Cotranslational folding of spectrin domains via partially structured states.
How do the key features of protein folding, elucidated from studies on native, isolated proteins, manifest in cotranslational folding on the ribosome? Using a well-characterized family of homologous α-helical proteins with a range of biophysical properties, we show that spectrin domains can fold vectorially on the ribosome and may do so via a pathway different from that of the isolated domain. We use cryo-EM to reveal a folded or partially folded structure, formed in the vestibule of the ribosome. Our results reveal that it is not possible to predict which domains will fold within the ribosome on the basis of the folding behavior of isolated domains; instead, we propose that a complex balance of the rate of folding, the rate of translation and the lifetime of folded or partly folded states will determine whether folding occurs cotranslationally on actively translating ribosomes.


Na(V)1.5 enhances breast cancer cell invasiveness by increasing NHE1-dependent H(+) efflux in caveolae.
Na(V)1.5 sodium channels enhance the invasiveness of breast cancer cells through the acidic-dependent activation of cysteine cathepsins. Here, we showed that the Na(+)/H(+) exchanger type 1 (NHE1) was an important regulator of H(+) efflux in breast cancer cells MDA-MB-231 and that its activity was increased by Na(V)1.5. Na(V)1.5 and NHE1 were colocalized in membrane rafts containing caveolin-1. The inhibition of Na(V)1.5 or NHE1 induced a similar reduction in cell invasiveness and extracellular matrix degradation; no additive effect was observed when they were simultaneously inhibited. Our study suggests that Na(V)1.5 and NHE1 are functionally coupled and enhance the invasiveness of cancer cells by increasing H(+) efflux.


GABA stimulates chemotaxis and chemokinesis of embryonic cortical neurons via calcium-dependent mechanisms.
During rat cortical development, when neurons migrate from the ventricular zone to the cortical plate, GABA localizes within the target destinations of migratory neurons. At this time, cells in germinal zones and along migratory pathways express GABA receptor subunit transcripts, implying that in vivo, GABA may be a chemoattractant. We used an in vitro strategy to study putative chemotropic effects of GABA on embryonic rat cortical cells. GABA stimulated neuronal migration in vitro at embryonic day 15 (E15). From E16 onward, two concentration ranges (fM and microM) induced motility. Femtomolar GABA primarily stimulated chemotaxis (migration along a chemical gradient), whereas micromolar GABA predominantly initiated chemokinesis (increased random movement). These effects were mimicked by structural analogs of GABA with relative specificity at GABAA (muscimol), GABAB (R-baclofen), and GABAC (trans- or cis-4-aminocrotonic acid) receptors. Antagonists of GABAB (saclofen) and GABAC (picrotoxin) receptors partially inhibited responses to both femto- and micromolar GABA; however, only responses to femtomolar GABA were partially blocked by bicuculline, a well established antagonist of GABA at GABAA receptors. Hence, chemotactic responses to femtomolar GABA seem to involve all three classes of GABA receptor proteins, whereas chemokinetic responses to micromolar GABA involve GABAB and GABAC receptor proteins. GABA-induced motility was blocked by loading the cells with the Ca(2+)-chelating molecule bis(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid, suggesting that intracellular Ca2+ mediates GABA-induced cell movement. Optical recordings of cells loaded with Ca2+ indicator dye revealed that both femto- and micromolar GABA evoked increases in intracellular Ca2+. Thus, GABA-stimulated increases in intracellular Ca2+ may mediate both chemotactic and chemokinetic responses in embryonic cortical cells.


Water dynamics in water/DMSO binary mixtures.
The dynamics of dimethyl sulfoxide (DMSO)/water solutions with a wide range of water concentrations are studied using polarization selective infrared pump-probe experiments, two-dimensional infrared (2D IR) vibrational echo spectroscopy, optical heterodyne detected optical Kerr effect (OHD-OKE) experiments, and IR absorption spectroscopy. Vibrational population relaxation of the OD stretch of dilute HOD in H(2)O displays two vibrational lifetimes even at very low water concentrations that are associated with water-water and water-DMSO hydrogen bonds. The IR absorption spectra also show characteristics of both water-DMSO and water-water hydrogen bonding. Although two populations are observed, water anisotropy decays (orientational relaxation) exhibit single ensemble behavior, indicative of concerted reorientation involving water and DMSO molecules. OHD-OKE experiments, which measure the orientational relaxation of DMSO, reveal that the DMSO orientational relaxation times are the same as orientational relaxation times found for water over a wide range of water concentrations within experimental error. The fact that the reorientation times of water and DMSO are basically the same shows that the reorientation of water is coupled to the reorientation of DMSO itself. These observations are discussed in terms of a jump reorientation model. Frequency-frequency correlation functions determined from the 2D IR experiments on the OD stretch show both fast and slow spectral diffusion. In analogy to bulk water, the fast component is assigned to very local hydrogen bond fluctuations. The slow component, which is similar to the slow water reorientation time at each water concentration, is associated with global hydrogen bond structural randomization.


Effect of partner violence in adolescence and young adulthood on blood pressure and incident hypertension.
Intimate partner violence has adverse health consequences, but little is known about its association with hypertension. This study investigates sex differences in the relationship between intimate partner violence and blood pressure outcomes. Data included 9,699 participants from waves 3 (2001-02) and 4 (2008-09) of the National Longitudinal Study of Adolescent Health (51% female). Systolic (SBP) and diastolic (DBP) blood pressure and incident hypertension (SBP≥140 mmHg, DBP≥90 mmHg, or taking antihypertensive medication) were ascertained at wave 4. Intimate partner violence was measured at wave 3 with 8 items from the revised Conflict Tactics Scales. Separate victimization and perpetration scores were calculated. Sex-specific indicators of severe victimization and perpetration were created using the 66th percentile among those exposed as a cut point. Sex-specific, linear and logistic regression models were developed adjusting for age, race, financial stress, and education. Thirty-three percent of men and 47% of women reported any intimate partner violence exposure; participants were categorized as having: no exposure, moderate victimization and / or perpetration only, severe victimization, severe perpetration, and severe victimization and perpetration. Men experiencing severe perpetration and victimization had a 2.66 mmHg (95% CI: 0.05, 5.28) higher SBP and a 59% increased odds (OR: 1.59, 95% CI: 1.07, 2.37) of incident hypertension compared to men not exposed to intimate partner violence. No other category of violence was associated with blood pressure outcomes in men. Intimate partner violence was not associated with blood pressure outcomes in women. Intimate partner violence may have long-term consequences for men's hemodynamic health. Screening men for victimization and perpetration may assist clinicians to identify individuals at increased risk of hypertension.


Physical activity and risk of revision total knee arthroplasty in individuals with knee osteoarthritis: a matched case-control study.
To determine if physical activity was a risk factor for revision arthroplasty after primary total knee arthroplasty (TKA) due to osteoarthritis (OA) within the previous 15 years. This was a matched case-control study. The cases had primary TKA followed by revision arthroplasty. Controls had primary TKA and no revision arthroplasty. Cases and controls were matched for age, sex, number of knees replaced, and date of primary TKA. Standardized telephone interviews were conducted to assess historical leisure activity, occupational activity, and instrumental activities of daily living after primary TKA in metabolic equivalent (MET)-hours per week. Conditional logistic regression was performed to identify the variables that predicted the need for revision arthroplasty. Seventeen female and 9 male pairs, aged 47 to 85 years, participated. Most of the reported activity was of low impact and low or moderate intensity. Cases reported a median of 44.5 (range 0 to 137) MET-hours of total historical physical activity per week compared with controls' 55.1 (range 0 to 278) MET-hours. Total historical physical activity was not associated with the risk of revision arthroplasty (OR 0.99, 95% CI 0.99-1.01). Participants with primary TKA (controls) consistently reported more MET-hours of leisure and occupational activity than those with revision arthroplasty (cases) regardless of the number of knees replaced or whether or not walking was accounted for. This study quantified and described patterns of physical activity in a population with TKA. Physical activity did not appear to be a risk factor for revision arthroplasty. Our results suggest that individuals undergoing primary TKA should be encouraged to remain active after surgery.


Comparative in vitro genotoxicity study of ZnO nanoparticles, ZnO macroparticles and ZnCl2 to MDCK kidney cells: Size matters.
In the present study, we evaluated the roles that ZnO particle size and Zn ion release have on cyto- and genotoxicity in vitro. The Madin-Darby canine kidney (MDCK) cells were treated with ZnO nanoparticles (NPs), ZnO macroparticles (MPs), and ZnCl2 as a source of free Zn ions. We first tested cytotoxicity to define sub-cytotoxic exposure concentrations and afterwards we performed alkaline comet and cytokinesis-block micronucleus assays. Additionally, the activities of both catalase (CAT) and glutathione S-transferase (GST) were evaluated in order to examine the potential impairment of cellular stress-defence capacity. The amount of dissolved Zn ions from ZnO NPs in the cell culture medium was evaluated by an optimized voltammetric method. The results showed that all the tested zinc compounds induced similar concentration-dependent cytotoxicity, but only ZnO NPs significantly elevated DNA and chromosomal damage, which was accompanied by a reduction of GST and CAT activity. Although Zn ion release from ZnO NPs in cell culture medium was significant, our results show that this reason alone cannot explain the ZnO genotoxicity seen in this experiment. We discuss that genotoxicity of ZnO NPs depends on the particle size, which determines the physical principles of their dissolution and cellular internalisation.


Investigating palygorskite's role in the development of mesothelioma in southern Nevada: Insights into fiber-induced carcinogenicity.
Similar to asbestos fibers, nonregulated mineral fibers can cause malignant mesothelioma (MM). Recently, increased proportions of women and young individuals with MM were identified in southern Nevada, suggesting that environmental exposure to carcinogenic fibers was causing the development of MM. Palygorskite, a fibrous silicate mineral with a history of possible carcinogenicity, is abundant in southern Nevada. In this study, our aim was to determine whether palygorskite was contributing to the development of MM in southern Nevada. While palygorskite, in vitro, displayed some cytotoxicity toward primary human mesothelial (HM) cells and reduced their viability, the effects were roughly half of those observed when using similar amounts of crocidolite asbestos. No Balb/c (0/19) or MexTAg (0/18) mice injected with palygorskite developed MM, while 3/16 Balb/c and 13/14 MexTAg mice injected with crocidolite did. Lack of MM development was associated with a decreased acute inflammatory response, as injection of palygorskite resulted in lower percentages of macrophages (p = .006) and neutrophils (p = .02) in the peritoneal cavity 3 d after exposure compared to injection of crocidolite. Additionally, compared to mice injected with crocidolite, palygorskite-injected mice had lower percentages of M2 (tumor-promoting) macrophages (p = .008) in their peritoneal cavities when exposed to fiber for several weeks. Our study indicates that palygorskite found in the environment in southern Nevada does not cause MM in mice, seemingly because palygorskite, in vivo, fails to elicit inflammation that is associated with MM development. Therefore, palygorskite is not a likely contributor to the MM cases observed in southern Nevada.


