, with the former victimizing more children (19.7%) and the latter adolescents (17.1%).


Detection of HCV-Specific IFN-γ Responses in HCV Antibody and HCV RNA Negative Injecting Drug Users.
Detectable HCV-specific cellular immune responses in HCV antibody and RNA negative people who inject drugs (PWID) raise the question of whether some are resistant to HCV infection. Immune responses from people who have been exposed to hepatitis C virus (HCV) and remain anti-HCV negative are of interest for HCV vaccine development; however, limited research addresses this area. In a cohort of HCV antibody and RNA negative PWID, we assessed whether the presence of HCV-specific IFN-γ responses or genetic associations provide any evidence of protection from HCV infection. One hundred and ninety-eight participants were examined longitudinally for clinical, behavioral, social, environmental and genetic characteristics (IFNL3 genotype [formally IL-28B] and HLA type). Sixty-one of the 198 participants were HCV antibody and RNA negative, with 53 able to be examined longitudinally for HCV-specific IFN-γ ELISpot T cell responses. Ten of the 53 HCV antibody and RNA negative participants had detectable HCV-specific IFN-γ responses at baseline (18%). The magnitude of IFN-γ responses averaged 131 +/- 96 SFC/106 PBMC and the breadth was mean 1 +/- 1 pool positive. The specificity of responses were mainly directed to E2, NS4b and NS5b. Participants with (10) and without (43) HCV-specific IFN-γ responses did not differ in behavioral, clinical or genetic characteristics (P > 0.05). There was a larger proportion sharing needles (with 70%, without 49%, P = 0.320) and a higher incidence of HCV (with 35.1 per 100 py, 95% CI 14.6, 84.4, without 16.0 per 100 py, 95% CI 7.2, 35.6, P = 0.212) in those with IFN-γ responses, although not statistically significant. Half the participants with baseline IFN-γ responses became HCV RNA positive (5/10), with one of these participants spontaneously clearing HCV. The spontaneous clearer had high magnitude and broad Th1 responses, favorable IFNL3 genotype and favorable HLA types. This study demonstrated the detection of HCV-specific IFN-γ responses in HCV antibody and RNA negative individuals, with a tendency for HCV-specific IFN-γ responses to be associated with HCV exposure. The potential role of HCV-specific IFN-γ responses in those who remained HCV RNA negative is of value for the development of novel HCV therapeutics.


The Toxin-Antitoxin System DarTG Catalyzes Reversible ADP-Ribosylation of DNA.
The discovery and study of toxin-antitoxin (TA) systems helps us advance our understanding of the strategies prokaryotes employ to regulate cellular processes related to the general stress response, such as defense against phages, growth control, biofilm formation, persistence, and programmed cell death. Here we identify and characterize a TA system found in various bacteria, including the global pathogen Mycobacterium tuberculosis. The toxin of the system (DarT) is a domain of unknown function (DUF) 4433, and the antitoxin (DarG) a macrodomain protein. We demonstrate that DarT is an enzyme that specifically modifies thymidines on single-stranded DNA in a sequence-specific manner by a nucleotide-type modification called ADP-ribosylation. We also show that this modification can be removed by DarG. Our results provide an example of reversible DNA ADP-ribosylation, and we anticipate potential therapeutic benefits by targeting this enzyme-enzyme TA system in bacterial pathogens such as M. tuberculosis.


Understanding hydrogelation processes through molecular dynamics.
Molecular dynamics (MD) is currently one of the preferred techniques employed to understand hydrogelation processes for its ability to include large amounts of atoms in computational calculations, since substantial amounts of solvent molecules are involved in gel formation. MD studies have helped to rationalize experimental outcomes that in many occasions were not well understood based on experimental observations. Additionally, MD has been used to study changes in gel physical properties triggered by variations in reaction conditions or gelator structures. Changes in many physical properties were understood using MD, including molecular diffusion, hydrogel swelling and volume transitions. All the examples gathered in this review might help the reader to discover the current state of the art in MD studies carried out to study hydrogelation processes as well as the pioneering studies that paved the way to introduce MD in the field of gels.


