Structures of and allelic diversity and relationships among the major outer membrane protein (ompA) genes of the four chlamydial species.
DNA sequences coding for 81% of the ompA gene from 24 chlamydial strains, representing all chlamydial species, were determined from DNA amplified by polymerase chain reactions. Chlamydial strains of serovars and strains with similar chromosomal restriction fragment length polymorphism had identical ompA DNA sequences. The ompA sequences were segregated into 23 different ompA alleles and aligned with each other, and phylogenetic relationships among them were inferred by neighbor-joining and maximum parsimony analyses. The neighbor-joining method produced a single phylogram which was rooted at the branch between two major clusters. One cluster included all Chlamydia trachomatis ompA alleles (trachoma group). The second cluster was composed of three major groups of ompA alleles: psittacosis group (alleles MN, 6BC, A22/M, B577, LW508, FEPN, and GPIC), pneumonia group (Chlamydia pneumoniae AR388 with the allele KOALA), and polyarthritis group (ruminant and porcine chlamydial alleles LW613, 66P130, L71, and 1710S with propensity for polyarthritis). These groups were distinguished through specific DNA sequence signatures. Maximum parsimony analysis yielded two equally most parsimonious phylograms with topologies similar to the ompA tree of neighbor joining. Two phylograms constructed from chlamydial genomic DNA distances had topologies identical to that of the ompA phylogram with respect to branching of the chlamydial species. Human serovars of C. trachomatis with essentially identical genomes represented a single taxonomic unit, while they were divergent in the ompA tree. Consistent with the ompA phylogeny, the porcine isolate S45, previously considered to be Chlamydia psittaci, was identified as C. trachomatis through biochemical characteristics. These data demonstrate that chlamydial ompA allelic relationships, except for human serovars of C. trachomatis, are cognate with chromosomal phylogenies.


Indirect involvement of delta opioid receptors in cholecystokinin octapeptide-induced analgesia in mice.
Sulfated cholecystokinin octapeptide (CCK-8; Asp-Tyr-SO3H-Met-Gly-Trp-Met-Asp-Phe-NH2) produced analgesia in mice when administered i.c.v. and tested in the acetic acid-induced writhing assay. The ED50 was found to be 0.03 nmol/mouse which was about 3, 24 and 714 times more potent than morphine. [D-Pen2,D-Pen5]enkephalin and U50,488H [trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1-pyrolidinyl)cyclohexyl] benzeneacetamidel], respectively. When administered i.t., CCK-8 produced partial analgesia of up to 22 to 23% at low doses ranging from 15 to 60 ng/mouse and hyperalgesia at doses over 120 ng/mouse. Naloxone, an opioid antagonist, inhibited the analgesia induced by CCK-8 (i.c.v. and i.t.) but potentiated hyperalgesia induced by CCK-8 (i.t.). Apparent pA2 value for CCK-8 (i.c.v.) against naloxone (s.c.) was 5.88 which was significantly different from those for morphine-naloxone and U50,488H-naloxone but was not significantly different from that for [D-Pen2,D-Pen5]enkephalin-naloxone. Studies using highly selective opioid antagonists showed that CCK-8-induced analgesia was significantly antagonized by the delta receptor antagonist, ICI154,129 [(Allyl)2-Tyr-gly-gly-psi-(CH2S)-Phe-Leu] but not by beta-funaltrexamine, a highly selective mu receptor antagonist or nor-binaltorphimine, a highly selective kappa receptor antagonist. Opioid receptor binding study using [3H]-[D-Ala2,D-Leu5]enkephalin (+unlabeled [D-Ala2,MePhe4,Gly-ol5]enkephalin) in mouse brain membrane preparations revealed that there were no changes in the maximum binding or Kd of delta opioid binding sites in the presence of CCK-8 (1 microM) in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)


Motor cortex stimulation for neuropathic pain.
Motor cortex stimulation is increasingly reported in the literature as a surgical option for the alleviation of neuropathic pain. The authors review the published literature and present their results including those demonstrated in a randomized controlled trial that confirmed the efficacy of the procedure. Patient selection and prediction of outcomes, however, remain difficult issues.


Positioning of a contextual implant along with a sinus lift anchored with a block of heterologous bone.
During a sinus lift procedure the main requirement in order to position an implant is to have a maxillary sinus floor cortical bone thick enough to guarantee a primary stability in the implant inserted. In this way, the healing process is facilitated and osseointegration of the titanium surface may occur simultaneously, thus reducing the waiting time for the engraftment of the implant into the body. Unfortunately, these conditions are not always present. Hence, the need of developing an alternative approach that could simultaneously allow to perform sinus floor elevation along with an implant placement. Here we present the case of a 62-year-old patient that requires implant-prosthetic rehabilitation from 1.2 to 1.6 at diagnosis. In this study, we reported a novel application derived from the use of a heterologous bone scaffold (SmartBone@) in a sinus lift procedure. We showed the successful implant along with sinus lift with SmartBone@, both at the time of the surgery and after follow-up of the patient at 10 months from the implant. The possibility to perform simultaneously the contextual implant along with sinus lift dramatically reduced the waiting time for the patient of minimum 5-6 months required for osseointegration of the grafted biomaterials, before performing the implant procedure. This surgery represents an advance both in terms of medical technique and as life-benefit for the patient.


Analysis of CYP2D6 genotype and response to tetrabenazine.
Tetrabenazine is effective in the treatment of the chorea associated with Huntington disease and other hyperkinetic movement disorders. Following oral administration, tetrabenazine is hepatically transformed into 2 active metabolites that are CYP2D6 substrates. There are 4 CYP2D6 genotypes: poor metabolizers, intermediate metabolizers, extensive metabolizers, and ultrarapid metabolizers. CYP2D6 genotyping was performed on sequential subjects treated with tetrabenazine, but results were not known at the time of titration. Duration of titration to a stable dose, total daily dose, response rating scores, and adverse events were retrospectively collected and subsequently analyzed. Of 127 patients, the majority (n = 100) were categorized as extensive metabolizers, 14 as intermediate metabolizers, 11 as poor metabolizers, and 2 as ultrarapid metabolizers. Ultrarapid metabolizer patients needed a longer titration (8 vs 3.3, 4.4, and 3 weeks, respectively; P < .01) to achieve optimal benefit and required a higher average daily dose than the other patients, but this difference did not reach statistical significance. The treatment response was less robust in the intermediate metabolizer group when compared with the extensive metabolizer patients (P = .013), but there were no statistically significant differences between the various groups with regard to adverse effects. Our findings demonstrate that, aside from the need for a longer titration in the ultrarapid metabolizers, there are no distinguishing features of patients with various CYP2D6 genotypes, and therefore the current recommendation to systematically genotype all patients prescribed more than 50 mg/day of tetrabenazine should be reconsidered.


[The trend of childhood bacterial meningitis in Japan (1997.7-2000.6)].
We surveyed the epidemiology of purulent meningitis in pediatrics for 3 years between July 1997 and June 2000 in Japan and obtained the following results. The number of cases of purulent meningitis was 428, which was equivalent of 1.1-1.7 children out of 1,000 hospitalized those in pediatrics per year. The age-distribution for the infections was the highest under 1 year of age and it decreased as the age increased. Under 1 year of age, the highest distribution was observed in one month of age and under 1 month of age, the highest distribution was observed in 7 days of or younger ages. Haemophilus influenzae was the most common pathogen causing the infections, followed by Streptococcus pneumoniae, group B streptococcus, and Escherichia coli. Relationship between causing pathogens and age-distribution was as follows: group B streptococcus and E. coli were major pathogens under 4 months of age and H. influenzae and S. pneumoniae were major pathogens over 3 months of age. Susceptibility tests performed at each facility demonstrated that 25.3% of H. influenzae isolates and 38.7% of S. pneumoniae isolates were drug-resistant. Analysis of resistant genes for H. influenzae and S. pneumoniae isolates, which were stored and sent, demonstrated higher rates of resistance than those observed in susceptibility tests. These results suggest that the increase in insufficient efficacy of usual treatment with combination of ampicillin and cefotaxime is predictable against the infections. Therefore, the treatment for the infections should be reconsidered.


Correlates of work-related stress among consultants and senior registrars in accident and emergency medicine.
A mail survey was conducted of consultants and senior registrars practising accident and emergency (A&E) medicine in the United Kingdom. The 201 respondents (72%) comprised 154 consultants (70.6%) and 47 senior registrars (77%), who provided demographic information and completed inventories measuring stress, depression, task and role clarity, work group functioning and overall satisfaction with work. The respondents did not report particularly high levels of stress or depression and generally evaluated aspects of their work environments favourably. Higher levels of stress were reported by consultants and respondents from district general hospitals. Levels of stress were similar to those reported by other groups of health care providers. Respondents generally considered tasks and roles to be clearly defined, work groups to be supportive, efficient units and work satisfying. There was no statistically significant correlation on the affective scales for the number of patient attendances, on call commitment or staffing numbers. Senior staff with more than 10 years experience in the specialty reported more satisfaction with work and work group functioning, and perceived their tasks and roles to be significantly clearer. Consultants over 45 evaluated their work groups favourably and were more likely to view them as cohesive, smoothly functioning units than senior registrars. The results probably reflect the ad hoc coping strategies adopted by a group of doctors, who have already demonstrated appropriate personality characteristics by completing a long training programme, with no realistic alternative late career opportunities. To prevent mid or late career attrition, however, A&E doctors should receive formal training in stress recognition and avoidance. Accessible counselling without stigma should be easily available. Senior A&E doctors have a role in detecting and managing stress amongst other staff in the department.


Cardiovascular aging and exercise in healthy older adults.
Physical inactivity in an aging population is a major contributing factor to the rising numbers of older persons with chronic illnesses and disabilities. The purpose of this article is to review the relationship between physical inactivity and age-associated changes to the cardiovascular system, and provide guidance on prescribing exercise to healthy older persons in order to mitigate the adverse effects of cardiovascular aging. Interpretive review of the literature. A number of structural and functional changes occur in the cardiovascular system with advancing age, many of which are mediated by changes in vascular stiffness. These changes lead not only to cardiovascular events and strokes, but also to frailty, functional decline, and cognitive impairment. A substantial proportion of the decline in aerobic capacity with age may result from physical inactivity. Guidelines for the prescription of aerobic, resistance, and balance training for otherwise healthy older persons are provided. Lack of physical activity is a major risk factor for the epidemic of chronic disease and disability facing an aging population. Many age-associated changes in cardiovascular function result from physical inactivity. The benefits of regular exercise include prevention of cardiovascular events, disability, and cognitive impairment. Age is not a contraindication to exercise, which can usually be initiated safely in older persons.


