Analysis of the local organization and dynamics of cellular actin networks.
Actin filaments, with the aid of multiple accessory proteins, self-assemble into a variety of network patterns. We studied the organization and dynamics of the actin network in nonadhesive regions of cells bridging fibronectin-coated adhesive strips. The network was formed by actin nodes associated with and linked by myosin II and containing the formin disheveled-associated activator of morphogenesis 1 (DAAM1) and the cross-linker filamin A (FlnA). After Latrunculin A (LatA) addition, actin nodes appeared to be more prominent and demonstrated drift-diffusion motion. Superresolution microscopy revealed that, in untreated cells, DAAM1 formed patches with a similar spatial arrangement to the actin nodes. Node movement (diffusion coefficient and velocity) in LatA-treated cells was dependent on the level and activity of myosin IIA, DAAM1, and FlnA. Based on our results, we developed a computational model of the dynamic formin-filamin-actin asters that can self-organize into a contractile actomyosin network. We suggest that such networks are critical for connecting distant parts of the cell to maintain the mechanical coherence of the cytoplasm.


National quality standards for neuro rehabilitation in a community setting: Do they achieve their purpose?
There is known variation in neuro-rehabilitation service provision, however, the extent of service variation and impact on people who experience an acquired brain injury (ABI) is not articulated in the literature. The aim of this study was to assess and determine the extent to which neuro-rehabilitation services in one part of the United Kingdom (UK) are meeting national quality standards. A mixed method design, across five community neuro-rehabilitation providers and six districts in South London, comprised of ABI population incidence data, web-based surveys to determine compliance with the National Institute for Health and Care Excellence (NICE) Head Injury Quality Standard, and focus groups to understand the patient perspective of community neuro-rehabilitation service provision. The population incidence of ABI amongst districts demonstrated differences between the datasets analyzed, resulting in an inability to determine whether service variation was based on population need. The web-based surveys revealed that five community neuro-rehabilitation providers have variations between the models of care provided, including clinical referral criteria, duration, intensity of therapy interventions, and overall cost per patient, which was correlated with workforce capacity and patient waiting times. Focus group discussion highlighted current key challenges of service restraints, disconnect between services and limited professional support, as well as improvement opportunities pertaining to access, flexible, local and timely health and social care services. This study indicates that despite the publication of the NICE Head Injury Quality Standard, there is variation in the local provision of community neuro-rehabilitation across six districts in South London. Each district partly meets the recommendations, highlighting variability in the model of care delivered, that impacts consumers/carers accessing quality neuro-rehabilitation services. A disconnect remains between evidence-based quality standards and implementation. No standardized ABI data set is available in the UK, which impacts planning for future clinical service delivery.


A reference genetic map of C. clementina hort. ex Tan.; citrus evolution inferences from comparative mapping.
Most modern citrus cultivars have an interspecific origin. As a foundational step towards deciphering the interspecific genome structures, a reference whole genome sequence was produced by the International Citrus Genome Consortium from a haploid derived from Clementine mandarin. The availability of a saturated genetic map of Clementine was identified as an essential prerequisite to assist the whole genome sequence assembly. Clementine is believed to be a 'Mediterranean' mandarin × sweet orange hybrid, and sweet orange likely arose from interspecific hybridizations between mandarin and pummelo gene pools. The primary goals of the present study were to establish a Clementine reference map using codominant markers, and to perform comparative mapping of pummelo, sweet orange, and Clementine. Five parental genetic maps were established from three segregating populations, which were genotyped with Single Nucleotide Polymorphism (SNP), Simple Sequence Repeats (SSR) and Insertion-Deletion (Indel) markers. An initial medium density reference map (961 markers for 1084.1 cM) of the Clementine was established by combining male and female Clementine segregation data. This Clementine map was compared with two pummelo maps and a sweet orange map. The linear order of markers was highly conserved in the different species. However, significant differences in map size were observed, which suggests a variation in the recombination rates. Skewed segregations were much higher in the male than female Clementine mapping data. The mapping data confirmed that Clementine arose from hybridization between 'Mediterranean' mandarin and sweet orange. The results identified nine recombination break points for the sweet orange gamete that contributed to the Clementine genome. A reference genetic map of citrus, used to facilitate the chromosome assembly of the first citrus reference genome sequence, was established. The high conservation of marker order observed at the interspecific level should allow reasonable inferences of most citrus genome sequences by mapping next-generation sequencing (NGS) data in the reference genome sequence. The genome of the haploid Clementine used to establish the citrus reference genome sequence appears to have been inherited primarily from the 'Mediterranean' mandarin. The high frequency of skewed allelic segregations in the male Clementine data underline the probable extent of deviation from Mendelian segregation for characters controlled by heterozygous loci in male parents.


Behavioral and other phenotypes in a cytoplasmic Dynein light intermediate chain 1 mutant mouse.
The cytoplasmic dynein complex is fundamentally important to all eukaryotic cells for transporting a variety of essential cargoes along microtubules within the cell. This complex also plays more specialized roles in neurons. The complex consists of 11 types of protein that interact with each other and with external adaptors, regulators and cargoes. Despite the importance of the cytoplasmic dynein complex, we know comparatively little of the roles of each component protein, and in mammals few mutants exist that allow us to explore the effects of defects in dynein-controlled processes in the context of the whole organism. Here we have taken a genotype-driven approach in mouse (Mus musculus) to analyze the role of one subunit, the dynein light intermediate chain 1 (Dync1li1). We find that, surprisingly, an N235Y point mutation in this protein results in altered neuronal development, as shown from in vivo studies in the developing cortex, and analyses of electrophysiological function. Moreover, mutant mice display increased anxiety, thus linking dynein functions to a behavioral phenotype in mammals for the first time. These results demonstrate the important role that dynein-controlled processes play in the correct development and function of the mammalian nervous system.


A simple artificial light-harvesting dyad as a model for excess energy dissipation in oxygenic photosynthesis.
Under excess illumination, plant photosystem II dissipates excess energy through the quenching of chlorophyll fluorescence, a process known as nonphotochemical quenching. Activation of nonphotochemical quenching has been linked to the conversion of a carotenoid with a conjugation length of nine double bonds (violaxanthin) into an 11-double-bond carotenoid (zeaxanthin). It has been suggested that the increase in the conjugation length turns the carotenoid from a nonquencher into a quencher of chlorophyll singlet excited states, but unequivocal evidence is lacking. Here, we present a transient absorption spectroscopic study on a model system made up of a zinc phthalocyanine (Pc) molecule covalently linked to carotenoids with 9, 10, or 11 conjugated carbon-carbon double bonds. We show that a carotenoid can act as an acceptor of Pc excitation energy, thereby shortening its singlet excited-state lifetime. The conjugation length of the carotenoid is critical to the quenching process. Remarkably, the addition of only one double bond can turn the carotenoid from a nonquencher into a very strong quencher. By studying the solvent polarity dependence of the quenching using target analysis of the time-resolved data, we show that the quenching proceeds through energy transfer from the excited Pc to the optically forbidden S(1) state of the carotenoid, coupled to an intramolecular charge-transfer state. The mechanism for excess energy dissipation in photosystem II is discussed in view of the insights obtained on this simple model system.


Hepatitis C virus infection of cholangiocarcinoma cell lines.
Hepatitis C virus (HCV) infects the liver and hepatocytes are the major cell type supporting viral replication. Hepatocytes and cholangiocytes derive from a common hepatic progenitor cell that proliferates during inflammatory conditions, raising the possibility that cholangiocytes may support HCV replication and contribute to the hepatic reservoir. We screened cholangiocytes along with a panel of cholangiocarcinoma-derived cell lines for their ability to support HCV entry and replication. While primary cholangiocytes were refractory to infection and lacked expression of several entry factors, two cholangiocarcinoma lines, CC-LP-1 and Sk-ChA-1, supported efficient HCV entry; furthermore, Sk-ChA-1 cells supported full virus replication. In vivo cholangiocarcinomas expressed all of the essential HCV entry factors; however, cholangiocytes adjacent to the tumour and in normal tissue showed a similar pattern of receptor expression to ex vivo isolated cholangiocytes, lacking SR-BI expression, explaining their inability to support infection. This study provides the first report that HCV can infect cholangiocarcinoma cells and suggests that these heterogeneous tumours may provide a reservoir for HCV replication in vivo.


[Educational methods in the prevention of smoking, among students of the Department of Herault, France].
To evaluate the efficacy of two educational strategies to prevent tobacco addiction in schoolchildren living in Herault, France. A study was conducted to deliver an intervention and make a concurrent comparison, among elementary school, fifth-grade students, during school years 1992 to 1995, in Herault Department, Montpellier, France. Three observation groups were included: Group 1 was subject to a cross-sectional educational intervention; Group 2 was only subject to a single health activity on World Day Against Smoking; and Group 3 or control was not subject to any intervention. The prognostic role of peers and the family setting on tobacco consumption was confirmed; after three years, the Group subject to the sustained cross-sectional intervention showed a prevalence of smokers significantly lower than that of the other two groups. Even though tobacco consumption seems to begin earlier in boys than in girls, in the former the prevalence increases from 0.7 to 7% after three years. Preventive educational interventions should be maintained and reinforced to be effective on the initiation and cessation of tobacco smoking among children.


Antimicrobial and antioxidant activities of three Mentha species essential oils.
The present study describes the antimicrobial activity and free radical scavenging capacity (RSC) of essential oils from Mentha aquatica L., Mentha longifolia L., and Mentha piperita L. The chemical profile of each essential oil was determined by GC-MS and TLC. All essential oils exhibited very strong antibacterial activity, in particularly against Esherichia coli strains. The most powerful was M. piperita essential oil, especially towards multiresistant strain of Shigella sonei and Micrococcus flavus ATTC 10,240. All tested oils showed significant fungistatic and fungicidal activity [expressed as minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) values, respectively], that were considerably higher than those of the commercial fungicide bifonazole. The essential oils of M. piperita and M. longifolia were found to be more active than the essential oil of M. aquatica. Especially low MIC (4 microL/mL) and MFC (4 microL/mL) were found with M. piperita oil against Trichophyton tonsurans and Candida albicans (both 8 microL/mL). The RSC was evaluated by measuring the scavenging activity of the essential oils on the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and OH radicals. All examined essential oils were able to reduce DPPH radicals into the neutral DPPH-H form, and this activity was dose-dependent. However, only the M. piperita oil reduced DPPH to 50 % (IC50 = 2.53 microg/mL). The M. piperita essential oil also exhibited the highest OH radical scavenging activity, reducing OH radical generation in the Fenton reaction by 24 % (pure oil). According to GC-MS and TLC (dot-blot techniques), the most powerful scavenging compounds were monoterpene ketones (menthone and isomenthone) in the essential oils of M. longifolia and M. piperita and 1,8-cineole in the oil of M. aquatica.