Mitochondrial alterations of skeletal muscle in a heat stress rat model.
The morphological and ultrastructural alterations of skeletal muscle in experimental rats with heat stress were investigated. Fifteen male Sprague-Dawley rats were exposed in a 42 degrees C constant temperature oven, resulting in a heat stress state; ten rats were used as controls. All treated rats had weakness of the 4 limbs associated with increased serum creatine kinase levels (p < 0.01). Soleus muscles were submitted to histological, histochemical, ultrastructural and quantitative-morphometric analysis. The group receiving heat stress showed many ragged-red fibers in Gomori trichrome stain and appeared hyper-reactive in succinate dehydrogenase and cytochrome C oxidase stains. The ultrastructure of ragged-red fibers showed increased mitochondrial aggregation as multiple small nests, which were particularly located in the subsarcolemmal space. The mitochondrial area was significantly increased in heat stress rats (p < 0.001). The consistently increased mitochondrial area and histochemical alterations of mitochondria are early pathological abnormalities in muscles with heat stress and indicate fundamental impairment of energy metabolism.


PI3K inhibition results in enhanced estrogen receptor function and dependence in hormone receptor-positive breast cancer.
Activating mutations of PIK3CA are the most frequent genomic alterations in estrogen receptor (ER)-positive breast tumors, and selective phosphatidylinositol 3-kinase α (PI3Kα) inhibitors are in clinical development. The activity of these agents, however, is not homogeneous, and only a fraction of patients bearing PIK3CA-mutant ER-positive tumors benefit from single-agent administration. Searching for mechanisms of resistance, we observed that suppression of PI3K signaling results in induction of ER-dependent transcriptional activity, as demonstrated by changes in expression of genes containing ER-binding sites and increased occupancy by the ER of promoter regions of up-regulated genes. Furthermore, expression of ESR1 mRNA and ER protein were also increased upon PI3K inhibition. These changes in gene expression were confirmed in vivo in xenografts and patient-derived models and in tumors from patients undergoing treatment with the PI3Kα inhibitor BYL719. The observed effects on transcription were enhanced by the addition of estradiol and suppressed by the anti-ER therapies fulvestrant and tamoxifen. Fulvestrant markedly sensitized ER-positive tumors to PI3Kα inhibition, resulting in major tumor regressions in vivo. We propose that increased ER transcriptional activity may be a reactive mechanism that limits the activity of PI3K inhibitors and that combined PI3K and ER inhibition is a rational approach to target these tumors.


Plasma concentrations of vitamin B12 and folate and global cognitive function in an older population: cross-sectional findings from The Irish Longitudinal Study on Ageing (TILDA).
The uncertainty surrounding high intakes of folic acid and associations with cognitive decline in older adults with low vitamin B12 status has been an obstacle to mandatory folic acid fortification for many years. We estimated the prevalence of combinations of low/normal/high vitamin B12 and folate status and compared associations with global cognitive function using two approaches, of individuals in a population-based study of those aged ≥50 years in the Republic of Ireland. Cross-sectional data from 3781 men and women from Wave 1 of The Irish Longitudinal Study on Ageing were analysed. Global cognitive function was assessed by the Mini Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA). Prevalence estimates for combinations of vitamin B12 (plasma vitamin B12 < or ≥258 pmol/l) and folate (plasma folate ≤ or >45·3 nmol/l) concentrations were generated. Negative binomial regression models were used to investigate the associations of vitamin B12 and folate status with global cognitive function. Of the participants, 1·5 % (n 51) had low vitamin B12 (<258 pmol/l) and high folate (>45·3 nmol/l) status. Global cognitive performance was not significantly reduced in these individuals when compared with those with normal status for both B-vitamins (n 2433). Those with normal vitamin B12/high folate status (7·6 %) had better cognitive performance (MMSE: incidence rate ratio (IRR) 0·82, 95 % CI 0·68, 0·99; P = 0·043, MoCA: IRR 0·89, 95 % CI 0·80, 0·99; P = 0·025). We demonstrated that high folate status was not associated with lower cognitive scores in older adults with low vitamin B12 status. These findings provide important safety information that could guide fortification policy recommendations in Europe.