Changes in stature following plyometric drop-jump and pendulum exercises.
The aim of this study was to compare the changes in stature following the performance of plyometric exercises using drop-jumps and a pendulum swing. Eight male participants aged 21.7 +/- 1.8 years with experience of plyometric training gave their informed consent to act as participants. Participants undertook two exercise regimens and a 15-min standing test in a random order. The exercises entailed the performance of 50 drop-jumps from a height of 0.28 m or 50 pendulum rebounds. Participants were instructed to perform maximal jumps or rebounds using a 'bounce' style. Measurements of stature were performed after a 20-min period of standing (pre-exercise), 2-min after exercise (post-exercise) and after a 20-min standing recovery (recovery). Back pain and muscle soreness were assessed using an analogue-visual scale, at each of the above times and also 24 and 36 h after the test. Peak torque during isokinetic knee extension at 1.04 rads-1 was measured immediately before and after the exercise bouts, to assess the degree of muscular fatigue. Ground/wall reaction force data were recorded using a Kistler force platform mounted in the floor for drop-jumps and vertically on the rebound wall for pendulum exercises. Drop-jumps resulted in the greatest (p < 0.05) change in stature (-2.71 +/- 0.8 mm), compared to pendulum exercises (-1.77 +/- 0.7 mm) and standing (-0.39 +/- 0.2 mm). Both exercise regimens resulted in a significant (p < 0.01) decrease in stature when compared to the standing condition. Drop-jumps resulted in significantly greater peak impact forces (p < 0.05) than pendulum exercises (drop-jumps = 3.2 +/- 0.5 x body weight, pendulum = 2.6 +/- 0.5 x body weight). The two exercise conditions both invoked a small degree of muscle soreness but there were no significant differences between conditions. Both exercise regimens resulted in a non-significant decrease in peak torque indicating a similar degree of muscular fatigue. Based on the lower shrinkage resulted and lower peak forces, it can be concluded that pendulum exercises pose a lower injury potential to the lower back than drop-jumps performed from a height of 28 cm.


NTP technical report on the toxicity and metabolism studies of chloral hydrate (CAS No. 302-17-0). Administered by gavage to F344/N rats and B6C3F1 mice.
Chloral hydrate is widely used as a sedative and a hypnotic in pediatric medicine. It is also a byproduct of water chlorination. Chloral hydrate has been shown to be genotoxic in numerous prokaryotic and eukaryotic assay systems including human lymphocytes in vitro. One of its metabolites, trichloroacetic acid, has demonstrated hepatocarcinogenic activity in mice. Trichloroethylene and perchloroethylene, both of which are metabolized to chloral hydrate, have been shown to be carcinogenic in rats and/or mice. Because of this evidence of carcinogenicity and because of the wide-spread use of chloral hydrate, 16- or 17-day range-finding toxicity studies and separate 16- or 17-day metabolism studies were performed in F344/N rats and B6C3F1 mice in preparation for further long-term rodent studies. In addition, in vitro studies of the metabolism and DNA-binding capacity of chloral hydrate and its metabolites were performed. Genetic toxicity studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, Drosophila melanogaster, and mouse bone marrow cells. For the range-finding studies, groups of eight male and eight female F344/N Nctr BR rats and B6C3F1/Nctr BR (C57BL/6N x C3H/HeN MTV-) mice were administered 0, 50, 100, 200, 400, or 800 mg chloral hydrate per kg body weight in water by gavage 5 days per week for 17 days (rats) or 16 days (mice) for a total of 12 doses. One male rat receiving 800 mg/kg died after five doses. Two 800 mg/kg female rats died after dosing ended but before study termination. One male mouse in each group except the 400 mg/kg group died before the end of the study. Two 800 mg/kg female mice also died before the end of the study. The final mean body weight of 800 mg/kg male rats and the mean body weight gains of 400 and 800 mg/kg males were significantly less than those of the vehicle controls. The mean body weight gains of all groups of dosed male mice were significantly greater than that of the vehicle control group. The only clinical finding in rats and mice attributed to chloral hydrate treatment was light sedation in the 400 mg/kg groups and heavy sedation in the 800 mg/kg groups; sedation subsided within 30 minutes or 3 hours, respectively. The liver weights of 400 mg/kg male mice and 800 mg/kg male and female mice were significantly greater than those of the vehicle control groups. No chemical-related lesions were observed in rats or mice. Male and female rats and mice were administered a single dose of 50 or 200 mg chloral hydrate per kg body weight in water by gavage, or 12 doses of 50 or 200 mg/kg over 17 days (rats) or 16 days (mice). Plasma concentrations of chloral hydrate and its metabolites were determined 15 minutes, 1, 3, 6, and 24 hours, and 2, 4, 8, and 16 days after receiving 1 or 12 doses. Maximum concentrations of chloral hydrate were observed at the initial sampling point of 15 minutes. By 1 hour, the concentrations had dropped substantially, and by 3 hours, chloral hydrate could not be detected in rats or mice. Trichloroacetic acid was the major metabolite detected in the plasma. In rats, the concentrations rose slowly, with the peaks occurring between 1 and 6 hours after treatment. In mice, the peak concentrations were found 1 hour after dosing. The concentrations then slowly decreased such that by 2 days the metabolite could no longer be detected in rats or mice. Trichloroethanol was assayed both as the free alcohol and its glucuronide. In rats, the maximum concentrations of free trichloroethanol occurred at 15 minutes, while the peak concentrations of trichloroethanol glucuronide were found at 1 hour; by 3 hours, concentrations of both metabolites approached background levels. In mice, the maximum concentrations of both metabolites occurred at 15 minutes, and by 1 to 3 hours concentrations approached background levels. The plasma concentrations of chloral hydrate and its metabolites were dose dependent in rats and mice. In mice, plasma concentrations of trichloroacetic acid were significantly higher after a single dose than after 12 doses. None of the metabolic parameters appears to account for species differences that may exist in hepatocarcinogenicity. The data from the study of metabolism and DNA adduct formation indicated that in vitro metabolism of 200 microM to 5 mM chloral hydrate by male B6C3F1 mouse liver microsomes (control microsomes) generated free radical intermediates that resulted in endogenous lipid peroxidation, forming malondialdehyde, formaldehyde, acetaldehyde, acetone, and propionaldehyde. Similar concentrations of trichloroacetic acid and trichloroethanol, the primary metabolites of chloral hydrate, also generated free radicals and induced lipid peroxidation. Lipid peroxidation induced by trichloroacetic acid nearly equaled that induced by chloral hydrate, while that from trichloroethanol was three- to fourfold less. Metabolism of 200 microM to 5 mM chloral hydrate, trichloroacetic acid, and trichloroethanol by liver microsomes of B6C3F1 mice pretreated with pyrazole (pyrazole-induced microsomes) yielded lipid peroxidation products at concentrations two- to threefold greater than those from liver microsomes of untreated mice. Additionally, chloral hydrate-induced lipid peroxidation catalyzed by control and pyrazole-induced microsomes was reduced significantly by 2,4-dichloro-6-phenylphenoxyethylamine, a general cytochrome P450 inhibitor. Human lymphoblastoid transgenic cells expressing cytochrome P(450)2E1 metabolized 200 to 5,000 micrograms/mL chloral hydrate to reactants inducing mutations, whereas the parental cell line was inactive. The malondialdehyde-modified DNA adduct, 3-(2-deoxy-beta-D-erythro-pentofuranosyl)pyrimido[1,2 alpha]purin-10(3H)-one (MDA-MG-1), formed from the metabolism of 1 mM chloral hydrate, trichloroacetic acid, and trichloroethanol by control B6C3F1 mouse liver microsomes, mouse pyrazole-induced microsomes, male F344/N rat liver microsomes, and human liver microsomes in the presence and absence of calf thymus DNA was also determined. When incubated in the absence of calf thymus DNA, the amount of malondialdehyde formed from metabolism by pyrazole-induced mouse microsomes was twice that from rat or human liver microsomes. Amounts of chloral hydrate-induced and trichloroacetic acid-induced lipid peroxidation products formed from metabolism by rat and human liver microsomes were similar, and these quantities were about twice those formed from the metabolism of trichloroethanol. The quantity of MDA-MG-1 formed from the metabolism of chloral hydrate, trichloroacetic acid, and trichloroethanol by mouse, rat, and human liver microsomes exhibited a linear correlation with the quantity of malondialdehyde formed under incubation conditions in the absence of calf thymus DNA. Chloral hydrate was shown to be mutagenic in vitro and in vivo. At doses from 1,000 to 10,000 micrograms/plate, it induced mutations in S. typhimurium strain TA100, with and without S9 activation; an equivocal response was obtained in S. typhimurium strain TA98 in the absence of S9, and no mutagenicity was detected with strain TA1535 or TA1537. Chloral hydrate at doses from 1,700 to 5,000 micrograms/mL induced sister chromatid exchanges; at doses from 1,000 to 3,000 micrograms/mL, chromosomal aberrations were induced in cultured Chinese hamster ovary cells, with and without S9. Results of a sex-linked recessive lethal test in D. melanogaster were unclear; administration of chloral hydrate by feeding produced an inconclusive increase in recessive lethal mutations, results of the injection experiment were negative. An in vivo mouse bone marrow micronucleus test with chloral hydrate at doses from 125 to 500 mg/kg gave a positive dose trend. In summary, due to the absence of chloral hydrate-induced histopathologic lesions in rats and mice, no-observed-adverse-effect levels (NOAELs) were based on body weights of rats and liver weights of mice. The NOAELs for rats and mice were 200 mg/kg. Chloral hydrate was rapidly metabolized by rats and mice, with trichloroacetic acid occurring as the major metabolite. Peak concentrations of trichloroacetic acid occurred more quickly in mice. Plasma concentrations of chloral hydrate were dose dependent, but metabolic rates were unaffected by dose or sex. Chloral hydrate was mutagenic in vitro and in vivo. Metabolism of chloral hydrate and its metabolites produced free radicals that resulted in lipid peroxidation in liver microsomes of mice, rats, and humans. Induction of cytochrome P(450)2E1 by pyrazole increased the concentrations of lipid peroxidation products; inhibition of cytochrome P(450)2E1 by 2,4-dinitrophenylhydrazine reduced these concentrations. Metabolism of chloral hydrate and its metabolites by mouse, rat, and human liver microsomes formed malondialdehyde, and in the presence of calf thymus DNA formed the DNA adduct MDA-MG-1.