Digital holographic profilometry of the inner surface of a pipe using a current-induced wavelength change of a laser diode.
Phase-shifting digital holography is applied to the measurement of the surface profile of the inner surface of a pipe for the detection of a hole in its wall. For surface contouring of the inner wall, a two-wavelength method involving an injection-current-induced wavelength change of a laser diode is used. To illuminate and obtain information on the inner surface, a cone-shaped mirror is set inside the pipe and moved along in a longitudinal direction. The distribution of a calculated optical path length in an experimental alignment is used to compensate for the distortion due to the misalignment of the mirror in the pipe. Using the proposed method, two pieces of metal sheet pasted on the inner wall of the pipe and a hole in the wall are detected. This shows that the three-dimensional profile of a metal plate on the inner wall of a pipe can be measured using simple image processing.


Lock-in detection of single photons after two-step frequency conversion.
We report on cascaded frequency conversion at the single-photon level, consisting of a downconversion from the red (738 nm) to a telecom band (1403 nm) and the reverse upconversion process in the same nonlinear crystal. Detecting the converted single photons suffers from a large noise floor. We were able to recover the signal by implementing a lock-in analyzer technique in the data postprocessing.


Onion extract and quercetin induce matrix metalloproteinase-1 in vitro and in vivo.
A scar is usually developed by an imbalance of collagen synthesis and degradation. It is believed that the flavonoids (quercetin and kaempferol) in onion extract play a role in reducing scar formation through inhibition of fibroblast activities. Even though several commercial products are composed of onion extract, the precise molecular mechanisms of onion extract in reduction of scar formation in skin are still largely unknown. In this study we investigated the effect both of onion extract and quercetin on the proliferation of fibroblasts, expression of type I collagen and matrix metalloproteinase-1 (MMP-1). Our data show that proliferation rates of fibroblasts were decreased in a dose-dependent manner of the onion extract and quercetin. The expression of type I collagen was not markedly changed by the onion extract and quercetin. Interestingly, the expression of MMP-1 was markedly increased by both onion extract and quercetin in vitro and in vivo. Thus, our data indicate that onion extract and quercetin play a role in the anti-scar effect in skin through up-regulation of MMP-1 expression, implying this agent is a promising material for reducing scar formation.


In Situ Powder Diffraction Studies of Electrode Materials in Rechargeable Batteries.
The ability to directly track the charge carrier in a battery as it inserts/extracts from an electrode during charge/discharge provides unparalleled insight for researchers into the working mechanism of the device. This crystallographic-electrochemical information can be used to design new materials or modify electrochemical conditions to improve battery performance characteristics, such as lifetime. Critical to collecting operando data used to obtain such information in situ while a battery functions are X-ray and neutron diffractometers with sufficient spatial and temporal resolution to capture complex and subtle structural changes. The number of operando battery experiments has dramatically increased in recent years, particularly those involving neutron powder diffraction. Herein, the importance of structure-property relationships to understanding battery function, why in situ experimentation is critical to this, and the types of experiments and electrochemical cells required to obtain such information are described. For each battery type, selected research that showcases the power of in situ and operando diffraction experiments to understand battery function is highlighted and future opportunities for such experiments are discussed. The intention is to encourage researchers to use in situ and operando techniques and to provide a concise overview of this area of research.


STD and Abortion Prevalence in Adolescent Mothers With Histories of Childhood Protection Involvement.
Early sexual debut and unprotected sexual activity place adolescents at risk of adverse sexual health outcomes. Adolescents involved with child protective services (CPS) may be a particularly vulnerable population. California birth records for 86,946 adolescents who became first-time mothers in 2008-2010 were probabilistically linked to statewide CPS records from 1998 and later. The prevalence of STDs at birth and of abortion history were explored by preconception CPS involvement. Generalized linear models, adjusted for health, socioeconomic and demographic characteristics, were used to assess correlates of current STDs and history of abortion. At the time they gave birth, 1% of adolescents had a documented STD, and 5% reported a previous abortion. After adjustment for other characteristics, CPS involvement was associated with a significantly elevated prevalence both of STDs (relative risk, 1.2) and of previous abortion (1.4). Other characteristics also were associated with both outcomes, but not always in the same direction. For example, delaying prenatal care until after the first trimester or getting none at all was associated with an increased prevalence of STDs (1.3), but a reduced prevalence of abortion (0.8-0.9); having public insurance coverage for the birth was associated with a reduced STD prevalence (0.9) and an elevated abortion history prevalence (1.2). To assess whether adolescents with a history of CPS involvement need targeted sexual health interventions, further research is needed on the mechanisms that underlie associations between CPS involvement and adverse sexual health outcomes.


Multilocus phylogenetic analysis of the genus Atherina (Pisces: Atherinidae).
Sand-smelts are small fishes inhabiting inshore, brackish and freshwater environments and with a distribution in the eastern Atlantic and Mediterranean Sea, extending south into the Indian Ocean. Here, we present a broad phylogenetic analysis of the genus Atherina using three mitochondrial (control region, 12S and 16S) and two nuclear markers (rhodopsin and 2nd intron of S7). Phylogenetic analyses fully support the monophyly of the genus. Two anti-tropical clades were identified, separating the South African Atherina breviceps from the north-eastern Atlantic and Mediterranean Atherina' species. In European waters, two groups were found. The first clade formed by a well supported species-pair: Atherina presbyter (eastern Atlantic) and Atherina hepsetus (Mediterranean), both living in marine waters; a second clade included Atherina boyeri (brackish and freshwater environments) and two independent lineages of marine punctated and non-punctated fishes, recently proposed as separate species. Sequence divergence values strongly suggest multiple species within the A. boyeri complex.


The speed of information in a 'fast-light' optical medium.
One consequence of the special theory of relativity is that no signal can cause an effect outside the source light cone, the space-time surface on which light rays emanate from the source. Violation of this principle of relativistic causality leads to paradoxes, such as that of an effect preceding its cause. Recent experiments on optical pulse propagation in so-called 'fast-light' media--which are characterized by a wave group velocity upsilon(g) exceeding the vacuum speed of light c or taking on negative values--have led to renewed debate about the definition of the information velocity upsilon(i). One view is that upsilon(i) = upsilon(g) (ref. 4), which would violate causality, while another is that upsilon(i) = c in all situations, which would preserve causality. Here we find that the time to detect information propagating through a fast-light medium is slightly longer than the time required to detect the same information travelling through a vacuum, even though upsilon(g) in the medium vastly exceeds c. Our observations are therefore consistent with relativistic causality and help to resolve the controversies surrounding superluminal pulse propagation.


Molecular markers that identify human astrocytomas and oligodendrogliomas.
The classification of human gliomas is currently based solely on neuropathological criteria. Prognostic and therapeutic parameters are dependent upon whether the tumors are deemed to be of astrocytic or oligodendroglial in origin. We sought to identify molecular reagents that might provide a more objective parameter to assist in the classification of these tumors. In order to identify mRNA transcripts for genes normally transcribed exclusively by oligodendrocytes. Northern blot analysis was carried out on RNA samples from 138 human gliomas. Transcripts encoding the myelin basic protein (MBP) were found in an equally high percentage of tumors that by neuropathological criteria were either astrocytic or oligodendroglial. In contrast, proteolipid protein (PLP) and cyclic nucleotide phosphodiesterase (CNP) mRNA molecules were found significantly more often in oligodendrogliomas than in astrocytomas. The strongest association with histological typing was found with the transcript for the myelin galactolipid biosynthetic enzyme UDP-galactose: ceramide galactosytransferase (CGT), which was about twice as frequently detected in tumors of oligodendroglial type. Results of glycolipid analyses were previously reported on a subset of the tumors studied herein. Statistical analyses of both molecular and biochemical data on this subset of astrocytomas, oligoastrocytomas, and oligodendrogliomas were performed to determine if a panel of markers could be used to separate astrocytic and oligodendroglial tumors. The presence of asialo GM1 (GA1) and the absence of paragloboside occurred most frequently in oligodendrogliomas. Ceramide monohexoside (CMH) levels correlated highly with the expression of mRNA for 4 myelin proteins: CGT, MBP, CNP, and PLP. The best combination of 2 markers of oligodendroglial tumors was CGT and GA1; the best combination of 3 markers was the presence of CGT, GA1, and the absence of paragloboside. We conclude that this combination of markers could be useful in distinguishing between astrocytic and oligodendroglial tumors.


Preparation and Characterization of Homopolymer Polyacrylonitrile-Based Fibrous Sorbents for Arsenic Removal.
This research investigated the modification of homopolymer polyacrylonitrile (PAN) fibers for use as an adsorbent for removing arsenic from drinking water. Fibers were chemically modified and cross-linked using combinations of hydrazine hydrate and sodium hydroxide (NaOH) before being loaded with ferric hydroxide using two different iron loading procedures. Effects of reagent concentrations and reaction times on degree of chemical modification and fiber properties were investigated using Fourier transform infrared spectroscopy and ion-exchange measurements. Arsenate adsorption was a function of both the iron loading and the properties of the underlying fiber. For fibers treated with only a single reagent, both Fe3+ and arsenate adsorption could be understood in terms of ion-exchange properties of the fiber surfaces. However, for fibers treated with both hydrazine and NaOH, the ion-exchange properties of the surface could not explain the Fe3+ and arsenate adsorption behavior. The best arsenate removal performance was obtained using the simplest pretreatment procedure of soaking in 10% NaOH at 95°C for 90 min, followed by precipitation coating of ferric hydroxide. This simple preparation procedure involves only two commonly available and inexpensive reagents and can be carried out without any specialized equipment. This suggests that adsorbents based on inexpensive homopolymer PAN fabric may be produced in developing areas of the world where commercial products may not be available.