Lipoprotein(a) and calcific aortic valve disease: current evidence and future directions.
Calcific aortic valve disease (CAVD), the most common cause of aortic stenosis (AS), is characterized by slowly progressive fibrocalcific remodelling of the valve cusps. Once symptomatic, severe AS is associated with poor survival unless surgical or transcatheter valve replacement is performed. Unfortunately, no pharmacological interventions have been demonstrated to alter the natural history of CAVD. Lipoprotein(a) [Lp(a)], a low-density lipoprotein-like particle, has been implicated in the pathophysiology of CAVD. The mechanisms by which Lp(a) results in CAVD are not well understood. However, the oxidized phospholipids carried by Lp(a) are considered a crucial mediator of the disease process. An increasing number of studies demonstrate a causal association between plasma Lp(a) levels and frequency of AS and need for aortic valve replacement, which is independent of inflammation, as measured by plasma C-reactive protein levels. However, not all studies show an association between Lp(a) and increased progression of calcification in individuals with established CAVD. Epidemiologic, genetic, and Mendelian randomization studies have collectively suggested that Lp(a) is a causal risk factor for CAVD. Whether Lp(a)-lowering can prevent initiation or slow progression of CAVD remains to be demonstrated.


The spread of EBV to ectopic lymphoid aggregates may be the final common pathway in the pathogenesis of ME/CFS.
According to the hypothesis presented here, myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) develops over 3 steps: Step 1 is characterized by the aggregation of lymphoid cells in dorsal root ganglia or other nervous structures. The cause of this formation of ectopic lymphoid aggregates may be an acute infection, asymptomatic reactivations of a common neurotropic virus, exposure to a neurotoxin, or physical injury to peripheral nerves. In step 2, Epstein-Barr virus (EBV)-infected lymphocytes or monocytes bring EBV from the circulation to one or several of these lymphoid aggregates, whereupon cell-to-cell transmission of EBV and proliferation of latently EBV-infected lymphocytes lead to the presence of many EBV-infected cells in the lymphoid aggregates. The EBV-infected cells in the aggregates ignite an inflammation in the surrounding nervous tissue. This local inflammation elicits, in turn, a wave of glial cell activation that spreads from the EBV-infected area to parts of the nervous system that are not EBV-infected, disturbing the neuron-glial interaction in both the peripheral - and central nervous system. In step 3, immune cell exhaustion contributes to a consolidation of the pathological processes. There might be a cure: Infusions of autologous EBV-specific T-lymphocytes can perhaps remove the EBV-infected cells from the nervous system.


Salvia (Sage): A Review of its Potential Cognitive-Enhancing and Protective Effects.
Genus Salvia, commonly known as sage, is the largest genus in the Lamiaceae family. It comprises many species traditionally used as brain-enhancing tonics. In vitro and animal studies have confirmed that several Salvia species contain a large array of active compounds that may enhance cognitive activity and protect against neurodegenerative disease. In this review, the active constituents in plants belonging to the genus Salvia are summarised, and their influence on pharmacodynamics pertinent to cognitive activity are detailed. In particular, the effects of plants belonging to the genus Salvia and their constituents on cognitive skills including memory, attention and learning are detailed. Their potential effects in dementia, including Alzheimer's disease, are also examined. Completed human trials are summarised, and factors influencing the potency of Salvia plants are covered. Finally, directions for future research are proposed to enhance our understanding of the potential health benefits of Salvia plants.


Effect of repetitive stimulation on lateral spreads and F-waves in hemifacial spasm.
The lateral spread (LS) response, which can be elicited in muscles innervated by other branches of the facial nerve, is electromyographycally specific for patients with hemifacial spasm (HFS), occurring about 10 ms after stimulus. The F-wave in facial muscles, which is a late response that antidromicaly propagates to the facial motonucleus and returns orthodromicaly down the same axon, revealed a trend toward enhancement in patients with HFS. The LSs were facilitated by repetitive stimulation during the microvascular decompression (MVD) operation, which has proved to be a successful treatment, and the F-waves were also facilitated by repetitive stimulation on the spasm side more than on the normal side. Greater facilitation of these responses was in direct proportion to higher stimulation rates and greater numbers of stimulations. The repetitive stimulation of the facial nerve may result in activation of the motoneuron pool and in the lowering of the threshold of somatic membranes. These results support the hypothesis that hemifacial spasm is caused by hyperexcitability of the facial motonucleus, which is increased by antidromic repetitive stimulation.


