Chimeric influenza viruses incorporating epitopes of outer membrane protein F as a vaccine against pulmonary infection with Pseudomonas aeruginosa.
Peptide 10 (NATAEGRAINRRVE, residues 305-318 of mature protein F) is one of two linear B-cell epitopes within outer membrane protein F of Pseudomonas aeruginosa both of which have been shown to elicit whole cell-reactive antibodies and to afford protection in animal models against P. aeruginosa infection. Influenza A virus was chosen as a vector to present this epitope in a human-compatible vaccine. Various lengths of the peptide 10 epitope ranging from a 5-mer (GRAIN), 7-mer (AINRRVE), 8-mer (TAEGRAIN), 9-mer (GRAINRRVE), 11-mer (AEGRAINRRVE) to a 12-mer (TAEGRAINRRVE) were attempted to be presented into the antigenic B-site of the hemagglutinin (HA) of live recombinant influenza virus. Using PCR, DNA sequences encoding these various peptide 10 lengths were inserted into the HA gene of influenza A/WSN/33 virus. By using a reverse-genetics transfection system, RNA transcribed in vitro from these chimeric HA genes was reassorted into infectious virus. To date chimeric viruses have been rescued and purified containing the peptide 10 5-mer, 7-mer, 8-mer, and 11-mer. RT-PCR and sequencing have confirmed the presence of P. aeruginosa sequences in the HA RNA segment of each chimeric virus. Each of the four chimeric viruses produced to date was used to immunize mice to determine the ability of each chimeric virus to elicit antibodies reactive with whole cells of P. aeruginosa. The immunization protocol consisted of a series of three intranasal inoculations, followed by two intramuscular injections of the chimeric virus. The chimeric virus incorporating the 11-mer elicited IgG antibodies that reacted with various immunotype strains of P. aeruginosa in a whole cell ELISA at titers of 80 to 2,560, whereas the chimeric virus incorporating the 8-mer elicited whole cell-reactive IgG antibodies at titers of 320 to 2,560. These data suggest that these two chimeric viruses may have vaccine efficacy against P. aeruginosa infection. These studies may result in the development of a chimeric influenza virus-protein F vaccine which would prove to be suitable for use in children with cystic fibrosis for the prevention of pulmonary colonization of these children with P. aeruginosa.


Virulence genes and previously unexplored gene clusters in four commensal Neisseria spp. isolated from the human throat expand the neisserial gene repertoire.
Commensal non-pathogenic Neisseria spp. live within the human host alongside the pathogenic Neisseria meningitidis and Neisseria gonorrhoeae and due to natural competence, horizontal gene transfer within the genus is possible and has been observed. Four distinct Neisseria spp. isolates taken from the throats of two human volunteers have been assessed here using a combination of microbiological and bioinformatics techniques. Three of the isolates have been identified as Neisseria subflava biovar perflava and one as Neisseria cinerea. Specific gene clusters have been identified within these commensal isolate genome sequences that are believed to encode a Type VI Secretion System, a newly identified CRISPR system, a Type IV Secretion System unlike that in other Neisseria spp., a hemin transporter, and a haem acquisition and utilization system. This investigation is the first to investigate these systems in either the non-pathogenic or pathogenic Neisseria spp. In addition, the N. subflava biovar perflava possess previously unreported capsule loci and sequences have been identified in all four isolates that are similar to genes seen within the pathogens that are associated with virulence. These data from the four commensal isolates provide further evidence for a Neisseria spp. gene pool and highlight the presence of systems within the commensals with functions still to be explored.


Virtual ward round.
"Virtual Ward Round" is a web-based blended learning tool. The program simulates hospital ward rounds. Within a virtual environment, students make diagnosis and order treatments. Tutors prepare cases easily to ensure realistic cases directly linked to the corresponding classes. The program "Virtual Ward Round" will hopefully be enrichment to the curriculum-based teaching.


Effects of Interspecific Chromosome Substitution in Upland Cotton on Cottonseed Micronutrients.
Micronutrients are essential for plant growth and development, and important for human health nutrition and livestock feed. Therefore, the discovery of novel germplasm with significant variability or higher micronutrients content in crop seeds is critical. Currently, there is no information available on the effects of chromosome or chromosome arm substitution in cotton on cottonseed micronutrients. Thus, the objective of this study was to evaluate the effects of chromosome or chromosome arm substitution on the variability and levels of micronutrients B, Fe, Cu, Zn, Mn, and Ni in cottonseed from chromosome substitution (CS) cotton lines. Our hypothesis was that interspecific chromosome substitution in cotton can affect cottonseed micronutrients content, resulting in significant differences and variabilities of these nutrients among CS lines and between CS lines and the controls. Nine CS lines were grown in two-field experiments at two locations (in 2013 in South Carolina, USA; and in 2014 in Mississippi, USA). TM-1 (the recurrent parent of the CS line) and AM UA48 (cultivar) were used as control. The results showed significant variability among CS lines compared to the controls AM UA48 and TM-1. For example, in South Carolina (SC), B concentration in cottonseed ranged from 10.35 mg kg-1 in CS-M02 to 13.67 mg kg-1 in CS-T04. The concentration of Cu ranged from 4.81 mg kg-1 in CS-B08sh to 7.65 mg kg-1 in CS-T02, and CS-T02 was higher than both controls. The concentration of Fe ranged from 36.09 mg kg-1 to 56.69 mg kg-1 (an increase up to 57%), and six CS lines (CS-B02, CS-B08sh, CS-M02, CS-M04, CS-T02, and CS-T04) had higher concentration than both controls in 2013. In 2014 at the Mississippi location (MS), similar observation was found with CS lines for micronutrients content. The CS lines with higher concentrations of these micronutrients can be used as a genetic tool toward QTL identification for desired seed traits because these lines are genetically similar with TM-1, except the substituted chromosome or chromosome segment pairs from the alien species. Chromosome substitution provides an effective means for upland cotton improvement by targeted interspecific introgression, yielding CS lines that facilitate trait discovery, such as seed micronutritional qualities, due to increased isogenicity and markedly reduced complexity from epistatic interactions with non-target alien chromosomes. The positive correlation between B, Cu, and Fe at both locations, between Ni and Mn, between Zn and Cu, and between Zn and Ni at both locations signify the importance of a good agricultural and fertilizer management of these nutrients to maintain higher cottonseed nutrient content.


[Prevention of experimental allergic encephalomyelitis in guinea pigs by treatment with magnetophores].
Subcutaneous application of magnetophores with a field intensity of 210 ersted prevented the development of experimental allergic encephalomyelitis (EAE) in guinea-pigs. The magnetophores appeared efficacious both at the beginning of the induction period and in the second half of the latent period, i.e. within the first two weeks after injection of encephalitogenic suspension. Application of magnetophores after the appearance of the neurological signs of EAE was ineffective. Magnetophores did not affect cellular or humoral response in animals. It may be assumed that the effect of magnetophores is related to the increased resistance of nerve cells to the immunopathological action of the immune response factors, a topic for further studies.


Heavy rainfall and waterborne disease outbreaks: the Walkerton example.
Recent research indicates that excessive rainfall has been a significant contributor to historical waterborne disease outbreaks. The Meteorological Service of Canada, Environment Canada, provided an analysis and testimony to the Walkerton Inquiry on the excessive rainfall events, including an assessment of the historical significance and expected return periods of the rainfall amounts. While the onset of the majority of the Walkerton, Ontario, Escherichia coli O157:H7 and Campylobacter outbreak occurred several days after a heavy rainfall on May 12, the accumulated 5-d rainfall amounts from 8-12 May were particularly significant. These 5-d accumulations could, on average, only be expected once every 60 yr or more in Walkerton and once every 100 yr or so in the heaviest rainfall area to the south of Walkerton. The significant link between excess rainfall and waterborne disease outbreaks, in conjunction with other multiple risk factors, indicates that meteorological and climatological conditions need to be considered by water managers, public health officials, and private citizens as a significant risk factor for water contamination. A system to identify and project the impacts of such challenging or extreme weather conditions on water supply systems could be developed using a combination of weather/climate monitoring information and weather prediction or quantitative precipitation forecast information. The use of weather monitoring and forecast information or a "wellhead alert system" could alert water system and water supply managers on the potential response of their systems to challenging weather conditions and additional requirements to protect health. Similar approaches have recently been used by beach managers in parts of the United States to predict day-to-day water quality for beach advisories.


Low back three-dimensional joint forces, kinematics, and kinetics during walking.
The purpose of this study was to examine the three-dimensional low back loads, spinal motions, and trunk muscular activity during gait. Specific objectives involved assessment of the effects of walking speed, and arm swing on spinal loads, lumbar spine motion, and muscular activation. An in vivo modeling experiment using five male participants. Thirty walking trials were performed by each participant yielding five repeats of each condition (3 walking cadences x 2 arm swing conditions). Walking is often prescribed as a rehabilitation task for individuals with low back injuries. However, there are few studies which have examined the joint loading, spinal motions, and muscular activity present when walking. Additionally, the majority of studies examining spine loading during gait have used an inverse dynamics model, commencing at the cranial aspect of the body, approach which does not include the impulsive phases of gait (i.e. heel strikes and toe offs). Low back joint forces (bone on bone) and moments were determined using an anatomically complex three-dimensional model (detailing 54 muscles and the passive structures acting at the low back) during three walking cadences and with free arm swing or restricted arm swing. In order to assess the influence of the transient factors such as heel contact on the joint forces a bottom up (from the feet to the lumbar spine) rigid link segment analyses approach was used as one input to the three-dimensional anatomic model. Lumbar spine motion and trunk muscle activation levels were also recorded to assist in partitioning forces amongst the active and passive tissues of the low back. Net joint anterior-posterior shear loading was the only variable significantly affected by walking cadence (fast versus slow P < 0.0003). No variable was significantly affected by the arm swing condition. Trends demonstrated an increase in all variables with increased walking cadence. Similarly, most variables, with the exception of axial twist and lateral bend lumbar spine motion and lateral joint shear, demonstrated increasing trends caused by the restriction of normal arm swing. Tissue loading during walking appears to be below levels caused by many specific rehabilitation tasks, suggesting that walking is a wise choice for general back exercise and rehabilitation programs. Slow walking with restricted arm swing produced more 'static' lumbar spine loading and motion patterns, which could be detrimental for certain injuries and tissues. Fast walking produced a more cyclic loading pattern.