The modular organization of projections from areas V1 and V2 to areas V4 and TEO in macaques.
In addition to the major anatomical pathways from V1 into the temporal lobe, there are other smaller, "bypass" routes that are poorly understood. To investigate the direct projection from V1 to V4 (bypassing V2) and from V2 to TEO (bypassing V4), we injected the foveal and parafoveal representations of V4 and TEO with different retrograde tracers in five hemispheres of four macaques and analyzed the distributions of labeled neurons in V1 and V2 using flattened preparations of the cortex. In V1, labeled neurons were seen after injections in V4 but not TEO. The V4-projecting neurons were located in the foveal representation of V1, in both cytochrome oxidase (CO)-rich blobs and CO-poor interblob regions. In V2, TEO-projecting neurons were intermingled with V4-projecting neurons, although the former were far sparser than the latter. Across the cases, 6-19% of the TEO-projecting neurons were double labeled, that is, also projected to area V4. Both V4- and TEO-projecting neurons formed bands that ran orthogonal to the V1/V2 border, and both were located in CO-rich thin stripes and CO-poor interstripe regions. In some cases, a continuous band of V4-projecting neurons was also found along the V1/V2 border in the foveal representation of V2. The results indicate that the pathways from V1 to V4 and from V2 to TEO involve anatomical subcompartments thought to be concerned with both color and form. These "bypass" routes may allow coarse information about color and form to arrive rapidly in the temporal lobe. The bypass route from V2 to TEO might explain the partial sparing of color and form vision that is seen after lesions of V4. By analogy, given the bypass route from the foveal representation of V1 to V4, lesions of V2 affecting the foveal visual field would also be insufficient to block color and form vision.


Cross-reactive human immunodeficiency virus type 1-neutralizing human monoclonal antibody that recognizes a novel conformational epitope on gp41 and lacks reactivity against self-antigens.
Broadly cross-reactive human immunodeficiency virus (HIV)-neutralizing antibodies are infrequently elicited in infected humans. The two best-characterized gp41-specific cross-reactive neutralizing human monoclonal antibodies, 4E10 and 2F5, target linear epitopes in the membrane-proximal external region (MPER) and bind to cardiolipin and several other autoantigens. It has been hypothesized that, because of such reactivity to self-antigens, elicitation of 2F5 and 4E10 and similar antibodies by vaccine immunogens based on the MPER could be affected by tolerance mechanisms. Here, we report the identification and characterization of a novel anti-gp41 monoclonal antibody, designated m44, which neutralized most of the 22 HIV type 1 (HIV-1) primary isolates from different clades tested in assays based on infection of peripheral blood mononuclear cells by replication-competent virus but did not bind to cardiolipin and phosphatidylserine in an enzyme-linked immunosorbent assay and a Biacore assay nor to any protein or DNA autoantigens tested in Luminex assays. m44 bound to membrane-associated HIV-1 envelope glycoproteins (Envs), to recombinant Envs lacking the transmembrane domain and cytoplasmic tail (gp140s), and to gp41 structures containing five-helix bundles and six-helix bundles, but not to N-heptad repeat trimers, suggesting that the C-heptad repeat is involved in m44 binding. In contrast to 2F5, 4E10, and Z13, m44 did not bind to any significant degree to denatured gp140 and linear peptides derived from gp41, suggesting a conformational nature of the epitope. This is the first report of a gp41-specific cross-reactive HIV-1-neutralizing human antibody that does not have detectable reactivity to autoantigens. Its novel conserved conformational epitope on gp41 could be helpful in the design of vaccine immunogens and as a target for therapeutics.


Protein tyrosine phosphatase-σ regulates hematopoietic stem cell-repopulating capacity.
Hematopoietic stem cell (HSC) function is regulated by activation of receptor tyrosine kinases (RTKs). Receptor protein tyrosine phosphatases (PTPs) counterbalance RTK signaling; however, the functions of receptor PTPs in HSCs remain incompletely understood. We found that a receptor PTP, PTPσ, was substantially overexpressed in mouse and human HSCs compared with more mature hematopoietic cells. Competitive transplantation of bone marrow cells from PTPσ-deficient mice revealed that the loss of PTPσ substantially increased long-term HSC-repopulating capacity compared with BM cells from control mice. While HSCs from PTPσ-deficient mice had no apparent alterations in cell-cycle status, apoptosis, or homing capacity, these HSCs exhibited increased levels of activated RAC1, a RhoGTPase that regulates HSC engraftment capacity. shRNA-mediated silencing of PTPσ also increased activated RAC1 levels in wild-type HSCs. Functionally, PTPσ-deficient BM cells displayed increased cobblestone area-forming cell (CAFC) capacity and augmented transendothelial migration capacity, which was abrogated by RAC inhibition. Specific selection of human cord blood CD34⁺CD38⁻CD45RA⁻lin⁻ PTPσ⁻ cells substantially increased the repopulating capacity of human HSCs compared with CD34⁺CD38⁻CD45RA⁻lin⁻ cells and CD34⁺CD38⁻CD45RA⁻lin⁻PTPσ⁺ cells. Our results demonstrate that PTPσ regulates HSC functional capacity via RAC1 inhibition and suggest that selecting for PTPσ-negative human HSCs may be an effective strategy for enriching human HSCs for transplantation.


MicroRNA-21 is induced early in pancreatic ductal adenocarcinoma precursor lesions.
Pancreatic ductal adenocarcinoma (PDAC) has the poorest overall prognosis among gastrointestinal cancers; however, curative resection in early-stage PDAC greatly improves survival rates, indicating the importance of early detection. Because abnormal microRNA production is commonly detected in cancer, we investigated noninvasive precursor pancreatic intraepithelial neoplasia (PanIN) lesions for microRNA production as a potential early biomarker of PDAC. Pathologists identified and classified ductal lesions. We extracted total RNA from laser-capture microdissected PanIN tissue samples from a conditional KRAS(G12D) mouse model (n = 29) or of human origin (n = 38) (KRAS is v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog). MicroRNA production was quantified by quantitative real-time PCR. Internal controls included 5S and U6 RNAs. Production of microRNAs miR-21, miR-205, and miR-200 paralleled PanIN progression in the KRAS(G12D) mouse model, compared with microRNA production in samples of nonpathologic ducts. miR-21 demonstrated the highest relative concentrations in the precursor lesions. Interestingly, miR-205 and miR-21 up-regulation preceded phenotypic changes in the ducts. The production of microRNAs miR-21, miR-221, miR-222, and let-7a increased with human PanIN grade, with peak production occurring in hyperplastic PanIN-2/3 lesions. In situ hybridization analysis indicated miR-21 production to be concentrated in pathologic ductal cells. miR-21 production was regulated by KRAS(G12D) and epidermal growth factor receptor in PDAC-derived cell lines. Aberrant microRNA production is an early event in the development of PanIN. Our findings indicate that miR-21 warrants further investigation as a marker for early detection of PDAC.


Uncoupling protein-2: evidence for its function as a metabolic regulator.
Uncoupling protein-2, discovered in 1997, belongs to a family of inner mitochondrial membrane proteins that, in general, function as carriers. The function(s) of uncoupling protein-2 have not yet been definitively described. However, mounting evidence suggests that uncoupling protein-2 could act in multiple tissues as a regulator of lipid metabolism. A role as a modulator of reactive oxygen species as a defence against infection is also postulated. In this review, a brief overview of the general and specific properties of uncoupling protein-2 is given and evidence for metabolic and immune regulatory functions is summarized. Uncoupling protein-2 could have particular importance in the regulation of lipid metabolism in adipose tissue and skeletal muscle. In addition, its ability to inhibit insulin secretion could also promote fat utilization over storage. Inhibition by uncoupling protein-2 of reactive oxygen species formation in macrophages and other tissues could have implications for regulation of immune function. The possibility of functions of uncoupling protein-2 in other tissues such as the brain are beginning to emerge.


Interhemispheric functional connectivity following prenatal or perinatal brain injury predicts receptive language outcome.
Early brain injury alters both structural and functional connectivity between the cerebral hemispheres. Despite increasing knowledge on the individual hemispheric contributions to recovery from such injury, we know very little about how their interactions affect this process. In the present study, we related interhemispheric structural and functional connectivity to receptive language outcome following early left hemisphere stroke. We used functional magnetic resonance imaging to study 14 people with neonatal brain injury, and 25 age-matched controls during passive story comprehension. With respect to structural connectivity, we found that increased volume of the corpus callosum predicted good receptive language outcome, but that this is not specific to people with injury. In contrast, we found that increased posterior superior temporal gyrus interhemispheric functional connectivity during story comprehension predicted better receptive language performance in people with early brain injury, but worse performance in typical controls. This suggests that interhemispheric functional connectivity is one potential compensatory mechanism following early injury. Further, this pattern of results suggests refinement of the prevailing notion that better language outcome following early left hemisphere injury relies on the contribution of the contralesional hemisphere (i.e., the "right-hemisphere-take-over" theory). This pattern of results was also regionally specific; connectivity of the angular gyrus predicted poorer performance in both groups, independent of brain injury. These results present a complex picture of recovery, and in some cases, such recovery relies on increased cooperation between the injured hemisphere and homologous regions in the contralesional hemisphere, but in other cases, the opposite appears to hold.