Chicory inulin enhances fermentation of 2'-fucosyllactose by infant fecal microbiota and differentially influences immature dendritic cell and T-cell cytokine responses under normal and Th2-polarizing conditions.
Scope: Non-digestible carbohydrates (NDCs) such as native chicory inulin and 2'-fucosyllactose (2'-FL) are added to infant formula to mimic some of the human milk oligosaccharide (HMO) functions. It is unknown whether combining inulin and 2'-FL influences their fermentation kinetics and whether the immune-modulatory effects of these NDCs are different under normal and inflammatory-prone Th2-polarizing conditions. Methods and results: We investigated the in vitro fermentation of 2'-FL and native chicory inulin, fermented individually and combined, using fecal inocula of 8-week-old infants. Native inulin was fermented in a size-dependent fashion and expedited the fermentation of 2'-FL. Fermentation of both native inulin and 2'FL increased the relative abundance of Bifidobacterium, which coincided with the production of acetate and lactate. The fermentation digesta of all fermentations differentially influenced both dendritic cell and T-cell cytokine responses under normal culture conditions or in presence of the Th2-polarizing cytokines IL-33 and TSLP, with the most pronounced effect for IL-1β in the presence of TSLP. Conclusions: Our findings show that native inulin can expedite the fermentation of 2'-FL by infant fecal microbiota and that these NDC fermentation digesta have different effects under normal and Th2-polarizing conditions, indicating that infants with different immune backgrounds might benefit from tailored NDC formulations.


Perturbation of epigenetic processes by doxorubicin in the mouse testis.
Epigenetic processes play a major role in normal mammalian development, particularly during gametogenesis and early embryogenesis. Thus, perturbation of epigenetic processes in the testis by xenobiotics could have a major impact on testicular function and fertility, and potentially affect the development and health of subsequent generations. There has been substantial research into the epigenetic toxicity of environmental exposures over the last decade. However, few studies have focussed on pharmaceutical drugs, which due to the nature of their use are typically found at much higher concentrations within exposed individuals than environmental chemicals. Here, we investigated genome-wide changes in testicular mRNA transcription, microRNA expression and DNA methylation to assess the contribution of epigenetic mechanisms to the testicular toxicity induced by doxorubicin (DOX) as a representative, widely used and well-characterised anti-cancer drug. We demonstrated that DOX is able to induce transcriptional, microRNA and DNA methylation changes, which perturb pathways involved in stress/cell death and survival and testicular function and lead to germ cell loss and reproductive organ damage. This identified potential novel mechanisms of DOX-induced testicular toxicity for further focussed investigations. Such work is required to fully assess the role of epigenetics in toxicity, determine whether single and/or multigenerational epigenetic toxicity is a real public health concern, and begin to develop and incorporate relevant epigenetic endpoints into regulatory toxicology.


The protective effect of the Mediterranean diet: focus on cancer and cardiovascular risk.
The lower occurrence of cancer and cardiovascular disease in the population around the Mediterranean basin has been linked to the dietary habits of the region. Indeed, this so-called Mediterranean diet is essentially different from the diets consumed in Western and Northern European countries and is rich in nuts, fruits, vegetables, legumes, whole-wheat bread, fish, and olive oil, with moderate amounts of red wine, which is mainly consumed during meals. Although a variety of cultural and religious traditions exist among the peoples of the Mediterranean area, olive oil, fish, and red wine hold a traditional and central position in the culinary routines of the region. The components of the diet contain an ample source of molecules with antioxidant and anti-inflammatory actions, among which omega-3 fatty acids, oleic acid, and phenolic compounds hold a prominent place. This review will summarize the results of important epidemiological studies that have investigated the protective effect of fish and olive oil on the risk of breast, prostate, and colorectal cancer and of wine on the risk of cardiovascular disease. The present review also aims to elucidate the various mechanisms by which various dietary components exhibit their beneficial action. In this respect, emphasis will be placed on the properties of omega-3 fatty acids from fish, oleic acid from olive oil, and phenolic compounds from olive oil and red wine.


Metabolomic Analyses of Leishmania Reveal Multiple Species Differences and Large Differences in Amino Acid Metabolism.
Comparative genomic analyses of Leishmania species have revealed relatively minor heterogeneity amongst recognised housekeeping genes and yet the species cause distinct infections and pathogenesis in their mammalian hosts. To gain greater information on the biochemical variation between species, and insights into possible metabolic mechanisms underpinning visceral and cutaneous leishmaniasis, we have undertaken in this study a comparative analysis of the metabolomes of promastigotes of L. donovani, L. major and L. mexicana. The analysis revealed 64 metabolites with confirmed identity differing 3-fold or more between the cell extracts of species, with 161 putatively identified metabolites differing similarly. Analysis of the media from cultures revealed an at least 3-fold difference in use or excretion of 43 metabolites of confirmed identity and 87 putatively identified metabolites that differed to a similar extent. Strikingly large differences were detected in their extent of amino acid use and metabolism, especially for tryptophan, aspartate, arginine and proline. Major pathways of tryptophan and arginine catabolism were shown to be to indole-3-lactate and arginic acid, respectively, which were excreted. The data presented provide clear evidence on the value of global metabolomic analyses in detecting species-specific metabolic features, thus application of this technology should be a major contributor to gaining greater understanding of how pathogens are adapted to infecting their hosts.


Effects of optical variables in immersion lens-based near-field optics.
We analyze the effects of optical variables, such as illumination state, focal position variation, near-field air-gap height, and refractive index mismatch, in immersion lens-based near-field optics on the resultant field propagation characteristics, including spot size, focal depth, and aberrations. First, to investigate the general behaviors of various incident polarization states, focused fields near the focal planes in simple two- or three-layered media structures are calculated under considerations of refractive index mismatch, geometric focal position variations, and air-gap height in a multi-layered medium. Notably, for solid immersion near-field optics, although purely TM polarized illumination generates a stronger and 15% smaller beam spot size in the focal region than in the case of circularly polarized incident light, the intensity of the focused field decreases sharply from the interface between air and the third medium. For the same optical configurations, we show that changes in geometric focal position to the recording or detecting medium increases focal depth. Finally, through focused field analysis on a ROM (read-only memory) and a RW (rewritable) medium, compound effects of considered variables are discussed. The resultant field propagation behaviors described in this study may be applicable to the design of either highly efficient reflection or transmission near-field optics for immersion lens based information storage, microscopy and lithographic devices.


The value of preoperative Glasgow Prognostic Score and the C-Reactive Protein to Albumin Ratio as prognostic factors for long-term survival in pathological T1N0 esophageal squamous cell carcinoma.
Plenty of studies have confirmed the prognostic values of inflammation-based prognostic scores in many malignant tumors. In present study, we aim to explore whether these indexes has same prognostic values in patients with stage T1N0 esophageal squamous cell carcinoma (ESCC). The clinicopathological data of 160 consecutive patients with pathological stage T1N0 ESCC from January 2005 to December 2012 were collected retrospectively. As prognostic factors, the inflammation-based prognostic scores, including C-reactive protein (CRP), Glasgow prognostic score (GPS), prognostic index (PI), neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR) and CRP to albumin ratio (CAR), were evaluated. The best cut-off values were determined by the receiver operating characteristic (ROC) curves. The median follow-up time was 71.8 months. During the follow-up period, 34 (21.3%) patients occurred postoperative recurrence and 30 (18.8%) tumor-related deaths were recorded. The best cut-off values of CRP, NLR, PLR and CAR were 1.090, 1.976, 103.200 and 0.023, respectively. After multivariate analysis, the GPS and CAR were identified as independently prognostic factors for overall survival (OS) (p=0.017 and 0.040, respectively). Of all 160 individuals, there were 86 (53.8%) and 85 (53.1%) patients classified into high GPS group (1-2) and elevated CAR group (>0.023), respectively. In addition, the GPS were positively associated with PI (p<0.000) and the levels of serum CRP (p<0.000), NLR (p=0.004), PLR (p=0.029) and CAR (p<0.000) and the above correlations were also observed between the CAR and other inflammation-based prognostic scores (all p<0.050, except for p=0.054 for PLR levels). The preoperative GPS and CAR were simple, inexpensive, readily available predictor for long-term survival in stage T1N0 ESCC patients who underwent esophagectomy.


Vocal development in a large-scale crosslinguistic corpus.
This study evaluates whether early vocalizations develop in similar ways in children across diverse cultural contexts. We analyze data from daylong audio recordings of 49 children (1-36 months) from five different language/cultural backgrounds. Citizen scientists annotated these recordings to determine if child vocalizations contained canonical transitions or not (e.g., "ba" vs. "ee"). Results revealed that the proportion of clips reported to contain canonical transitions increased with age. Furthermore, this proportion exceeded 0.15 by around 7 months, replicating and extending previous findings on canonical vocalization development but using data from the natural environments of a culturally and linguistically diverse sample. This work explores how crowdsourcing can be used to annotate corpora, helping establish developmental milestones relevant to multiple languages and cultures. Lower inter-annotator reliability on the crowdsourcing platform, relative to more traditional in-lab expert annotators, means that a larger number of unique annotators and/or annotations are required, and that crowdsourcing may not be a suitable method for more fine-grained annotation decisions. Audio clips used for this project are compiled into a large-scale infant vocalization corpus that is available for other researchers to use in future work.


Proton pump inhibitor-responsive oesophageal eosinophilia: an entity challenging current diagnostic criteria for eosinophilic oesophagitis.
Consensus diagnostic recommendations to distinguish GORD from eosinophilic oesophagitis (EoE) by response to a trial of proton pump inhibitors (PPIs) unexpectedly uncovered an entity called 'PPI-responsive oesophageal eosinophilia' (PPI-REE). PPI-REE refers to patients with clinical and histological features of EoE that remit with PPI treatment. Recent and evolving evidence, mostly from adults, shows that patients with PPI-REE and patients with EoE at baseline are clinically, endoscopically and histologically indistinguishable and have a significant overlap in terms of features of Th2 immune-mediated inflammation and gene expression. Furthermore, PPI therapy restores oesophageal mucosal integrity, reduces Th2 inflammation and reverses the abnormal gene expression signature in patients with PPI-REE, similar to the effects of topical steroids in patients with EoE. Additionally, recent series have reported that patients with EoE responsive to diet/topical steroids may also achieve remission on PPI therapy. This mounting evidence supports the concept that PPI-REE represents a continuum of the same immunological mechanisms that underlie EoE. Accordingly, it seems counterintuitive to differentiate PPI-REE from EoE based on a differential response to PPI therapy when their phenotypic, molecular, mechanistic and therapeutic features cannot be reliably distinguished. For patients with symptoms and histological features of EoE, it is reasonable to consider PPI therapy not as a diagnostic test, but as a therapeutic agent. Due to its safety profile, ease of administration and high response rates (up to 50%), PPI can be considered a first-line treatment before diet and topical steroids. The reasons why some patients with EoE respond to PPI, while others do not, remain to be elucidated.