The role of clinical practice guidelines in enhancing quality and reducing racial/ethnic disparities in paediatrics.
Paediatricians believe that clinical practice guidelines (CPGs) can improve patient care but only when the CPGs are flexible, rigorously tested, effective, not used punitively and are motivated by desires to improve quality, not reduce costs. Paediatricians may avoid using CPGs because few have been subjected to rigorous evaluation to demonstrate improved quality and reduced costs. Effective physician adherence to a CPG can be viewed as a multi-step process with potential barriers at each step. Barriers to adherence can be classified as internal (lack of awareness, familiarity, agreement, self-efficacy, outcome expectancy regarding a given CPG and inability to overcome the inertia of current practice) or external (deficits in the required CPG implementation infrastructure). Effective interventions for overcoming barriers to CPG adherence can include multiple modes of disseminating CPG information to practitioners, continuing medical education, feedback to physicians and establishing a solid infrastructure for CPG implementation. Adherence to evidence-based asthma CPGs may be a potentially powerful tool for reducing or eliminating racial/ethnic disparities in asthma treatment because CPGs consist of strict, explicit protocols based on symptoms, not race, ethnicity or socioeconomic status.


Capillary isoelectric focusing and high-performance cation-exchange chromatography compared for qualitative and quantitative analysis of hemoglobin variants.
We have developed two assays for complete analysis of hemoglobins (Hbs) in the field of hemoglobinopathies: a high-performance cation-exchange liquid chromatography (HPLC) assay on the weak cation-exchanger Poly Cat A and a two-step capillary isoelectric focusing (CIEF) assay on the neutral-coated capillary from Beckman in a narrow pH gradient. The resolution was satisfactory for both HPLC and CIEF and allowed separation of normal and common abnormal Hbs, i.e., Hb A, Hb F, Hb A2, Hb S, Hb C, and Hb E; slight differences were shown for the resolution of unusual variants such as Hb C-Harlem and Hb D-Punjab. The reproducibility of retention times was satisfactory as well for HPLC (CV 3.3%) and CIEF (CV 4.9%). The imprecision of quantification of Hb A2, evaluated at two concentrations, and of Hb F and Hb S was < 5%, except for low concentrations of Hb A2 quantified by CIEF. Quantitative data obtained for these three Hb forms were highly correlated between the two assays. These results suggest that the new CIEF assay can be competitive with HPLC for complete routine analysis of Hb variants.


Differentiation of mouse embryonic palatal epithelium in culture: selective cytokeratin expression distinguishes between oral, medial edge and nasal epithelial cells.
During normal murine palatogenesis, regional specific differentiation of the epithelium results in three cell phenotypes: nasal (ciliated pseudostratified columnar cells), oral (stratified squamous cells) and medial edge (migratory, epithelio-mesenchymally transformed cells). We have developed a defined, serum-free, culture system which supports the growth and differentiation of isolated murine embryonic palatal epithelia in vitro. Using immunofluorescence microscopy, an established panel of antibodies was used to characterise the cytokeratin intermediate filament profile of palatal epithelial sheets at a precise developmental stage, following culture in serum-free medium with and without either transforming growth factor alpha (TGF alpha) or 10% donor calf serum (DCS). The morphologically discernable oral, medial edge and nasal phenotypes exhibited distinctive cytokeratin profiles, which remained consistent for all culture conditions, and which correlated with the known differentiation states of the epithelial types. The oral epithelia stained positively for cytokeratin 19 and cytokeratins characteristic of multilayered epithelia (1, 5, 14). Nasal epithelia stained similarly but in addition expressed the simple-epithelial cytokeratin pair, 8 and 18. Medial edge epithelia also expressed cytokeratins 1, 5 and 14 but with the exception of a few isolated cells there was no staining for cytokeratins 8 and 18. Cytokeratin 19 was absent specifically from the medial edge epithelial cells: this result may be related to the loss of cytokeratin expression observed during epithelial-mesenchymal transformations. By exhibiting a complexity of expression linked to differentiation state and independent of culture conditions, cytokeratins constitute useful markers of palatal epithelial differentiation in vitro as well as in vivo.


[Mini-invasive fixation of proximal femoral fractures: what benefit for elderly patients?].
Fixation is used to treat more than two-thirds of proximal femur fractures. The mortality rate is about 25% at one year in these patients, who have an average age of about 80 years. This is mainly due to aging, but also to a gradual deterioration of general health (especially if the operation has been delayed, or after a long stay in the surgical ward) and to local complications (displacement, infection, hematoma). Two fixation devices (a sliding screw plate and a trochanteric nail) have been designed for mini-invasive treatment with fluoroscopic guidance and an incision smaller than 50 mm. The aim is to respect the soft tissues and thereby to avoid local complications, diminish pain, and facilitate early weight-bearing Hospital discharge is possible after 3 or 4 days. Laboratory experiments have shown the satisfactory resistance of the implant and bone at full weight bearing A preliminary series of 30 patients showed the feasibility of these techniques. Primary fusion was achieved in 27 cases. There were no infections and no bleeding, despite antiplatelet treatment. The techniques have now been optimized and multicenter studies are held to determine their real benefit. Fracture fusion and hip motion should be at least as good as with open surgery (90 to 96% fusions, albeit influenced by the precise position of the implants and by osteoporosis). Mortality may be slightly reduced, thanks to immediate operation, early discharge, and fewer local complications. The cost of treatment could also be significantly reduced by the shorter hospital stay. In a few years' time, mini-invasive treatment may become the standard for elderly patients with proximal femur fractures.


Marginal adaptation of class V restorations with current-generation dentin-bonding agents: effect of different dentin surface treatments.
This study sought to assess the use of chlorhexidine with several excipients as a dentin surface treatment and its effect on marginal adaptation of class V restorations with current-generation dentin bonding agents. A total of 120 human third molars were selected and allocated into 12 groups, with standardized buccal class V restorations randomly divided into preconditioned dentin rinsed with: water; water + chlorhexidine; ethanol; or ethanol + chlorhexidine. After rinsing of dentin (previously conditioned with 35% phosphoric acid) with the test solutions, the Adper single bond 2, prime and bond 2.1, and Excite bonding systems were applied randomly. Restorations were performed with Filtek™ Z350 XT composite resin. The resulting specimens were subjected to thermal and mechanical load cycling. Quantitative analysis of marginal adaptation was performed on epoxy replicas by means of scanning electron microscopy. Results were assessed by means of the Kruskal-Wallis test (percentages of continuous margins) and Wilcoxon test (differences between percentages of continuous margins before and after thermal cycling and mechanical loading), at a significance level of p < 0.05. Outcomes in the chlorhexidine-treated groups were not superior to those obtained with other treatments.


Tapered Fiber-Optic Mach-Zehnder Interferometer for Ultra-High Sensitivity Measurement of Refractive Index.
A Mach-Zehnder interferometer (MZI) based fiberoptic refractive index (RI) sensor is constructed by uniformly tapering standard single mode fiber (SMF) for RI measurement. A custom flame-based tapering machine is used to fabricate microfiber MZI sensors directly from SMFs. The fabricated MZI device does not require any splicing of fibers and shows excellent RI sensitivity. The sensor with a cladding diameter of 35.5 µm and length of 20 mm exhibits RI sensitivity of 415 nm/RIU for RI range of 1.332 to 1.384, 1103 nm/RIU for RI range of 1.384 to 1.4204 and 4234 nm/RIU for RI range of 1.4204 to 1.4408, respectively. The sensor reveals a temperature sensitivity of 0.0097 nm/°C, which is relatively low in comparison to its ultra-high RI sensitivity. The proposed inexpensive and highly sensitive optical fiber RI sensors have numerous applications in chemical and biochemical sensing fields.


Endogenous ethylene and abscisic Acid relative to phytogerontology.
Endogenous production of ethylene and endogenous levels of abscisic acid were measured from Hibiscus rosa-sinensis L. abscission zone explants at six stages of development: tight bud, open flower, closed flower, petal abscission, calyx abscission, and peduncle abscission.Explants acropetal and basipetal to the abscission zone produced less ethylene than the abscission zone explants. Ethylene production increased with time both prior to and during abscission, reaching a peak in the later stages of senescence after abscission was complete.Bound abscisic acid was greatest in segments acropetal to the abscission zone at the closed flower stage. Free abscisic acid was double that of bound abscisic acid in the tight bud stage with the basipetal level exceeding that of the acropetal level until flower closure. Acropetal-free abscisic acid began to rise at petal abscission increasing sharply to a peak at calyx abscission. Both free and bound abscisic acid were greatly reduced at peduncle abscission. A relationship of ethylene and abscisic acid to abscission and senescence appears to exist.


A new murine model of allergic conjunctivitis and effectiveness of nedocromil sodium.
Allergic conjunctivitis is the most common atopic disease affecting the eye. To study the pathophysiology and effectiveness of antiallergic drugs, it is necessary to develop animal models that closely mimic human allergic conjunctivitis. The study was performed to develop an experimental murine model of ocular allergic conjunctivitis to an airborne allergen. SWR/J mice were divided into the following groups: group 1, untreated, experimental; group 2, phosphate-buffered saline-treated; group 3, nedocromil sodium-treated; and group 4, unmanipulated controls. Groups 1, 2, and 3 were exposed to ragweed by topical contact with the nasal and conjunctival mucosae. Allergic conjunctivitis was evaluated by scoring of clinical signs, serum IgE levels, and histologic findings. Mice exposed to ragweed had clinicopathologic signs of allergic conjunctivitis and specific anti-ragweed IgE. Allergic conjunctivitis was modulated by nedocromil sodium. Treated mice had fewer clinical signs of allergy, lower levels of ragweed-specific IgE, reduction of conjunctival eosinophil infiltration, decrease in the number of intact and degranulating mast cells, and reduction of cytokine release. This is the first report of a murine model of allergic conjunctivitis to an airborne allergen that can be used to study the disease pathophysiology and its response to treatment.


Barriers for progress in salt reduction in the general population. An international study.
Salt reduction is important for reducing hypertension and the risk of cardiovascular events, nevertheless worldwide salt intakes are above recommendations. Consequently strategies to reduce intake are required, however these require an understanding of salt intake behaviours to be effective. As limited information is available on this, an international study was conducted to derive knowledge on salt intake and associated behaviours in the general population. An online cohort was recruited consisting of a representative sample from Germany, Austria, United States of America, Hungary, India, China, South Africa, and Brazil (n=6987; aged 18-65 years; age and gender stratified). Participants completed a comprehensive web-based questionnaire on salt intake and associated behaviours. While salt reduction was seen to be healthy and important, over one third of participants were not interested in salt reduction and the majority were unaware of recommendations. Salt intake was largely underestimated and people were unaware of the main dietary sources of salt. Participants saw themselves as mainly responsible for their salt intake, but also acknowledged the roles of others. Additionally, they wanted to learn more about why salt was bad for health and what the main sources in the diet were. As such, strategies to reduce salt intake must raise interest in engaging in salt reduction through improving understanding of intake levels and dietary sources of salt. Moreover, while some aspects of salt reduction can be globally implemented, local tailoring is required to match level of interest in salt reduction. These findings provide unique insights into issues surrounding salt reduction and should be used to develop effective salt reduction strategies and/or policies.