Release of secretory phospholipase A2 from rat neuronal cells and its possible function in the regulation of catecholamine secretion.
Here we show that secretory phospholipase A2 (sPLA2) that is immunochemically indistinguishable from type II sPLA2 is (i) stored in neuroendocrine cells, (ii) released in response to neurotransmitters or depolarization, and (iii) involved in the regulation of catecholamine secretion by these cells. Rat brain synaptic vesicle fractions contained PLA2 activity, which was neutralized completely by an antibody raised against rat type II sPLA2. sPLA2 immunoreactive with anti-(type II sPLA2) antibody was released from synaptosomes in response to depolarization evoked by a high concentration of potassium in the presence of Ca2+. Rat pheochromocytoma PC12 cells, which differentiated into adherent cells similar to sympathetic neurons in response to nerve growth factor, were used for the detailed analysis of the dynamics and function of sPLA2 in neuronal cells. Antibody against rat type II sPLA2 precipitated approximately 80% of the PLA2 activity in PC12 cell lysates. Transcript for type II sPLA2 was detected in PC12 cells by reverse transcriptase-PCR. When neuronally differentiated PC12 cells were stimulated with carbamylcholine or potassium, sPLA2 was released into the medium and reached a maximal approximately 40% release by 15 min. Inhibitors specific to type II sPLA2 suppressed catecholamine secretion by PC12 cells which had been activated by carbamylcholine. Furthermore, treatment of PC12 cells with exogenous type II sPLA2 alone elicited catecholamine secretion. These observations indicate that sPLA2 released from neuronal cells may regulate the degranulation process leading to release of neurotransmitters and are compatible with our earlier finding that this enzyme is involved in the degranulation of rat mast cells.


Ibero⁻American Consensus on Low- and No-Calorie Sweeteners: Safety, Nutritional Aspects and Benefits in Food and Beverages.
International scientific experts in food, nutrition, dietetics, endocrinology, physical activity, paediatrics, nursing, toxicology and public health met in Lisbon on 2⁻4 July 2017 to develop a Consensus on the use of low- and no-calorie sweeteners (LNCS) as substitutes for sugars and other caloric sweeteners. LNCS are food additives that are broadly used as sugar substitutes to sweeten foods and beverages with the addition of fewer or no calories. They are also used in medicines, health-care products, such as toothpaste, and food supplements. The goal of this Consensus was to provide a useful, evidence-based, point of reference to assist in efforts to reduce free sugars consumption in line with current international public health recommendations. Participating experts in the Lisbon Consensus analysed and evaluated the evidence in relation to the role of LNCS in food safety, their regulation and the nutritional and dietary aspects of their use in foods and beverages. The conclusions of this Consensus were: (1) LNCS are some of the most extensively evaluated dietary constituents, and their safety has been reviewed and confirmed by regulatory bodies globally including the World Health Organisation, the US Food and Drug Administration and the European Food Safety Authority; (2) Consumer education, which is based on the most robust scientific evidence and regulatory processes, on the use of products containing LNCS should be strengthened in a comprehensive and objective way; (3) The use of LNCS in weight reduction programmes that involve replacing caloric sweeteners with LNCS in the context of structured diet plans may favour sustainable weight reduction. Furthermore, their use in diabetes management programmes may contribute to a better glycaemic control in patients, albeit with modest results. LNCS also provide dental health benefits when used in place of free sugars; (4) It is proposed that foods and beverages with LNCS could be included in dietary guidelines as alternative options to products sweetened with free sugars; (5) Continued education of health professionals is required, since they are a key source of information on issues related to food and health for both the general population and patients. With this in mind, the publication of position statements and consensus documents in the academic literature are extremely desirable.


[Logopedic diagnosis and therapy in psychiatric treatment].
The paper outlines real and potential possibilities of logopedic activities within the framework of psychiatric care. The account is based on the experience in clinical logopedics and work published abroad in the field of diagnosis and therapy. The author presents the results of comprehensive logopedic examinations as well as its possible perfection and the contribution of logopedic diagnosis to the differential diagnosis in psychiatry. The author also focuses attention on possible therapeutic action and areas attended for logopedic care. In the conclusion he emphasizes the importance and necessity to include the clinical logopedist in the psychiatric in-patient team and the necessity of close cooperation of ambulatory departments concerned with psychiatric and logopedic problems.


The emerging role of Mule and ARF in the regulation of base excision repair.
The ARF (Alternative Reading Frame) protein is encoded in the Ink4a locus of human chromosome 9 that is frequently mutated in cancer cells. It was recently demonstrated that ARF is induced in response to DNA damage and inhibits, by direct interaction, the E3 ubiquitin ligase Mule that regulates p53 protein levels. Mule inhibition leads to p53 accumulation and activates cellular DNA damage responses. Mule has also recently been identified as a major E3 ubiquitin ligase involved in the regulation of DNA base excision repair. In this review, we will summarise the major properties of Mule and ARF and their roles in the coordination of DNA repair and DNA replication.


A tumor mRNA-triggered photodynamic molecular beacon based on oligonucleotide hairpin control of singlet oxygen production.
We report a new class of photodynamic molecular beacon (PMB) with tumor specific mRNA-triggered control of singlet oxygen ((1)O(2)) production. The beacon contains a single-stranded oligonucleotide linker that forms a stem-loop structure (hairpin) in which the sequence is an antisense oligonucleotide (AS-ON) complementary to a target mRNA. The stem is formed by the annealing of two complementary arm sequences that are on either side of the loop sequence. A photosensitizer molecule (PS) is attached to the end of one arm and a quencher (Q) is similarly attached to the other end. The conformationally-restricted hairpin forces Q to efficiently silence the photoreactivity of PS. In the presence of target mRNA, the hairpin opens and the PS is no longer silenced. Upon irradiating with light, the PS then emits fluorescence and generates cytotoxic (1)O(2). To show proof of concept, we have synthesized a c-raf-1 mRNA-triggered PMB using pyropheophorbide (Pyro) as PS, carotenoid as Q and c-raf-1 mRNA-targeted AS-ON as the loop sequence. We show that the (1)O(2) production of Pyro is quenched in its native state by 15-fold and is restored 9-fold by the addition of the target RNA. Comparing this to our recently reported self-folding peptide linker-based PMB, the hairpin effect results in an enhanced (1)O(2) quenching efficiency that decreases the residual (1)O(2) production by over 3-fold, thus providing enhanced control of (1)O(2) production upon target-linker interactions. When incubated with c-raf-1 expressing MDA-MB-231 cancer cells, the PMB displayed efficient cellular uptake and subsequently effective PDT activation in targeted cells.


The Green Berry Consortia of the Sippewissett Salt Marsh: Millimeter-Sized Aggregates of Diazotrophic Unicellular Cyanobacteria.
Microbial interactions driving key biogeochemical fluxes often occur within multispecies consortia that form spatially heterogeneous microenvironments. Here, we describe the "green berry" consortia of the Sippewissett salt marsh (Falmouth, MA, United States): millimeter-sized aggregates dominated by an uncultured, diazotrophic unicellular cyanobacterium of the order Chroococcales (termed GB-CYN1). We show that GB-CYN1 is closely related to Crocosphaera watsonii (UCYN-B) and "Candidatus Atelocyanobacterium thalassa" (UCYN-A), two groups of unicellular diazotrophic cyanobacteria that play an important role in marine primary production. Other green berry consortium members include pennate diatoms and putative heterotrophic bacteria from the Alphaproteobacteria and Bacteroidetes. Tight coupling was observed between photosynthetic oxygen production and heterotrophic respiration. When illuminated, the green berries became supersaturated with oxygen. From the metagenome, we observed that GB-CYN1 encodes photosystem II genes and thus has the metabolic potential for oxygen production unlike UCYN-A. In darkness, respiratory activity rapidly depleted oxygen creating anoxia within the aggregates. Metagenomic data revealed a suite of nitrogen fixation genes encoded by GB-CYN1, and nitrogenase activity was confirmed at the whole-aggregate level by acetylene reduction assays. Metagenome reads homologous to marker genes for denitrification were observed and suggest that heterotrophic denitrifiers might co-occur in the green berries, although the physiology and activity of facultative anaerobes in these aggregates remains uncharacterized. Nitrogen fixation in the surface ocean was long thought to be driven by filamentous cyanobacterial aggregates, though recent work has demonstrated the importance of unicellular diazotrophic cyanobacteria (UCYN) from the order Chroococcales. The green berries serve as a useful contrast to studies of open ocean UCYN and may provide a tractable model system to investigate microbial dynamics within phytoplankton aggregates, a phenomenon of global importance to the flux of particulate organic carbon and nitrogen in surface waters.


Evaluation of efficacy- versus affinity-driven agonism with biased GLP-1R ligands P5 and exendin-F1.
The glucagon-like peptide-1 receptor (GLP-1R) is an important regulator of glucose homeostasis and has been successfully targeted for the treatment of type 2 diabetes. Recently described biased GLP-1R agonists with selective reductions in β-arrestin versus G protein coupling show improved metabolic actions in vivo. However, two prototypical G protein-favouring GLP-1R agonists, P5 and exendin-F1, are reported to show divergent effects on insulin secretion. In this study we aimed to resolve this discrepancy by performing a side-by-side characterisation of these two ligands across a variety of in vitro and in vivo assays. Exendin-F1 showed reduced acute efficacy versus P5 for several readouts, including recruitment of mini-G proteins, G protein-coupled receptor kinases (GRKs) and β-arrestin-2. Maximal responses were also lower for both GLP-1R internalisation and the presence of active GLP-1R-mini-Gs complexes in early endosomes with exendin-F1 treatment. In contrast, prolonged insulin secretion in vitro and sustained anti-hyperglycaemic efficacy in mice were both greater with exendin-F1 than with P5. We conclude that the particularly low acute efficacy of exendin-F1 and associated reductions in GLP-1R downregulation appear to be more important than preservation of endosomal signalling to allow sustained insulin secretion responses. This has implications for the ongoing development of affinity- versus efficacy-driven biased GLP-1R agonists as treatments for metabolic disease.


Methods and Protocols for Reliable Electrochemical Testing in Post-Li Batteries (Na, K, Mg, and Ca).
While less mature than the Li-ion battery, technologies based on Na, K, Mg, and Ca are attracting more and more attention from the battery community. New material (cathode, anode, or electrolyte) testing for these post-Li systems commonly involves the use of an electrochemical setup called a half-cell in which metal counter and reference electrodes are used. Here we first describe the different issues that become critical when moving away from Li with respect to the cell hardware (cell design, current collector, separator, insulator) and the nature of the counter and reference electrodes. Workarounds are given, and a versatile setup is proposed to run reliable electrochemical tests for post-Li battery materials in general, in a broad range of electrolyte compositions.