Delineating wetland catchments and modeling hydrologic connectivity using lidar data and aerial imagery.
In traditional watershed delineation and topographic modeling, surface depressions are generally treated as spurious features and simply removed from a digital elevation model (DEM) to enforce flow continuity of water across the topographic surface to the watershed outlets. In reality, however, many depressions in the DEM are actual wetland landscape features with seasonal to permanent inundation patterning characterized by nested hierarchical structures and dynamic filling-spilling-merging surface-water hydrological processes. Differentiating and appropriately processing such ecohydrologically meaningful features remains a major technical terrain-processing challenge, particularly as highresolution spatial data are increasingly used to support modeling and geographic analysis needs. The objectives of this study were to delineate hierarchical wetland catchments and model their hydrologic connectivity using high-resolution lidar data and aerial imagery. The graph-theory-based contour tree method was used to delineate the hierarchical wetland catchments and characterize their geometric and topological properties. Potential hydrologic connectivity between wetlands and streams were simulated using the least-cost-path algorithm. The resulting flow network delineated potential flow paths connecting wetland depressions to each other or to the river network on scales finer than those available through the National Hydrography Dataset. The results demonstrated that our proposed framework is promising for improving overland flow simulation and hydrologic connectivity analysis.


Effect of carboxymethyl cellulose concentration on physical properties of biodegradable cassava starch-based films.
Cassava starch, the economically important agricultural commodity in Thailand, can readily be cast into films. However, the cassava starch film is brittle and weak, leading to inadequate mechanical properties. The properties of starch film can be improved by adding plasticizers and blending with the other biopolymers. Cassava starch (5%w/v) based films plasticized with glycerol (30 g/100 g starch) were characterized with respect to the effect of carboxymethyl cellulose (CMC) concentrations (0, 10, 20, 30 and 40%w/w total solid) and relative humidity (34 and 54%RH) on the mechanical properties of the films. Additionally, intermolecular interactions were determined by Fourier transform infrared spectroscopy (FT-IR), melting temperature by differential scanning calorimetry (DSC), and morphology by scanning electron microscopy (SEM). Water solubility of the films was also determined. Increasing concentration of CMC increased tensile strength, reduced elongation at break, and decreased water solubility of the blended films. FT-IR spectra indicated intermolecular interactions between cassava starch and CMC in blended films by shifting of carboxyl (C = O) and OH groups. DSC thermograms and SEM micrographs confirmed homogeneity of cassava starch-CMC films. The addition of CMC to the cassava starch films increased tensile strength and reduced elongation at break of the blended films. This was ascribed to the good interaction between cassava starch and CMC. Cassava starch-CMC composite films have the potential to replace conventional packaging, and the films developed in this work are suggested to be suitable for low moisture food and pharmaceutical products.


[Occupational accidents in an oil refinery in Brazil].
Work in oil refineries involves the risk of minor to major accidents. National data show the impact of accidents on this industry. A study was carried out to describe accident profile and evaluate the adequacy of accident reporting system. Data on all accidents reported in an oil refinery in the state of Rio de Janeiro for the year 1997 were organized and analyzed. The study population consisted of 153 injury cases, 83 hired and 69 contracted workers. The variables were: type of accident, operation mode and position of the worker injured. Among hired workers, minor accidents predominated (54.2%) and they occurred during regular operation activities (62.9%). Among contracted workers, there also predominated minor accidents (75.5%) in a higher percentage, but they occurred mainly during maintenance activities (96.8%). The study results showed that there is a predominance of accidents in lower hierarchy workers, and these accidents occur mainly during maintenance activities. There is a need to improve the company's accident reporting system and accident investigation procedures.


In vitro and in vivo biological responses to a novel radiopacifying agent for bone cement.
Iodixanol (IDX) and iohexol (IHX) have been investigated as possible radiopacification agents for polymethylmethacrylate (PMMA) bone cement, to replace the currently used barium sulphate and zirconia. IDX and IHX are both water-soluble iodine-based contrast media and for the last 20 years have been used extensively in clinical diagnostic procedures such as contrast media enhanced computed tomography, angiography and urography. One of the major reasons to remove the current radiopacifying agents is their well-documented cytotoxicity and their potential to increase bone resorption. Using in vitro bone resorption assays, the effect of PMMA particles plus IDX or IHX to induce osteoclast formation and lacunar resorption on dentine slices has been investigated. These responses have been compared with the in vitro response to PMMA particles containing the conventional radiopacifying agents, that is, barium sulphate and zirconia. In parallel, the in vivo reaction, in terms of new bone formation, to particles of these materials has been tested using a bone harvest chamber in rabbit tibiae. In vitro cell culture showed that PMMA containing IHX resulted in significantly less bone resorption than PMMA containing the conventional opacifiers. In vivo testing, however, showed no significant differences between the amounts of new bone formed around cement samples containing the two iodine-based opacifying agents in particulate form, although both led to fewer inflammatory cells than particles of PMMA containing zirconia. Our results suggest that a non-ionic radiopacifier could be considered as an alternative to the conventional radiopacifying agents used in biomaterials in orthopaedic surgery.


Can Pay-for Performance Incentive Levels be Determined Using a Cost-Effectiveness Framework?
Healthcare payers in the United States are increasingly tying provider payments to quality and value using pay-for-performance policies. Cost-effectiveness analysis quantifies value in healthcare but is not currently used to design or prioritize pay-for-performance strategies or metrics. Acute ischemic stroke care provides a useful application to demonstrate how simulation modeling can be used to determine cost-effective levels of financial incentives used in pay-for-performance policies and associated challenges with this approach. Our framework requires a simulation model that can estimate quality-adjusted life years and costs resulting from improvements in a quality metric. A monetary level of incentives can then be back-calculated using the lifetime discounted quality-adjusted life year (which includes effectiveness of quality improvement) and cost (which includes incentive payments and cost offsets from quality improvements) outputs from the model. We applied this framework to an acute ischemic stroke microsimulation model to calculate the difference in population-level net monetary benefit (willingness-to-pay of $50 000 to $150 000/quality-adjusted life year) accrued under current Medicare policy (stroke payment not adjusted for performance) compared with various hypothetical pay-for-performance policies. Performance measurement was based on time-to-thrombolytic treatment with tPA (tissue-type plasminogen activator). Compared with current payment, equivalent population-level net monetary benefit was achieved in pay-for-performance policies with 10-minute door-to-needle time reductions (5057 more acute ischemic stroke cases/y in the 0-3-hour window) incentivized by increasing tPA payment by as much as 18% to 44% depending on willingness-to-pay for health. Cost-effectiveness modeling can be used to determine the upper bound of financial incentives used in pay-for-performance policies, although currently, this approach is limited due to data requirements and modeling assumptions. For tPA payments in acute ischemic stroke, our model-based results suggest financial incentives leading to a 10-minute decrease in door-to-needle time should be implemented but not exceed 18% to 44% of current tPA payment. In general, the optimal level of financial incentives will depend on willingness-to-pay for health and other modeling assumptions around parameter uncertainty and the relationship between quality improvements and long-run quality-adjusted life expectancy and costs.


Functional expression of the GABA(A) receptor α2 and α3 subunits at synapses between intercalated medial paracapsular neurons of mouse amygdala.
In the amygdala, GABAergic neurons in the intercalated medial paracapsular cluster (Imp) have been suggested to play a key role in fear learning and extinction. These neurons project to the central (CE) amygdaloid nucleus and to other areas within and outside the amygdala. In addition, they give rise to local collaterals that innervate other neurons in the Imp. Several drugs, including benzodiazepines (BZ), are allosteric modulators of GABA(A) receptors. BZ has both anxiolytic and sedative actions, which are mediated through GABA(A) receptors containing α2/α3 and α1 subunits, respectively. To establish whether α1 or α2/α3 subunits are expressed at Imp cell synapses, we used paired recordings of anatomically identified Imp neurons and high resolution immunocytochemistry in the mouse. We observed that a selective α3 subunit agonist, TP003 (100 nM), significantly increased the decay time constant of the unitary IPSCs. A similar effect was also induced by zolpidem (10 μM) or by diazepam (1 μM). In contrast, lower doses of zolpidem (0.1-1 μM) did not significantly alter the kinetics of the unitary IPSCs. Accordingly, immunocytochemical experiments established that the α2 and α3, but not the α1 subunits of the GABA(A) receptors, were present at Imp cell synapses of the mouse amygdala. These results define, for the first time, some of the functional GABA(A) receptor subunits expressed at synapses of Imp cells. The data also provide an additional rationale to prompt the search of GABA(A) receptor α3 selective ligands as improved anxiolytic drugs.


Unveiling N-protonation and anion-binding effects on Fe/N/C-catalysts for O2 reduction in PEM fuel cells.
The high cost of proton-exchange-membrane fuel cells would be considerably reduced if platinumbased catalysts were replaced by iron-based substitutes, which have recently demonstrated comparable activity for oxygen reduction, but whose cause of activity decay in acidic medium has been elusive. Here, we reveal that the activity of Fe/N/C-catalysts prepared through a pyrolysis in NH3 is mostly imparted by acid-resistant FeN4-sites whose turnover frequency for the O2 reduction can be regulated by fine chemical changes of the catalyst surface. We show that surface N-groups protonate at pH 1 and subsequently bind anions. This results in decreased activity for the O2 reduction. The anions can be removed chemically or thermally, which restores the activity of acid-resistant FeN4-sites. These results are interpreted as an increased turnover frequency of FeN4-sites when specific surface N-groups protonate. These unprecedented findings provide new perspective for stabilizing the most active Fe/N/C-catalysts known to date.


