Rational redesign of the 4-chlorobenzoate binding site of 4-chlorobenzoate: coenzyme a ligase for expanded substrate range.
Environmental aromatic acids are transformed to chemical energy in bacteria that possess the requisite secondary pathways. Some of these pathways rely on the activation of the aromatic acid by coenzyme A (CoA) thioesterification catalyzed by an aromatic acid: CoA ligase. Adaptation of such pathways to the bioremediation of man-made pollutants such as polychlorinated biphenyl (PCB) and dichlorodiphenyltrichloroethane (DDT) requires that the chlorinated benzoic acid byproduct that is formed be able to be eliminated by further degradation. To take advantage of natural benzoic acid degrading pathways requiring initial ring activation by thioesterification, the pathway aromatic acid:CoA ligase must be an effective catalyst with the chlorinated benzoic acid. This study, which focuses on the 4-chlorobenzoate:CoA ligase (CBL) of the 4-monochlorobiphenyl degrading bacterium Alcaligenes sp. strain ALP83, was carried out to determine if the 4-chlorobenzoate binding site of this enzyme can be transformed by rational design to recognize the chlorobenzoic acids formed in the course of breakdown of other environmental PCB congeners. The fundamental question addressed in this study is whether it is possible to add or subtract space from the substrate-binding pocket of this ligase (to complement the topology of the unnatural aromatic substrate) without causing disruption of the ligase catalytic machinery. Herein, we report the results of a substrate specificity analysis that, when interpreted within the context of the X-ray crystal structures, set the stage for the rational design of the ligase for thioesterification of two PCB-derived chlorobenzoic acids. The ligase was first optimized to catalyze CoA thioesterification of 3,4-dichlorobenzoic acid, a poor substrate, by truncating Ile303, a large hydrophobic residue that packs against the ring meta-C(H) group. The structural basis for the approximately 100-fold enhancement in the rate of 3,4-dichlorobenzoate thioesterification catalyzed by the I303A and I303G CBL mutants was validated by determination of the crystal structure of the 3,4-dichlorobenzoate-bound enzymes. Determinations of the structures of I303 mutant complexes of 3-chlorobenzoate, a very poor substrate, revealed nonproductive binding as a result of the inability of the substrate ring C(4)H group to fill the pocket that binds the C(4)Cl group of the native substrate. The C(4)Cl pocket of the CBL I303A mutant was then reduced in size by strategic amino acid replacement. A 54-fold improvement in catalytic efficiency was observed for the CBL F184W/I303A/V209T triple mutant. The results of this investigation are interpreted as evidence that the plasticity of the ligase catalytic scaffold is sufficient to allow expansion of substrate range by rational design. The combination of structural and kinetic analyses of the constructed mutants proved to be an effective approach to engineering the ligase for novel substrates.


Functional consequences of the creation of an Asp-His-Fe triad in a 3/3 globin.
The proximal side of dehaloperoxidase-hemoglobin A (DHP A) from Amphitrite ornata has been modified via site-directed mutagenesis of methionine 86 into aspartate (M86D) to introduce an Asp-His-Fe triad charge relay. X-ray crystallographic structure determination of the metcyano forms of M86D [Protein Data Bank (PDB) entry 3MYN ] and M86E (PDB entry 3MYM ) mutants reveal the structural origins of a stable catalytic triad in DHP A. A decrease in the rate of H(2)O(2) activation as well as a lowered reduction potential versus that of the wild-type enzyme was observed in M86D. One possible explanation for the significantly lower activity is an increased affinity for the distal histidine in binding to the heme Fe to form a bis-histidine adduct. Resonance Raman spectroscopy demonstrates a pH-dependent ligation by the distal histidine in M86D, which is indicative of an increased trans effect. At pH 5.0, the heme Fe is five-coordinate, and this structure resembles the wild-type DHP A resting state. However, at pH 7.0, the distal histidine appears to form a six-coordinate ferric bis-histidine (hemichrome) adduct. These observations can be explained by the effect of the increased positive charge on the heme Fe on the formation of a six-coordinate low-spin adduct, which inhibits the ligation and activation of H(2)O(2) as required for peroxidase activity. The results suggest that the proximal charge relay in peroxidases regulate the redox potential of the heme Fe but that the trans effect is a carefully balanced property that can both activate H(2)O(2) and attract ligation by the distal histidine. To understand the balance of forces that modulate peroxidase reactivity, we studied three M86 mutants, M86A, M86D, and M86E, by spectroelectrochemistry and nuclear magnetic resonance spectroscopy of (13)C- and (15)N-labeled cyanide adducts as probes of the redox potential and of the trans effect in the heme Fe, both of which can be correlated with the proximity of negative charge to the N(δ) hydrogen of the proximal histidine, consistent with an Asp-His-Fe charge relay observed in heme peroxidases.