Is walkability associated with a lower cardiometabolic risk?
Walkability of residential environments has been associated with more walking. Given the health benefits of walking, it is expected that people living in locations with higher measured walkability should have a lower risk of cardiometabolic diseases. This study tested the hypothesis that higher walkability was associated with a lower cardiometabolic risk (CMR) for two administrative spatial units and three road buffers. Data were from the North West Adelaide Health Study first wave of data collected between 2000 and 2003. CMR was expressed as a cumulative sum of six clinical risk markers, selected to reflect components of the metabolic syndrome. Walkability was based on an established methodology and operationalised as dwelling density, intersection density, land-use mix and retail footprint. Walkability was associated with lower CMR for the three road buffer representations of the built environment but not for the two administrative spatial units. This may indicate a limitation in the use of administrative spatial units for analyses of walkability and health outcomes.


Survival Analysis of COPD Patients in a 13-Year Nationwide Cohort Study of the Brazilian National Health System.
Chronic obstructive pulmonary disease (COPD) has an appreciable socioeconomical impact in low- and middle-income countries, but most epidemiological data originate from high-income countries. For this reason, it is especially important to understand survival and factors associated with survival in COPD patients in these countries. To assess survival of COPD patients in Brazil, to identify risk factors associated with overall survival, including treatment options funded by the Brazilian National Health System (SUS). We built a retrospective cohort study of patients dispensed COPD treatment in SUS, from 2003 to 2015 using a National Database created from the record linkage of administrative databases. We further matched patients 1:1 based on sex, age and year of entry to assess the effect of the medicines on patient survival. We used the Kaplan-Meier method to estimate overall survival of patients, and Cox's model of proportional risks to assess risk factors. Thirty seven thousand and nine hundred and thirty eight patients were included. Patient's survival rates at 1 and 10 years were 97.6% (CI 95% 97.4-97.8) and 83.1% (CI 95% 81.9-84.3), respectively. The multivariate analysis showed that male patients, over 65 years old and underweight had an increased risk of death. Therapeutic regimens containing a bronchodilator in a free dose along with a fixed-dose combination of corticosteroid and bronchodilator seem to be a protective factor when compared to other regimens. Our findings contribute to the knowledge of COPD patients' profile, survival rate and related risk factors, providing new evidence that supports the debate about pharmacological therapy and healthcare of these patients.


Combining enzymes and organometallic complexes: novel artificial metalloenzymes and hybrid systems for C-H activation chemistry.
This review describes the recent advances in the design of novel artificial metalloenzymes and their application in C-H activation reactions. The combination of enzymes and metal or organometallic complexes for the creation of new artificial metalloenzymes has represented a very exciting research line. In particular, the development of proteins with the ability to perform C-H functionalization presents a significant challenge. Here we discuss the development of these processes on natural metalloenzymes by using directed evolution, biotin-(strept)avidin technologies, photocatalytic hybrids or reconstitution of heme-protein technology.


Utilization of medicinal cannabis for pain by individuals with spinal cord injury.
A cross-sectional multi-center study using an on-line survey addressing utilization, knowledge, and perceptions of medicinal cannabis (MC) by people with spinal cord injury (SCI). To characterize differences between current (CU), past (PU), and never users (NU) of MC with SCI; to determine why people with SCI use MC; to examine reports of MCs' efficacy and tolerability by individuals with SCI. Three academic medical centers in the United States. Comparison of demographic and attitudinal differences between CU, PU, and NU and differences in the groups' reports of pain, health, and quality of life (QOL). Evaluation of utilization patterns and perceived efficacy of MC among CU and PU and reports of side effects of MC versus prescription medications. Data were analyzed using either Chi Square, distribution-free exact statistics, or t-tests for continuous data. Among a nationwide sample (n = 353) of individuals with SCI, NU were less likely than CU and PU to believe that cannabis ought to be legalized and more likely to endorse risks of use. Current users and PU reported greater pain interference in daily life than did NU, but there were no between group differences in QOL or physical or emotional health. Current users and PU took MC to address pain (65.30%), spasms (63.30%), sleeplessness (32.70%), and anxiety (24.00%), and 63.30% reported it offered "great relief" from symptoms. Participants reported that MC is more effective and carries fewer side effects than prescription medications. Medicinal cannabis is an effective and well-tolerated treatment for a number of SCI-related symptoms.


A Randomized Controlled Trial of Tong Len Meditation Practice in Cancer Patients: Evaluation of a Distant Psychological Healing Effect.
Tong Len meditation is an important therapeutic tool in the Tibetan medicine, and it can be used for self-healing and/or to heal others. Currently, in the West, there is no scientific study concerning the efficacy of a Tong Len distant healing effect on psychological disorders in cancer patients. To evaluate a distant healing effect of Tong Len meditation on stress, anxiety, depression, fatigue, and self-perceived quality of life in cancer patients. These psychological objectives were chosen as a consequence of the limited scientific literature of present day. We performed a double-blind randomized controlled trial on 103 cancer patients with tumors. Overall, 12 meditators used Tong Len in aid of 52 patients randomly selected as experimental group, while the remaining 51 patients constituted the control group. Patients and meditators did not know each other. All patients completed profile of mood states (POMS) and European Quality of Life-5 dimensions (EQ-5D) questionnaires before treatment (T0), after two (T1) and three months of treatment (T2), and one month after treatment cessation (T3). With regard to the parameters related to depression, a statistically significant improvement (P = .003) was observed in the treatment group compared to controls. On the other hand, the vigor/activity parameter saw significant improvements in the control group (P = .009). Both groups exhibited significant improvements in the other factors assessed in the POMS and EQ-5D questionnaires. This study did not provide sufficient evidence supporting an efficacy of Tong Len meditation in distant psychological healing as compared to a control condition. The research highlighted some psychological improvements through Tong Len distant meditation in a group of patients unknown to meditators. Therefore, the enhancement detected in most parameters in both treatment and control groups raises interest on in-depth analysis and evaluation of distant meditation on cancer patients to mitigate psychological problems caused by the disease.


The quality of dietary intake methodology and reporting in child and adolescent obesity intervention trials: a systematic review.
Assessing dietary intake is important in evaluating childhood obesity intervention effectiveness. The purpose of this review was to evaluate the dietary intake methods and reporting in intervention studies that included a dietary component to treat overweight or obese children. A systematic review of studies published in the English language, between 1985 and August 2010 in health databases. The search identified 2,295 papers, of which 335 were retrieved and 31 met the inclusion criteria. Twenty-three studies reported energy intake as an outcome measure, 20 reported macronutrient intakes and 10 studies reported food intake outcomes. The most common dietary method employed was the food diary (n = 13), followed by 24-h recall (n = 5), food frequency questionnaire (FFQ) (n = 4) and dietary questionnaire (n = 4). The quality of the dietary intake methods reporting was rated as 'poor' in 15 studies (52%) and only 3 were rated as 'excellent'. The reporting quality of FFQs tended to be higher than food diaries/recalls. Deficiencies in the quality of dietary intake methods reporting in child obesity studies were identified. Use of a dietary intake methods reporting checklist is recommended. This will enable the quality of dietary intake results to be evaluated, and an increased ability to replicate study methodology by other researchers.


Bif-1 suppresses breast cancer cell migration by promoting EGFR endocytic degradation.
Dysregulation of EGFR expression and signaling is well documented to contribute to disease progression and metastasis in many types of cancer including breast cancer. EGF-stimulated EGFR activation leads to receptor internalization and endocytic degradation to control EGFR-mediated signaling. This process is frequently deregulated in cancer cells, leading to increased EGFR expression and mitogenic signaling. Here, we demonstrate that Bif-1, a tumor suppressor, plays a role in EGFR endocytic degradation and chemotactic migration in MDA-MB-231 breast cancer cells. Our data reveal that suppression of Bif-1 expression delays EGFR degradation and sustains Erk1/2 activation in response to EGF stimulation. Mechanistically, loss of Bif-1 sequesters internalized EGF in Rab5-positive endosomes and delays EGFR trafficking to lysosomes. Recruitment of Rab7 to EGF-positive vesicles and the activation of Rab7 are impaired in Bif-1 knockdown cells. Additionally, intracellular pH and the localization of acidic vesicles are altered by suppression of Bif-1. Furthermore, inhibition of Bif-1 increases chemotactic cell migration in response to EGF or serum, which correlates with prolonged cytoskeletal reorganization. Importantly, the effect of Bif-1 on EGF-induced cell migration is abolished by gefitinib, an EGFR-specific inhibitor. Taken together, these data suggest a novel function for Bif-1 as a suppressor of breast cancer cell migration by promoting EGFR degradation through the regulation of endosome maturation.


Stereotactic radiosurgery for glossopharyngeal neuralgia: preliminary report of 5 cases.
Glossopharyngeal neuralgia (GPN) is a rare pain disorder characterized by severe, stabbing pain of the ear, posterior tongue, and throat. The authors report their early experience of using stereotactic radiosurgery (SRS) as an alternative to posterior fossa surgery for patients with medically resistant GPN. Five patients (3 men, 2 woman) with medically resistant GPN underwent Gamma Knife surgery. The radiosurgical target was the distal portion of the glossopharyngeal and vagus nerves at the jugular foramen (glossopharyngeal meatus). The maximum radiation dose in all cases was 80 Gy. The median follow-up after radiosurgery was 13 months (range 2-19 months). Three patients became pain-free at 2 days, 3 days, and 2 weeks, respectively, and were able to discontinue the medications taken preoperatively for their pain. None of these patients have suffered recurrent pain since becoming pain free. Two patients experienced no benefit from the procedure and underwent posterior fossa surgery 2 and 5 months after SRS. Both of these patients continued to have pain after posterior fossa surgery. One of these patients was later discovered to have a head and neck cancer as the cause of his pain. No patient developed hoarseness or dysphagia after radiosurgery. This preliminary experience demonstrates that SRS is an option for patients with medically resistant GPN. Additional follow-up and a larger number of patients are needed to demonstrate the long-term safety and optimal radiation dosimetry for this indication.