Can ultrasound measures of intrinsic foot muscles and plantar soft tissues predict future diabetes-related foot disease? A systematic review.
Diabetes mellitus (DM) is associated with hyperglycaemia and advanced glycosylation end-products. In the foot, the consequences of chronic or uncontrolled diabetes are micro and macrovascular disease, neuropathy, reduced joint mobility and structural and soft tissue changes that increase the risk of ulcer development and amputation. Diabetes foot assessment currently includes a comprehensive history, neurological and vascular assessments and examination focussed on dermatological and musculoskeletal abnormalities. Whilst these assessments are helpful for predicting ulceration risk, direct identifiers that enable early therapeutic intervention are lacking. The intention of this review was to ascertain if B-mode ultrasound could be clinically applied to identify structural change in the diabetic foot and be utilised as an early predictor of ulceration risk. Primary databases and grey literature sources were systematically searched. Selection criteria were that the study included a diabetic sample and used B-mode ultrasound to assess soft tissue structures of the foot (plantar skin, plantar fat pad or intrinsic muscles). Fifteen studies were identified for inclusion (combined diabetic sample of 773). Ultrasound demonstrated reductions in tissue thickness in diabetics compared to non-diabetics under first (p = 0.01) and second (p = 0.03) metatarsal heads, but not the third (p = 0.24). Statistical heterogeneity was high for ultrasound thickness measures under metatarsal heads four/five (I2 65%, 81%) and very high for plantar skin (I2 98%), heel pad (I2 76%) and intrinsic muscles (I2 91%, 81%). Extensor digitorum brevis (EDB) ultrasound measures were significantly thinner in diabetics for all dimension measures compared to healthy controls except one study, which reported no significant differences in EDB thickness. No direct evidence was found to indicate B-mode ultrasound measures can predict soft tissue changes in the plantar foot in diabetes, although low level studies indicate ultrasound has the potential to identify structural change. Clinical, methodological and statistical heterogeneity limit result applicability. This review highlights the need for robust prospective longitudinal research to examine the predictive validity of this method.


Aspirational pursuit of mates in online dating markets.
Romantic courtship is often described as taking place in a dating market where men and women compete for mates, but the detailed structure and dynamics of dating markets have historically been difficult to quantify for lack of suitable data. In recent years, however, the advent and vigorous growth of the online dating industry has provided a rich new source of information on mate pursuit. We present an empirical analysis of heterosexual dating markets in four large U.S. cities using data from a popular, free online dating service. We show that competition for mates creates a pronounced hierarchy of desirability that correlates strongly with user demographics and is remarkably consistent across cities. We find that both men and women pursue partners who are on average about 25% more desirable than themselves by our measures and that they use different messaging strategies with partners of different desirability. We also find that the probability of receiving a response to an advance drops markedly with increasing difference in desirability between the pursuer and the pursued. Strategic behaviors can improve one's chances of attracting a more desirable mate, although the effects are modest.


An analysis of the citation climate in neurosurgical literature and description of an interfield citation metric.
The citation climate in neurosurgical literature is largely undefined. To study the patterns of citation of articles in neurosurgery as a scientific field and to evaluate the performance of neurosurgery journals vis-à-vis journals in other fields. References cited in articles published in neurosurgery journals during a specified time period were analyzed to determine the age of articles cited in neurosurgical literature. In the next analysis, articles published in neurosurgical journals were followed up for 13 years after publication. The postpublication citation patterns were analyzed to determine the time taken to reach the maximally cited state and the time when articles stopped being cited. The final part of the study dealt with the evolution of a new interfield citation metric, which was then compared with other standardized citation indexes. The mean ± SD age of articles cited in neurosurgical literature was 11.6 ± 11.7 years (median, 8 years). Citations received by articles gradually increased to a peak (at 6.25 years after publication in neurosurgery) and then reached a steady state; articles were still cited well into the late postpublication period. Neurosurgical articles published in nonneurosurgical high-impact journals were cited more highly than those in neurosurgical journals, although they took approximately the same time to reach the maximally cited state (7.2 years). The most cited pure neurosurgery journal was Neurosurgery. The citation climate for neurosurgery was adequately described. The interfield citation metric was able to ensure cross-field comparability of journal performance. G1, group 1G2, group 2G3, group 3G4, group 4IFCM, interfield citation metric.


Human skull from the Taza locality, Jijel, Algeria.
Archaeological excavations were carried out in 1990 in prehistorically inhabited caves sites in the Taza locality along the eastern Algerian coast. Recovered materials included stone and bone artifacts, faunal remains and a single human skull (Taza 1). A radiocarbon date for the upper cave level from which the skull was found was 16,100 B.P. While the skull, which is likely female, is very small overall, its shape compared closely with other North African cranial series. The obtained date and archaeological materials are characteristic of the Iberomaurusian industry of the Late Paleolithic period of North Africa.


Efficacy of postoperative epidural analgesia: a meta-analysis.
Whether epidural analgesia is a better method than parenteral opioids for postoperative pain control remains controversial. To systematically review the efficacy of postoperative epidural analgesia vs parenteral opioids, the primary alternative technique. Studies were identified primarily by searching the National Library of Medicine's PubMed database (1966 to April 25, 2002) and other sources for studies related to postoperative epidural analgesia. Inclusion criteria were a comparison of epidural therapy vs parenteral opioids for postoperative analgesia, measurement of pain using a visual analog scale (VAS) or numeric rating scale, randomization of patients to either therapy, and adult patients (> or =18 years). A total of 1404 abstracts were identified, 100 of which met all inclusion criteria. Each article was reviewed and data extracted from tables, text, or extrapolated from figures as needed. Weighted mean pain scores, weighted mean differences in pain score, and weighted incidences of complications were determined by using a fixed-effect model. Epidural analgesia provided better postoperative analgesia compared with parenteral opioids (mean [SE], 19.40 mm [0.17] vs 29.40 mm [0.20] on the VAS; P<.001). When analyzed by postoperative day, epidural analgesia was better than parenteral opioids on each postoperative day (P<.001 for each day after surgery). For all types of surgery and pain assessments, all forms of epidural analgesia provided significantly better postoperative analgesia compared with parenteral opioid analgesia (P<.001 for all), with the exception of thoracic epidural analgesia vs opioids for rest pain after thoracic surgery (weighted mean difference, 0.6 mm; 95% confidence interval, -0.3 to 1.5 mm; P =.12). The complication rates were lower than expected for nausea or vomiting and pruritus but comparable with existing data for lower extremity motor block. Epidural analgesia, regardless of analgesic agent, location of catheter placement, and type and time of pain assessment, provided better postoperative analgesia compared with parenteral opioids.


Quartet-based phylogeny reconstruction with answer set programming.
In this paper, a new representation is presented for the Maximum Quartet Consistency (MQC) problem, where solving the MQC problem becomes searching for an ultrametric matrix that satisfies a maximum number of given quartet topologies. A number of structural properties of the MQC problem in this new representation are characterized through formulating into answer set programming, a recent powerful logic programming tool for modeling and solving search problems. Using these properties, a number of optimization techniques are proposed to speed up the search process. The experimental results on a number of simulated data sets suggest that the new representation, combined with answer set programming, presents a unique perspective to the MQC problem.


Online health information seeking behaviors of Hispanics in New York City: a community-based cross-sectional study.
The emergence of the Internet has increased access to health information and can facilitate active individual engagement in health care decision making. Hispanics are the fastest-growing minority group in the United States and are also the most underserved in terms of access to online health information. A growing body of literature has examined correlates of online health information seeking behaviors (HISBs), but few studies have included Hispanics. The specific aim of this descriptive, correlational study was to examine factors associated with HISBs of Hispanics. The study sample (N=4070) was recruited from five postal zip codes in northern Manhattan for the Washington Heights Inwood Informatics Infrastructure for Comparative Effectiveness Research project. Survey data were collected via interview by bilingual community health workers in a community center, households, and other community settings. Data were analyzed using bivariate analyses and logistic regression. Among individual respondents, online HISBs were significantly associated with higher education (OR 3.03, 95% CI 2.15-4.29, P<.001), worse health status (OR 0.42, 95% CI 0.31-0.57, P<.001), and having no hypertension (OR 0.60, 95% CI 0.43-0.84, P=.003). Online HISBs of other household members were significantly associated with respondent factors: female gender (OR 1.60, 95% CI 1.22-2.10, P=.001), being younger (OR 0.75, 95% CI 0.62-0.90, P=.002), being married (OR 1.36, 95% CI 1.09-1.71, P=.007), having higher education (OR 1.80, 95% CI 1.404-2.316, P<.001), being in worse health (OR 0.59, 95% CI 0.46-0.77, P<.001), and having serious health problems increased the odds of their household members' online HISBs (OR 1.83, 95% CI 1.29-2.60, P=.001). This large-scale community survey identified factors associated with online HISBs among Hispanics that merit closer examination. To enhance online HISBs among Hispanics, health care providers and policy makers need to understand the cultural context of the Hispanic population. Results of this study can provide a foundation for the development of informatics-based interventions to improve the health of Hispanics in the United States.


Use of oseltamivir in lung transplant recipients with suspected or proven influenza infection: a 1-year observational study of outcomes and safety.
Influenza virus infections in lung transplant recipients (LTRs) have an increased risk of unfavourable outcomes. Early initiation of treatment is associated with improved outcomes. In clinical practice, empirical oseltamivir treatment is therefore commonly started prior to diagnostic microbiological confirmation. There is limited data on the patient characteristics, outcomes and safety of this practice. This study investigated outcomes and safety of this pre-emptive treatment strategy using oseltamivir. Descriptive analysis of LTRs who received oseltamivir for ≥2 days for suspected influenza infection between July 2011 and June 2012. Analyses were based on data from electronic medical records and our standardized LTR database with prospective documentation of clinical information including medication, laboratory and radiological results, outcomes and adverse events. We included 133 patients with a total of 261 oseltamivir treatment episodes (87.4% as outpatients). Median duration of oseltamivir treatment was 4 days (range 2 to 67) and 98.5% had concomitant antibiotic pharmacotherapy. Indications for oseltamivir included acute respiratory infection (66.7%), non-distinctive inflammatory reaction (51.3%) and influenza-like illness (2.7%). Influenza virus infection was confirmed by PCR in only 7%. Rhinovirus was the most frequent pathogen detected (14.9%). We discovered a wide range of adverse events but none occurred in >5%, and most were mild and of questionable causal relationship to oseltamivir administration. This non-controlled retrospective analysis suggests that the pre-emptive use of oseltamivir for respiratory tract infections pending microbiological results is safe in LTRs.