Landscape-scaled strategies can outperform Lévy random searches.
Information on the relevant global scales of the search space, even if partial, should conceivably enhance the performance of random searches. Here we show numerically and analytically that the paradigmatic uninformed optimal Lévy searches can be outperformed by informed multiple-scale random searches in one (1D) and two (2D) dimensions, even when the knowledge about the relevant landscape scales is incomplete. We show in the low-density nondestructive regime that the optimal efficiency of biexponential searches that incorporate all key scales of the 1D landscape of size L decays asymptotically as η_{opt}∼1/sqrt[L], overcoming the result η_{opt}∼1/(sqrt[L]lnL) of optimal Lévy searches. We further characterize the level of limited information the searcher can have on these scales. We obtain the phase diagram of bi- and triexponential searches in 1D and 2D. Remarkably, even for a certain degree of lack of information, partially informed searches can still outperform optimal Lévy searches. We discuss our results in connection with the foraging problem.


Pluronic-based micelle encapsulation potentiates myricetin-induced cytotoxicity in human glioblastoma cells.
As one of the natural herbal flavonoids, myricetin has attracted much research interest, mainly owing to its remarkable anticancer properties and negligible side effects. It holds great potential to be developed as an ideal anticancer drug through improving its bioavailability. This study was performed to investigate the effects of Pluronic-based micelle encapsulation on myricetin-induced cytotoxicity and the mechanisms underlying its anticancer properties in human glioblastoma cells. Cell viability was assessed using a methylthiazol tetrazolium assay and a real-time cell analyzer. Immunoblotting and quantitative reverse transcriptase polymerase chain reaction techniques were used for determining the expression levels of related molecules in protein and mRNA. The results indicated that myricetin-induced cytotoxicity was highly potentiated by the encapsulation of myricetin. Mitochondrial apoptotic pathway was demonstrated to be involved in myricetin-induced glioblastoma cell death. The epidermal growth factor receptor (EGFR)/PI3K/Akt pathway located in the plasma membrane and cytosol and the RAS-ERK pathway located in mitochondria served as upstream and downstream targets, respectively, in myricetin-induced apoptosis. MiR-21 inhibitors interrupted the expression of EGFR, p-Akt, and K-Ras in the same fashion as myricetin-loaded mixed micelles (MYR-MCs) and miR-21 expression were dose-dependently inhibited by MYR-MCs, indicating the interaction of miR-21 with MYR-MCs. This study provided evidence supportive of further development of MYR-MC formulation for preferentially targeting mitochondria of glioblastoma cells.


Comparison of the effects of chronic desmethylimipramine administration on alpha 2- and beta-adrenoceptors in different regions of rat brain.
The effects of ten once-daily injections of desmethylimipramine (10 mg/kg i.p.) on alpha 2- and beta-adrenoceptor binding were investigated in the rat cortex, hippocampus and hypothalamus. [3H]clonidine and [3H]dihydroalprenolol were used as ligands for alpha 2- and beta-adrenoceptors, respectively. Twenty-four hours after the last injection, the density of beta-adrenoceptors was reduced in all regions studied. In contrast, there was no change in alpha 2-adrenoceptor binding in the cortex, while increased binding was found in the hypothalamus. The effects of desmethylimipramine are discussed in relation to those of other antidepressant treatments.


Incidence of heat-labile enterotoxin-producing Escherichia coli detected by means of polymerase chain reaction amplification.
Diarrhoea can be caused by many different organisms, some of which are notoriously difficult to identify. One of these is enterotoxin-producing Escherichia coli. Recently a new diagnostic technique that uses polymerase chain reaction DNA amplification was developed for detection of the 'A' subunit of the labile enterotoxin-producing E. coli gene. This technique was used to evaluate the incidence of heat-labile (LT+) enterotoxin-producing E. coli in the causation of diarrhoea. The results from this study showed that LT+ E. coli is a cause of diarrhoea in the western Cape and that 5.3% of non-diagnosed diarrhoea patients in Tygerberg Hospital were infected with this pathogen. This represented less than 1% of the total number of cases of diarrhoea investigated in this hospital. The peak coincides with the wetter months in this locality and the infection rate is lower than that reported in most other countries. Given the low incidence of occurrence of this organism we do not recommend routine implementation of the diagnostic procedure. However, this test may be useful at times, e.g. to ascertain the source of a diarrhoea epidemic.