Evidence for expression of estrogen receptor cofactor messenger ribonucleic acid in the ovary and uterus of domesticated animals (sheep, cow and pig).
Expression levels of estrogen receptor cofactors (coactivators or corepressors) in specific tissue compartments and cells are thought to influence the expression of estrogen responsive genes and thereby influence overall hormonal responsiveness of target tissues. To date, the presence of cofactors has been reported in tissues from humans, rats and mice. We analyzed the presence and distribution of messenger ribonucleic acids (mRNAs) encoding several transcriptional cofactors in the ovary and uterus of three domestic animal species, the sheep, cow and pig. Northern analysis for cofactors SRC-1, GRIP1, RAC3, p300, RIP140, and SPA showed expression in ovaries from all three species. In addition, lower expression of SRC-1, GRIP1, RAC3, p300, and RIP140 mRNAs was observed during the luteal phase (day 10-12 of the estrous cycle) than at estrus (day 0); however, SPA transcript levels remained unchanged. We then examined expression of mRNAs for changing (SRC-1, RIP140) and constitutively expressed (SPA) cofactors in ovine ovaries. SRC-1 and RIP140 transcripts in corpus luteum were lower compared to the surrounding ovarian tissue. SPA mRNA expression, however, was similar in corpus luteum and surrounding tissues. To determine which ovarian cell types express SRC-1, RIP140, and SPA, in situ hybridization was performed on sheep ovaries. Silver grains corresponding to these cofactors were seen in ovarian granulosa, theca and stromal cells, but appeared to be most abundant in the granulosa cells. Expression of SRC-1 and RIP140 in corpus luteum, however, was reduced compared to expression in follicular cells. Finally, we examined cofactor expression in ovine, bovine, and porcine uterus. Northern blot analysis for SRC-1, GRIP1, RAC3, p300, and RIP140 mRNAs showed higher expression in extracts of the endometrium compared to whole uterus. We provide the first evidence for the presence of estrogen receptor cofactor mRNAs in the ovary and uterus of three domestic animal species. We suggest that coactivators are conserved among species and associated with hormonal responsiveness of reproductive tract tissues in sheep, cow and pig.


Emotionally focused therapy with couples facing breast cancer: a theoretical foundation and descriptive case study.
Women with breast cancer experience a range of distressing emotions, and their partners experience increased distress as well. As a result, their relationship and the patient's health can be negatively affected. A theoretical foundation and specific techniques for applying Emotionally Focused Therapy in couples counseling for breast cancer is presented. A descriptive case study is used to illustrate the process.


Mechanistic insights into type III restriction enzymes.
Type III restriction-modification (R-M) enzymes need to interact with two separate unmethylated DNA sequences in indirectly repeated, head-to-head orientations for efficient cleavage to occur at a defined location next to only one of the two sites. However, cleavage of sites that are not in head-to-head orientation have been observed to occur under certain reaction conditions in vitro. ATP hydrolysis is required for the long-distance communication between the sites prior to cleavage. Type III R-M enzymes comprise two subunits, Res and Mod that form a homodimeric Mod2 and a heterotetrameric Res2Mod2 complex. The Mod subunit in M2 or R2M2 complex recognizes and methylates DNA while the Res subunit in R2M2 complex is responsible for ATP hydrolysis, DNA translocation and cleavage. A vast majority of biochemical studies on Type III R-M enzymes have been undertaken using two closely related enzymes, EcoP1I and EcoP15I. Divergent opinions about how the long-distance interaction between the recognition sites exist and at least three mechanistic models based on 1D- diffusion and/or 3D- DNA looping have been proposed.


Andrographolide Inhibits Proliferation and Metastasis of SGC7901 Gastric Cancer Cells.
To explore the mechanisms by which andrographolide inhibits gastric cancer cell proliferation and metastasis, we employed the gastric cell line SGC7901 to investigate the anticancer effects of andrographolide. The cell survival ratio, cell migration and invasion, cell cycle, apoptosis, and matrix metalloproteinase activity were assessed. Moreover, western blotting and real-time PCR were used to examine the protein expression levels and the mRNA expression levels, respectively. The survival ratio of cells decreased with an increasing concentration of andrographolide in a dose-dependent manner. Consistent results were also obtained using an apoptosis assay, as detected by flow cytometry. The cell cycle was blocked at the G2/M2 phase by andrographolide treatment, and the proportion of cells arrested at G1/M was enhanced as the dose increased. Similarly, wound healing and Transwell assays showed reduced migration and invasion of the gastric cancer cells at various concentrations of andrographolide. Andrographolide can inhibit cell proliferation, invasion, and migration, block the cell cycle, and promote apoptosis in SGC7901 cells. The mechanisms may include upregulated expression of Timp-1/2, cyclin B1, p-Cdc2, Bax, and Bik and downregulated expression of MMP-2/9 and antiapoptosis protein Bcl-2.


Symmetric cyanine dyes for detecting nucleic acids.
A novel approach to the design of sensitive fluorescent probes for nucleic acids detection is proposed. Suitable modifications of tri- and pentamethine cyanine dyes in the polymethine chain and/or in the heterocyclic residues can result in a significant decrease in unbound dye fluorescence intensity and an increase in dye emission intensity in the presence of DNA compared to the unsubstituted dye. The sharp enhancement in the fluorescence intensity upon dye interaction with double-stranded DNA permits the application of the modified tri- and pentamethine dyes as fluorescent probes in double-stranded DNA detection in homogeneous assays.


Room temperature two terminal tunnel magnetoresistance in a lateral graphene transistor.
We investigate the behavior of both pure spin and spin-polarized currents measured with four-probe non-local and two probe local configurations up to room temperature and under an external gate voltage in a lateral graphene transistor, produced using a standard large-scale microfabrication process. The high spin diffusion length of pristine graphene in the channel, measured both directly and by the Hanle effect, and the tuning of the relationship between the electrode resistance areas present in the device architecture allowed us to observe local tunnel magnetoresistance at room temperature, a new finding for this type of device. The results also indicate that while pure spin currents are less sensitive to temperature variations, spin-polarized current switching by an external voltage is more efficient, due to a combination of the Rashba effect and a change in carrier mobility by a Fermi level shift.


Effect of 20% in vitro haemodilution with warmed buffered salt solution and cerebrospinal fluid on coagulation.
We have conducted an in vitro coagulation study consisting of two separate groups of 20 subjects using the thrombelastograph. In the first group, haemodilution was performed with a physiological balanced salt solution similar to plasma, with the exception of calcium, and buffered to a normal pH (Plasmalyte B) at 37 degrees C on blood obtained from consenting volunteers. In the second group, a protein-poor body fluid (cerebrospinal fluid (CSF)) obtained from parturient patients undergoing spinal anaesthesia for Caesarean section was used as the diluent. There were statistically significant differences between the warmed Plasmalyte B treated samples and their untreated controls for all variables measured by the thrombelastograph, except for maximum amplitude, and between the CSF treated samples and their untreated controls for all variables. We conclude that electrolyte and acid-base composition of the diluent fluid had no effect on the observation that crystalloid haemodilution produces hypercoagulability. The marked increase in coagulability produced by addition of CSF cannot be explained on a simple haemodilution basis and confirms previous suggestions of the presence of a procoagulant factor in CSF.


Topical tretinoin (retinoic acid) treatment for liver spots associated with photodamage.
The hyperpigmented lesions commonly called liver spots distress patients, in part because such lesions are associated with aging. We investigated their treatment with topical 0.1 percent tretinoin (retinoic acid). Fifty-eight patients completed a 10-month randomized, double-blind study in which they applied either 0.1 percent tretinoin (n = 28) or vehicle (n = 30) cream daily to the face, upper extremities, or both. Fifteen patients who responded well were than randomly assigned to continue tretinoin therapy or use vehicle alone for six more months. Patients were evaluated by physical examination every month and by analysis of biopsy specimens of lesions obtained at base line and at the end of the 10-month trial. After one month of treatment the patients treated with tretinoin had significant lightening of hyperpigmented lesions as compared with the patients who received vehicle (P less than 0.002). After 10 months, 20 (83 percent) of the 24 patients with facial lesions who were treated with tretinoin had lightening of these lesions, as compared with 8 (29 percent) of the 28 patients with facial lesions who received vehicle. The results for lesions of the upper extremities were similar. As compared with vehicle, tretinoin caused a significant decrease in the degree of epidermal pigmentation and increases in the degree of compaction of stratum corneum, thickness of the granular cell layer, and epidermal thickness. Reductions in epidermal pigmentation evident on histologic analysis were significantly correlated with the degree of clinical lightening of lesions (r = -0.53, P less than 0.0001). During the 6-month follow-up study, specifically identified lesions that had disappeared during the first 10 months of tretinoin treatment did not return in any patient, and six of seven patients who continued to use tretinoin had further improvement. Topical 0.1 percent tretinoin significantly improves both clinical and microscopical manifestations of liver spots; these lesions do not return for at least six months after therapy is discontinued.


Cortisol response under low intensity exercise during cognitive-behavioral therapy is associated with therapeutic outcome in panic disorder-an exploratory study.
Patients with Panic Disorder (PD) show an abnormal stress-induced functioning of the Hypothalamic-Pituitary-adrenal (HPA)-axis. Different protocols for stress induction are of rather low relevance for the psychotherapeutic treatment. In practice, interoceptive exposure is often realized as Low Intensity Exercise (LIE), as compared to an incremental cycle exercise test to exhaustion. Currently, it is not known, whether LIE displays an effective interoceptive stressor 1.) leading to a significant anxiety induction; 2.) a comparable HPA- and Sympathetic-Adreno-Medullar (SAM)-axis response in both patients and healthy controls; 3.) stress responses under LIE are associated with treatment outcomes. N = 20 patients with PD and n = 20 healthy controls were exposed to ten minutes of LIE on an exercise bike. LIE was applied as part of the interoceptive exposure, during an intensive Cognitive-Behavioral Therapy (CBT) in a day clinic. Heart rate was monitored and salivary cortisol samples collected. Before and after the LIE, state anxiety/ arousal were assessed. In order to evaluate psychopathology, the Panic and Agoraphobia Scale, Mobility Inventory, Agoraphobic Cognitions Questionnaire and Body Sensations Questionnaire were applied, before (T1) and after five weeks (T2) of an intensive CBT. LIE led to a significant and similar heart rate increase in both groups. Cortisol decreased over time in both groups, especially in male patients. A higher psychopathology before, and after CBT, was associated with a significantly lower cortisol response under LIE. In the present study, LIE led to a divergent stress response: while there was a significant heart rate increase, cortisol decreased over time, particularly in male patients. A lower reactivity of the HPA-axis seems to be associated with a lower treatment outcome, which may affect extinction based learning. The findings suggest, that interoceptive stimuli should be designed carefully in order to be potent stressors.