Natural orifice transluminal endoscopic radical prostatectomy: initial perioperative and pathologic results.
To describe the first clinical experience, pathologic, and perioperative outcomes of natural orifice transluminal endoscopic surgery (NOTES) radical prostatectomy. NOTES represents the evolution of minimally invasive surgery. The conceptual feasibility has been shown in careful laboratory and animal studies, but a scarcity of information regarding clinical applications exists. After institutional review board approval, 2 patients agreed to undergo NOTES radical prostatectomy for localized prostate cancer. The prostate was radically resected using a 26F resectoscope, 550-μm laser fiber, and holmium laser. The prostate was delivered into the bladder and removed at the conclusion of the procedure through a suprapubic cystotomy for histopathologic analysis. The vesicourethral anastomosis was completed using a cannula scope, urethral-vesical suturing device, and titanium knot applier. Cystograms were taken immediately postoperatively and at catheter removal. Both patients tolerated the procedure without operative complications. All intraoperative cystograms showed watertight anastomoses. The pathologic examination revealed Gleason score 3 + 3 and Stage pT2aNxMx for 1 patient and Gleason score 3 + 4 and Stage pT2cNxMx for 1 patient, with negative margins for both. No blood transfusions were required. Patient 2 experienced some left-sided gluteal and suprapubic pain postoperatively. NOTES radical prostatectomy appears to be a safe and feasible option for the management of carefully selected, organ-confined prostate cancer. The perioperative and pathologic outcomes show promise with this new technique; however, the high standards of oncologic and functional outcomes demand close and longer follow-up before adoption into the surgical armamentarium can be recommended.


Different molecular signatures in magnetic resonance imaging-staged facioscapulohumeral muscular dystrophy muscles.
Facioscapulohumeral muscular dystrophy (FSHD) is one of the most common muscular dystrophies and is characterized by a non-conventional genetic mechanism activated by pathogenic D4Z4 repeat contractions. By muscle Magnetic Resonance Imaging (MRI) we observed that T2-short tau inversion recovery (T2-STIR) sequences identify two different conditions in which each muscle can be found before the irreversible dystrophic alteration, marked as T1-weighted sequence hyperintensity, takes place. We studied these conditions in order to obtain further information on the molecular mechanisms involved in the selective wasting of single muscles or muscle groups in this disease. Histopathology, gene expression profiling and real time PCR were performed on biopsies from FSHD muscles with different MRI pattern (T1-weighted normal/T2-STIR normal and T1-weighted normal/T2-STIR hyperintense). Data were compared with those from inflammatory myopathies, dysferlinopathies and normal controls. In order to validate obtained results, two additional FSHD samples with different MRI pattern were analyzed. Myopathic and inflammatory changes characterized T2-STIR hyperintense FSHD muscles, at variance with T2-STIR normal muscles. These two states could be easily distinguished from each other by their transcriptional profile. The comparison between T2-STIR hyperintense FSHD muscles and inflammatory myopathy muscles showed peculiar changes, although many alterations were shared among these conditions. At the single muscle level, different stages of the disease correspond to the two MRI patterns. T2-STIR hyperintense FSHD muscles are more similar to inflammatory myopathies than to T2-STIR normal FSHD muscles or other muscular dystrophies, and share with them upregulation of genes involved in innate and adaptive immunity. Our data suggest that selective inflammation, together with perturbation in biological processes such as neoangiogenesis, lipid metabolism and adipokine production, may contribute to the sequential bursts of muscle degeneration that involve individual muscles in an asynchronous manner in this disease.


Biosafety of non-surface modified carbon nanocapsules as a potential alternative to carbon nanotubes for drug delivery purposes.
Carbon nanotubes (CNTs) have found wide success in circuitry, photovoltaics, and other applications. In contrast, several hurdles exist in using CNTs towards applications in drug delivery. Raw, non-modified CNTs are widely known for their toxicity. As such, many have attempted to reduce CNT toxicity for intravenous drug delivery purposes by post-process surface modification. Alternatively, a novel sphere-like carbon nanocapsule (CNC) developed by the arc-discharge method holds similar electric and thermal conductivities, as well as high strength. This study investigated the systemic toxicity and biocompatibility of different non-surface modified carbon nanomaterials in mice, including multi-walled carbon nanotubes (MWCNTs), single-walled carbon nanotubes (SWCNTs), carbon nanocapsules (CNCs), and C ₆₀ fullerene (C ₆₀). The retention of the nanomaterials and systemic effects after intravenous injections were studied. MWCNTs, SWCNTs, CNCs, and C ₆₀ were injected intravenously into FVB mice and then sacrificed for tissue section examination. Inflammatory cytokine levels were evaluated with ELISA. Mice receiving injection of MWCNTs or SWCNTs at 50 µg/g b.w. died while C ₆₀ injected group survived at a 50% rate. Surprisingly, mortality rate of mice injected with CNCs was only at 10%. Tissue sections revealed that most carbon nanomaterials retained in the lung. Furthermore, serum and lung-tissue cytokine levels did not reveal any inflammatory response compared to those in mice receiving normal saline injection. Carbon nanocapsules are more biocompatible than other carbon nanomaterials and are more suitable for intravenous drug delivery. These results indicate potential biomedical use of non-surface modified carbon allotrope. Additionally, functionalization of the carbon nanocapsules could further enhance dispersion and biocompatibility for intravenous injection.


Dynamic transcription factor activity and networks during ErbB2 breast oncogenesis and targeted therapy.
Tissue development and disease progression are multi-stage processes controlled by an evolving set of key regulatory factors, and identifying these factors necessitates a dynamic analysis spanning relevant time scales. Current omics approaches depend on incomplete biological databases to identify critical cellular processes. Herein, we present TRACER (TRanscriptional Activity CEll aRrays), which was employed to quantify the dynamic activity of numerous transcription factor (TFs) simultaneously in 3D and networks for TRACER (NTRACER), a computational algorithm that allows for cellular rewiring to establish dynamic regulatory networks based on activity of TF reporter constructs. We identified major hubs at various stages of culture associated with normal and abnormal tissue growth (i.e., ELK-1 and E2F1, respectively) and the mechanism of action for a targeted therapeutic, lapatinib, through GATA-1, which were confirmed in human ErbB2 positive breast cancer patients and human ErbB2 positive breast cancer cell lines that were either sensitive or resistant to lapatinib.


Mass spectrometry of nicotinic acetylcholine receptors and associated proteins as models for complex transmembrane proteins.
Studies were conducted to optimize matrix-assisted laser desorption/ionization, time-of-flight mass spectrometry (MALDI TOF MS) in analyzing the composition of nicotinic acetylcholine receptors (nAChR) from Torpedo californica electric tissue in their membrane-bound, detergent-solubilized, and affinity-purified states. Mass spectra obtained from nAChR-rich membrane fractions gave reasonably good representations of protein compositions indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of those same samples. Efficiency of extraction of nAChR from membranes was not markedly different for most detergents, but quality and signal size of mass spectra were clearly influenced by detergent composition and concentration, protein concentration, and MALDI matrix composition. The best spectra, allowing detection and accurate size determinations for samples containing as little as 10 fmol of pure nAChR, were obtained for samples solubilized in Triton X-100 and assayed by use of a sinapinic acid matrix. Although informative spectra could be obtained for nAChR affinity purified on alpha-cobratoxin (Naja naja siamensis) columns and extracted using sinapinic acid, superior spectra with much higher signal:noise were obtained if extraction media contained Triton X-100 or sodium dodecyl sulfate. nAChR subunit masses determined were similar regardless of the membrane-associated, detergent-solubilized, or affinity-purified state of the preparation. These studies illustrate how masses can be determined for nAChR subunits and for other protein components in Torpedo membrane preparations, such as RAPsyn and Na(+)-K(+)-ATPase alpha and beta subunits. They also provide an underpinning for streamlined analysis of the composition of complex transmembrane proteins using MALDI TOF MS.


The last innovation in achalasia treatment; per-oral endoscopic myotomy.
Per-oral endoscopic myotomy (POEM) is a minimally invasive endoscopic treatment option for patients with achalasia and has been performed since 2010. It is less invasive than Heller myotomy and its use is spreading rapidly worldwide. We present our results of POEM that, to the best of our knowledge, are the first cases in Turkey. We enrolled patients between May 2014 and September 2014; 8 patients with achalasia whose complaints recurred after pneumatic balloon dilatation underwent POEM. The procedure was performed under general anesthesia at the endoscopy unit of the gastroenterology clinic. Demographic data was recorded before the procedure, and the results of the procedure were recorded prospectively. The median age of the patients was 42.5 (30-72) years. Preoperative and postoperative median Eckardt scores were 10 (8-12) and 1 (0-2), respectively. The median total duration of the procedure was 101 (71-158) min, and the median myotomy length was 13.5 (10-16) cm. Postoperative oral intake started on median day 1 (1-2) and the length of hospital stay was 4 (3-6) days. In 2 patients, capnoperitoneum developed during the procedure and was treated with a Veress needle. POEM is a safe endoscopic treatment modality for patients with achalasia in centers that are experienced in advanced endoscopic techniques.


Probiotics for the treatment or prevention of atopic dermatitis: a review of the evidence from randomized controlled trials.
Probiotics are defined as live microorganisms which, when administered in adequate amounts, confer a health benefit on the host. To synthesize the evidence for the effectiveness of probiotics in the treatment or prevention of atopic dermatitis (AD) in children, we reviewed the results of 13 relevant randomized (placebo)-controlled trials (RCTs), 10 of which evaluated probiotics as treatment and 3 for prevention of AD. The main outcome measure in 9 RCTs was the change in SCORAD (SCORing Atopic Dermatitis). Four RCTs suggested that there was a statistically significant decrease in SCORAD after probiotic administration to infants or children with AD for 1 or 2 months compared with that after placebo, while in two RCTs SCORAD was significantly reduced after treatment with lactobacilli only in children with IgE-associated AD. In four of these six RCTs, clinical improvement was associated with a change in some inflammatory markers. In three RCTs, the change in SCORAD was not statistically significant between probiotic- and placebo-treated children, although in one of these trials SCORAD was significantly lower after probiotic than with placebo treatment in food-sensitized children. In most RCTs, probiotics did not cause a statistically significant change in interferon-gamma, interleukin-4, tumor necrosis factor-alpha, eosinophil cationic protein or transforming growth factor-beta compared with placebo. Regarding the effectiveness of probiotics in the prevention of AD, in two RCTs, infants at high risk for atopy who received probiotics developed AD significantly less frequently during the first 2 years of life than infants who received placebo. In these studies, mothers were administered Lactobacillus rhamnosus GG with or without other probiotics perinatally, followed by treatment of the infants with the same probiotics for the first 6 months of life. However, in another trial, neither the frequency nor the severity of AD during the first year of life were significantly different between infants with atopic mothers who received L. acidophilus for the first 6 months of life compared with infants who received placebo.Probiotics, especially L. rhamnosus GG, seem to be effective for the prevention of AD. They were also found to reduce the severity of AD in approximately half of the RCTs evaluated, although they were not found to change significantly most of the inflammatory markers measured in the majority of the RCTs evaluated. More RCTs need to be conducted to elucidate whether probiotics are useful for the treatment or prevention of AD.