CD4+T Cell Subset Profiling in Biliary Atresia Reveals ICOS- Regulatory T Cells as a Favorable Prognostic Factor.
Biliary atresia (BA) is a destructive pediatric liver disease and CD4+T cell activation is demonstrated to play an important role in BA. However, a comprehensive scenario regarding the involvement of CD4+T cell subsets to the development of BA remains unclear. Here, we aim to explore the infiltration of CD4+T cell subsets and their clinical significance in BA. In the present study, thirty BA liver samples were collected during surgery and were divided into good (BA1, n = 16) and poor prognosis (BA2, n = 14), with samples from choledochal cyst patients (n = 8) as control. By using multiplex immunohistochemistry, we evaluated the infiltration level of CD4+T cell subsets in the portal areas. RT-qPCR and flow cytometry were further applied to explore detailed features of Treg subsets. We revealed that hepatic infiltrating Th1, Th2, Th17, and ICOS+Treg cells were significantly increased in BA patients compared to controls and were negatively associated with prognosis, while high infiltrating ICOS-Tregs showed a favorable outcome. Phenotypic analysis indicated that, in contrast to ICOS+Tregs, ICOS-Tregs were mainly CD45RAhiCD45ROlow, and preferentially expressed more CD73. Besides, RT-qPCR revealed elevated expression of CD25, CD73, TGF-β, and BCL-2 genes in ICOS-Tregs. Finally, functional assay confirmed that ICOS-Tregs had a higher suppressive capacity to cytokine secretion and were more resistant to apoptosis in vitro. Collectively, we demonstrate that a mixed immune response is involved in BA pathogenesis, and the globally enhanced effector CD4+T cell response is associated with unfavorable prognosis, highly suppressive ICOS-Tregs is a protective factor and may serve an important reference to predict prognosis.


Xanthine oxidoreductase and cardiovascular disease: molecular mechanisms and pathophysiological implications.
There is substantial evidence that oxidative stress participates in the pathophysiology of cardiovascular disease. Biochemical, molecular and pharmacological studies further implicate xanthine oxidoreductase (XOR) as a source of reactive oxygen species in the cardiovascular system. XOR is a member of the molybdoenzyme family and is best known for its catalytic role in purine degradation, metabolizing hypoxanthine and xanthine to uric acid with concomitant generation of superoxide. Gene expression of XOR is regulated by oxygen tension, cytokines and glucocorticoids. XOR requires molybdopterin, iron-sulphur centres, and FAD as cofactors and has two interconvertible forms, xanthine oxidase and xanthine dehydrogenase, which transfer electrons from xanthine to oxygen and NAD(+), respectively, yielding superoxide, hydrogen peroxide and NADH. Additionally, XOR can generate superoxide via NADH oxidase activity and can produce nitric oxide via nitrate and nitrite reductase activities. While a role for XOR beyond purine metabolism was first suggested in ischaemia-reperfusion injury, there is growing awareness that it also participates in endothelial dysfunction, hypertension and heart failure. Importantly, the XOR inhibitors allopurinol and oxypurinol attenuate dysfunction caused by XOR in these disease states. Attention to the broader range of XOR bioactivity in the cardiovascular system has prompted initiation of several randomised clinical outcome trials, particularly for congestive heart failure. Here we review XOR gene structure and regulation, protein structure, enzymology, tissue distribution and pathophysiological role in cardiovascular disease with an emphasis on heart failure.


Apraxia and motor dysfunction in corticobasal syndrome.
Corticobasal syndrome (CBS) is characterized by multifaceted motor system dysfunction and cognitive disturbance; distinctive clinical features include limb apraxia and visuospatial dysfunction. Transcranial magnetic stimulation (TMS) has been used to study motor system dysfunction in CBS, but the relationship of TMS parameters to clinical features has not been studied. The present study explored several hypotheses; firstly, that limb apraxia may be partly due to visuospatial impairment in CBS. Secondly, that motor system dysfunction can be demonstrated in CBS, using threshold-tracking TMS, and is linked to limb apraxia. Finally, that atrophy of the primary motor cortex, studied using voxel-based morphometry analysis (VBM), is associated with motor system dysfunction and limb apraxia in CBS. Imitation of meaningful and meaningless hand gestures was graded to assess limb apraxia, while cognitive performance was assessed using the Addenbrooke's Cognitive Examination - Revised (ACE-R), with particular emphasis placed on the visuospatial subtask. Patients underwent TMS, to assess cortical function, and VBM. In total, 17 patients with CBS (7 male, 10 female; mean age 64.4+/- 6.6 years) were studied and compared to 17 matched control subjects. Of the CBS patients, 23.5% had a relatively inexcitable motor cortex, with evidence of cortical dysfunction in the remaining 76.5% patients. Reduced resting motor threshold, and visuospatial performance, correlated with limb apraxia. Patients with a resting motor threshold <50% performed significantly worse on the visuospatial sub-task of the ACE-R than other CBS patients. Cortical function correlated with atrophy of the primary and pre-motor cortices, and the thalamus, while apraxia correlated with atrophy of the pre-motor and parietal cortices. Cortical dysfunction appears to underlie the core clinical features of CBS, and is associated with atrophy of the primary motor and pre-motor cortices, as well as the thalamus, while apraxia correlates with pre-motor and parietal atrophy.


Effects of differing pedalling speeds on the power-duration relationship of high intensity cycle ergometry.
The aim of this experiment was to investigate the effects of differing pedalling speeds on the power-duration relationship during high intensity cycle ergometry with pedal cadences of 50 (low), 90 (intermediate) and 110 (high) r.min-1. This hyperbolic power-duration relationship can be described as: (P - phiPA).t = W', where P = power output, t = time to exhaustion, and phiPA and W' are constants. Eight volunteer male subjects, aged 24 +/- 2.6 yr, with no competitive cycling training took part in this study and each undertook thirteen tests on a Lode BV Excalibur Sport V1.52 cycle ergometer over an eight week period. The first exercise bout was a 30 W.min-1 incremental cycle at 50 r.min-1 to volitional fatigue. This allowed the identification of a range of power outputs that would be used to construct and examine the power-duration relationships for each subject at 50, 90 and 110 r.min-1. At both 50 and 90 r.min-1, power outputs of 30 W above and below and 60 W above the highest work rate, as well as the maximum work rate achieved during the incremental exercise test were chosen, while at 110 r.min-1, the power outputs chosen were 25 W above and below as well as 50 W above the highest work rate achieved during the incremental exercise test and also the maximum work rate achieved during the incremental exercise test were chosen. These four work rates for each pedalling frequency were chosen because they would have exercise times to exhaustion in the range of 1-10 minutes. Each exercise bout was preceded by four minutes of unloaded cycling and then the work rate was adjusted quickly to the desired load setting by the previously programmed computerised ergometer. The results of this work indicate that for the group of subjects studied, pedalling a cycle ergometer at 50 r.min-1 allows subjects to pedal for a significantly greater time than when pedalling at either 90 or 110 r.min-1. phiPA at 50 r.min-1 was significantly greater than when pedalling at either 90 (F(1,21) = 7.47, p < 0.01) or 110 r.min-1 (F(1,21) = 10.83, p < 0.0005). There was no significant (p > 0.22) difference between phiPA at 90 and 110 r.min-1, F(1,21) = 1.36. W' however, was not significantly different when the data for 50 r.min-1, 90 r.min-1 and 110 r.min-1 were compared (F50 r.min-1 (1,21) = 0.95; p > 0.41; F90 r.min-1(1,21) = 0.79, p > 0.53; F110 r.min-1 (1,21) = 0.78, p > 0.53). Our hypothesis, that endurance performance was reduced when recreational cyclists pedal at a high cadence when compared to a low cadence was correct. Maximum sustainable power output during cycle ergometry was higher at 50 r.min-1 than at either 90 or 110 r.min-1. At the intermediate cadence endurance was better than at the high but worse than at the low cadence. In conclusion, during endurance cycling, recreaional cyclists should pedal at lower rather than higher cadences.


[Diagnostic Test Requests in the Family Medicine Consultation: An Ethical Perspective Concerning Requests for Maxillofacial Computerized Tomography Scan Prior to Dental Implants].
In the Portuguese National Health Service, little attention has been paid to oral health care. The almost nonexistence of a dentistry network raises concern about accessibility to services, and justifies the need to call on a predominantly private provision of services. The coexistence between the public and private settings is not always easy, especially when services need to interact and actively collaborate in order to find answers to the patient's problems. Dental implant procedures and the need to perform a previous maxillofacial computerized tomography to study the bone bed where osseointegrated dental implants are placed are a common situation. The current governmental regulation, blinded to the clinical context, may limit the accessibility to the tests. Based on this scenario, we discuss the possible options from an ethical point of view, framing the patient's and the physician's perspective and the relation between both. We conclude that the medical decision can't be disregarded from the clinical evaluation, in the intimacy of the medical consultation. This is an ethical duty that overrules the administrative and bureaucratic constraints. A good management of this apparent dichotomy may enhance better health and greater empowerment for the patient.