Domain structure and function of dynamin probed by limited proteolysis.
Dynamin is a 100-kDa GTPase with multiple domains. Some of these have known functions, namely, the N-terminal GTPase domain, the PH domain that binds phosphatidylinositol lipids, and the C-terminal proline-arginine-rich domain (PRD) that binds to several SH3 domain-containing dynamin partners. Others, for example, the "middle" located between the GTPase domain and the PH domain and a predicted alpha-helical domain located between the PH domain and PRD, have unknown functions. Dynamin exists as a homotetramer in solution and self-assembles into higher-order structures resembling rings and helical stacks of rings. Dynamin self-assembly stimulates its GTPase activity. We used limited proteolysis to dissect dynamin's domain structure and to gain insight into intradomain interactions that regulate dynamin self-assembly and stimulate GTPase activity. We found that the PH domain functions as a negative regulator of dynamin self-assembly and stimulates GTPase activity and that the alpha-helical domain, termed GED for GTPase effector domain, is required for stimulated GTPase activity.


Tibia-hindfoot osteomusculocutaneous rotationplasty with calcaneopelvic arthrodesis for extensive loss of bone from the proximal part of the femur. A report of two cases.
We report a new technique to create an effective lower extremity weight-bearing stump for two patients who had extensive segmental loss of femoral bone proximal to the distal femoral condyles. One patient had previously had complete resection of the proximal part of the femur because of an infection following the insertion of a custom femoral replacement and hip arthroplasty prosthesis. The other patient had had débridement of the femur from the subcapital line to the femoral condyles because of post-traumatic osteomyelitis after failure of a reconstruction with a massive allograft. Both patients were managed with a tibia-hindfoot osteomusculocutaneous rotationplasty after transtarsal (Chopart) amputation, with calcaneopelvic arthrodesis to create stable fixation of the extremity to the pelvis; this fixation allowed flexion, extension, abduction, and adduction of the hip by means of the retained tibiotalar and subtalar joints. At the time of the latest follow-up (at thirty-three and forty-four months), both patients were bearing full weight, without pain, with the use of a standard above-the-knee-amputation prosthesis. We report this procedure as a useful alternative to disarticulation at the level of the hip in patients who have massive loss of femoral bone and destruction of the hip joint in association with scarred and previously infected soft tissues and are not considered to be candidates for other forms of limb-preservation reconstruction. The patient must be willing to accept the equivalent of a low above-the-knee amputation and recognize the potential value of a weight-bearing stump.


Neglected children, shame-proneness, and depressive symptoms.
Neglected children may be at increased risk for depressive symptoms. This study examines shame-proneness as an outcome of child neglect and as a potential explanatory variable in the relation between neglect and depressive symptoms. Participants were 111 children (52 with a Child Protective Services [CPS] allegation of neglect) seen at age 7. Neglected children reported more shame-proneness and more depressive symptoms than comparison children. Guilt-proneness, in contrast, was unrelated to neglect and depressive symptoms, indicating specificity for shame-proneness. The potential role of shame as a process variable that can help explain how some neglected children exhibit depressive symptoms is discussed.


A residue in MutY important for catalysis identified by photocross-linking and mass spectrometry.
MutY is an adenine glycosylase in the base excision repair (BER) superfamily that is involved in the repair of 7,8-dihydro-8-oxo-2'-deoxyguanosine (OG):A and G:A mispairs in DNA. MutY contains a [4Fe-4S]2+ cluster that is part of a novel DNA binding motif, referred to as the iron-sulfur cluster loop (FCL) motif. This motif is found in a subset of members of the BER glycosylase superfamily, defining the endonuclease III-like subfamily. Site-specific cross-linking was successfully employed to investigate the DNA-protein interface of MutY. The photoreactive nucleotide 4-thiothymidine (4ST) incorporated adjacent to the OG:A mismatch formed a specific cross-link between the substrate DNA and MutY. The amino acid participating in the cross-linking reaction was characterized by positive ion electrospray ionization (ESI) tandem mass spectrometry. This analysis revealed Arg 143 as the site of modification in MutY. Arg 143 and nearby Arg 147 are conserved throughout the endo III-like subfamily. Replacement of Arg 143 and Arg 147 with alanine by site-directed mutagenesis reduces adenine glycosylase activity of MutY toward OG:A and G:A mispairs. In addition, the R143A and R147A enzymes exhibit a reduced affinity for duplexes containing the substrate analogue 2'-deoxy-2'-fluoroadenosine opposite OG and G. Modeling of MutY bound to DNA using an endonuclease III-DNA complex structure shows that these two conserved arginines are located within close proximity to the DNA backbone. The insight from mass spectrometry experiments combined with functional mutagenesis results indicate that these two amino acids in the [4Fe-4S]2+ cluster-containing subfamily play an important role in recognition of the damaged DNA substrate.


Is the risk of refeeding syndrome a problem in reaching nutritional requirements? A cohort of patients on enteral nutrition support.
Background: refeeding syndrome (RS) is a metabolic complication associated with nutritional support. The lack of management protocols for patients on nutritional support may lead to the development of RS or undernourishment. Objective: to evaluate adherence to the algorithm for total enteral nutritional support (ASNET), and the efficacy of reaching nutrient requirements in patients at risk of RS. Methods: a cohort study: screening and nutritional evaluation of patients and risk of RS were performed. Adults admitted to hospital for noncritical illness who received exclusively enteral nutrition (EN) were eligible. Patients with RS, intestinal failure or chronic diarrhea were excluded. Adherence to ASNET was evaluated along with the efficacy of nutritional support to reach the optimal protein and energy requirement (OPER) and any associated complications. Patient follow-up ended when the feeding route was changed or upon discharge. Results: a total of 73 patients were included (mean age, 62 ± 16 yrs; BMI, 18.5 ± 4.2 kg/m2), and 55 % were men. All had nutritional risk of developing RS (57, low; 15, high; 1, very high); 34 % had adequate adherence to ASNET, and 33 % managed to cover the total requirement between 4 and 6 days. OPER was reached by 38 % by the fourth day of EN, and adequate adherence to ASNET increased the probability of achieving it (RR, 2.2; 95 % CI, 1.6-3.2, p < 0.0001) without increasing the associated complications. Nonetheless, 36 % developed complications, of whom 96 % did not adhere to ASNET. Conclusion: adherence to ASNET in patients at risk of RS allowed the achievement of OPER safely by day four with fewer associated complications.


Phylogeny and taxonomy of powdery mildew caused by Erysiphe species on Lupinus hosts.
The genus Lupinus (Fabaceae) consists of over 250 plant species located throughout the world. Powdery mildew, caused by Erysiphe species, is a common disease infecting these ecologically, ornamentally, and agriculturally important plants. In the present work, we conducted phylogenetic and taxonomic analyses on Erysiphe species colonizing hosts of the leguminous genus Lupinus, using sequences from the internal transcribed spacer (ITS) and 28S genomic regions. Powdery mildews of the genus Erysiphe on Fabaceae are taxonomically intricate and challenging. Therefore, it is necessary to phylogenetically analyze the DNA retrieved from powdery mildew on lupines in a broad context that includes common and allied powdery mildew species that occur on a range of leguminous plants such as Erysiphe astragali, E. baeumleri, E. pisi, and E. trifoliorum. A new species Erysiphe lupini, found in the USA on Lupinus lepidus, L. polyphyllus, and Lupinus sp., is described. Additionally, Erysiphe intermedia (≡ Microsphaera trifolii var. intermedia) has been confirmed as a North American lupine powdery mildew that is a sister species to E. astragali on Astragalus spp. European Erysiphe collections on lupines were often referred to as E. intermedia, but our analyses have shown that they pertain to E. trifoliorum. The E. trifoliorum clade is composed of several species (i.e. E. baeumleri, E. euonymi, E. hyperici, and E. trifoliorum), that cannot be sufficiently resolved based solely on ITS+28S sequences. Morphological and biological differences between the species are discussed and provide evidence that the species concerned should be maintained. Finally, a sequence obtained from a powdery mildew collected in Portugal on the native Lupinus micranthus pertained to the Erysiphe guarinonii clade. This collection is tentatively treated as Erysiphe sp. To fix the application of the species names E. astragali, E. baeumleri (including its synonym E. marchica), and E. intermedia, epitypes have been designated with ex-epitype sequences.


[The neurochemistry and neuropharmacology of diffuse Lewy body disease].
The main neurochemical alteration in diffuse Lewy body disease (DLBD) is the cholinergic deficit in the cerebral cortex, which involves mainly cholin-acetyl-transferase. There have been also described dopamine deficiency and alterations affecting other neurotransmitters and neuromodulators, such as serotonin, noradrenaline, neuropeptides, etc. Cerebral perfusion and glucose metabolism studies usually show diffuse hypoperfusion or hypometabolism, with higher alteration of associative cortex, including occipital involvement. Several studies have shown increased markers of oxidative stress in brain and other tissues, suggesting its possible role in the pathogenesis of DLBD. Acetylcholinesterase inhibitors seem to improve cognitive and conductual symptoms, although their usefulness according evidence-based medicine criteria is weak. Some patients need atypical neuroleptics at low doses to get the symptomatic control of conductual alterations.


Desmin expression in fibroblasts of murine periovular granuloma during liver Schistosoma mansoni infection.
We have studied the expression of the desmin gene, a muscle-specific intermediate filament protein in the granuloma cells of mouse liver infected with Schistosoma mansoni. In situ hybridization using a desmin DNA probe showed that fibroblastic cells in the granuloma strongly expressed desmin mRNAs, while in normal liver these cells did not express this mRNA to a detectable degree. The quantitative analysis of total RNAs demonstrated that the proportion of specific desmin mRNA increased from 14 to 18 weeks after infection and decreased at 20 weeks. The analysis of collagen gene expression indicated that the amount of type III collagen mRNAs was still increasing after 18 weeks from infection; in contrast, the amount of type I collagen mRNAs remained unchanged at that stage. A good correlation was observed between the detection of the specific mRNAs and the detection of both desmin and collagen molecules. Therefore, these data point to a coordinate induction of desmin and collagen gene expression during Schistosomal granuloma formation. They also suggest that the expression of the myofibroblast phenotype involves the induction of both genes.