Development of the University Food Environment Assessment (Uni-Food) Tool and Process to Benchmark the Healthiness, Equity, and Environmental Sustainability of University Food Environments.
Globally, there is increasing interest in monitoring actions to create healthy, equitable and environmentally sustainable food environments. Currently, there is a lack of detailed tools for monitoring and benchmarking university food environments. This study aimed to develop the University Food Environment Assessment (Uni-Food) tool and process to benchmark the healthiness, equity, and environmental sustainability of food environments in tertiary education settings, and pilot test its implementation in three Australian universities in 2021. The Uni-Food tool development was informed by a review of the literature and input from an expert advisory panel. It comprises three components: (1) university systems and governance, (2) campus facilities and environments, and (3) food retail outlets. The process for implementing the tool is designed for universities to self-assess the extent to which they have implemented recommended practice in 68 indicators, across 16 domains, weighted based on their relative importance. The pilot implementation of the tool identified moderate diversity in food environments across universities and highlighted several opportunities for improvements at each institution. The assessment process was found to be reliable, with assessors rating the tool as easy to use, requiring minimal resources. Broad application of the tool has the potential to increase accountability and guide best practice in tertiary education and other complex institutional settings.


Zinc Source and Concentration Altered Physiological Responses of Beef Heifers during a Combined Viral-Bacterial Respiratory Challenge.
To determine the effects of zinc supplementation on the immune response to a combined viral-bacterial respiratory disease challenge, thirty-two beef heifers (255 ± 15 kg) were subjected to a 30-d period of Zn depletion, then randomly assigned to one of three treatment diets fed for 30 d before the challenge: (1) supplementation with 100 mg of Zn from Zn sulfate/kg of DM (Zn100), (2) supplementation with 200 mg of Zn from Zn sulfate/kg of DM (Zn200), and (3) supplementation with 80 mg of Zn/kg of DM from zinc methionine and 20 mg of Zn from Zn sulfate/kg of DM (ZinMet). After the 30-d supplementation period, all heifers were fitted with indwelling vaginal temperature (VT) devices and intra-nasally challenged with 1 × 108 PFU bovine herpesvirus-1 on d -3, and then allowed to rest in outdoor pens for 3 d. On d 0, each heifer was challenged intra-tracheally with an average dose of 2.38 × 107 CFU Mannheimia haemolytica (MH), fitted with an indwelling jugular catheter, and then moved into individual stalls in an environmentally-controlled enclosed barn. Whole blood samples were collected at 1-h (serum) and 2-h (complete blood counts) intervals from 0 to 8 h, and at 12, 24, 36, 48, 60, 72, 168, and 360 h relative to MH challenge. Data were analyzed using the MIXED procedure of SAS specific for repeated measures with fixed effects of treatment, time, and their interaction. There was a treatment effect (p < 0.01) for VT such that Zn200 heifers had greater VT than Zn100 and ZinMet heifers. There was a trend (p = 0.10) for a serum cortisol treatment effect with Zn100 heifers having greater cortisol than ZinMet heifers. Total leukocytes and lymphocytes were greater (p ≤ 0.01) in Zn100 heifers than Zn200 and ZinMet heifers, whereas monocytes were less (p = 0.05) in ZinMet heifers than Zn100 and Zn200 heifers. Concentrations of IL-6 were greater (p = 0.02) in ZinMet heifers than Zn100 and Zn200 heifers. Concentrations of IFN-γ were greater in Zn200 heifers than ZinMet heifers at 0 h, and Zn100 heifers from 0 to 12 h post-MH challenge (treatment x time p = 0.02). Serum haptoglobin was not affected by treatment or treatment x time (p ≥ 0.36) but increased over time (p < 0.01) in all groups. There was a trend (p = 0.11) for ZinMet heifers to have less severe nasal lesion scores than Zn100 heifers. The observed differential physiological responses in this study indicate that zinc source and concentration may alter the response to a bovine respiratory challenge in heifers.


Single-Access Fetal Endoscopy (SAFE) for myelomeningocele in sheep model I: amniotic carbon dioxide gas approach.
This study aimed to assess the feasibility of single-access fetal endoscopy (SAFE) for the management of myelomeningocele (MMC) using intrauterine carbon dioxide as a distension medium in a sheep model. This prospective experimental case-control study investigated 12 lamb fetuses that had a myelomeningocele-like defect surgically created on the 75th day of gestation. Four fetuses remained untreated (control group), and eight fetuses had MMC repair using two fetoscopic approaches with carbon dioxide used to distend the amniotic cavity. A collagen patch was placed over the defect and secured with surgical sealant. Four animals had a two-port fetoscopic procedure, and four animals had SAFE. Clinical and pathologic studies were performed after delivery. This study confirmed the validity of the animal MMC model. None of the control animals was able to stand or walk, and all had a significant defect in the lumbar area with continuous leakage of cerebrospinal fluid, ventriculomegaly, and a Chiari-II malformation. All the treated animals, independently of the number of ports used in the repair, were able to walk and had a closed defect with resolution of the Chiari malformation. The SAFE patch and glue coverage of surgically created fetal MMC is feasible and effective in restoring gross neurologic function in the fetal lamb model.


Comparing Waste-to-Energy technologies by applying energy system analysis.
Even when policies of waste prevention, re-use and recycling are prioritised, a fraction of waste will still be left which can be used for energy recovery. This article asks the question: How to utilise waste for energy in the best way seen from an energy system perspective? Eight different Waste-to-Energy technologies are compared with a focus on fuel efficiency, CO(2) reductions and costs. The comparison is carried out by conducting detailed energy system analyses of the present as well as a potential future Danish energy system with a large share of combined heat and power as well as wind power. The study shows potential of using waste for the production of transport fuels. Biogas and thermal gasification technologies are hence interesting alternatives to waste incineration and it is recommended to support the use of biogas based on manure and organic waste. It is also recommended to support research into gasification of waste without the addition of coal and biomass. Together the two solutions may contribute to alternate use of one third of the waste which is currently incinerated. The remaining fractions should still be incinerated with priority to combined heat and power plants with high electric efficiency.


Concordancing: use of language-based research in medical communication.
The available literature on medical communication reports almost exclusively on observational, qualitative studies. We aimed to apply a novel approach to the analysis of doctor-patient consultation by means of computer concordancing. This methodology, established in linguistic research but rarely applied to professional language, allows both the quantitative and qualitative study of language. We analysed the language of 40 doctors and their patients during 373 complete primary-care consultations. We examined the use of jargon by doctors, the language of power and absence of power, and ways in which language was used to diminish the potential threat of the presenting disorder. There was no evidence that the doctors used medical jargon. Some doctors used language associated with social power, and some patients used language associated with absence of power. There was substantial evidence that the doctors used language to express emotions (eg, anxiety), to diminish threats (eg, words such as "little"), and to reassure patients. Concordancing is a valuable resource to study the consultation. The finding that doctors do not use jargon suggests that they are aware of the need to avoid it, but it does not follow that they are easily understood by patients. The use of some elements of the language of power may imply that consultations may be less democratic than is appropriate. The language of emotion and diminution is a major part of the primary-care doctor's repertoire and denotes a therapeutic use of language.


Large-Scale Topological Changes Restrain Malignant Progression in Colorectal Cancer.
Widespread changes to DNA methylation and chromatin are well documented in cancer, but the fate of higher-order chromosomal structure remains obscure. Here we integrated topological maps for colon tumors and normal colons with epigenetic, transcriptional, and imaging data to characterize alterations to chromatin loops, topologically associated domains, and large-scale compartments. We found that spatial partitioning of the open and closed genome compartments is profoundly compromised in tumors. This reorganization is accompanied by compartment-specific hypomethylation and chromatin changes. Additionally, we identify a compartment at the interface between the canonical A and B compartments that is reorganized in tumors. Remarkably, similar shifts were evident in non-malignant cells that have accumulated excess divisions. Our analyses suggest that these topological changes repress stemness and invasion programs while inducing anti-tumor immunity genes and may therefore restrain malignant progression. Our findings call into question the conventional view that tumor-associated epigenomic alterations are primarily oncogenic.


Principles of electroanatomic mapping.
Electrophysiologic testing and radiofrequency ablation have evolved as curative measures for a variety of rhythm disturbances. As experience in this field has grown, ablation is progressively being used to address more complex rhythm disturbances. Paralleling this trend are technological advancements to facilitate these efforts, including electroanatomic mapping (EAM). At present, several different EAM systems utilizing various technologies are available to facilitate mapping and ablation. Use of these systems has been shown to reduce fluoroscopic exposure and radiation dose, with less significant effects on procedural duration and success rates. Among the data provided by EAM are chamber reconstruction, tagging of important anatomic landmarks and ablation lesions, display of diagnostic and mapping catheters without using fluoroscopy, activation mapping, and voltage (or scar) mapping. Several EAM systems have specialized features, such as enhanced ability to map non-sustained or hemodynamically unstable arrhythmias, ability to display diagnostic as well as mapping catheter positions, and wide compatibility with a variety of catheters. Each EAM system has its strengths and weaknesses, and the system chosen must depend upon what data is required for procedural success (activation mapping, substrate mapping, cardiac geometry), the anticipated arrhythmia, the compatibility of the system with adjunctive tools (i.e. diagnostic and ablation catheters), and the operator's familiarity with the selected system. While EAM can offer significant assistance during an EP procedure, their incorrect or inappropriate application can substantially hamper mapping efforts and procedural success, and should not replace careful interpretation of data and strict adherence to electrophysiologic principles.


Characterization of a complex glucocorticoid response unit in the phosphoenolpyruvate carboxykinase gene.
The minimal DNA sequence required for glucocorticoid induction of the phosphoenolpyruvate carboxykinase (PEPCK) gene in H4IIE rat hepatoma cells was defined. This novel glucocorticoid response unit (GRU) spans about 110 base pairs (bp) and includes two receptor-binding elements plus two accessory factor-binding elements. Purified glucocorticoid receptor bound to two regions (GR1 and GR2) between -395 and -349 bp relative to the transcription start site. Factors in crude rat liver nuclear extract bound to DNA in the regions -455 to -431 and -420 to -403 bp, which are designated accessory factor 1 (AF1) and accessory factor 2 (AF2) elements, respectively. Gel retardation analysis revealed that at least two proteins bound to AF1 and that they were distinct from the protein(s) that bound to AF2. Various combinations of GR1, GR2, AF1, and AF2 were fused to the chloramphenicol acetyltransferase (CAT) reporter gene and cotransfected with a glucocorticoid receptor expression plasmid (pSVGR1) into H4IIE cells to identify the functional GRU. Neither the glucocorticoid receptor binding region nor the accessory factor binding region alone was sufficient to confer glucocorticoid responsiveness. The two components of the glucocorticoid receptor binding region functioned independently, and each accounted for half of the maximal response, provided the accessory factor elements were present. Similarly, deletion of either AF1 or AF2 diminished glucocorticoid induction of the PEPCK gene to approximately half of the maximum. We propose that the complex PEPCK gene GRU provides the stringent regulation required of this critical enzyme in liver.