Light regulation of the abundance of mRNA encoding a nucleolin-like protein localized in the nucleoli of pea nuclei.
A cDNA encoding a nucleolar protein was selected from a pea (Pisum sativum) plumule library, cloned, and sequenced. The translated sequence of the cDNA has significant percent identity to Xenopus laevis nucleolin (31%), the alfalfa (Medicago sativa) nucleolin homolog (66%), and the yeast (Saccharomyces cerevisiae) nucleolin homolog (NSR1) (28%). It also has sequence patterns in its primary structure that are characteristic of all nucleolins, including an N-terminal acidic motif, RNA recognition motifs, and a C-terminal Gly- and Arg-rich domain. By immunoblot analysis, the polyclonal antibodies used to select the cDNA bind selectively to a 90-kD protein in purified pea nuclei and nucleoli and to an 88-kD protein in extracts of Escherichia coli expressing the cDNA. In immunolocalization assays of pea plumule cells, the antibodies stained primarily a region surrounding the fibrillar center of nucleoli, where animal nucleolins are typically found. Southern analysis indicated that the pea nucleolin-like protein is encoded by a single gene, and northern analysis showed that the labeled cDNA binds to a single band of RNA, approximately the same size and the cDNA. After irradiation of etiolated pea seedlings by red light, the mRNA level in plumules decreased during the 1st hour and then increased to a peak of six times the 0-h level at 12 h. Far-red light reversed this effect of red light, and the mRNA accumulation from red/far-red light irradiation was equal to that found in the dark control. This indicates that phytochrome may regulate the expression of this gene.


Susceptibility of Salmonella isolated from fish feed factories to disinfectants and air-drying at surfaces.
The objective of this work was to determine whether persistence of certain Salmonella isolates in fish feed factories involved enhanced resistance to disinfectants or air-drying. Salmonella isolates known to be persistent in fish feed factories and Salmonella isolates from other origins were tested for their sensitivity to nine disinfectants by using a suspension test. More than 5log(10)reduction in viable count for all isolates was only achieved by two of the disinfectants at 80% of the lowest recommended user concentration. However, Salmonella isolates from fish feed factories were not more resistant to disinfectants compared to Salmonella from other origins. For four of the disinfectants, presence of fish feed had a more adverse effect on disinfection efficacy compared to the protein bovine serum albumin (BSA). In general, the Salmonella isolates were more resistant to disinfection than Escherichia coli DSM 682, a strain recommended in testing of disinfectants. The Salmonella isolates were also tested for their ability to survive air-drying at stainless steel surfaces, but there were no differences in survival of isolates from fish feed factories compared to isolates of other origin. In conclusion the Salmonella isolates from fish feed factories were not particularly resistant to disinfection or air-drying at surfaces. The data show that disinfectants to be used against Salmonella should be thoroughly tested and selected, since not all disinfectants appear to be effective against Salmonella.


Hema e-Chart: Italian Registry for prospective analysis of epidemiology, management and outcome of febrile events in patients with hematological malignancies.
The aim of the study was to create a prospective computerized registry to collect and analyze febrile events, particularly due to fungal infections, in patients with hematological malignancies. A systematic approach that starts from the registration of new diagnosis and complete follow-up can be of help for the study of treatment and evolution of these complications. The software allows several concurrent users to create and manage medical information in a website. Its aim is to improve the speed, quality and integration of information related to subjects with febrile event, ultimately resulting in improving patients' care.Patients included adults and children with acute and chronic myeloid or lymphoid leukemia, Hodgkin's and non-Hodgkin's lymphoma, myelodysplastic syndrome, or multiple myeloma. The registry also included data regarding event onset in hematopoietic stem cell transplants (HSCTs). In order to evaluate the incidence of febrile events, all new diagnoses of hematological malignancy and all HSCTs were reported.The Hema e-Chart can be a very useful network collecting information about febrile events in patients with hematological malignancy and HSCTs. Significant trends and treatment practices are expected to be observed. As enrollment continues, data will be analyzed and published, which will provide valuable information concerning the epidemiology, therapy, and outcome of infectious complications.


Bifunctional antimicrobial conjugates and hybrid antimicrobials.
Covering: up to the end of 2016Novel antimicrobial drugs are continuously needed to counteract bacterial resistance development. An innovative molecular design strategy for novel antibiotic drugs is based on the hybridization of an antibiotic with a second functional entity. Such conjugates can be grouped into two major categories. In the first category (antimicrobial hybrids), both functional elements of the hybrid exert antimicrobial activity. Due to the dual targeting, resistance development can be significantly impaired, the pharmacokinetic properties can be superior compared to combination therapies with the single antibiotics, and the antibacterial potency is often enhanced in a synergistic manner. In the second category (antimicrobial conjugates), one functional moiety controls the accumulation of the other part of the conjugate, e.g. by mediating an active transport into the bacterial cell or blocking the efflux. This approach is mostly applied to translocate compounds across the cell envelope of Gram-negative bacteria through membrane-embedded transporters (e.g. siderophore transporters) that provide nutrition and signalling compounds to the cell. Such 'Trojan Horse' approaches can expand the antibacterial activity of compounds against Gram-negative pathogens, or offer new options for natural products that could not be developed as standalone antibiotics, e.g. due to their toxicity.


The ontogeny of central melatonin binding sites in the rat.
The maternal transfer of circadian rhythmicity and photoperiodic information to the fetus has been clearly demonstrated in several species, as has the importance of the pineal hormone, melatonin, in conveying this information. Maternal melatonin is known to readily cross the placenta and be taken up by the fetal brain. Specific melatonin receptors have been demonstrated in the rodent brain and pituitary from the 21st day of gestation. To better understand the mechanisms by which melatonin brings about the transfer of information to the fetus and to define the receptivity of the fetus to the melatonin signal, we have followed the ontogeny of central melatonin binding sites in the rat from the 13th day of gestation to 5 days after birth. The use of in vitro autoradiography allows for the precise localization of binding sites as well as their quantification. In the present study melatonin binding sites were first found on the 15th day of gestation, at which time specific binding was limited to the pituitary. At birth it was possible to identify a strong label in the pars tuberalis of the pituitary, whereas the pars distalis appeared to be less intensely and more unevenly labeled. Neuronal melatonin binding sites became apparent from day 17 of gestation in an area of the dorsal brain that in older fetuses and in neonatal rats appeared to be the paraventricular nucleus of the thalamus. Moreover, melatonin binding sites are identifiable over the suprachiasmatic nuclei from day 18 of gestation. These data show that the fetal pituitary may have the potential to respond to the maternal melatonin signal as early as the 15th day of gestation, and that the brain may attain that potential at the 17th or 18th day of gestation.


Evaluation of the effect of oleuropein on alveolar bone loss, inflammation, and apoptosis in experimental periodontitis.
The present study aimed to evaluate the effects of oleuropein on ligature-induced alveolar bone loss. In this respect, osteoblastic activity, osteoclastic activity, inflammatory markers, and apoptosis were evaluated. Oleuropein is a flavonoid, which has potent anti-inflammatory and bone-protective effects. Thirty-two Wistar rats were divided into four experimental groups as following: control (C, n = 8) group; periodontitis (P, n = 8) group; periodontitis and low-dose oleuropein group (12 mg/kg/day oleuropein, LDO group, n = 8); and periodontitis and high-dose oleuropein group (24 mg/kg/day oleuropein, HDO group, n = 8). Periodontitis was induced via ligatures. Study period was 14 days, and animals were sacrificed at end of this period. Mandibles were examined via a stereomicroscope and underwent histological procedures. Osteoblast, tartrate-resistant acid phosphatase (TRAP)-positive osteoclast, and inflammatory cell counts were determined in hematoxylin-eosin stained sections. Inducible nitric oxide synthase (iNOS), bone morphogenetic protein-4, the cluster of differentiation (CD)-68, cysteine-aspartic proteases-3 (Caspase 3), and B-cell lymphoma-2 (Bcl-2) expressions were evaluated via immunohistochemistry. Periodontitis group had highest alveolar bone loss, and these levels significantly decreased in LDO and HDO groups. Both 12 and 24 mg/kg oleuropein groups significantly increased osteoblast cell counts and decreased TRAP-positive osteoclast and inflammatory cell counts. BMP-4 and bcl-2 expressions were elevated in oleuropein groups while caspase-3 expressions decreased. iNOS and CD68 were higher in periodontitis group compared to control group, but there was no significant difference between other groups. Oleuropein successfully decreased alveolar bone loss as a result of decreased osteoclastic activity, inflammation, and apoptosis and increased osteoblastic activity.


Multifocal Electroretinography in the Presence of Temporal and Spatial Correlations and Eye Movements.
Releasing patients from the fixation task, and permitting them to view natural stimuli such as movies, would provide increased comfort, and potentially additional signs of retinal function, when recording multifocal electroretinograms (mfERGs). Techniques must be developed to handle the difficulties that arise from these alternative stimulation strategies. Multifocal stimuli were presented to volunteer human subjects with and without fixation. Retinocentric analyses were performed to deal with shifts of the stimulus across the retina in the presence of eye movements. Artificial scotomas that moved with the eyes to simulate local retinal defects were presented to assess whether such defects might be detectable in the presence of eye movements. Temporal and spatial correlations in the stimulus can be discounted, permitting retinal kernels to be measured in response to natural stimuli. Responses to temporally natural stimuli tend to have slightly stronger amplitudes because of the presence of low temporal frequencies in these stimuli. The effects of eye movement artifacts can be reduced, permitting similar kernels to be obtained in the absence and presence of eye movements. Convergence to stable kernels took slightly longer in the presence of temporal correlations or eye movements. Artificial scotomas can be localized with these methods. It may be possible to perform multifocal ERG recordings in the clinic using more flexible, natural techniques. However, work is needed to achieve results comparable to those routinely obtained with conventional methods.


Risk Factors for the Development and Progression of Thoracic Aorta Calcification: The Multi-Ethnic Study of Atherosclerosis.
Vascular calcification independently predicts cardiovascular disease (CVD), and computed tomography (CT) is a useful tool to evaluate and quantify not only coronary but also thoracic aortic calcification (TAC). Previous TAC progression reports were limited to dialysis and renal transplant patients. This is the first study to evaluate TAC progression in a large multiethnic cohort without clinically evident CVD at entry. Non-contrast-enhanced cardiac CTs were obtained in 5886 of 6814 Multi-Ethnic Study of Atherosclerosis (MESA) participants (mean age, 62 years; 48% males; 40% white, 27% black, 21% Hispanic, and 12% Chinese). Baseline and follow-up TAC scores were derived. Overall, 4308 (73%) participants had no detectable baseline TAC. Mean follow-up duration was 2.4 ± 0.8 years, during which 12% developed TAC. The overall incidence rate was 4.8%/year and was greater with age across gender and ethnic groups; TAC incidence was significantly lower in blacks than whites. After adjustment for follow-up duration, regression analyses showed age, systolic blood pressure, antihypertensives, and smoking were associated with incident TAC. A total of 1578 (27%) participants had TAC at baseline with a positive association between average annual TAC change and baseline age. Although the overall median change was 32.9 (-1.4 to 112.2) Agatston units, 27% showed an annual score change of ≥100 and blacks showed the lowest median across ethnic groups; 22.7 (-3 to 86.8). Age, systolic blood pressure, lipid-lowering medication, diabetes, and smoking were associated with TAC progression. In MESA, traditional CV risk factors were related to both TAC incidence and progression. Blacks had the lowest incidence and median change across ethnic groups, consistent with previous findings for coronary calcification.