Effects of landscape features on population genetic variation of a tropical stream fish, Stone lapping minnow, Garra cambodgiensis, in the upper Nan River drainage basin, northern Thailand.
Spatial genetic variation of river-dwelling freshwater fishes is typically affected by the historical and contemporary river landscape as well as life-history traits. Tropical river and stream landscapes have endured extended geological change, shaping the existing pattern of genetic diversity, but were not directly affected by glaciation. Thus, spatial genetic variation of tropical fish populations should look very different from the pattern observed in temperate fish populations. These data are becoming important for designing appropriate management and conservation plans, as these aquatic systems are undergoing intense development and exploitation. This study evaluated the effects of landscape features on population genetic diversity of Garra cambodgiensis, a stream cyprinid, in eight tributary streams in the upper Nan River drainage basin (n = 30-100 individuals/location), Nan Province, Thailand. These populations are under intense fishing pressure from local communities. Based on 11 microsatellite loci, we detected moderate genetic diversity within eight population samples (average number of alleles per locus = 10.99 ± 3.00; allelic richness = 10.12 ± 2.44). Allelic richness within samples and stream order of the sampling location were negatively correlated (P < 0.05). We did not detect recent bottleneck events in these populations, but we did detect genetic divergence among populations (Global FST = 0.022, P < 0.01). The Bayesian clustering algorithms (TESS and STRUCTURE) suggested that four to five genetic clusters roughly coincide with sub-basins: (1) headwater streams/main stem of the Nan River, (2) a middle tributary, (3) a southeastern tributary and (4) a southwestern tributary. We observed positive correlation between geographic distance and linearized FST (P < 0.05), and the genetic differentiation pattern can be moderately explained by the contemporary stream network (STREAMTREE analysis, R2 = 0.75). The MEMGENE analysis suggested genetic division between northern (genetic clusters 1 and 2) and southern (clusters 3 and 4) sub-basins. We observed a high degree of genetic admixture in each location, highlighting the importance of natural flooding patterns and possible genetic impacts of supplementary stocking. Insights obtained from this research advance our knowledge of the complexity of a tropical stream system, and guide current conservation and restoration efforts for this species in Thailand.


Effects of alendronate on the proliferation and osteogenic differentiation of MG-63 cells.
Previous studies of the direct actions of bisphosphonates on bone have mainly been limited to their effects on bone-resorbing osteoclasts and little is known about the direct effects of bisphosphonates on osteoblasts. Here we report the direct effects of alendronate on the proliferation and osteogenic differentiation of the MG-63 osteoblast-like cell line. Cell proliferation was determined with the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay, osteogenic differentiation was evaluated with an alkaline phosphatase bioassay and by analysis of gene expression by reverse transcription-polymerase chain reaction, and the extent of calcium deposition was measured using Alizarin Red S staining. Alendronate significantly increased cell numbers over control values, with the greatest effect at 10(-8) M. Alkaline phosphatase activity and gene expression of bone morphogenetic protein 2, type I collagen and osteocalcin were increased after alendronate treatment. Alendronate also stimulated calcium deposition. We conclude that alendronate, apart from inhibiting osteoclastic bone resorption, is also a promoter of osteoblast proliferation and maturation.


Fast operando spectroscopy tracking in situ generation of rich defects in silver nanocrystals for highly selective electrochemical CO2 reduction.
Electrochemical CO2 reduction (ECR) is highly attractive to curb global warming. The knowledge on the evolution of catalysts and identification of active sites during the reaction is important, but still limited. Here, we report an efficient catalyst (Ag-D) with suitable defect concentration operando formed during ECR within several minutes. Utilizing the powerful fast operando X-ray absorption spectroscopy, the evolving electronic and crystal structures are unraveled under ECR condition. The catalyst exhibits a ~100% faradaic efficiency and negligible performance degradation over a 120-hour test at a moderate overpotential of 0.7 V in an H-cell reactor and a current density of ~180 mA cm-2 at -1.0 V vs. reversible hydrogen electrode in a flow-cell reactor. Density functional theory calculations indicate that the adsorption of intermediate COOH could be enhanced and the free energy of the reaction pathways could be optimized by an appropriate defect concentration, rationalizing the experimental observation.


Prevalence and evolutionary analyses of human T-cell lymphotropic virus in Guangdong province, China: Transcontinental and Japanese subtype lineages dominate the prevalence.
To systematically characterize the prevalence and evolution of human T-cell lymphotropic virus (HTLV) infection among voluntary blood donors (BDs) in Guangdong province, China. A three-year survey for HTLV epidemiology among BDs was performed in Guangdong during 2016-2018. Anti-HTLV-1/2 was screened by ELISA and ECLIA, and subsequently confirmed by western blot (WB) and nucleic acid testing (NAT). The prevalence of HTLV in donors from different cities was calculated. The identified HTLV-positive cases were phylogenetically genotyped and analyzed in a Bayesian phylogenetic framework. Among 3,262,271 BDs, 59 were confirmed positive for HTLV-1 (1.81 per 100,000) and no HTLV-2 infection was found. The prevalence of HTLV-1 varied significantly among 21 cities in Guangdong province, China. The highest prevalence was found in donors from Shanwei (13.94 per 100,000), which is a coastal city in eastern Guangdong. Viral genomic sequences genotyped from 55 HTLV-1 carriers showed that 39 were transcontinental subtype and 16 were Japanese subtype. Specially, 13 out of 39 transcontinental subtype sequences were characterized with L55P mutation and 21 out of 55 sequences were characterized with L19F mutation in viral gp46 protein. The L55P mutation seemed be specific to eastern Asia since it only presented in the sequences from Japan, mainland China, and Taiwan. Phylogenetic analysis of gp46 gene shows that HTLV-1a may have been introduced to Guangdong through four different introduction events and formed major transmission clusters: clades I(13,602 years ago), II(16, 010 years ago), III(15,639 years ago) and IV(16,517 years ago). In general, Guangdong is considered to be a low-prevalence region for HTLV-1 infection, but the prevalence is significantly higher in Shanwei city. Transcontinental and Japanese subtype lineages dominate the prevalence in Guangdong. In terms of blood safety, HTLV antibody screening for first-time blood donors can effectively reduce the risk of HTLV transmission.


Diagnostic Value of Single-Photon Emission Tomography Stress Test in Patients With Suspected Coronary Artery Disease in Saudi Arabia.
Background Invasive coronary angiography (ICA) is the gold standard procedure for the diagnosis of coronary artery disease (CAD). ICA allows for clear visualization of the coronary arterial blood flow. Single-photon emission computed tomography (SPECT) is currently in widespread use to non-invasively evaluate patients known or suspected of coronary artery disease (CAD). This study aimed to examine the association between (SPECT) stress test and elective ICA in terms of diagnostic value in patients suspected of coronary artery disease at the King Faisal Cardiac Center (KFCC), Jeddah, Saudi Arabia. Methods This study is a retrospective diagnostic validation study using a consecutive sampling technique to select the study sample at KFCC. The study included all patients who presented with chest pain that were investigated with either exercise or pharmacologic myocardial perfusion SPECT study followed by elective ICA within six months from January 2015 to January 2020. Results A total of 207 patients met the inclusion criteria, where 43% (n = 90) of patients were females and 57% (n = 117) were males; 68% (n = 141) of the patients had both test results concordant (both SPECT and ICA results were in agreement). In 32% of the patients (n = 66), there was a discordant result (discrepant result between SPECT and ICA). SPECT had a sensitivity of 92.4% and a specificity of 26.3%. SPECT had a negative predictive value of 0.68 and a positive predictive value of 0.66 compared to ICA. There was a low degree of reliability between SPECT and ICA. Conclusion Reliability between the SPECT and ICA in exclusion of significant CAD is high. The rate of false-positive tests was high while the accuracy of SPECT in detecting CAD in patients with diabetes and hypertension was high. The overall reliability of SPECT to ICA in the Saudi population was low.


Chronic hypoxia upregulates pulmonary arterial ASIC1: a novel mechanism of enhanced store-operated Ca2+ entry and receptor-dependent vasoconstriction.
Acid-sensing ion channel 1 (ASIC1) is a newly characterized contributor to store-operated Ca(2+) entry (SOCE) in pulmonary vascular smooth muscle (VSM). Since SOCE is implicated in elevated basal VSM intracellular Ca(2+) concentration ([Ca(2+)](i)) and augmented vasoconstriction in chronic hypoxia (CH)-induced pulmonary hypertension, we hypothesized that ASIC1 contributes to these responses. To test this hypothesis, we examined effects of the specific pharmacologic ASIC1a inhibitor, psalmotoxin 1 (PcTX1), on vasoconstrictor and vessel wall [Ca(2+)](i) responses to UTP and KCl (depolarizing stimulus) in fura-2-loaded, pressurized small pulmonary arteries from control and CH (4 wk at 0.5 atm) Wistar rats. PcTX1 had no effect on basal vessel wall [Ca(2+)](i), but attenuated vasoconstriction and increases in vessel wall [Ca(2+)](i) to UTP in arteries from control and CH rats; normalizing responses between groups. In contrast, responses to the depolarizing stimulus, KCl, were unaffected by CH exposure or PcTX1. Upon examining potential Ca(2+) influx mechanisms, we found that PcTX1 prevented augmented SOCE following CH. Exposure to CH resulted in a significant increase in pulmonary arterial ASIC1 protein. This study supports a novel role of ASIC1 in elevated receptor-stimulated vasoconstriction following CH which is likely mediated through increased ASIC1 expression and SOCE.


Mast cells in the human testis and epididymis from birth to adulthood.
Mast cells are a constant cell-type in the connective tissues of the human testis and epididymis from birth to adulthood. Ultrastructural study shows that these cells are similar to those found in other connective tissues. Histometric studies revealed that the number of mast cells in the interstitium, mediastinum and albuginea of the testis as well as in the epididymal connective tissue increases slightly during infancy, decreases during childhood, and then increases again at puberty. Increases at puberty are particularly evident in both the testicular interstitium and the epididymis. During adulthood, the number of mast cells progressively decreases in all testicular and epididymal connective tissues. Changes in mast cell number may be related to changes observed in the development of testicular connective tissue which occurs primarily during infancy and puberty.


Targeted disruption of carcinoembryonic antigen-related cell adhesion molecule 1 promotes diet-induced hepatic steatosis and insulin resistance.
Carcinoembryonic antigen-related cell adhesion molecule 1 (CC1) is a cell adhesion molecule within the Ig superfamily. The Tyr-phosphorylated isoform of CC1 (CC1-L) plays an important metabolic role in the regulation of hepatic insulin clearance. In this report, we show that CC1-deficient (Cc1(-/-)) mice are prone to hepatic steatosis, as revealed by significantly elevated hepatic triglyceride and both total and esterified cholesterol levels compared with age-matched wild-type controls. Cc1(-/-) mice were also predisposed to lipid-induced hepatic steatosis and dysfunction as indicated by their greater susceptibility to store lipids and express elevated levels of enzymatic markers of liver damage after chronic feeding of a high-fat diet. Hepatic steatosis in the Cc1(-/-) mice was linked to a significant increase in the expression of key lipogenic (fatty acid synthase, acetyl CoA carboxylase) and cholesterol synthetic (3-hydroxy-3-methylglutaryl-coenzyme A reductase) enzymes under the control of sterol regulatory element binding proteins-1c and -2 transcription factors. Cc1(-/-) mice also exhibited impaired insulin clearance, glucose intolerance, liver insulin resistance, and elevated hepatic expression of the key gluconeogenic transcriptional activators peroxisome proliferator-activated receptor-gamma coactivator-1 and Forkhead box O1. Lack of CC1 also exacerbated both glucose intolerance and hepatic insulin resistance induced by high-fat feeding, but insulin clearance was not further deteriorated in the high-fat-fed Cc1(-/-) mice. In conclusion, our data indicate that CC1 is a key regulator of hepatic lipogenesis and that Cc1(-/-) mice are predisposed to liver steatosis, leading to hepatic insulin resistance and liver damage, particularly when chronically exposed to dietary fat.