Second-Look Colonoscopies and the Impact on Capacity in FIT-Based Colorectal Cancer Screening.
Fecal immunochemical testing (FIT) and colonoscopy are tandem procedures in colorectal cancer (CRC) screening. A positive FIT predicts advanced neoplasia (AN) that requires endoscopic detection and removal. En bloc or piecemeal resection of AN is associated with a significant rate of residual or recurrent neoplasia. Second-look colonoscopies are indicated to assess completeness of removal of AN. These colonoscopies can make a substantial demand on colonoscopy capacity and health-care system. This study is the first to evaluate the demand and risk factors for second-look colonoscopy in FIT CRC screening. All colonoscopies after a positive FIT, in subjects aged 50-74 years approached for 3 rounds of FIT screening, were prospectively registered. Second-look colonoscopies were defined as any colonoscopy within 1 year following a colonoscopy after positive FIT. Out of 1,215 FIT-positive screenees undergoing colonoscopy, 105 (8.6%) patients underwent a second-look colonoscopy, of whom 30 (2.5%) underwent more than one colonoscopy (range 2-9), leading to a total of 149 (12.3%) additional colonoscopies. Main reasons for second-look colonoscopies were assessment of complete AN removal (41.9%) and need for additional polypectomy (34.3%). Risk factors were advanced adenomas and poor bowel preparation (P<0.001). High fecal hemoglobin concentration was the only predictor of a second-look colonoscopy before index colonoscopy (P<0.001). Second-look colonoscopies have substantial impact on colonoscopy resources, increasing the demand with 12%. The main reasons for these second-look colonoscopies were previous incomplete polypectomy and control of completeness of removal of neoplastic lesions. A high fecal hemoglobin concentration as measured by FIT can help to identify patients at risk of a second-look colonoscopy.


The Role of Family for Youth Friendships: Examining a Social Anxiety Mechanism.
The quality of family relationships and youth friendships are intricately linked. Previous studies have examined different mechanisms of family-peer linkage, but few have examined social anxiety. The present study examined whether parental rejection and family climate predicted changes in youth social anxiety, which in turn predicted changes in friendship quality and loneliness. Possible bidirectional associations also were examined. Data for mothers, fathers, and youth (M age at Time 1 = 11.27; 52.3% were female) from 687 two-parent households over three time points are presented. Results from autoregressive, cross-lagged analyses revealed that father rejection (not mother rejection or family climate) at Time 1 (Fall of 6th Grade) predicted increased youth social anxiety at Time 2 (Spring of 7th Grade), which in turn, predicted increased loneliness at Time 3 (Spring of 8th Grade). The indirect effect of father rejection on loneliness was statistically significant. Mother rejection, father rejection, and a poor family climate were associated with decreased friendship quality and increased loneliness over time. Finally, there was some evidence of transactional associations between father rejection and youth social anxiety as well as between social anxiety and loneliness. This study's findings underscore the important role of fathers in youth social anxiety and subsequent social adjustment.


Quantifying neutralising antibody responses against SARS-CoV-2 in dried blood spots (DBS) and paired sera.
The ongoing SARS-CoV-2 pandemic was initially managed by non-pharmaceutical interventions such as diagnostic testing, isolation of positive cases, physical distancing and lockdowns. The advent of vaccines has provided crucial protection against SARS-CoV-2. Neutralising antibody (nAb) responses are a key correlate of protection, and therefore measuring nAb responses is essential for monitoring vaccine efficacy. Fingerstick dried blood spots (DBS) are ideal for use in large-scale sero-surveillance because they are inexpensive, offer the option of self-collection and can be transported and stored at ambient temperatures. Such advantages also make DBS appealing to use in resource-limited settings and in potential future pandemics. In this study, nAb responses in sera, venous blood and fingerstick blood stored on filter paper were measured. Samples were collected from SARS-CoV-2 acutely infected individuals, SARS-CoV-2 convalescent individuals and SARS-CoV-2 vaccinated individuals. Good agreement was observed between the nAb responses measured in eluted DBS and paired sera. Stability of nAb responses was also observed in sera stored on filter paper at room temperature for 28 days. Overall, this study provides support for the use of filter paper as a viable sample collection method to study nAb responses.


Human transmissible spongiform encephalopathies in eleven countries: diagnostic pattern across time, 1993-2002.
The objective of this study was to describe the diagnostic panorama of human transmissible spongiform encephalopathies across 11 countries. From data collected for surveillance purposes, we describe annual proportions of deaths due to different human transmissible spongiform encephalopathies in eleven EUROCJD-consortium countries over the period 1993-2002, as well as variations in the use of diagnostic tests. Using logistic models we quantified international differences and changes across time. In general, pre-mortem use of diagnostic investigations increased with time. International differences in pathological confirmation of sporadic Creutzfeldt-Jakob disease, stable over time, were evident. Compared to their counterparts, some countries displayed remarkable patterns, such as: 1) the high proportion, increasing with time, of variant Creutzfeldt-Jakob disease in the United Kingdom, (OR 607.99 95% CI 84.72-4363.40), and France (OR 18.35, 95% CI 2.20-152.83); 2) high, decreasing proportions of iatrogenic Creutzfeldt-Jakob disease in France, (OR 5.81 95% CI 4.09-8.24), and the United Kingdom, (OR 1.54 95% CI 1.03-2.30); and, 3) high and stable ratios of genetic forms in Slovakia (OR 21.82 95% CI 12.42-38.33) and Italy (OR 2.12 95% CI 1.69-2.68). Considerable international variation in aetiological subtypes of human transmissible spongiform encephalopathies was evident over the observation period. With the exception of variant Creutzfeldt-Jakob disease and iatrogenic Creutzfeldt-Jakob disease in France and the United Kingdom, these differences persisted across time.


Unintentional miRNA ablation is a risk factor in gene knockout studies: a short report.
One of the most powerful techniques for studying the function of a gene is to disrupt the expression of that gene using genetic engineering strategies such as targeted recombination or viral integration of gene trap cassettes. The tremendous utility of these tools was recognized this year with the awarding of the Nobel Prize in Physiology or Medicine to Capecchi, Evans, and Smithies for their pioneering work in targeted recombination mutagenesis in mammals. Another noteworthy discovery made nearly a decade ago was the identification of a novel class of non-coding genes called microRNAs. MicroRNAs are among the largest known classes of regulatory elements with more than 1000 predicted to exist in the mouse genome. Over 50% of known microRNAs are located within introns of coding genes. Given that currently about half of the genes in mouse have been knocked out, we investigated the possibility that intronic microRNAs may have been coincidentally deleted or disrupted in some of these mouse models. We searched published murine knockout studies and gene trap embryonic stem cell line databases for cases where a microRNA was located within or near the manipulated genomic loci, finding almost 200 cases where microRNA expression may have been disrupted along with another gene. Our results draw attention to the need for careful planning in future knockout studies to minimize the unintentional disruption of microRNAs. These data also raise the possibility that many knockout studies may need to be reexamined to determine if loss of a microRNA contributes to the phenotypic consequences attributed to loss of a protein-encoding gene.


Estimation of copper in saliva and areca nut products and its correlation with histological grades of oral submucous fibrosis.
The purpose of this study was to estimate the copper levels in saliva of patients with oral submucous fibrosis (OSF) and different areca nut products and its correlation with different histological grades of OSF. The study comprised 60 individuals, 30 OSF patients and 30 non-OSF individuals. Unstimulated whole saliva was collected, and copper analysis was performed using colorimetric method. The commercial areca nut products used by the patients were acquired and subjected to copper analysis through the atomic absorption spectrophotometer method. Oral biopsies were performed for OSF patients for histopathological correlation. The mean salivary copper level was 27.023 μg/dl in OSF patients when compared with 8.393 μg/dl in non-OSF individuals (P < 0.005). The mean copper content in different areca nut products was 13.313 ppm (P < 0.005). Comparison of copper content in different areca nut products with salivary copper levels of OSF patients showed negative correlation (P < 0.853). Comparison of salivary copper levels between different histological grades of OSF yielded a statistically significant association between grades I and III (P < 0.005) and grades II and III OSF (P < 0.019). Comparison of copper content in areca nut products and different histological grades of OSF yielded weak negative statistical correlation (r = -0.116). Despite high copper content in areca nut products, the observations yielded a negative correlation with different histological grades of OSF. This further raises a doubt about the copper content in areca nut as an etiological factor for this crippling disease.


Photonic band structures for surface waves on structured metal surfaces.
Photonic band structures of surface modes on structured metal surfaces with periodic holes in them are obtained using the full-vectorial finite-difference time-domain method. Surface modes do exist for almost all investigated lattice types with any hole size, shape and depth. The results for a square lattice of wax-filled box holes in brass are also in very good agreement with the experimental results. We also show that, for structured surfaces with holes of finite depth, the holes might act as cavities. Thus there exist propagating coupled cavity modes with low group velocity, confined at the surface and decaying exponentially into the dielectric above.


Chemically etched fiber tips for near-field optical microscopy: a process for smoother tips.
An improved method for producing fiber tips for scanning near-field optical microscopy is presented. The improvement consists of chemically etching quartz optical fibers through their acrylate jacket. This new method is compared with the previous one in which bare fibers were etched. With the new process the meniscus formed by the acid along the fiber does not move during etching, leading to a much smoother surface of the tip cone. Subsequent metallization is thus improved, resulting in better coverage of the tip with an aluminum opaque layer. Our results show that leakage can be avoided along the cone, and light transmission through the tip is spatially limited to an optical aperture of a 100-nm dimension.