Human breast cancer invasion and aggression correlates with ECM stiffening and immune cell infiltration.
Tumors are stiff and data suggest that the extracellular matrix stiffening that correlates with experimental mammary malignancy drives tumor invasion and metastasis. Nevertheless, the relationship between tissue and extracellular matrix stiffness and human breast cancer progression and aggression remains unclear. We undertook a biophysical and biochemical assessment of stromal-epithelial interactions in noninvasive, invasive and normal adjacent human breast tissue and in breast cancers of increasingly aggressive subtype. Our analysis revealed that human breast cancer transformation is accompanied by an incremental increase in collagen deposition and a progressive linearization and thickening of interstitial collagen. The linearization of collagen was visualized as an overall increase in tissue birefringence and was most striking at the invasive front of the tumor where the stiffness of the stroma and cellular mechanosignaling were the highest. Amongst breast cancer subtypes we found that the stroma at the invasive region of the more aggressive Basal-like and Her2 tumor subtypes was the most heterogeneous and the stiffest when compared to the less aggressive luminal A and B subtypes. Intriguingly, we quantified the greatest number of infiltrating macrophages and the highest level of TGF beta signaling within the cells at the invasive front. We also established that stroma stiffness and the level of cellular TGF beta signaling positively correlated with each other and with the number of infiltrating tumor-activated macrophages, which was highest in the more aggressive tumor subtypes. These findings indicate that human breast cancer progression and aggression, collagen linearization and stromal stiffening are linked and implicate tissue inflammation and TGF beta.


Comparison of metabolic profiles and bioactivities of the leaves of three edible Congolese Hibiscus species.
Methanolic and dichloromethane extracts from the leaves of Congolese Hibiscus species were characterised by chromatographic and spectroscopic techniques and their in vitro biochemical activities against ROS production were evaluated in cellular models and on an enzyme, myeloperoxidase (MPO), involved in inflammation. Hibiscus acetosella has a chemical fingerprint different from Hibiscus cannabinus and Hibiscus sabdariffa both having similar fingerprints. Major compounds were polyphenols, represented mainly by caffeoyl-hydroxycitric acid for H. acetosella and neochlorogenic acid for the two other species. All extracts displayed high cellular antioxidant activity with IC50 values ranging from 0.5 to 3 μg mL-1 using lucigenin on neutrophils. Dichloromethane extracts showed more efficient effects on extracellular ROS production and MPO activity. Antioxidant and anti-inflammatory activities of caffeoyl-hydroxycitric acid were significantly higher than those of neochlorogenic acid. The bioactivities of Hibiscus species were positively correlated with their phytochemical content and could justify their use as local nutraceutical resources and medicines.


[Dose optimization of anticancer drugs in the elderly].
Age-related physiological changes affect pharmacokinetics and pharmacodynamics in elderly individuals. Cancer treatment in the elderly requires a careful and multidisciplinary assessment of each patient prior to therapy initiation with respect to treatment decisions and dose optimization considerations, given the wide interpatient variability in this population. As renal function declines with age, renal function assessments are necessary when drugs increasing exposure in renal impairment will be administered to elderly patients. Serum creatinine is an insufficient marker for evaluating renal function in elderly patients because muscle mass decreases with age. Renal function should be assessed according to the index used for dosing recommendation at drug administration planning; in most of drugs, does adjustment is recommended according to the creatinine clearance (Ccr) calculated using the Cockcroft-Gault equation. Elderly patients are apparently more sensitive to cytotoxic agent-induced neutropenia and therefore should be monitored closely. Following the first cycle, a proper assessment of tolerability should be conducted before the second cycle; such assessments are required to discuss the appropriateness of the initiation dose and determine doses during the following cycles for each patient.