Collective States in Molecular Monolayers on 2D Materials.
Collective excited states form in organic two-dimensional layers through Coulomb coupling of the molecular transition dipole moments. They manifest as characteristic strong and narrow peaks in the excitation and emission spectra that are shifted to lower energies compared with the monomer transition. We study experimentally and theoretically how robust the collective states are against homogeneous and inhomogeneous broadening, as well as spatial disorder that occurs in real molecular monolayers. Using a microscopic model for a two-dimensional dipole lattice in real space, we calculate the properties of collective states and their extinction spectra. We find that the collective states persist even for 1-10% random variation in the molecular position and in the transition frequency, with a peak position and integrated intensity similar to those for the perfectly ordered system. We measured the optical response of a monolayer of the perylene derivative MePTCDI on two-dimensional materials. On the wide-band-gap insulator hexagonal boron nitride, it shows strong emission from the collective state with a line width that is dominated by the inhomogeneous broadening of the molecular state. When the semimetal graphene is used as a substrate, however, the luminescence is completely quenched. By combining optical absorption, luminescence, and multiwavelength Raman scattering, we verify that the MePTCDI molecules form very similar collective monolayer states on hexagonal boron nitride and graphene substrates, but on graphene the line width is dominated by nonradiative excitation transfer from the molecules to the substrate. Our study highlights the transition from the localized molecular state of the monomer to a delocalized collective state in the two-dimensional molecular lattice that is entirely based on Coulomb coupling between optically active excitations of the electrons and molecular vibrations. The excellent properties of organic monolayers make them promising candidates for components of soft-matter optoelectronic devices.


Prevalence of hypothyroidism in infertile women and evaluation of response of treatment for hypothyroidism on infertility.
The aim was to study the prevalence of hypothyroidism in infertile women and evaluation of response of treatment for hypothyroidism on infertility. A total of 95 infertile women were investigated for thyroid stimulating hormone (TSH). Infertile women with clinical/subclinical hypothyroidism were given thyroxine ranges from 25 to 150 μg. Of 95 infertile women, 53.7% were hypothyroid (TSH > 4.6 μIU/ml). After the treatment with thyroxine, 33.3% of subclinical hypothyroid women conceived within 6 weeks to 2-year period. The mean time to conception was 14.56 ± 4.83 months. Thyroid profile should be done in infertility work up. Women with normal TSH levels who are positive for thyroid antibodies should also be treated with levothyroxine.


Experiences of Dutch families who parent an adopted Romanian child.
The pilot phase of a longitudinal study of 83 children (response 86%) adopted from Romania is reported; the present status of the children is compared with the past status on the basis of parental recollection. At placement, the average age of the children was 2.9 years; at the time of the interview of the adoptive parents, the average age of the children was 6.8 years. At placement, 67% of the adoptive parents needed professional help, and 10% of the children were infected with Hepatitis B. Only 13% did not show any important psychosocial problems. The number of psychosocial problems reported by the parents at placement predicts 69% of the lag of development observed 4 years later; the age of arrival adds up to 72%. Most problems still exist 4 years after placement; however, no disruptions of the adoption have taken place yet.


Determination of kinetic parameters of enantiomerization of benzothiadiazines by DCXplorer.
Benzothiadiazines differently substituted at the sulfonamidic nitrogen atom, at the stereogenic carbon atom and at the anilinic nitrogen atom have been synthesized and fully characterized. Enantioseparation of these compounds has revealed rapid on-column enantiomerization. The recently developed software DCXplorer has been successfully applied to calculate enantiomerization kinetic parameters. Enantiomerization barriers of 3-phenyl substituted benzothiadiazines, calculated in this work, have indicated a higher enantiomerization rate suggesting that the aromatic substituent exerts a strong effect on the enantiomerization process. Methyl substitution on N(2) position led to higher free energy barriers of enantiomerization, suggesting negative influence of methyl in the N(2) position on enantiomerization kinetics. However, methylation on N(4) position increases the enantiomerization rates significantly. The results obtained have been employed to postulate an enantiomerization mechanism for chiral benzothiadiazine type compounds.


Association of chronic obstructive pulmonary, coronary artery, or peripheral artery disease with abdominal aortic aneurysm rupture.
Chronic obstructive pulmonary disease (COPD), coronary artery disease (CAD), and peripheral artery disease (PAD) are positively associated with abdominal aortic aneurysm (AAA) presence. It remains unclear, however, whether these 3 comorbidities are associated with AAA rupture. To assess the association of COPD, CAD, or PAD with AAA rupture, we reviewed currently available studies with a systematic literature search and meta-analytic estimates. Databases including MEDLINE and EMBASE were searched through December 2015 using PubMed and OVID. For each study, data regarding prevalence of COPD, CAD, or PAD in both the ruptured and non-ruptured groups were used to generate odds ratios (ORs) for rupture and 95% confidence intervals (CIs). Study-specific estimates were combined using inverse variance-weighted averages of logarithmic ORs in a random-effects model. Our search identified 9 relevant studies including data on a total of 8873 AAA patients (rupture, 1241; non-rupture, 7632). Pooled analyses demonstrated that COPD was significantly and positively (OR, 1.51; 95% CI, 1.06 to 2.16; P=0.02), CAD was not significantly (OR, 0.83; 95% CI, 0.43 to 1.60; P=0.58), and PAD tended to be negatively (though non-significantly) associated with AAA rupture (OR, 0.44; 95% CI, 0.16 to 1.23; P=0.12). Our analysis suggests that COPD is positively associated with AAA rupture, CAD is not associated with it, and PAD tends to be negatively associated with it. Further investigations would be required to understand precise mechanisms regarding the association of COPD, CAD, and PAD with AAA presence, growth, and rupture.


Insights into the membrane interactions of the saposin-like proteins Na-SLP-1 and Ac-SLP-1 from human and dog hookworm.
Saposin-like proteins (SAPLIPs) from soil-transmitted helminths play pivotal roles in host-pathogen interactions and have a high potential as targets for vaccination against parasitic diseases. We have identified two non-orthologous SAPLIPs from human and dog hookworm, Na-SLP-1 and Ac-SLP-1, and solved their three-dimensional crystal structures. Both proteins share the property of membrane binding as monitored by liposome co-pelleting assays and monolayer adsorption. Neither SAPLIP displayed any significant haemolytic or bactericidal activity. Based on the structural information, as well as the results from monolayer adsorption, we propose models of membrane interactions for both SAPLIPs. Initial membrane contact of the monomeric Na-SLP-1 is most likely by electrostatic interactions between the membrane surface and a prominent basic surface patch. In case of the dimeric Ac-SLP-1, membrane interactions are most likely initiated by a unique tryptophan residue that has previously been implicated in membrane interactions in other SAPLIPs.


Transperitoneal laparoscopic simple nephrectomy for giant hydronephrosis: Tips and tricks to make it easier.
We report our experience with transperitoneal laparoscopic nephrectomy (LN) for giant hydronephrosis (GH) and compare the outcome data with open nephrectomy (ON). The retrospective data of 88 patients (52 males and 36 females) who underwent LN or ON for treatment of GH in the period between October 2015 and December 2019 were investigated. LN was performed in 38 patients, while 50 patients underwent ON. We compared the two groups for success, operative time, and intraoperative and postoperative complications. The mean age of the patients in the LN group was 45.8 ± 11.4 years, and it was 44.7 ± 10.8 years in the ON group. The mean operative time in the LN group was statistically significantly longer when compared with the ON group195 ± 18 min versus 127 ± 22 min (p = 0.01).The estimated blood loss was significantly greater in the ON group (p = 0.01). However, no patients required blood transfusions in either group. The visual analog pain (VAP) scores were significantly higher on both day 1 and day 2 in the ON group 3.6 ± 0.9 and 2 ± 0.7 versus 2.7 ± 0.6 and 1.4 ± 0.5 in LN group, (p = 0.01). LN for GH is feasible, safe, and efficacious. Compared to open surgery, the laparoscopic approach resulted in significantly shorter hospital stays, decreased morbidity, and quicker recovery. Some tips and tricks could help to do it in an easier way and reduce the operative time.


Chemosensitization of carmustine with maitake beta-glucan on androgen-independent prostatic cancer cells: involvement of glyoxalase I.
To improve the poor efficacy (< 10%) of chemotherapy for patients with hormone-refractory prostate cancer, we investigated a possible cytotoxic effect of carmustine/beta-glucan combination on prostatic cancer PC-3 cells, focusing on a glutathione-dependent detoxifying enzyme, glyoxalase I (Gly-I). Carmustine (BCNU) is an anticancer agent and a putative inhibitor of Gly-I, while beta-glucan is a unique, nontoxic polysaccharide extracted from maitake mushrooms. The cytotoxic effects of BCNU or other anticancer agents with beta-glucan on PC-3 cells were assessed by cell-viability testing and Gly-I activity was measured using the spectrophotometric method. BCNU, 5-fluorouracil (5-FU), and methotrexate (MTX) were capable of inducing approximately a 50% reduction in cell viability at 72 hours, while etoposide, cisplatin, and mitomycin C were all ineffective. Only the combination of BCNU (50 micro ;mol) and beta-glucan (60 micro g/mL) exhibited an enhanced cytotoxicity with an approximate 90% cell viability reduction, but little improvement was seen with any combinations of 5-FU, MTX, or beta-glucon. Gly-I assays revealed that such a profound (approximately 90%) cell death was accompanied by an approximate 80% reduction in Gly-I activity by 6 hours. This study demonstrates a sensitized cytotoxic effect of BCNU with beta-glucan in PC-3 cells, which was associated with a drastic (approximately 80%) inactivation of Gly-I. Therefore, the BCNU/beta-glucan combination may help to improve current treatment efficacy by targeting Gly-I, which appears to be critically involved in prostate cancer viability.