Tardive Dystonia.
Tardive dyskinesia is a movement disorder that develops after exposure to dopamine receptor blocking agents. Less well-appreciated are other, more recently described tardive syndromes that are phenomenologically distinct from tardive dyskinesia and respond to different treatments. Patients may simultaneously have more than one tardive syndrome. Major subtypes of tardive syndromes include tardive dyskinesia, characterized by orobuccolingual, truncal, or appendicular, choreiform movements; tardive dystonia, characterized by sustained, stereotyped muscle spasms of a twisting or turning character; and tardive akathisia, characterized by an inner sense of restlessness or unease. The sensation often is unpleasant and may be accompanied by repetitive, purposeless movements (stereotypies), such as pacing. Less common tardive syndromes include tardive myoclonus, tardive tourettism, and tardive tremor. Tardive syndromes often are a source of great distress and disability to patients and may be permanent, despite discontinuing the responsible medication. Prevention, early detection, and prompt management are the major clinical focus. When a patient develops a tardive syndrome appropriate actions include 1) review of the primary diagnosis that prompted starting a dopamine receptor blocking agent; 2) characterization of the movement disorder(s); 3) where possible, discontinuation of dopamine blocking agent or replacement with a less potent alternative agent; 4) gradual withdrawl of the offending drug because some patients have an exacerbation of a tardive syndrome after abrupt withdrawal; and 5) assessment of the severity of symptoms and development of a treatment plan based on the phenomenology, with the goal of maximizing patient comfort and function. Although tardive dyskinesia typically develops after chronic exposure to dopamine receptor blocking agents, it, and other variants (such as tardive dystonia) can develop very rapidly after treatment. There seems to be no minimal safe duration of exposure for the development of a tardive syndrome. It is important to recognize that anti-emetics, which are dopamine receptor blockers, such as prochlorperazine, promethazine and metoclopramide, can cause tardive syndromes. Clinicians should become familiar with antipsychotic agents that have a lower risk of causing tardive syndromes, such as clozapine, quetiapine, and olanzapine. We review treatment options for tardive dystonia.


Predicting oral disintegrating tablet formulations by neural network techniques.
Oral disintegrating tablets (ODTs) are a novel dosage form that can be dissolved on the tongue within 3 min or less especially for geriatric and pediatric patients. Current ODT formulation studies usually rely on the personal experience of pharmaceutical experts and trial-and-error in the laboratory, which is inefficient and time-consuming. The aim of current research was to establish the prediction model of ODT formulations with direct compression process by artificial neural network (ANN) and deep neural network (DNN) techniques. 145 formulation data were extracted from Web of Science. All datasets were divided into three parts: training set (105 data), validation set (20) and testing set (20). ANN and DNN were compared for the prediction of the disintegrating time. The accuracy of the ANN model have reached 85.60%, 80.00% and 75.00% on the training set, validation set and testing set respectively, whereas that of the DNN model were 85.60%, 85.00% and 80.00%, respectively. Compared with the ANN, DNN showed the better prediction for ODT formulations. It is the first time that deep neural network with the improved dataset selection algorithm is applied to formulation prediction on small data. The proposed predictive approach could evaluate the critical parameters about quality control of formulation, and guide research and process development. The implementation of this prediction model could effectively reduce drug product development timeline and material usage, and proactively facilitate the development of a robust drug product.


Nuclear ribosomal pseudogenes resolve a corroborated monophyly of the eucalypt genus Corymbia despite misleading hypotheses at functional ITS paralogs.
Divergent paralogs can create both obstacles and opportunities for phylogenetic reconstruction. Phylogenetic relationships among eucalypt genera have been incongruent among datasets in previous studies, where morphological characters supported monophyly of the genus Corymbia, while intergenic spacers of the nuclear ribosomes (ITS) and chloroplast loci (trnL, trnH, psbA) showed Corymbia as either equivocal or paraphyletic. Ribosomal DNA occurs in multiple copies in a genome. We cloned and sequenced the nrITS to investigate if gene duplication was the cause of incongruence among trees in the eucalypts. Three ITS riboforms, two of them widespread, were recovered within some genomes. One of the ITS riboforms recovered a robust phylogeny showing Corymbia as a monophyletic genus, corroborating the evidence from morphology, fossil data, a recent ITS/ETS dataset and microsatellites (SSRs). Compelling evidence suggested that this divergent riboform is a pseudogene, i.e., non-functional paralog: comparatively lower GC content suggesting lower structural stability, deamination-like mutations at potential methylation sites, lack of conserved helices and hairpins and conspicuously lower thermodynamic stability in secondary structures. Phylogenies from the apparently functional riboform retained Corymbia as paraphyletic. We show here that pseudogenes can recover a well-corroborated phylogeny whereas their functional paralogs show misleading hypotheses. We explain that phylogenetic signals may be obscured when functional constraints in ITS necessitate compensatory mutations in the secondary structure helices involved in RNA transcription, whereas pseudogenes mutate under neutrality.


Fracture Resistance and Failure Mode of Custom-made Post-and-cores of Polyetheretherketone and Nano-ceramic Composite.
Custom-made post-and-cores of polyetheretherketone (PEEK) and nano-ceramic composite showed good fracture resistance and failure mode. Clinicians should be aware that these materials might be an efficient alternative for post-and-cores, because they promote esthetics while reducing the risk of root fractures. Purpose: The purpose of this study was to evaluate the fracture resistance and failure mode of custom-made post-and-cores manufactured with different esthetic materials.Methods and Materials: A total of 48 mandibular premolar extracted teeth were selected, endodontically treated, and prepared to receive the posts. Specimens were randomly divided into four groups (n=12/group): group 1, polyetheretherketone (PEEK, Degos; G1); group 2, nano-ceramic composite (Lava Ultimate, 3M ESPE; G2); group 3, cast metal post (NiCr alloy, control; G3); group 4, fiberglass post (Whitepost, FGM, control; G4) customized with a nano-hybrid resin composite (Z250, 3M ESPE). Experimental post-and-cores (G1 and G2) were manufactured with a computer-aided design/computer-aided manufacturing system. All posts were cemented with self-adhesive resin cement (Rely X U200, 3M ESPE), and specimens were stored in distilled water at 37°C for 60 days. A universal testing machine was used to measure the fracture resistance (0.5 mm/min at an angle of 45° to the long axis of the tooth). Fractures were classified as repairable or catastrophic. One-way analysis of variance with Tukey post hoc and χ2 tests for independence and Bonferroni adjustments were applied (α=0.05).Results: The fracture resistance values (mean ± SD, in newtons) were 379.4 ± 119.8 (G1), 506.4 ± 138 (G2), 939.6 ± 146.5 (G3), and 449.6 ± 66.5 (G4). Only G3 exhibited a significant difference (p<0.05). The χ2 test showed an association between failure mode and post-and-core material [χ2(3) = 23.65; p<0.001]. After Bonferroni adjustment, only G3 presented a higher than expected incidence of catastrophic failures (p<0.001). In the remaining groups, most failures were repairable and related to debonding.Conclusions: Customized post-and-cores of PEEK and nano-ceramic composites exhibited good mechanical performance. Their fracture resistance was comparable to that observed for fiberglass customized posts, yet lower than that for cast metal posts. For PEEK post-and-cores, in particular, additional studies are needed to improve adhesiveness and reduce the risk of debonding.


Cardiomyocyte ageing and cardioprotection: consensus document from the ESC working groups cell biology of the heart and myocardial function.
Advanced age is a major predisposing risk factor for the incidence of coronary syndromes and comorbid conditions which impact the heart response to cardioprotective interventions. Advanced age also significantly increases the risk of developing post-ischaemic adverse remodelling and heart failure after ischaemia/reperfusion (IR) injury. Some of the signalling pathways become defective or attenuated during ageing, whereas others with well-known detrimental consequences, such as glycoxidation or proinflammatory pathways, are exacerbated. The causative mechanisms responsible for all these changes are yet to be elucidated and are a matter of active research. Here, we review the current knowledge about the pathophysiology of cardiac ageing that eventually impacts on the increased susceptibility of cells to IR injury and can affect the efficiency of cardioprotective strategies.


Sequence-specific DNA binding activity of the cross-brace zinc finger motif of the piggyBac transposase.
The piggyBac transposase (PB) is distinguished by its activity and utility in genome engineering, especially in humans where it has highly promising therapeutic potential. Little is known, however, about the structure-function relationships of the different domains of PB. Here, we demonstrate in vitro and in vivo that its C-terminal Cysteine-Rich Domain (CRD) is essential for DNA breakage, joining and transposition and that it binds to specific DNA sequences in the left and right transposon ends, and to an additional unexpectedly internal site at the left end. Using NMR, we show that the CRD adopts the specific fold of the cross-brace zinc finger protein family. We determine the interaction interfaces between the CRD and its target, the 5'-TGCGT-3'/3'-ACGCA-5' motifs found in the left, left internal and right transposon ends, and use NMR results to propose docking models for the complex, which are consistent with our site-directed mutagenesis data. Our results provide support for a model of the PB/DNA interactions in the context of the transpososome, which will be useful for the rational design of PB mutants with increased activity.


Creating and using real-world evidence to answer questions about clinical effectiveness.
New forms of evidence are needed to complement evidence generated from randomised controlled trials (RCTs). Real-World Evidence (RWE) is a potential new form of evidence, but remains undefined. This paper sets to fill that gap by defining RWE as the output from a rigorous research process which: (1) includes a clear a priori statement of a hypothesis to be tested or research question to be answered; (2) defines the data sources that will be used and critically appraises their strengths and weaknesses; and (3) applies appropriate methods, including advanced analytics. These elements should be set out in advance of the study commencing, ideally in a published protocol.The strengths of RWE studies are that they are more inclusive than RCTs and can enable an evidence base to be developed around real-world effectiveness and to start to address the complications of managing other real-world problems such as multimorbidity. Computerised medical record systems and big data provide a rich source of data for RWE studies. However, guidance is needed to help assess the rigour of RWE studies so that the strength of recommendations based on their output can be determined. Additionally, RWE advanced analytics methods need better categorisation and validation.We predict that the core role of RCTs will shift towards assessing safety and achieving regulatory compliance. RWE studies, notwithstanding their limitations, may become established as the best vehicle to assess efficacy.