Ultraviolet sunlight exposure during adolescence and adulthood and breast cancer risk: a population-based case-control study among Ontario women.
Recent studies suggest that vitamin D may be associated with reduced breast cancer risk, but most studies have evaluated only dietary vitamin D intake. The associations among ultraviolet radiation from sunlight, factors related to cutaneous vitamin D production, and breast cancer risk were evaluated in a population-based case-control study conducted in Ontario, Canada, between 2003 and 2004 (n = 3,101 cases and n = 3,471 controls). Time spent outdoors was associated with reduced breast cancer risk during 4 periods of life (>21 vs. ≤6 hours/week age-adjusted odds ratio (OR) = 0.71, 95% confidence interval (CI): 0.60, 0.85 in the teenage years; OR = 0.64, 95% CI: 0.53, 0.76 in the 20s-30s; OR = 0.74, 95% CI: 0.61, 0.88 in the 40s-50s; and OR = 0.50, 95% CI: 0.37, 0.66 in the 60s-74 years). Sun protection practices and ultraviolet radiation were not associated with breast cancer risk. A combined solar vitamin D score, including all the variables related to vitamin D production, was significantly associated with reduced breast cancer risk. These associations were not confounded or modified by menopausal status, dietary vitamin D intake, or physical activity. This study suggests that factors suggestive of increased cutaneous production of vitamin D are associated with reduced breast cancer risk.


Comparative effectiveness of 5-fluorouracil with and without oxaliplatin in the treatment of colorectal cancer in clinical practice.
First-line chemotherapeutic treatment of colorectal cancer (CRC) typically comprises oral (capecitabine) or intravenous 5-fluorouracil (5-FU) plus leucovorin (LV), in combination with oxaliplatin (XELOX or FOLFOX, respectively), although debate exists regarding the best course of treatment by modality in clinical practice. Evidence from practice comparisons is important in considering the net benefit of alternative chemotherapy regimens, given expected differences in survival associated with compliance and age of patients treated in real life versus controlled trial settings. Practice variation in 5-FU treatment (i.e. 5-FU/leucovorin, FOLFOX, capecitabine and XELOX) of patients with CRC from an Australian area health service (n=636) was analyzed between modalities by patient age, tumour stage and site using non-parametric tests. Survival analyses (n=434) were conducted over a three-year follow-up period using Cox regression, adjusting for observed confounders. FOLFOX was the most commonly administered regimen. 5-FU modality was significantly associated with patient age (p<0.001), tumour stage (p<0.001) and site (p<0.001). Cox regression analyses found no significant difference in survival with the addition of oxaliplatin to 5-FU regimens. Our findings suggested no survival benefit with the addition of oxaliplatin to 5-FU modalities in treating CRC in practice. This raises questions as to the net benefit of oxaliplatin, given its known toxicity profile and expense.


Co-culture supernatants from Vibrio vulnificus-infected INT-407 cells induce IL-8 production in intestinal epithelial cells: crucial role of V. vulnificus rtxE.
In a previous study, we reported that a gene mutation of repeat in toxin E (RtxE), a transporter of cytotoxic factors, resulted in a significant impairment of epithelial cell cytotoxicity in Vibrio vulnificus, and that the expression of the rtxE gene was induced by the exposure to the host cells. In this study, we evaluated and compared the effects of co-culture supernatants from V. vulnificus-infected INT-407 cells and either the V. vulnificus wild-type or rtxE mutant on the production of interleukin (IL)-8, a pro-inflammatory cytokine, as well as its underlying mechanisms in human intestinal epithelial cells. INT-407 cells were co-cultured with the wild-type V. vulnificus or the rtxE mutant strain to obtain the conditioned supernatants. IL-8 production and nuclear factor (NF)-κB activation from the INT-407 cells treated with each supernatant, were investigated. The co-culture supernatants from the rtxE mutant V. vulnificus-infected INT-407 cells significantly induced lower levels of IL-8 production and promoter activation, NF-κB DNA binding activity, and NF-κB minimal promoter activation in human intestinal epithelial cells, than those from the wild-type V. vulnificus-infected INT-407 cells. Importantly, the reduced IL-8 production and NF-κB activity of the V. vulnificus rtxE mutant, were restored by co-culture supernatants from the rtxE-complemented V. vulnificus. On the whole, these results show that the rtxE gene of V. vulnificus performs a critical role in the secretion of factors from bacteria and host cells, which are involved in IL-8 production via the NF-κB activation pathway in host cells.


Longitudinal coherence measurements of an extreme-ultraviolet free-electron laser.
We have measured the average single-pulse longitudinal coherence characteristics of FLASH, a self amplified spontaneous emission free electron laser, at extreme UV wavelengths. Electric field autocorrelation measurements in the time domain were enabled by a wavefront division beam splitter applied to a tunable delay Mach-Zehnder interferometer. These data agree with the spectral bandwidth measurements made in the frequency domain. They exhibit two correlation time scales and the measured coherence curves have relevant implications for single-shot measurements.


Chronic musculoskeletal pain among elderly in rural Tamil Nadu: Mixed-method study.
(1) To determine the prevalence of chronic pain of musculoskeletal origin at different body sites among elderly (≥60 years). (2) To study the determinants of chronic pain and the healthcare seeking behavior among elderly with chronic pain. (3) To explore the perceptions of chronic pain among elderly. This study was done in 55 field practice villages of the Rural Health Training Centre (RHTC) of the Department of Community Medicine, SMVMCH, Puducherry. An Exploratory Mixed-Method study design, where a qualitative phase (in-depth interviews [IDI]) followed the quantitative phase (Survey). A representative sample of 850 respondents was selected by two-stage cluster sampling. A trained investigator did a house-to-house visit and interviewed selected elderly respondents by using predesigned and pretested questionnaire. The intensity of chronic pain was measured using the Functional Rating Scale and Numeric Rating Scale. The manual content analysis was done for qualitative data. Multiple logistic regression was performed on quantitative data. Qualitative data were described under two broad themes: (1) perceptions of elderly people about chronic pain and 2) healthcare seeking and coping mechanisms. Prevalence of chronic pain among the elderly respondents was found to be 47.6%. Most common site for chronic pain was knee joint (64.5%) followed by a low backache (21.7%). Most of the older people with chronic pain had mild-to-moderate and tolerable pain. Two predictors for chronic pain were the age of the respondent (adjusted OR -1.03, 95% CI = 1.01-1.05) and the presence of at least one chronic morbidity (adjusted OR -1.37, 95% CI = 1.03-1.82). Since the chronic pain was found to be associated with aging and the presence of at least one of the chronic morbidities, it is crucial for treating community physicians to consider the comorbid conditions while managing chronic pain in elderly. Considering the complex nature of chronic pain in old age, the primary care physician should be trained in drug and context-specific nondrug interventions to address the biomedical causes and other cognitive-behavior factors, respectively, through active support from their family and neighborhood.


Effectiveness of postoperative physical therapy for upper-limb impairments after breast cancer treatment: a systematic review.
To systematically review the effectiveness of various postoperative physical therapy modalities and timing of physical therapy after treatment of breast cancer on pain and impaired range of motion (ROM) of the upper limb. We searched the following databases: PubMed/MEDLINE, Cumulative Index to Nursing and Allied Health Literature, Embase, Physiotherapy Evidence Database, and Cochrane. Articles published until October 2012 were included. Only (pseudo) randomized controlled trials and nonrandomized experimental trials investigating the effectiveness of passive mobilization, manual stretching, myofascial therapy, and/or exercise therapy and timing of physical therapy after treatment for breast cancer are reviewed. Primary outcomes are pain of the upper limb and/or ROM of the shoulder. Secondary outcomes are decreased shoulder strength, arm lymphedema, limitations in activities of daily living, decreased quality of life, and wound drainage volume. Physical therapy modalities had to be started in the first 6 weeks after surgery. Articles were selected by 2 independent researchers in 3 phases and compared for consensus. First the titles were analyzed, and then the selected abstracts and finally the full texts were reviewed. Eighteen randomized controlled trials were included in the review. Three studies investigated the effect of multifactorial therapy: 2 studies confirmed that the combination of general exercises and stretching is effective for the treatment of impaired ROM another study showed that passive mobilization combined with massage had no beneficial effects on pain and impaired ROM. Fifteen studies investigated the effectiveness of a single physical therapy modality. One study of poor quality found evidence supporting the beneficial effects of passive mobilization. The only study investigating the effect of stretching did not find any beneficial effects. No studies were found about the effectiveness of myofascial therapy in the postoperative phase. Five studies found that active exercises were more effective than no therapy or information on the treatment of impairments of the upper limb. Three studies supported the early start of exercises for recovery of shoulder ROM, whereas 4 studies supported the delay of exercises to avoid prolonged wound healing. Multifactorial physical therapy (ie, stretching, exercises) and active exercises were effective to treat postoperative pain and impaired ROM after treatment for breast cancer. High-quality studies are necessary to determine the effectiveness of passive mobilization, stretching, and myofascial therapy as part of the multifactorial treatment. In addition, the appropriate timing and content of the exercise programs need to be further investigated.


Identifying epitopes responsible for neutralizing antibody and DC-SIGN binding on the spike glycoprotein of the severe acute respiratory syndrome coronavirus.
The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) uses dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN) to facilitate cell entry via cellular receptor-angiotensin-converting enzyme 2. For this project, we used recombinant baculoviruses expressing different lengths of SARS-CoV spike (S) protein in a capture assay to deduce the minimal DC-SIGN binding region. Our results identified the region location between amino acid (aa) residues 324 to 386 of the S protein. We then generated nine monoclonal antibodies (MAbs) against the S protein to map the DC-SIGN-binding domain using capture assays with pseudotyped viruses and observed that MAb SIa5 significantly blocked S protein-DC-SIGN interaction. An enhancement assay using the HKU39849 SARS-CoV strain and human immature dendritic cells confirmed our observation. Data from a pepscan analysis and M13 phage peptide display library system mapped the reactive MAb SIa5 epitope to aa residues 363 to 368 of the S protein. Results from a capture assay testing three pseudotyped viruses with mutated N-linked glycosylation sites of the S protein indicate that only two pseudotyped viruses (N330Q and N357Q, both of which lost glycosylation sites near the SIa5 epitope) had diminished DC-SIGN-binding capacity. We also noted that MAb SIb4 exerted a neutralizing effect against HKU39849; its reactive epitope was mapped to aa residues 435 to 439 of the S protein. We offer the data to facilitate the development of therapeutic agents and preventive vaccines against SARS-CoV infection.