Gender differences in autoantibodies to oxidative DNA base damage in cigarette smokers.
Oxidative DNA damage and antibodies to that damage have been implicated in lung, breast, and colorectal cancer. In this observational validation study, the relationship between anti-5-hydroxymethyl-2'-deoxyuridine (HMdU) autoantibody (aAb) and plasma micronutrients was assessed in 140 heavy smokers by ELISA. Anti-HMdU aAbs were 50% higher in women after adjustment for cigarettes/day (CPD; P = 0.002), although men smoked more and had higher plasma cotinine levels. The women reported taking more vitamin C (P < 0.005) and had higher plasma levels of alpha-carotene and beta-carotene (P < 0.001) and cryptoxanthin (P < 0.01) than men. Neither CPD nor cotinine was associated with aAb titers. Anti-HMdU aAbs were associated inversely with alpha-tocopherol (P = 0.10), retinol (P = 0.06), and age (P = 0.04) in women but not in men. In contrast to the men, women <or=50 years of age had significantly higher aAbs than those >50 years of age (P = 0.05). Given the same duration of exposure, women had higher anti-HMdU aAbs and also reached peak levels at a lower cumulative smoking exposure (30 years) compared with male smokers (40 years). Subjects smoked an average of 28.9 +/- 0.81 CPD and initiated smoking at 17.2 +/- 0.33 (SE) years of age. Therefore, smokers who reported smoking for 30 years were typically <50 years old. Women <or=50 years in the younger age group that smoked 21-30 years had significantly higher levels of aAbs than did men of the same age and smoking history (P = 0.012). Gender difference in aAbs was also evident in 29 persons who gave serial samples before and after quitting smoking (P < 0.028). In women, aAbs remained elevated for 14 months after smoking cessation but decreased significantly by 20.5 months (P < 0.032) by paired t tests. In men, aAbs increased with time since quitting smoking but not significantly. The finding of significantly elevated aAbs to oxidized DNA in females <or=50 compared with male smokers of the same age and exposure suggests a possible interaction with hormones (e.g., estrogens) and may explain a heightened risk of smoking-induced lung cancer in women compared with men.


Analysis of the Consumption of Caffeinated Energy Drinks among Polish Adolescents.
Energy drinks (EDs) are extremely popular among adults and adolescents. Regular intake of EDs may lead to an overdose of caffeine, loss of bone mass, overweight, hypertension and, in older age, osteoporosis and cardiovascular diseases. Some people mix EDs with alcohol, which adversely affects their health. The objective of this study was to analyze the consumption of EDs by adolescents. The study consisted of a questionnaire surveying amounts of drinks, preferences and product awareness among younger consumers. The study was carried out in junior and senior high schools in Poland (n = 2629). EDs were consumed by 67% of students (quite frequently by 16%). Students who practiced sports were more willing to drink EDs. Also, boys drank them more often than girls. When selecting a particular ED, young people looked at the taste, price and effect. Most respondents consumed one ED (250 mL) daily, although there were individuals consuming two or more drinks daily. Most respondents knew the ingredients of EDs, and 24% admitted to mixing EDs with alcohol. EDs are extremely popular among adolescents. Young people drinking EDs every day are potentially at risk of taking an overdose of caffeine.


Development and intracellular distribution of enzymes of the oxidative pentose phosphate cycle in radish cotyledons.
Developmental and compartmentation studies were used to evaluate the relative roles of the oxidative pentose phosphate cycle, the Calvin cycle, and the glycolysis in cotyledons of radish (Raphanus sativus L.).Glucose-6-P dehydrogenase, 6-P-gluconate dehydrogenase, glucose-6-P isomerase, and the NAD-dependent glyceraldehyde-3-P dehydrogenase were present in high activity in ungerminated seeds, increased about 2-fold during germination in the dark, and were slightly enhanced by light. In contrast, NADP-dependent glyceraldehyde-3-P dehydrogenase was developed to only a small degree in the dark, but increased severalfold in continuous white or far red light. The activity of phosphofructokinase was low throughout germination.The separation of cell compartment-specific isoenzymes showed that, except in ungerminated seeds, the plastid enzyme accounted for 40 to 45% of the total activity of 6-P-gluconate dehydrogenase and for 15 to 20% of glucose-6-P isomerase. The remaining activity was due to the cytosolic isoenzymes. The presence of glucose-6-P dehydrogenase and 6-P-gluconate dehydrogenase in plastids was also established by their presence in the isolated organelle. The NAD-dependent glyceraldehyde-3-P dehydrogenase was mostly due to the cytosolic isoenzyme, whereas the NAD-dependent activity associated with the NADP-dependent glyceraldehyde-3-P dehydrogenase was very small.The data indicate that the enzymes of the oxidative pentose phosphate cycle are present in the cytosol throughout germination. In the plastids these enzymes already became fully developed during early germination in the dark, whereas enzymes of the Calvin cycle increased only in the light. Glycolysis seemed to be of minor importance.


Histogram-based models on non-thin section chest CT predict invasiveness of primary lung adenocarcinoma subsolid nodules.
109 pathologically proven subsolid nodules (SSN) were segmented by 2 readers on non-thin section chest CT with a lung nodule analysis software followed by extraction of CT attenuation histogram and geometric features. Functional data analysis of histograms provided data driven features (FPC1,2,3) used in further model building. Nodules were classified as pre-invasive (P1, atypical adenomatous hyperplasia and adenocarcinoma in situ), minimally invasive (P2) and invasive adenocarcinomas (P3). P1 and P2 were grouped together (T1) versus P3 (T2). Various combinations of features were compared in predictive models for binary nodule classification (T1/T2), using multiple logistic regression and non-linear classifiers. Area under ROC curve (AUC) was used as diagnostic performance criteria. Inter-reader variability was assessed using Cohen's Kappa and intra-class coefficient (ICC). Three models predicting invasiveness of SSN were selected based on AUC. First model included 87.5 percentile of CT lesion attenuation (Q.875), interquartile range (IQR), volume and maximum/minimum diameter ratio (AUC:0.89, 95%CI:[0.75 1]). Second model included FPC1, volume and diameter ratio (AUC:0.91, 95%CI:[0.77 1]). Third model included FPC1, FPC2 and volume (AUC:0.89, 95%CI:[0.73 1]). Inter-reader variability was excellent (Kappa:0.95, ICC:0.98). Parsimonious models using histogram and geometric features differentiated invasive from minimally invasive/pre-invasive SSN with good predictive performance in non-thin section CT.


Inhalation of ozone produces a decrease in superoxide anion radical production in mouse alveolar macrophages.
The potentiation of fatal bacterial pneumonia in mice by prior inhalation of ozone occurs at levels of this oxidant pollutant that are frequently present in ambient air. A likely mechanism for this effect is an ozone-induced inhibition in the ability of pulmonary alveolar macrophages (PAM) to produce superoxide anion radical (O2-) demonstrated in the present study. A 25% decrease in PAM O2- production, as measured by nitroblue tetrazolium reduction, occurred after exposure of Swiss-Webster mice to 0.11 ppm ozone for 3 h (p less than 0.05). After 1 ppm there was almost complete inhibition of O2- release. In contrast, the rat, which is highly resistant to the potentiation of bacterial infections by ozone, was less sensitive to inhibition of PAM O2- production, as measured by cytochrome c reduction (mouse IC50, 0.41 ppm; rat IC50, 3.0 ppm ozone for 3 h). The observed decrement in mouse PAM O2- production was not associated with any change in phagocytic ability, as measured by both latex bead ingestion and 51Cr-labeled sheep red blood cell ingestion. This decrease in O2- production in the presence of normal phagocytic activity is analogous to certain of the findings in the neutrophils of children with chronic granulomatous disease. A decrease in rat PAM membrane cytochrome b558 levels was observed after ozone exposure of 3 ppm for 3 h, preliminarily suggesting that the mechanism by which ozone interferes with PAM O2- production may be through interaction with this heme-containing electron carrier.


CH···π interactions do not contribute to hydrogen transfer catalysis by glycerol dehydratase.
The role of the nonbonded CH···π interaction in the hydrogen abstraction from glycerol by the coenzyme B(12)-independent glycerol dehydratase (GDH) was examined using the QM/MM (ONIOM), MP2, and CCSD(T) methods. The studied CH···π interaction included the hydrogen atom of the -C(2)H(OH)- group of the glycerol substrate and the tyrosine-339 residue of the dehydratase. A contribution of this interaction to the stabilization of the transition state for the transfer of a hydrogen atom from the adjacent terminal C(1)H(2)(OH) group to cysteine 433 was determined by ab initio HF, MP2, and CCSD(T) calculations with the aug-cc-pvDZ basis set for the corresponding methane/benzene, methanol/phenol, and glycerol radical/phenol subsystems. The calculated CH···π distance, defined as the distance between the H atom and the center of the phenol ring, shortened from 2.62 to 2.52 Å upon going from the ground- to the transition-state of the GDH-catalyzed reaction. However, this shortening was not accompanied by the expected lowering of the CH···π interaction free energy. Instead, this interaction remained weak (about -1 kcal/mol) along the entire reaction coordinate. Additionally, the mutual orientation of the CH group and the phenol ring did not change significantly during the reaction. These results suggest that the phenol group of the tyrosine-339 does not contribute to lowering the activation barrier in the enzyme, but do not exclude the possibility that tyrosine 339 facilitates proper orientation of glycerol for the electrostatic catalysis, or inhibits side-reactions of the reactive glycerol radical intermediate.


Redox state of the endoplasmic reticulum is controlled by Ero1L-alpha and intraluminal calcium.
Formation of intra- and intermolecular disulfide bonds is an essential step in the synthesis of secretory proteins. In eukaryotic cells, this process occurs in the endoplasmic reticulum (ER) and requires an oxidative environment with the action of several chaperones and folding catalysts. During protein folding, Ero1p oxidizes protein disulfide isomerase (PDI), which then directly catalyzes the formation of disulfide bonds in folding proteins. Recent cell-free studies suggest that the terminal electron acceptor in the pathway is molecular oxygen, with the resulting formation of hydrogen peroxide (H(2)O(2)). We report for the first time the measurement of ER H(2)O(2) level in live cells. By targeting a fluorescent protein-based H(2)O(2) sensor to various intracellular compartments, we show that the ER has the highest level of H(2)O(2), and this high concentration is well confined to the lumen of the organelle. Manipulation of the Ero1-Lalpha level--either by overexpression or by siRNA-mediated inhibition--caused parallel changes in luminal H(2)O(2), proving that the activity of Ero1-Lalpha results in H(2)O(2) formation in the ER. We also found that calcium mobilization from intracellular stores induces a decrease in ER H(2)O(2) level, suggesting a complex interplay between redox and calcium signaling in the mammalian ER.