Eudiplozoon nipponicum (Monogenea, Diplozoidae) and its adaptation to haematophagy as revealed by transcriptome and secretome profiling.
Ectoparasites from the family Diplozoidae (Platyhelminthes, Monogenea) belong to obligate haematophagous helminths of cyprinid fish. Current knowledge of these worms is for the most part limited to their morphological, phylogenetic, and population features. Information concerning the biochemical and molecular nature of physiological processes involved in host-parasite interaction, such as evasion of the immune system and its regulation, digestion of macromolecules, suppression of blood coagulation and inflammation, and effect on host tissue and physiology, is lacking. In this study, we report for the first time a comprehensive transcriptomic/secretome description of expressed genes and proteins secreted by the adult stage of Eudiplozoon nipponicum (Goto, 1891) Khotenovsky, 1985, an obligate sanguivorous monogenean which parasitises the gills of the common carp (Cyprinus carpio). RNA-seq raw reads (324,941 Roche 454 and 149,697,864 Illumina) were generated, de novo assembled, and filtered into 37,062 protein-coding transcripts. For 19,644 (53.0%) of them, we determined their sequential homologues. In silico functional analysis of E. nipponicum RNA-seq data revealed numerous transcripts, pathways, and GO terms responsible for immunomodulation (inhibitors of proteolytic enzymes, CD59-like proteins, fatty acid binding proteins), feeding (proteolytic enzymes cathepsins B, D, L1, and L3), and development (fructose 1,6-bisphosphatase, ferritin, and annexin). LC-MS/MS spectrometry analysis identified 721 proteins secreted by E. nipponicum with predominantly immunomodulatory and anti-inflammatory functions (peptidyl-prolyl cis-trans isomerase, homolog to SmKK7, tetraspanin) and ability to digest host macromolecules (cathepsins B, D, L1). In this study, we integrated two high-throughput sequencing techniques, mass spectrometry analysis, and comprehensive bioinformatics approach in order to arrive at the first comprehensive description of monogenean transcriptome and secretome. Exploration of E. nipponicum transcriptome-related nucleotide sequences and translated and secreted proteins offer a better understanding of molecular biology and biochemistry of these, often neglected, organisms. It enabled us to report the essential physiological pathways and protein molecules involved in their interactions with the fish hosts.


Gene therapy for malignant mesothelioma: beyond the infant years.
Mesothelioma may be particularly well suited for gene therapy treatment owing to its accessibility, allowing both intrapleural and intratumoral gene delivery. At least four gene therapy trials have been carried out in mesothelioma patients, using different vector systems (adenovirus, vaccinia virus, irradiated tumor cells), and different transgenes (herpes simplex virus thymidine kinase (HSVtk) combined with ganciclovir, IL-2, IFN-beta). Although small in scale, these trials have given an inkling of hope for therapeutic efficacy. However, it is clear that gene therapy protocols need to be optimized further. This paper will review progress made in (i) vector development, (ii) defining optimal transgenes, and (iii) gene delivery. Adenoviruses are the most commonly used vectors for gene therapy, and are continuously being improved. With respect to the nature of the transgenes, five categories can be distinguished: (i) 'suicide' or sensitivity genes (e.g., HSVtk), (ii) cytokines and other immune modulators, (iii) replacements for mutant tumor suppressor genes (e.g., p53), (iv) antiangiogenic proteins and (v) tumor antigens. It seems clear that expression of a single transgene is unlikely to be sufficient to eradicate a tumor, such as mesothelioma, that is diagnosed late in disease progression. Hence, multimodality therapy, including conventional therapy (chemo- and radiotherapy, surgery) with one or more transgenes has a higher chance of success.


Control of immune pathology by regulatory T cells.
There is now compelling evidence that immune responses for both foreign and self antigens are downregulated by T cells that are specialised for this function; these are known as regulatory T (T reg) cells. This review describes progress in the characterisation of the T reg cells that mediate both mucosal tolerance and tolerance to self antigens. The recent work on the antigen specificity, generation and mode of action of T reg cells is also reviewed.


Fruit and vegetable consumption is inversely associated with having pancreatic cancer.
Studies on fruit, vegetable, fiber, and grain consumption and pancreatic cancer risk are inconclusive. We used a clinic-based case-control study specifically designed to address limitations of both cohort and case-control studies to examine the relationship. Participants were excluded who reported changing their diet within 5 years prior to study entry. And 384 rapidly ascertained cases and 983 controls (frequency matched on age (±5 years), race, sex, and residence) completed epidemiologic surveys and 144-item food frequency questionnaires. Odds ratios (OR) and 95% confidence intervals were calculated using logistic regression adjusted for age, sex, smoking, body mass index, energy intake, and alcohol consumption. Comparing highest to lowest quintiles, we observed significant inverse associations (OR < 0.8) with significant trends (p (trend) < 0.05) for citrus, melon, and berries, other fruits, dark green vegetables, deep yellow vegetables, tomato, other vegetables, dry bean and pea, insoluble fiber, soluble fiber, whole grains, and orange/grapefruit juice, and an increased association with non-whole grains. Results were similar after adjusting for diabetes or total sugar intake. We provide evidence that lower consumption of fruits, vegetables, whole grains, and fiber is associated with having pancreatic cancer. This may have a role in developing prevention strategies.