Electronic structure and spectroscopy of oxyallyl: a theoretical study.
Electronic structure of the oxyallyl diradical and the anion is investigated using high-level ab initio methods. Converged theoretical estimates of the energy differences between low-lying electronic states of oxyallyl (OXA) as well as detachment energies of the anion are reported. Our best estimates of the adiabatic energy differences between the anion (2)A(2) and the neutral (3)B(2) and (3)B(1) states are 1.94 and 2.73 eV, respectively. The (1)A(1) state lies above (3)B(2) vertically, but geometric relaxation brings it below the triplet. The two-dimensional scan of the singlet (1)A(1) potential energy surface (PES) reveals that there is no minimum corresponding to a singlet diradical structure. Thus, singlet OXA undergoes prompt barrierless ring closure. However, a flat shape of the PES results in the resonance trapping in the Franck-Condon region, giving rise to the experimentally observable features in the photoelectron spectrum. By performing reduced-dimensionality wave packet calculations, we estimated that the wave packet lingers in the Franck-Condon region for about 170 fs, which corresponds to the spectral line broadening of about 200 cm(-1). We also present calculations of the photodetachment spectrum and compare it with experimental data. Our calculations lend strong support to the assignment of the photoelectron spectrum of the OXA anion reported in Ichino et al. (Angew. Chem., Int. Ed. Engl. 2009, 48, 8509).


Regulation of mammalian autophagy by class II and III PI 3-kinases through PI3P synthesis.
Synthesis of phosphatidylinositol-3-phosphate (PI3P) by Vps34, a class III phosphatidylinositol 3-kinase (PI3K), is critical for the initial steps of autophagosome (AP) biogenesis. Although Vps34 is the sole source of PI3P in budding yeast, mammalian cells can produce PI3P through alternate pathways, including direct synthesis by the class II PI3Ks; however, the physiological relevance of these alternate pathways in the context of autophagy is unknown. Here we generated Vps34 knockout mouse embryonic fibroblasts (MEFs) and using a higher affinity 4x-FYVE finger PI3P-binding probe found a Vps34-independent pool of PI3P accounting for (~)35% of the total amount of this lipid species by biochemical analysis. Importantly, WIPI-1, an autophagy-relevant PI3P probe, still formed some puncta upon starvation-induced autophagy in Vps34 knockout MEFs. Additional characterization of autophagy by electron microscopy as well as protein degradation assays showed that while Vps34 is important for starvation-induced autophagy there is a significant component of functional autophagy occurring in the absence of Vps34. Given these findings, class II PI3Ks (α and β isoforms) were examined as potential positive regulators of autophagy. Depletion of class II PI3Ks reduced recruitment of WIPI-1 and LC3 to AP nucleation sites and caused an accumulation of the autophagy substrate, p62, which was exacerbated upon the concomitant ablation of Vps34. Our studies indicate that while Vps34 is the main PI3P source during autophagy, class II PI3Ks also significantly contribute to PI3P generation and regulate AP biogenesis.


The effects of prostaglandin F2alpha on the fine structure of the corpus luteum of the pregnant hamster.
Corpora lutea (C.L.) from normal and prostaglandin-treated pregnant hamsters were examined by light and electron microscopy. Luteal cells from pregnant control animals were hypertrophied and contained an abundance of agranular endoplasmic reticulum (AER) suggesting steroidogenic activity. Animals which were injected on day 4 of pregnancy had ovulated by the third day following the injection, hence, two generations of corpora lutea were present in the ovary. Luteal cells from the "old" C.L. of pregnancy underwent a progressive degeneration during the four days following the single prostaglandin injection, whereas, cells from "new" C.L. appeared viable in most respects. The presence of aggregated platelets in both "old" and "new" C.L. was noted. It is suggested that the "new" C.L. are responsible for the rebound of plasma progesterone reported in previous investigations three days following prostaglandin treatment in pregnant hamsters.


Nanoscale imaging of RNA with expansion microscopy.
The ability to image RNA identity and location with nanoscale precision in intact tissues is of great interest for defining cell types and states in normal and pathological biological settings. Here, we present a strategy for expansion microscopy of RNA. We developed a small-molecule linker that enables RNA to be covalently attached to a swellable polyelectrolyte gel synthesized throughout a biological specimen. Then, postexpansion, fluorescent in situ hybridization (FISH) imaging of RNA can be performed with high yield and specificity as well as single-molecule precision in both cultured cells and intact brain tissue. Expansion FISH (ExFISH) separates RNAs and supports amplification of single-molecule signals (i.e., via hybridization chain reaction) as well as multiplexed RNA FISH readout. ExFISH thus enables super-resolution imaging of RNA structure and location with diffraction-limited microscopes in thick specimens, such as intact brain tissue and other tissues of importance to biology and medicine.


Passing the baton--to whom?
Scientific discovery occasionally occurs as a sudden and dramatic leap ahead but more often proceeds at a subtler and steadier pace. Each small step forward may escape public notice but is ultimately vital to the journey's success. Indeed, such gradual advancement represents the collective contributions of many workers in the field, some new to the journey. While the notion of combined effort and multiple contributors is honorable, it poses an inherent danger. In our society, unproven, unorthodox, or unnoticed researchers may not receive the funding or support needed to make their contributions. Furthermore, even if they have the potential to make a leap, a hostile environment may preclude their doing so. This article concentrates on the looming crisis in diabetes research, but the principles pertain to all fields of clinical and biomedical science.


Use of 111-Indium-labeled autologous eosinophils to establish the in vivo kinetics of human eosinophils in healthy subjects.
Eosinophils are the major cellular effectors of allergic inflammation and represent an important therapeutic target. Although the genesis and activation of eosinophils have been extensively explored, little is known about their intravascular kinetics or physiological fate. This study was designed to determine the intravascular life span of eosinophils, their partitioning between circulating and marginated pools, and sites of disposal in healthy persons. Using autologous, minimally manipulated 111-Indium-labeled leukocytes with blood sampling, we measured the eosinophil intravascular residence time as 25.2 hours (compared with 10.3 hours for neutrophils) and demonstrated a substantial marginated eosinophil pool. γ camera imaging studies using purified eosinophils demonstrated initial retention in the lungs, with early redistribution to the liver and spleen, and evidence of recirculation from a hepatic pool. This work provides the first in vivo measurements of eosinophil kinetics in healthy volunteers and shows that 111-Indium-labeled eosinophils can be used to monitor the fate of eosinophils noninvasively.


An outbreak of necrotizing enterocolitis. Association with transfusions of packed red blood cells.
Of 187 newborns admitted to a 33-bed, level III neonatal intensive care unit between January 1, 1985 and June 23, 1985, 33 developed necrotizing enterocolitis during their hospital stay. Twenty of the 33 newborns (61%) had onset of symptoms between April 1 and June 23, suggesting clustering during this period. A case-control study, with matching on birth weight class, approximate date of admission to the unit and approximate duration of stay, failed to reveal any association of the syndrome with type or timing of feeding, perinatal hypoxic events, as determined by apgar scores and labor history, or specific microbial organisms. By contrast, however, transfusion of packed red blood cells was highly and significantly associated with the syndrome (odds ratio = 15.1, 95% confidence interval = 2.59-92.51). In addition, therapy with caffeine, with theophylline, and with furosemide were moderately associated with the syndrome, although not significantly so. During this outbreak period, the incidence of necrotizing enterocolitis by birth weight was 30.6% in infants less than 1,500 gm, 10.8% in infants 1,500-2,500 gm, and 11.9% in infants 2,500 gm or more. These findings confirm the importance of low birth weight as a risk factor for development of the syndrome and suggest that insults to volume homeostasis, such as transfusion and use of diuretics, need to be considered as possible mechanisms whereby necrotizing enterocolitis is initiated.


Plasma IAPP-Autoantibody Levels in Alzheimer's Disease Patients Are Affected by APOE4 Status.
Pancreas-derived islet amyloid polypeptide (IAPP) crosses the blood-brain barrier and co-deposits with amyloid beta (Aβ) in brains of type 2 diabetes (T2D) and Alzheimer's disease (AD) patients. Depositions might be related to the circulating IAPP levels, but it warrants further investigation. Autoantibodies recognizing toxic IAPP oligomers (IAPPO) but not monomers (IAPPM) or fibrils have been found in T2D, but studies on AD are lacking. In this study, we have analyzed plasma from two cohorts and found that levels of neither immunoglobulin (Ig) M, nor IgG or IgA against IAPPM or IAPPO were altered in AD patients compared with controls. However, our results show significantly lower IAPPO-IgA levels in apolipoprotein E (APOE) 4 carriers compared with non-carriers in an allele dose-dependent manner, and the decrease is linked to the AD pathology. Furthermore, plasma IAPP-Ig levels, especially IAPP-IgA, correlated with cognitive decline, C-reactive protein, cerebrospinal fluid Aβ and tau, neurofibrillary tangles, and brain IAPP exclusively in APOE4 non-carriers. We speculate that the reduction in IAPPO-IgA levels may be caused by increased plasma IAPPO levels or masked epitopes in APOE4 carriers and propose that IgA and APOE4 status play a specific role in clearance of circulatory IAPPO, which may influence the amount of IAPP deposition in the AD brain.


Multiple insulin degrading enzyme variants alter in vitro reporter gene expression.
The insulin degrading enzyme (IDE) variant, v311 (rs6583817), is associated with increased post-mortem cerebellar IDE mRNA, decreased plasma β-amyloid (Aβ), decreased risk for Alzheimer's disease (AD) and increased reporter gene expression, suggesting that it is a functional variant driving increased IDE expression. To identify other functional IDE variants, we have tested v685, rs11187061 (associated with decreased cerebellar IDE mRNA) and variants on H6, the haplotype tagged by v311 (v10; rs4646958, v315; rs7895832, v687; rs17107734 and v154; rs4646957), for altered in vitro reporter gene expression. The reporter gene expression levels associated with the second most common haplotype (H2) successfully replicated the post-mortem findings in hepatocytoma (0.89 fold-change, p = 0.04) but not neuroblastoma cells. Successful in vitro replication was achieved for H6 in neuroblastoma cells when the sequence was cloned 5' to the promoter (1.18 fold-change, p = 0.006) and 3' to the reporter gene (1.29 fold change, p = 0.003), an effect contributed to by four variants (v10, v315, v154 and v311). Since IDE mediates Aβ degradation, variants that regulate IDE expression could represent good therapeutic targets for AD.