Elevated coronary whole blood viscosity in acute coronary syndrome patients.
As most clinical studies measure whole blood viscosity (WBV) from peripheral samples, potential differences in WBV obtained from the coronary arteries are often ignored. This study investigated differences in WBV measured from coronary artery specimens in patients with and without acute coronary syndrome (ACS). Consecutive patients with chest pain who underwent diagnostic coronary angiography were divided into two groups [non-ACS (n = 16), ACS (n = 22)]. The ACS group consisted of unstable angina (n = 13) and acute myocardial infarction (n = 9) patients. Two blood samples were obtained from each patient at the both coronary artery ostia prior to coronary angiography. Low-shear and high-shear blood viscosities (BVs) were measured at shear rates of 1 and 300 s-1, respectively, by a scanning capillary tube viscometer (Bio-Visco Inc., South Korea). Both low-shear and high-shear BVs obtained from peripheral, left and right coronary arteries were not different in both groups. Mean coronary low-shear WBV values obtained in ACS group were 29.2% higher than those in non-ACS group (295.3 ± 87.2 mP vs. 228.5 ± 69.2 mP, p = 0.016). Mean coronary high-shear WBV values obtained in ACS group were 15.6% higher than those in non-ACS group (42.9 ± 10.0 mP vs. 37.1 ± 4.6 mP, p = 0.036). Direct measurement of WBV from the coronary artery showed no differences with peripheral samples. Future larger studies are needed to clarify our results.


Diversity-oriented generation and biological evaluation of new chemical scaffolds bearing a 2,2-dimethyl-2H-chromene unit: Discovery of novel potent ANO1 inhibitors.
Chemical territory bearing a 2,2-dimethyl-2H-chromene motif was expanded by utilizing an o-hydroxy aldehyde group of 5-hydroxy-2,2-dimethyl-2H-chromene-6-carbaldehyde as a synthetic handle to install distinctive morphology and functionality of each scaffold. Cell based assays and in silico docking analysis led us to discover that these new compounds exhibit inhibitory effect on anoctamin1 (ANO1). ANO1 is amplified and highly expressed in various carcinomas including prostate cancer, esophageal cancer, breast cancer, and pancreatic cancer. Biological assays revealed that (E)-1-(7,7-dimethyl-7H-furo[2,3-f]chromen-2-yl)-3-(1H-pyrrol-2-yl)prop-2-en-1-one (3n, Ani-FCC) is a novel, potent and selective ANO1 inhibitor with an IC50 value of 1.23 μM. 3n showed 144 times stronger activity on ANO1 inhibition than ANO2 inhibition and did not alter the chloride channel activity of CFTR and the intracellular calcium signaling. Notably, 3n strongly decreased cell viability of PC-3 and FaDu cells expressing high levels of ANO1 with a decrease in ANO1 protein levels. In addition, 3n significantly enhanced apoptosis via activation of caspase 3 and cleavage of PARP in PC-3 and FaDu cells. This study shows that a novel ANO1 inhibitor, 3n, can be a potential candidate for the treatment of cancers overexpressing ANO1, such as prostate cancer and esophageal cancer.


Effect of anaerobic digestion and liming on plant availability of phosphorus in iron- and aluminium-precipitated sewage sludge from primary wastewater treatment plants.
More efficient plant utilisation of the phosphorus (P) in sewage sludge is required because rock phosphate is a limited resource. To meet environmental legislation thresholds for P removal from wastewater (WW), primary treatment with iron (Fe) or aluminium (Al) coagulants is effective. There is also a growing trend for WW treatment plants (WWTPs) to be coupled to a biogas process, in order to co-generate energy. The sludge produced, when stabilised, is used as a soil amendment in many countries. This study examined the effects of anaerobic digestion (AD), with or without liming as a post-treatment, on P release from Fe- and Al-precipitated sludges originating from primary WWTPs. Plant uptake of P from Fe- and Al-precipitated sludge after lime treatment but without AD was also compared. Chemical characterisation with sequential extraction of P and a greenhouse experiment with barley (Hordeum vulgare) were performed to assess the treatment effects on plant-available P. Liming increased the P-labile fraction in all cases. Plant P uptake increased from 18.5 mg pot-1 to 53 mg P pot-1 with liming of Fe-precipitated sludge and to 35 mg P pot-1 with liming of the digestate, while it increased from 18.7 mg pot-1 to 39 and 29 mg P pot-1 for the Al-precipitated substrate and digestate, respectively. Thus, liming of untreated Fe-precipitated sludge and its digestate resulted in higher P uptake than liming its Al-precipitated counterparts. AD had a negative impact on P mobility for both sludges.


Sequence and genome organization of a human small round-structured (Norwalk-like) virus.
Small round-structured viruses (SRSVs), also known as Norwalk or Norwalk-like viruses, are the major worldwide cause of acute, epidemic nonbacterial gastroenteritis in humans. These viruses, which contain a single-stranded RNA genome, have remained refractory to molecular characterization because of the small amounts of virus in clinical samples and the absence of an animal model and an in vitro culture system. The complete genomic nucleotide sequence of an SRSV, Southampton virus, was determined. The 7696-nucleotide RNA genome encodes three open reading frames whose sequences and organization strongly support proposals that SRVSs are members of the Caliciviridae.


The diversion and injection of a buprenorphine-naloxone soluble film formulation.
We compared the diversion and injection of a new formulation of buprenorphine, a buprenorphine-naloxone film product (BNX film), with buprenorphine-naloxone tablets (BNX tablets), mono-buprenorphine (BPN) and methadone (MET) in Australia. Surveys were conducted with people who inject drugs regularly (PWID) (2004-2012) and opioid substitution treatment (OST) clients (2012, N=543). Key outcome measures: the unsanctioned removal of supervised doses, diversion, injection, motivations, drug liking and street price. Levels of injection among PWID were adjusted for background availability of medication using sales data. Doses not taken as directed by OST clients were adjusted by total number of daily doses dispensed. Among out-of-treatment PWID, levels of injection for BNX film were comparable to those for MET and BNX tablet formulations, adjusting for background availability; BPN injecting levels were higher. Among OST clients, recent injecting of one's medication was similar among clients in all OST types; weekly or more frequent injection of prescribed doses was reported by fewer BNX film clients (3%; 95% CI: 1-6) than BPN clients (11%; 95% CI: 3-17), but at levels similar to those observed among MET and BNX tablet clients. The proportion of BNX film doses injected was lower than that for BPN and BNX tablets, and equivalent to that for MET. The majority of BNX film doses injected by OST clients were unsupervised doses, although some injection of supervised doses of BNX film did occur. The median price of all buprenorphine forms on the illicit market was the same. Non-adherence and diversion of the BNX film formulation was similar to MET and BNX tablet formulations; BPN had higher levels of all indicators of non-adherence and diversion.


Which patients pursue fertility preservation treatments? A multicenter analysis of the predictors of fertility preservation in women with breast cancer.
To evaluate predictors of undergoing fertility preservation treatment (FPT) in women with breast cancer. Secondary analysis of a clinical database. Three academic fertility preservation centers. One hundred eight patients with breast cancer undergoing FPT and 77 patients with breast cancer not undergoing FPT from 2005 to 2010. None. Patients' demographic and medical information. Women who had FPT were older, wealthier, and had lower cancer stage compared with women who did not have FPT. The rate of the administration of neoadjuvant chemotherapy (NAC) was significantly lower in women who underwent FPT. After adjusting for age, body mass index (BMI), income, cancer stage, and center, a negative correlation persisted between NAC and FPT (odds ratio 0.091, 95% confidence interval 0.009-0.904). When we stratified the women by center, women at center 1 had a significantly lower FPT rate, lower parity, higher BMI, more advanced cancer stage, and lower income compared with centers 2 and 3. The rates of NAC were significantly higher in center 1. Although age, BMI, income, cancer stage, center, and NAC seem to be associated with undergoing FPT, NAC is the only modifiable variable. Because NAC restricts the time available for FPT, oncologists may consider offering adjuvant chemotherapy, except in cases in which NAC clearly improves survival, in women who are interested in FPT.


High-Density Peptide Microarray Analysis of IgG Autoantibody Reactivities in Serum and Cerebrospinal Fluid of Multiple Sclerosis Patients.
Intrathecal immunoglobulin G (IgG) synthesis and oligoclonal IgG bands in cerebrospinal fluid (CSF) are hallmarks of multiple sclerosis (MS), but the antigen specificities remain enigmatic. Our study is the first investigating the autoantibody repertoire in paired serum and CSF samples from patients with relapsing-remitting MS (RRMS), primary progressive MS (PPMS), and other neurological diseases by the use of high-density peptide microarrays. Protein sequences of 45 presumed MS autoantigens (e.g.MOG, MBP, and MAG) were represented on the microarrays by overlapping 15mer peptides. IgG reactivities were screened against a total of 3991 peptides, including also selected viral epitopes. The measured antibody reactivities were highly individual but correlated for matched serum and CSF samples. We found 54 peptides to be recognized significantly more often by serum or CSF antibodies from MS patients compared with controls (pvalues <0.05). The results for RRMS and PPMS clearly overlapped. However, PPMS patients presented a broader peptide-antibody signature. The highest signals were detected for a peptide mapping to a region of the Epstein-Barr virus protein EBNA1 (amino acids 392-411), which is homologous to the N-terminal part of human crystallin alpha-B. Our data confirmed several known MS-associated antigens and epitopes, and they delivered additional potential linear epitopes, which await further validation. The peripheral and intrathecal humoral immune response in MS is polyspecific and includes antibodies that are also found in serum of patients with other diseases. Further studies are required to assess the pathogenic relevance of autoreactive and anti-EBNA1 antibodies as well as their combinatorial value as biomarkers for MS.


Cooperative Game-Based Energy Efficiency Management over Ultra-Dense Wireless Cellular Networks.
Ultra-dense wireless cellular networks have been envisioned as a promising technique for handling the explosive increase of wireless traffic volume. With the extensive deployment of small cells in wireless cellular networks, the network spectral efficiency (SE) is improved with the use of limited frequency. However, the mutual inter-tier and intra-tier interference between or among small cells and macro cells becomes serious. On the other hand, more chances for potential cooperation among different cells are introduced. Energy efficiency (EE) has become one of the most important problems for future wireless networks. This paper proposes a cooperative bargaining game-based method for comprehensive EE management in an ultra-dense wireless cellular network, which highlights the complicated interference influence on energy-saving challenges and the power-coordination process among small cells and macro cells. Especially, a unified EE utility with the consideration of the interference mitigation is proposed to jointly address the SE, the deployment efficiency (DE), and the EE. In particular, closed-form power-coordination solutions for the optimal EE are derived to show the convergence property of the algorithm. Moreover, a simplified algorithm is presented to reduce the complexity of the signaling overhead, which is significant for ultra-dense small cells. Finally, numerical simulations are provided to illustrate the efficiency of the proposed cooperative bargaining game-based and simplified schemes.