BREC: an R package/Shiny app for automatically identifying heterochromatin boundaries and estimating local recombination rates along chromosomes.
Meiotic recombination is a vital biological process playing an essential role in genome's structural and functional dynamics. Genomes exhibit highly various recombination profiles along chromosomes associated with several chromatin states. However, eu-heterochromatin boundaries are not available nor easily provided for non-model organisms, especially for newly sequenced ones. Hence, we miss accurate local recombination rates necessary to address evolutionary questions. Here, we propose an automated computational tool, based on the Marey maps method, allowing to identify heterochromatin boundaries along chromosomes and estimating local recombination rates. Our method, called BREC (heterochromatin Boundaries and RECombination rate estimates) is non-genome-specific, running even on non-model genomes as long as genetic and physical maps are available. BREC is based on pure statistics and is data-driven, implying that good input data quality remains a strong requirement. Therefore, a data pre-processing module (data quality control and cleaning) is provided. Experiments show that BREC handles different markers' density and distribution issues. BREC's heterochromatin boundaries have been validated with cytological equivalents experimentally generated on the fruit fly Drosophila melanogaster genome, for which BREC returns congruent corresponding values. Also, BREC's recombination rates have been compared with previously reported estimates. Based on the promising results, we believe our tool has the potential to help bring data science into the service of genome biology and evolution. We introduce BREC within an R-package and a Shiny web-based user-friendly application yielding a fast, easy-to-use, and broadly accessible resource. The BREC R-package is available at the GitHub repository https://github.com/GenomeStructureOrganization .


Could glutamate spectroscopy differentiate bipolar depression from unipolar?
Accurate differentiation of bipolar and unipolar depression is a key clinical challenge. A biological measure that could differentiate bipolar and unipolar depression might supplement clinical assessment. Magnetic Resonance Spectroscopy measurements of total glutamate and glutamine (Glx) in anterior cingulate cortex are one potential measure. The objective of this study was to assess the potential performance of this measure. Meta-analysis of data from eleven studies where anterior cingulate Glx of depressed patients has been compared to that of healthy controls was performed. Effect sizes for bipolar and unipolar depression were calculated as Standardised Mean Differences. The best estimate of test classification performance on the basis of observed effects was calculated. People with unipolar depression had on average lower levels of Glx than healthy controls (effect size -1.05; 95% CI -058 to -1.53). People with bipolar depression tended towards higher Glx than healthy controls (effect size 0.40; 95% CI -0.04 to 0.85). This yielded a difference in Glx between unipolar and bipolar depression of effect size 1.46 (95% CI 0.80-2.11). Based on this difference, a test differentiating bipolar from unipolar depression by whether Glx was higher or lower than the average in healthy population would have sensitivity 0.66 and specificity 0.85. There is an absence of studies directly comparing unipolar and bipolar depressed patients. On available data, measurement of anterior cingulate Glx is a promising potential tool for differentiation of bipolar and unipolar depression. This potential effect requires direct validation within mixed clinical cohorts.


Historical underpinnings of the term essential tremor in the late 19th century.
The term essential tremor has been in regular use since the second half of the 20th century. To modern neurologists, the word "essential" may seem cryptic. The historical underpinnings of this term have not been examined. To bring to attention early medical reports using the term essential tremor and examine the characteristics of the disorder that contributed to the proposed use of the term. Review of 19th and early 20th century medical literature on essential tremor. The term tremore semplice essenziale (simple essential tremor) was first used by Burresi (Italy, 1874) to describe an 18-year-old man with severe, isolated action tremor. Several years later, Maragliano (Italy, 1879), Nagy (Austria, 1890), and Raymond (France, 1892) described similar cases and proposed the terms tremore essenziale congenito (essential congenital tremor), essentieller Tremor (essential tremor), and tremblement essentiel héréditaire (hereditary essential tremor) to define the illness. Mirroring contemporaneous views of constitutional and inherited disease, the key ingredients of the disorder were viewed as the constant presence of tremor in the absence of other neurologic signs and its heritable nature. By the early 20th century, the term began to appear in the medical literature with greater frequency. Toward the end of the 19th century, several clinicians attempted to provide a nosologic separation for a tremor diathesis that was often familial and occurred in isolation of other neurologic signs. This disorder, which was termed essential tremor, was later recognized as one of the most common neurologic disorders.


Dietary Fat Patterns and Outcomes in Acute Pancreatitis in Spain.
Background/Objective: Evidence from basic and clinical studies suggests that unsaturated fatty acids (UFAs) might be relevant mediators of the development of complications in acute pancreatitis (AP). Objective: The aim of this study was to analyze outcomes in patients with AP from regions in Spain with different patterns of dietary fat intake. Materials and Methods: A retrospective analysis was performed with data from 1,655 patients with AP from a Spanish prospective cohort study and regional nutritional data from a Spanish cross-sectional study. Nutritional data considered in the study concern the total lipid consumption, detailing total saturated fatty acids, UFAs and monounsaturated fatty acids (MUFAs) consumption derived from regional data and not from the patient prospective cohort. Two multivariable analysis models were used: (1) a model with the Charlson comorbidity index, sex, alcoholic etiology, and recurrent AP; (2) a model that included these variables plus obesity. Results: In multivariable analysis, patients from regions with high UFA intake had a significantly increased frequency of local complications, persistent organ failure (POF), mortality, and moderate-to-severe disease in the model without obesity and a higher frequency of POF in the model with obesity. Patients from regions with high MUFA intake had significantly more local complications and moderate-to-severe disease; this significance remained for moderate-to-severe disease when obesity was added to the model. Conclusions: Differences in dietary fat patterns could be associated with different outcomes in AP, and dietary fat patterns may be a pre-morbid factor that determines the severity of AP. UFAs, and particulary MUFAs, may influence the pathogenesis of the severity of AP.


Mercury and drought along the lower Carson River, Nevada: I. Snowy egret and black-crowned night-heron annual exposure to mercury, 1997-2006.
The dynamic nature of the annual volume of water discharged down the Carson River over a 10-year period, which included a century flood and drought, was examined in order to gain a better understanding of mercury movement, biological availability, and exposure to waterbirds nesting at Lahontan Reservoir. Total annual water discharge directly influenced total mercury (THg) in unfiltered water above the reservoir and downstream of a mining area, whereas methyl mercury (MeHg) at the same site was negatively related to annual discharge. Annual water storage at Lahontan Reservoir in the spring and early summer, as expected, was directly related to annual Carson River discharge. In contrast to the findings from above the reservoir, annual MeHg concentrations in water sampled below the reservoir were positively correlated with the total discharge and the amount of water stored in the reservoir on 1 July; that is, the reservoir is an important location for mercury methylation, which agrees with earlier findings. However, unfiltered water MeHg concentrations were about 10-fold higher above than below the reservoir, which indicated that much MeHg that entered as well as that produced in the reservoir settled out in the reservoir. Avian exposure to mercury at Lahontan Reservoir was evaluated in both eggs and blood of young snowy egrets (Egretta thula) and black-crowned night-herons (Nycticorax nycticorax). Annual MeHg concentrations in unfiltered water below the reservoir, during the time period (Julian Days 90-190) when birds were present, correlated significantly with mercury concentrations in night-heron blood (r(2 )= 0.461, p = 0.027), snowy egret blood (r(2 )= 0.474, p = 0.024), and night-heron eggs (r(2) = 0.447, p = 0.029), but not snowy egret eggs. A possible reason for lack of an MeHg water correlation with snowy egret eggs is discussed and relates to potential exposure differences associated with the food habits of both species. THg concentrations in water collected below the reservoir were not related to egg or blood mercury concentrations for either species.


Changes in vitamin D and matrix metalloproteinase-9 in submariners during a submerged patrol.
To quantify changes in vitamin D and matrix metalloproteinase-9 (MMP9) in submariners over a single long patrol and compare the data to a group of non-deploying servicemen from their base port. A prospective time-series analysis was performed. Blood samples were taken from 49 submariners deploying on patrol and 43 shore-side controls from the base port (naval officers from base or non-deploying submariners), following a winter ashore at latitude 56° north. Samples were drawn immediately before the submarine sailed, in January, and again in the final week of patrol 85 days later. Paired pre-patrol and late samples from each individual were assayed together and changes in vitamin D and MMP9 were assessed. Mean pre-patrol vitamin D concentrations were 58 and 49 nmol/L for the controls and submariners, respectively. Mean vitamin D concentrations increased in controls as expected (mean increase 12.6 nmol/L), but not in the submariners (mean decrease 1.6 nmol/L). MMP9 levels were significantly higher in submariners pre-patrol, and increased significantly during the patrol. There was a significant inverse correlation between MMP9 and vitamin D levels (r=-0.41, p=0.01). This is the first study to quantify vitamin D and MMP levels in submariners. Circulating vitamin D concentrations on board were insufficient to prevent a rise in MMP. This has potential for adverse health effects and requires further study.