Post-translational polymodification of β1-tubulin regulates motor protein localisation in platelet production and function.
In specialised cells, the expression of specific tubulin isoforms and their subsequent post-translational modifications drive and coordinate unique morphologies and behaviours. The mechanisms by which β1-tubulin, the platelet and megakaryocyte (MK) lineage restricted tubulin isoform, drives platelet production and function remains poorly understood. We investigated the roles of two key post-translational tubulin polymodifications (polyglutamylation and polyglycylation) on these processes using a cohort of thrombocytopenic patients, human induced pluripotent stem cell (iPSC) derived MKs, and healthy human donor platelets. We find distinct patterns of polymodification in MKs and platelets, mediated by the antagonistic activities of the cell specific expression of Tubulin Tyrosine Ligase Like (TTLLs) and Cytosolic Carboxypeptidase (CCP) enzymes. The resulting microtubule patterning spatially regulates motor proteins to drive proplatelet formation in megakaryocytes, and the cytoskeletal reorganisation required for thrombus formation. This work is the first to show a reversible system of polymodification by which different cell specific functions are achieved.


Food Bioactive HDAC Inhibitors in the Epigenetic Regulation of Heart Failure.
Approximately 5.7 million U.S. adults have been diagnosed with heart failure (HF). More concerning is that one in nine U.S. deaths included HF as a contributing cause. Current HF drugs (e.g., β-blockers, ACEi) target intracellular signaling cascades downstream of cell surface receptors to prevent cardiac pump dysfunction. However, these drugs fail to target other redundant intracellular signaling pathways and, therefore, limit drug efficacy. As such, it has been postulated that compounds designed to target shared downstream mediators of these signaling pathways would be more efficacious for the treatment of HF. Histone deacetylation has been linked as a key pathogenetic element for the development of HF. Lysine residues undergo diverse and reversible post-translational modifications that include acetylation and have historically been studied as epigenetic modifiers of histone tails within chromatin that provide an important mechanism for regulating gene expression. Of recent, bioactive compounds within our diet have been linked to the regulation of gene expression, in part, through regulation of the epi-genome. It has been reported that food bioactives regulate histone acetylation via direct regulation of writer (histone acetyl transferases, HATs) and eraser (histone deacetylases, HDACs) proteins. Therefore, bioactive food compounds offer unique therapeutic strategies as epigenetic modifiers of heart failure. This review will highlight food bio-actives as modifiers of histone deacetylase activity in the heart.


Multi-omics systems toxicology study of mouse lung assessing the effects of aerosols from two heat-not-burn tobacco products and cigarette smoke.
Cigarette smoke (CS) causes adverse health effects and, for smoker who do not quit, modified risk tobacco products (MRTPs) can be an alternative to reduce the risk of developing smoking-related diseases. Standard toxicological endpoints can lack sensitivity, with systems toxicology approaches yielding broader insights into toxicological mechanisms. In a 6-month systems toxicology study on ApoE-/- mice, we conducted an integrative multi-omics analysis to assess the effects of aerosols from the Carbon Heated Tobacco Product (CHTP) 1.2 and Tobacco Heating System (THS) 2.2-a potential and a candidate MRTP based on the heat-not-burn (HnB) principle-compared with CS at matched nicotine concentrations. Molecular exposure effects in the lungs were measured by mRNA/microRNA transcriptomics, proteomics, metabolomics, and lipidomics. Integrative data analysis included Multi-Omics Factor Analysis and multi-modality functional network interpretation. Across all five data modalities, CS exposure was associated with an increased inflammatory and oxidative stress response, and lipid/surfactant alterations. Upon HnB aerosol exposure these effects were much more limited or absent, with reversal of CS-induced effects upon cessation and switching to CHTP 1.2. Functional network analysis revealed CS-induced complex immunoregulatory interactions across the investigated molecular layers (e.g., itaconate, quinolinate, and miR-146) and highlighted the engagement of the heme-Hmox-bilirubin oxidative stress axis by CS. This work exemplifies how multi-omics approaches can be leveraged within systems toxicology studies and the generated multi-omics data set can facilitate the development of analysis methods and can yield further insights into the effects of toxicological exposures on the lung of mice.


Improving treatment times for patients with in-hospital stroke using a standardized protocol.
Previous reports have shown significant delays in treatment of in-hospital stroke (IHS). We developed and implemented our IHS alert protocol in April 2014. We aimed to determine the influence of implementation of our IHS alert protocol. Our implementation processes comprise the following four main steps: IHS protocol development, workshops for hospital staff to learn about the protocol, preparation of standardized IHS treatment kits, and obtaining feedback in a monthly hospital staff conference. We retrospectively compared protocol metrics and clinical outcomes of patients with IHS treated with intravenous thrombolysis and/or endovascular therapy between before (January 2008-March 2014) and after implementation (April 2014-December 2016). Fifty-five patients were included (pre, 25; post, 30). After the implementation, significant reductions occurred in the median time from stroke recognition to evaluation by a neurologist (30 vs. 13.5min, p<0.01) and to first neuroimaging (50 vs. 26.5min, p<0.01) and in the median time from first neuroimaging to intravenous thrombolysis (45 vs. 16min, p=0.02). The median time from first neuroimaging to endovascular therapy had a tendency to decrease (75 vs. 53min, p=0.08). There were no differences in the favorable outcomes (modified Rankin scale score of 0-2) at discharge or the incidence of symptomatic intracranial hemorrhage between the two periods. Our IHS alert protocol implementation saved time in treating patients with IHS without compromising safety.


Validity of a new abdominal bioelectrical impedance device to measure abdominal and visceral fat: comparison with MRI.
Abdominal fat, and in particular, visceral adipose tissue (VAT), is the critical fat depot associated with metabolic aberrations. At present, VAT can only be accurately measured by computed tomography or magnetic resonance imaging (MRI). This study was designed to compare a new abdominal bioelectrical impedance (BIA) device against total abdominal adipose tissue (TAAT) and VAT area measurements made from an abdominal MRI scan, and to assess its reliability and accuracy. One-hundred twenty participants were recruited, stratified by gender and BMI. Participants had triplicate measures of abdominal fat and waist circumference (WC) with the AB-140 (Tanita, Tokyo, Japan) and WC measurements using a manual tape measure. A single abdominal MRI scan was performed as the reference method. Triplicate measures with the AB-140 showed excellent precision for "visceral fat level," trunk fat %, and WC. AB-140 "visceral fat level" showed significantly stronger correlations with MRI TAAT area than with MRI VAT area (r = 0.94 vs. 0.65 in men and 0.92 vs. 0.64 in women). AB-140 WC showed good correlation with manual WC measurements (r = 0.95 in men and 0.90 in women). AB-140 and manual WCs showed comparable correlations with MRI TAAT area (r = 0.92 and 0.96 in men and 0.88 and 0.88 in women). AB-140 is a simple, quick, and precise technique to measure abdominal fat and WC in healthy adults. It provides a useful proxy for TAAT measured by MRI, comparable to the correlation obtained with manual WC measurements. Neither the AB-140 abdominal fat measures nor WC measurement appear to provide a useful proxy measure of VAT.


Apolipoprotein E genotype has a modest impact on the postprandial plasma response to meals of varying fat composition in healthy men in a randomized controlled trial.
Apolioprotein E (APOE) genotype is reported to influence a person's fasting lipid profile and potentially the response to dietary fat manipulation. The impact of APOE genotype on the responsiveness to meals of varying fat composition is unknown. We examined the effect of meals containing 50 g of fat rich in saturated fatty acids (SFAs), unsaturated fatty acids (UNSATs), or SFAs with fish oil (SFA-FO) on postprandial lipemia. A randomized, controlled, test meal study was performed in men recruited according to the APOE genotype (n = 10 APOE3/3, n = 11 APOE3/E4). For the serum apoE response (meal × genotype interaction P = 0.038), concentrations were on average 8% lower after the UNSAT than the SFA-FO meal in APOE4 carriers (P = 0.015) only. In the genotype groups combined, there was a delay in the time to reach maximum triacylglycerol (TG) concentration (mean ± SEM: 313 ± 25 vs. 266 ± 27 min) and higher maximum nonesterified fatty acid (0.73 ± 0.05 vs. 0.60 ± 0.03 mmol/L) and glucose (7.92 ± 0.22 vs. 7.25 ± 0.22 mmol/L) concentrations after the SFA than the UNSAT meal, respectively (P ≤ 0.05). In the Svedberg flotation rate 60-400 TG-rich lipoprotein fraction, meal × genotype interactions were observed for incremental area under the curve (IAUC) for the TG (P = 0.038) and apoE (P = 0.016) responses with a 58% lower apoE IAUC after the UNSAT than the SFA meal (P = 0.017) in the E4 carriers. Our data indicate that APOE genotype had a modest impact on the postprandial response to meals of varying fat composition in normolipidemic men. The physiologic importance of greater apoE concentrations after the SFA-rich meals in APOE4 carriers may reflect an impact on TG-rich lipoprotein clearance from the circulation. This trial was registered at clinicaltrials.gov as NCT01522482.


Cell-penetrating peptides: mechanisms and applications.
A major obstacle in the development of new therapeutic agents is the low bioavailability of hydrophilic substances. Drugs that bind to intracellular targets must penetrate the lipid bilayer surrounding the cell in order to exert their effect. A relatively new research area that addresses this problem by introducing novel transport peptides that facilitate the cellular penetration of potential drugs has emerged. These peptides predominantly have a positive net charge and/or an amphipathic nature, but can otherwise have very different characteristics. This group of peptides, although sometimes called protein transduction domains (PTDs), is here referred to as cell-penetrating peptides (CPPs). For many years it was believed that these peptides were internalized into cells via a non-endocytotic, receptor-independent pathway. However, recent publications have suggested that an endocytotic pathway cannot be ruled out, and that earlier results might be based on artifacts associated with fixation of cells and membrane association of peptides. Although the mechanism of cellular uptake remains unclear, there is an increasing amount of reports on biological effects of CPPs and their cargos. Thus, CPPs are an attractive pharmaceutical and biochemical tool that needs more attention. This review will discuss some recent results in this research field with focus on the cell-penetrating peptide transportan.