Evaluation of emerging technologies for coronary revascularization. Participants in the National Heart, Lung, and Blood Institute Conference on the Evaluation of Emerging Coronary Revascularization Technologies.
Despite significant advances in coronary angioplasty, the problems of restenosis, chronic total occlusion, diffuse disease, and abrupt closure after the procedure remain to be solved. New devices that address these problems continue to evolve. Although a controlled clinical trial is the ultimate test of any device, rapid device design changes and resource limitations require evaluating which of the new devices justify a full clinical trial. A registry mechanism for this initial evaluation is proposed and detailed. Payment for procedures using new devices is a problem for industry, insurers, hospitals, and physicians. It is proposed that reimbursement be based on the service performed rather than the device used to perform it. Costs beyond the clinical costs should be borne by the sponsor of the device. Financial conflict of interest is a recognized problem in new device evaluation. It is not always practical to insist on exclusion from the evaluation process of individuals with a financial stake in the outcome, but disclosure of the presence of such a financial interest is recommended in the reporting of results.


Global Regulatory Pathways Converge To Control Expression of Pseudomonas aeruginosa Type IV Pili.
The opportunistic pathogen Pseudomonas aeruginosa relies upon type IV pili (Tfp) for host colonization and virulence. Tfp are retractile surface appendages that promote adherence to host tissue and mediate twitching motility, a form of surface-associated translocation. Tfp are composed of a major structural pilin protein (PilA), several less abundant, fiber-associated pilin-like proteins (FimU, PilV, PilW, PilX, and PilE), and a pilus-associated tip adhesin and surface sensor (PilY1). Several proteins critical for Tfp biogenesis and surface sensing are encoded by the fimU-pilVWXY1Y2E operon. Tfp biogenesis is regulated by the global transcription factor Vfr and its allosteric effector, cyclic AMP (cAMP). Our investigation into the basis for reduced Tfp production in cAMP/vfr mutants revealed a defect in the expression of the fimU operon. We found that cAMP/Vfr activation of the fimU operon occurs via direct binding of Vfr to a specific fimU promoter sequence. We also refined the role of the AlgZ/AlgR two-component system in fimU regulation by demonstrating that phosphorylation of the response regulator AlgR is required for maximal binding to the fimU promoter region in vitro. Vfr also regulates expression of the algZR operon, revealing an indirect regulatory loop affecting fimU operon transcription. Overall, these results demonstrate that two linked but independent regulatory systems couple the expression of Tfp biogenesis and surface sensing genes and highlight the regulatory complexity governing expression of P. aeruginosa virulence factors. IMPORTANCE Pseudomonas aeruginosa is an opportunistic pathogen responsible for a wide range of infections. An extensive repertoire of virulence factors aid in P. aeruginosa pathogenesis. Type IV pili (Tfp) play a critical role in host colonization and infection by promoting adherence to host tissue, facilitating twitching motility and mediating surface-associated behaviors. The fimU operon encodes several pilus-associated proteins that are essential for proper Tfp function and surface sensing. In this study, we report that linked but independent regulatory systems dictate Tfp biogenesis. We also demonstrated the importance of different phosphorylation states of the AlgZ/AlgR two-component system and its role in Tfp biogenesis. Overall, this study furthers our understanding of the complex regulatory mechanisms that govern the production of a critical and multifaceted virulence factor.


Sexual health of women aged 40 and over attending an inner city integrated sexual health clinic.
Sexual health policy remains focussed on younger adults. However, rates of sexually transmitted infections (STIs) in older people continue to increase. We explored the sexual healthcare needs of women aged 40 and over attending an integrated sexual health clinic in South London. We conducted a retrospective case note review and found that almost 20% of these women had STIs. These included genital herpes, trichomoniasis, genital warts, chlamydia and gonorrhoea. Less than a quarter of women reported use of condoms during most recent sexual contact, indicating sexual risk-taking behaviour. 38% of women attended for contraception. The sexual health needs of older people can only continue to increase, given our rapidly ageing population. Age-specific health promotion strategies are needed.


Design and Validation of a Torque Dense, Highly Backdrivable Powered Knee-Ankle Orthosis.
This paper presents the mechatronic design and experimental validation of a novel powered knee-ankle orthosis for testing torque-driven rehabilitation control strategies. The modular actuator of the orthosis is designed with a torque dense motor and a custom low-ratio transmission (24:1) to provide mechanical transparency to the user, allowing them to actively contribute to their joint kinematics during gait training. The 4.88 kg orthosis utilizes frameless components and light materials, such as aluminum alloy and carbon fiber, to reduce its mass. A human subject experiment demonstrates accurate torque control with high output torque during stance and low backdrive torque during swing at fast walking speeds. This work shows that backdrivability, precise torque control, high torque output, and light weight can be achieved in a powered orthosis without the high cost and complexity of variable transmissions, clutches, and/or series elastic components.


Posterior vertebral column resection in fixed lumbosacral deformity.
Retrospective study. To report the results and the technique of posterior vertebral column resection (PVCR) in fixed lumbosacral deformity. Fixed lumbosacral deformity has gross imbalance as well as progressive compensatory thoracolumbar deformity because of the absence of a mobile spine caudally. There is no consensus of the surgical treatment of fixed lumbosacral deformity. Fixed lumbosacral deformity can be managed by PVCR, which not only corrects the deformity but also restores the trunk balance by translation of spinal column. Twenty-five consecutive fixed lumbosacral deformity patients subjected to PVCR were reviewed after a minimum follow-up of 2 years. The apical vertebra and the ultimately resected vertebra were below L4 in most cases. Etiologic diagnoses were congenital scoliosis in 6, congenital kyphoscoliosis in 3, post-traumatic kyphosis in 2, and postinfectious kyphosis in 14. The average age at the operation was 38 years (range, 2.5-61 years) with a male:female ratio of 7:18. The indication for PVCR was fixed lumbosacral deformities that could not be brought to a reasonable balance on traction or forced side bending. Before surgery, 12 patients (48%) presented with neurologic compromise and 20 patients (80%) presented with intractable back pain. The surgical technique consisted of segmental pedicle screw fixation, decompression of neural structures, and resection of the vertebral column at the apex of the deformity via the posterior route, followed by deformity correction and global fusion. During the resection, all the nerve roots were preserved. Two and one tenth (2.1) vertebrae removed on average (range, 1-5 vertebrae), and 52 in all. The average fusion extent was 4.5 vertebrae (range, 2-8 vertebrae). The anterior column reconstruction was carried out with autogenous bone graft in all patients and additional insertion of titanium mesh in 12. The distal anchor went down to L5 in 4 patients, S1 in 12, and S2 in 9. Preoperative scoliosis of 38 degrees +/- 12 degrees was corrected to 15 degrees +/- 8 degrees (60% correction) at most recent follow-up, and preoperative kyphosis of 35 degrees +/- 25 degrees was corrected to -5 degrees +/- 11 degrees (40 degrees correction). The compensatory coronal curve of 26 degrees was spontaneously corrected to 12 degrees at most recent follow-up. The preoperative thoracic lordosis of -16 degrees was corrected to 4 degrees at most recent follow-up, and the preoperative lumbar kyphosis of 20 degrees was corrected to -17 degrees . Preoperative coronal imbalance of 2.0 cm was improved to 0.9 cm at most recent follow-up, and preoperative sagittal imbalance of 9.3 cm was improved to 4.6 cm. Mean operation time was 280 minutes with a blood loss of 2,810 mL. The preoperative neurologic compromise was improved in all patients, and pain was reduced to less than half of preoperative visual analog analog scale in 17 patients. The following complications were encountered in 5 patients: 2 with transient neurology that spontaneously improved without a sequel within 6 months, 2 with compression fractures at proximal adjacent vertebra, and 1 with pseudarthrosis. PVCR is an effective procedure for the management of fixed lumbosacral deformity. It provides satisfactory correction and improved functional outcomes. However, it is a technically demanding and exhausting procedure with possible risks for complications.


Bayesian spatio-temporal analysis of joint patterns of male and female lung cancer risks in Yorkshire (UK).
Recent advances in disease mapping have focused first on including the time dimension, thus giving rise to spatio-temporal analysis of the variation of disease risk and, secondly, on carrying out joint analysis of two diseases that share common environmental risk factors and are, therefore, related. Here, we try to combine both issues and present a joint analysis of the spatio-temporal variation of the risks of two related diseases processes-male and female lung cancer incidence-in a region of England. To do so, we use a Bayesian hierarchical model that splits the risk of disease into two spatio-temporal components: a shared component and a specific component that calibrates the differential between the two diseases.


[In vitro susceptibilities of Mycobacterium avium and Mycobacterium intracellulare to various drugs].
Mycobacterium avium and M. intracellulare isolated from patients infected with M. avium complex (MAC), which were identified by Gen-Probe Rapid Diagnostic System for the MAC, were studied for susceptibility to various antimicrobial agents, including rifampicin, rifabutin, kanamycin, streptomycin, amikacin, ethambutol, clofazimine, isoniazid, ofloxacin, ciprofloxacin and cycloserine. Ratio of resistant strains to test strains to a given agent at prescribed concentration in cases of M. avium and M. intracellulare was compared with each other. Test strains of M. avium were more resistant to rifampicin, rifabutin, kanamycin, streptomycin, amikacin, ethambutol and clofazimine than test strains of M. intracellulare. Conversely, the M. avium strains were more susceptible to ofloxacin, ciprofloxacin and cycloserine than M. intracellulare strains. The difference in the drug susceptibility between M. avium and M. intracellulare was statistically significant by chi 2-test (P less than 0.005-0.05). There was no statistically significant difference between the two MAC species with respect to the susceptibility to isoniazid.


Novel ferromagnetic materials of SmCo5 nanoparticles in single-nanometer size: chemical syntheses and characterizations.
We first succeeded in preparing ferromagnetic materials of SmCo(5) nanoparticles in sizes of single nanometers by a chemical route using a polyol process. In order to produce ferromagnetic SmCo(5) nanoparticles with a CaCu(5)-type structure, the chemical preparation was conducted under a molar ratio of 1:1.3 of Sm:Co as compositions of metallic precursors when tetraethylene glycol (TEG), and oleic acid and oleylamine were used as a reductant and protectant, respectively. However, XRD profiles of the products showed the presence of oxides and other by-products besides the CaCu(5)-type structured nanoparticles. The best metallic precursors and protective reagent to prevent oxidization under reaction were SmCl(3) and Co(acac)(3), and 1-adamantanecarboxylic acid (ACA) and poly(N-vinyl-2-pyrrolidone) (PVP), respectively when TEG was used as the reducing reagent. We found that oxides and other by-products were not detected and pure SmCo(5) nanoparticles with the CaCu(5)-type structure were prepared under the chemical condition. The SmCo(5) nanoparticles coated by ACA and PVP were 6.2 nm in diameter and showed a coercivity of 1500 Oe at 300 K.