Attitudes toward opioid use for chronic pain: a Canadian physician survey.
To measure chronic pain patient volumes seen in primary care practice; to determine what medications physicians choose for the treatment of moderate to severe chronic pain; to identify barriers to the use of opioids in the treatment of chronic pain; and to assess physicians' attitudes toward the current management of chronic pain in Canada. A computer-assisted telephone survey of 100 regionally representative Canadian physicians with a defined interest in palliative care (PC, n=30) or noncancer pain (GP, n=70). A survey was conducted by Ipsos-Reid in June 2001. Only physicians who met the eligibility criteria of having written 20 or more prescriptions for moderate to severe pain in the preceding four weeks or having devoted 20% of time to palliative care were eligible to participate. In one month, the average number of patients with moderate to severe chronic pain seen by PCs was 94.2; the average seen by GPs was 44.7. The pain experienced by 83.3% of GP patients was noncancer related. For chronic cancer pain, an opioid analgesic was the treatment of choice of 79% of physicians (48% preferred morphine, 21% codeine, 10% other). For moderate to severe chronic noncancer pain, opioids were the first-line treatment of only 32% of physicians (16% preferred codeine, 16% major opioids) because a significant number preferred either non-steroidal anti-inflammatory drugs (29%) or acetaminophen (16%). Thirty-five per cent of GPs and 23% of PCs would never use opioids for noncancer pain, even when described as severe. Chronic pain was deemed by 68% of physicians to be inadequately managed. Almost 60% thought that pain management could be enhanced by improved physician education. Identified barriers to opioid use included addiction potential (37%) and side effects (25%). Seventeen per cent of GPs and 10% of PCs thought that regulatory sanctions limited opioid prescribing. Even among physicians experienced in chronic pain treatment, there is a reluctance to use opioids for severe nonmalignant pain. One-half of the survey participants believed that there was a need for improved physician education in pain management, including the use of opioids.


The Mycobacterium tuberculosis TrcR response regulator represses transcription of the intracellularly expressed Rv1057 gene, encoding a seven-bladed beta-propeller.
The Mycobacterium tuberculosis TrcR response regulator binds and regulates its own promoter via an AT-rich sequence. Sequences within this AT-rich region determined to be important for TrcR binding were used to search the M. tuberculosis H37Rv genome to identify additional related TrcR binding sites. A similar AT-rich sequence was identified within the intergenic region located upstream of the Rv1057 gene. In the present work, we demonstrate that TrcR binds to a 69-bp AT-rich sequence within the Rv1057 intergenic region and generates specific contacts on the same side of the DNA helix. An M. tuberculosis trcRS deletion mutant, designated STS10, was constructed and used to determine that TrcR functions as a repressor of Rv1057 expression. Additionally, identification of the Rv1057 transcriptional start site suggests that a SigE-regulated promoter also mediates control of Rv1057 expression. Using selective capture of transcribed sequences (SCOTS) analysis as an evaluation of intracellular expression, Rv1057 was shown to be expressed during early M. tuberculosis growth in human macrophages, and the Rv1057 expression profile correlated with a gene that would be repressed by TrcR. Based on structural predictions, motif analyses, and molecular modeling, Rv1057 consists of a series of antiparallel beta-strands which adopt a beta-propeller fold, and it was determined to be the only seven-bladed beta-propeller encoded in the M. tuberculosis genome. These results provide evidence of TrcR response regulator repression of the Rv1057 beta-propeller gene that is expressed during growth of M. tuberculosis within human macrophages.


E-cadherin promotor methylation and mutation are inversely related to motility capacity of breast cancer cells.
Inactivation of the tumor suppressor E-cadherin is an important event during breast tumorigenesis, as its decreased expression is linked to aggressiveness and metastasis. However, the relationship between the different modes of E-cadherin inactivation (mutation versus promotor hypermethylation) and breast cancer cell behavior is incompletely understood. The high correlation between E-cadherin inactivation status and cell morphology in vitro suggests different biological roles for the two inactivation modes during breast tumorigenesis. Because E-cadherin has been linked to cell invasion and metastasis, and cell motility is a crucial prerequisite to form metastases, we here compared the cell motility capacities of breast cancer cell lines with known E-cadherin status. Using barrier migration assays and time-lapse microscopy, we analyzed the migratory capacity of nine well-characterized human breast cancer cell lines (MDA-MB-231, MCF-7, T47D, BT549, MPE600, CAMA-1, SUM159PT, SUM52PE, and SK-BR-3). This subset was chosen based on E-cadherin gene status (wild-type, mutated, and promotor hypermethylated): three cell lines of each group. In addition, cell proliferation assays were performed for all conditions, to dissect migratory from proliferative effects. In this study, we demonstrate an overt association between the mode of E-cadherin inactivation and cell migration. Promotor hypermethylated E-cadherin cell lines showed a higher migration capacity, while cell lines with mutated E-cadherin were less motile compared to wild-type E-cadherin cell lines. Migration induction by fibronectin and basic fibroblast growth factor did not alter the cell motility association differences. Cell proliferation assays showed that the associations found were not caused by proliferation differences. Inhibition and overexpression of E-cadherin as well as DNA demethylation confirmed the relationship between E-cadherin and breast cancer cell motility. Our results demonstrate an association between the mode of E-cadherin inactivation and migration of breast cancer cells, which justifies more detailed research on the role of E-cadherin inactivation in cell migration and metastasis.


Central CO2 chemoreception: a mechanism involving P2 purinoceptors localized in the ventrolateral medulla of the anaesthetized rat.
1. The involvement of P2 purinoceptors in chemosensory function in the ventrolateral regions of the medulla oblongata was investigated in the anaesthetized rat. We have investigated the effect of antagonizing, or desensitizing, P2 receptors in the retrofacial area of the ventrolateral medulla on factors modifying respiratory activity. 2. Bilateral microinjection of suramin (50 nl, 0.02 M), a P2 purinoceptor antagonist, into the retrofacial area in the artificially ventilated rat reduced resting phrenic nerve discharge. It also markedly affected the response of the phrenic nerve to increases in arterial CO2. Under conditions of hyperoxic, hypocapnic apnoea, the mean threshold for inducing phrenic nerve activity was raised significantly (from an end-tidal CO2 of 2.5 % to 4.5 %, n = 9). 3. In addition, the slope of the respiratory response curve to increases in CO2 was reduced after suramin. A similar effect was observed after desensitization of certain P2X receptors with alphabeta-methyleneATP. As arterial levels of O2 were greater than 100 mmHg, and an equivalent pattern of response was observed in sino-aortically denervated and vagotomized animals, we believe any contribution of the peripheral chemoreceptors to be minimal. 4. Our data suggest that respiratory neurones within the retrofacial area (Botzinger complex) represent part of the central site of action of CO2 on respiration. Moreover, our observations lead us to suggest that CO2-evoked changes in respiration are mediated at least in part by P2X purinoceptors.


Difference in aftereffects following prolonged Achilles tendon vibration on muscle activity during maximal voluntary contraction among plantar flexor synergists.
It has been suggested that a suppression of maximal voluntary contraction (MVC) induced by prolonged vibration is due to an attenuation of Ia afferent activity. The purpose of the present study was to test the hypothesis that aftereffects following prolonged vibration on muscle activity during MVC differ among plantar flexor synergists owing to a supposed difference in muscle fiber composition. The plantar flexion MVC torque and surface electromyogram (EMG) of the medial head of gastrocnemius (MG), the lateral head of gastrocnemius (LG), and the soleus (Sol) were recorded in 13 subjects before and after prolonged vibration applied to the Achilles tendon at 100 Hz for 30 min. The maximal H reflexes and M waves were also determined from the three muscles, and the ratio between H reflexes and M waves (H/Mmax) was calculated before and after the vibration. The MVC torque was decreased by 16.6 +/- 3.7% after the vibration (P < 0.05; ANOVA). The H/Mmax also decreased for all three muscles, indicating that Ia afferent activity was successfully attenuated by the vibration in all plantar flexors. However, a reduction of EMG during MVC was observed only in MG (12.7 +/- 4.0%) and LG (11.4 +/- 3.9%) (P < 0.05; ANOVA), not in Sol (3.4 +/- 3.0%). These results demonstrated that prolonged vibration-induced MVC suppression was attributable mainly to the reduction of muscle activity in MG and LG, both of which have a larger proportion of fast-twitch muscle fibers than Sol. This finding suggests that Ia-afferent activity that reinforces the recruitment of high-threshold motor units is necessary to enhance force exertion during MVC.


Stretching DNA in polymer nanochannels fabricated by thermal imprint in PMMA.
We present results regarding the fast and inexpensive fabrication of polymer biochips for investigating the statics and dynamics of DNA confined in nanochannels. The biochips have been fabricated by means of nanoimprint lithography (NIL) in low molecular weight polymethyl methacrylate (PMMA) using a 4 inch diameter two-level hybrid stamp. The fluidic structures were sealed using thermal polymer fusion bonding. The stamp has nanometer- and micrometer-sized protrusions defined in a thermally grown SiO(2) layer and the sol-gel process derived duromeric hybrid polymer Ormocomp, respectively. The stamp is compatible with molecular vapor deposition (MVD), used for applying a durable chlorosilane based antistiction coating, and allows for imprint up to a temperature of 270 °C. The extension of YOYO-1 stained T4 GT7 bacteriophage DNA inside the PMMA nanochannels has been experimentally investigated using epi-fluorescence microscopy. The measured average extension length amounts to 20% of the full contour length with a standard deviation of 4%. These results are in good agreement with results obtained by stretching DNA in conventional fused silica nanochannels.


Fluorescence enhancement from single gold nanostars: towards ultra-bright emission in the first and second near-infrared biological windows.
Gold nanostars (AuNSs) are promising agents for the development of high-performance diagnostic devices, by enabling metal enhanced fluorescence (MEF) in the physiological near-infrared (NIR) and second near-infrared (NIR-II) windows. The local electric field near their sharp tips and between their branches can be enhanced by several orders of magnitude, holding great promise for large fluorescence enhancements from single AuNS particles, rather than relying on interparticle coupling in nanoparticle substrates. Here, guided by electric field simulations, two different types of AuNSs with controlled morphologies and plasmonic responses in the NIR and NIR-II regions are used to investigate the mechanism of MEF from colloidal AuNSs. Fluorophore conjugation to AuNSs allows significant fluorescence enhancement of up to 30 times in the NIR window, and up to 4-fold enhancement in the NIR-II region. Together with other inherent advantages of AuNSs, including their multispike morphology offering easy access to cell membranes and their large surface area providing flexible multifunctionality, AuNS are promising for the development of in vivo imaging applications. Using time-resolved fluorescence measurements to deconvolute semi-quantitatively excitation enhancement from emission enhancement, we show that a combination of enhanced excitation and an increased radiative decay rate, both contribute to the observed large enhancement. In accordance to our electric field modelling, however, excitation enhancement is the component that varies most with particle morphology. These findings provide important insights into the mechanism of MEF from AuNSs, and can be used to further guide particle design for high contrast enhancement, enabling the development of MEF biodetection technologies.