Single incision laparoscopic hepatectomy: A systematic review.
Single incision laparoscopic surgery is a rather innovative surgical technique. A systematic literature review was performed with the intention to evaluate the till now clinical evidence regarding the application of single incision technique on liver resections as a method of management in hepatic lesions. Twelve relative studies were found in the field including 30 patients with a age range from 29 to 90 years and a body mass index from 20.1 to 36.5 kg/m(2). Primary hepatic carcinoma (40%), metastatic nodules (26.7%), hepatic cysts (16.7%), hepatic haemangiomas (13.3%) and hepatic adenoma (3.3%) were the most common indications of the lesions resected. The types of hepatectomy performed included partial hepatectomy (43.3%), segmentectomy (30%) and lobectomy (26.7%). In the majority of the patients, left lateral segments (II-III-IV) (76.7%) were resected. The median operative time was 110 min (range: 55-235) while the median quantity of blood loss was 50 ml (range: 0-100). No conversion to open surgery and no transfusion were needed. The duration of hospital stay ranged between 2 and 11 days. No complications, no cases of disease recurrence or death of patients were reported. None of the studies included described data on the cosmesis of the application of single incision laparoscopic technique on hepatic resections. Moreover, the surgical technique, as well as the different type of ports used is also presented in this review. Single site port laparoscopic surgery is a promising minimally invasive procedure for liver resections.


Immunomodulatory role of vascular endothelial growth factor and angiopoietin-1 in airway remodeling.
The blood vessels formed in asthmatic airways are involved in inflammatory and airway remodeling processes in chronic asthma. Vascular endothelial cell growth factor (VEGF) and angiopoietin-1 (Ang-1) are primary angiogenic growth factors, involved in the formation of such blood vessels. VEGF has been reported to contribute to non-specific airway hyper-responsiveness, have chemotactic effects on eosinophils, and enhance airway smooth muscle cell proliferation. Furthermore, Th2 cells have receptors for VEGF, and Th2-associated cytokines increase VEGF production. There are reports that elevated levels of VEGF correlates with the severity of asthma. Ang-1 has been shown to induce pro-inflammatory effects such as eosinophil chemotaxis via tie-2 receptors. Reports indicate ang-1 contribution to increased secretion of matrix metalloproteinase-2 (MMP-2) and decreased secretion of tissue inhibitors of metalloproteinase-2 (TIMP-2). However, Ang-1 has also been shown to exhibit several anti-inflammatory properties such as suppressing expression of adhesion molecules, blocking vascular permeability and eosinophil chemotaxis induced by VEGF. These findings support the notion that apart from their roles in blood vessels formation, these angiogenic growth factors are directly involved in the pathogenesis of chronic asthma. This paper reviews individual and combined roles of VEGF and Ang-1. The potential therapeutic applications involving these factors are also discussed.


A genome scan for quantitative trait loci affecting the Salmonella carrier-state in the chicken.
Selection for increased resistance to Salmonella colonisation and excretion could reduce the risk of foodborne Salmonella infection. In order to identify potential loci affecting resistance, differences in resistance were identified between the N and 6(1) inbred lines and two QTL research performed. In an F2 cross, the animals were inoculated at one week of age with Salmonella enteritidis and cloacal swabs were carried out 4 and 5 wk post inoculation (thereafter called CSW4F2 and CSW4F2) and caecal contamination (CAECF2) was assessed 1 week later. The animals from the (N x 6(1)) x N backcross were inoculated at six weeks of age with Salmonella typhimurium and cloacal swabs were studied from wk 1 to 4 (thereafter called CSW1BC to CSW4BC). A total of 33 F2 and 46 backcross progeny were selectively genotyped for 103 and 135 microsatellite markers respectively. The analysis used least-squares-based and non-parametric interval mapping. Two genome-wise significant QTL were observed on Chromosome 1 for CSW2BC and on Chromosome 2 for CSW4F2, and four suggestive QTL for CSW5F2 on Chromosome 2, for CSW5F2 and CSW2BC on chromosome 5 and for CAECF2 on chromosome 16. These results suggest new regions of interest and the putative role of SAL1.


A novel indirect sequence readout component in the E. coli cyclic AMP receptor protein operator.
The cyclic AMP receptor protein (CRP) from Escherichia coli has been extensively studied for several decades. In particular, a detailed characterization of CRP interaction with DNA has been obtained. The CRP dimer recognizes a consensus sequence AANTGTGANNNNNNTCACANTT through direct amino acid nucleobase interactions in the major groove of the two operator half-sites. Crystal structure analyses have revealed that the interaction results in two strong kinks at the TG/CA steps closest to the 6-base-pair spacer (N6). This spacer exhibits high sequence variability among the more than 100 natural binding sites in the E. coli genome, but the exact role of the N6 region in CRP interaction has not previously been systematic examined. Here we employ an in vitro selection system based on a randomized N6 spacer region to demonstrate that CRP binding to the lacP1 site may be enhanced up to 14-fold or abolished by varying the N6 spacer sequences. Furthermore, on the basis of sequence analysis and uranyl (UO2(2+)) probing data, we propose that the underlying mechanism relies on N6 deformability.


Gender, body, biomedicine: how some feminist concerns dragged reproduction to the center of social theory.
This article tracks the growth of medical anthropology in the United States in the decades since the 1970s, as it has intersected the expansion of feminist activism and scholarship. I argue that feminist attention to embodied inequalities quickly focused on reproduction as a site of investigation and intervention. Medical anthropology has benefited from feminist concern with stratified reproduction, especially its interrogation of nonnormative and stigmatized fertility and childbearing. When reproduction becomes problematic, it provides a lens through which cultural norms, struggles, and transformations can be viewed. Examples drawn from prenatal diagnosis are particularly revelatory of the diverse interests and stakes we all hold in reproduction.


The evolution to stool DNA testing for colorectal cancer.
Despite a variety of screening strategies and recent trends showing death rate stabilization, colorectal cancer still remains the second leading cause of overall cancer death. Current screening tools suffer from performance limitations, low patient acceptability, and marginal reliable access within the health care system. Noninvasive strategies present the lowest risk with the highest potential for patient satisfaction. However, serious implementation barriers exist requiring consistent programmatic screening, strict patient adherence, and poor sensitivity for adenomas. Colonoscopy remains an invasive screening test with the best sensitivity and specificity, but faces large financial costs, manpower requirements, patient access and adherence. Development of advanced molecular techniques identifying altered DNA markers in exfoliated colonocytes signify early or precancerous growth. Stool-based DNA testing provides an entirely noninvasive population-based screening strategy which patients can perform easier than faecal occult blood testing (FOBT). Large-scale prospective randomized control trials currently pending should help characterize accurate test performance, screening intervals, cost-effectiveness, direct comparison to FOBT and analysis of patient adherence. As tumour development pathways and potential target genes are further elucidated, refinements in multi-assay stool-based DNA testing portend enhanced test characteristics to detect and treat this genetically heterogeneous disease.


Access to therapy services for people with disability in rural Australia: a carers' perspective.
In Australia and around the world, people with disability and their carers living in rural areas face additional challenges compared to their urban-dwelling counterparts. This cross-sectional study explored current access to therapy services for people with disability living in western New South Wales as reported by their primary carers. Data were collected using an online and paper survey distributed to carers, from December 2012 until June 2013. Information was sought about the carers, the person they care for, access to physiotherapy, speech pathology, occupational therapy and psychology services. A total of 166 carers participated. Respondents were mostly the carers of a son or daughter (83.6%) , the person they care for had an average age of 17.6 years (range 1-69 years) and more than half were males (56%). The majority of people with disability (73.5%) had received therapy services in the last 2 years. Waiting time and distance travelled to access physiotherapy, speech and occupational therapy services varied. People with disability had to wait an average of 6.6 months to receive speech pathology and had to travel an average of 2.6 hours to receive physiotherapy. The main provider of all services was the specialist disability government department, except for psychology, which is mainly provided in the private sector. From the carers' perspective, availability and accessibility to services are limited. Carers noted that availability of services was more restricted once people with disability reached adulthood. Lack of choice and frustration with the lack of availability of specialist disability services was reported frequently. Carers believed that timely allied health intervention prevented the development of more severe or complicated conditions that had a greater impact on carers, families, communities and the person with disabilities.


Experimental study of structure-borne sound transmission loss of mechanical joints.
A mechanical joint is one of the most effective ways to reduce the transmission of structure-borne sound. In order to increase the transmission loss, heavily damped joints are often used, which, in many cases, will reduce the structure integrity and hence can only be used in limited cases. In this study attention is focused on a type of resonant joint, i.e., a joint which will increase the transmission loss but will not reduce the structure integrity. The study is based on experiments in a one-dimensional structure. It is found that by adjusting the overlap of the joint, the transmission loss of 30 dB can be obtained at a certain frequency range without adding any dissipative materials. The mechanism of this high transmission loss is the cantilever-type resonance. The resonant frequency can be predicted precisely. The influence of extra dissipative material is investigated. The performance of the same joint in a finite structure is also examined by using the concept of vibrational insertion loss. When there is a certain damping in a finite system, a rather high insertion loss can still be achieved by using the above-mentioned joint, but the resonant frequency is shifted to higher end. It seems that the effective length of the cantilever is shortened by the finiteness.


A microfluidic reciprocating intracochlear drug delivery system with reservoir and active dose control.
Reciprocating microfluidic drug delivery, as compared to steady or pulsed infusion, has unique features which may be advantageous in many therapeutic applications. We have previously described a device, designed for wearable use in small animal models, that periodically infuses and then withdraws a sub-microliter volume of drug solution to and from the endogenous fluid of the inner ear. This delivery approach results in zero net volume of liquid transfer while enabling mass transport of compounds to the cochlea by means of diffusion and mixing. We report here on an advanced wearable delivery system aimed at further miniaturization and complex dosing protocols. Enhancements to the system include the incorporation of a planar micropump to generate reciprocating flow and a novel drug reservoir that maintains zero net volume delivery and permits programmable modulation of the drug concentration in the infused bolus. The reciprocating pump is fabricated from laminated polymer films and employs a miniature electromagnetic actuator to meet the size and weight requirements of a head-mounted in vivo guinea pig testing system. The reservoir comprises a long microchannel in series with a micropump, connected in parallel with the reciprocating flow network. We characterized in vitro the response and repeatability of the planar pump and compared the results with a lumped element simulation. We also characterized the performance of the reservoir, including repeatability of dosing and range of dose modulation. Acute in vivo experiments were performed in which the reciprocating pump was used to deliver a test compound to the cochlea of anesthetized guinea pigs to evaluate short-term safety and efficacy of the system. These advances are key steps toward realization of an implantable device for long-term therapeutic applications in humans.


Severe unilateral bronchospasm mimicking inadvertent endobronchial intubation: a complication of the use of a topical lidocaine Laryngojet injector.
A healthy young woman is described in whom the left chest was unable to be inflated after intubation. The differential diagnosis and management are discussed. Severe unilateral bronchospasm was probably caused by topical lidocaine injected at the vocal cords and, inadvertently, into the left main bronchus with a Laryngojet device.