Aldosterone deficiency in mice burdens respiration and accentuates diet-induced hyperinsulinemia and obesity.
Aldosterone synthase inhibitors (ASIs) should alleviate obesity-related cardiovascular and renal problems resulting partly from aldosterone excess, but their clinical use may have limitations. To improve knowledge for the use of ASIs, we investigated physiology in aldosterone synthase-knockout (ASKO) mice. On regular chow diet (CD), ASKO mice ate more and weighed less than WT mice, largely because they hyperventilated to eliminate acid as CO2. Replacing CD with high-fat diet (HFD) lessened the respiratory burden in ASKO mice, as did 12- to 15-hour fasting. The latter eliminated the genotype differences in respiratory workload and energy expenditure (EE). Thus, aldosterone deficiency burdened the organism more when the animals ate carbohydrate-rich chow than when they ate a HFD. Chronic HFD exposure further promoted hyperinsulinemia in ASKO mice that contributed to visceral fat accumulation accompanied by reduced lipolysis, thermogenic reprogramming, and the absence of weight-gain-related EE increases. Intracerebroventricular aldosterone supplementation in ASKO mice attenuated the HFD-induced hyperinsulinemia, but did not affect EE, suggesting that the presence of aldosterone increased the body's energetic efficiency, thus counteracting the EE-increasing effect of low insulin. ASIs may therefore cause acid-overload-induced respiratory burden and promote obesity. Their use in patients with preexisting renal and cardiopulmonary diseases might be contraindicated.


Polyamine depletion with two different polyamine analogues causes DNA damage in human breast cancer cell lines.
It is well known that the positively charged polyamines have a DNA-stabilizing function and that polyamine depletion alters chromatin function. We have previously shown that polyamine depletion causes an S phase prolongation, and others have shown that there is an accumulation of Okazaki-like fragments in polyamine-depleted cells. In the present study, we have used the comet assay to investigate polyamine depletion-induced DNA strand breaks. Three breast cancer cell lines and one normal-like breast cell line were treated with the polyamine analogue N(1),N(11)-diethylnorspermine or with the polyamine biosynthesis inhibitor 4-amidinoindan-1-one 2'-amidinohydrazone (CGP 48664). The comet assay showed that polyamine depletion resulted in DNA strand breaks. We also show that these DNA strand breaks occurred in cells where there was no expression of gamma-H2AX, which is a marker of DNA double-strand breaks. Thus, our conclusion is that polyamine depletion causes DNA single-strand breaks, which may be the cause for the observed delay in S phase progression.


Structural organization of interphase 3T3 fibroblasts studied by total internal reflection fluorescence microscopy.
We studied the laminar organization of 3T3 fibroblast cells growing on glass slides by use of total internal reflection illumination to excite fluorescence emission (TIRF) from labeled molecules and stained cellular compartments that are very close to the cell-substrate contact region. Mitochondria, distant from the contact regions and stained with the water-soluble cationic dye, dil-C3-(3), fluoresced only as the glass/cytoplasm critical angle was approached. A similar result was obtained when the nuclei were stained with Hoechst dye 33342. From this measured angle a cytoplasmic refractive index in the range 1.358-1.374 was computed. The plasma membrane of 3T3 cells was stained with dil-C18-(3), and the cytoplasmic compartment was stained with fluoresceinyl-dextran (FTC-dextran) or with carboxyfluorescein. We have demonstrated a high degree of correspondence between the low-reflectance zones in the reflection interference image of a live cell and the TIRF images of both the plasma membrane and cytoplasmic compartment. TIRF photometry of selected contact regions of cells provided data from which the absolute separation of cell and substrate was computed. From a population of 3T3 cells microinjected with fluorescein-labeled actin, motile and adherent interphase cells were selected for study. For adherent cells, which displayed fluorescent stress fibers, the TIRF image was composed of intense patches and less intense regions that corresponded, respectively, to the focal contact and close-contact zones of the reflection-interference image. The intense patches corresponded to the endpoints of the stress fibers. Cells of motile morphology, which formed some focal contacts and extensive close-contact zones, gave AF-actin TIRF images of relatively even intensity. Thin lamellar regions of the cytoplasm were found to contain concentrations of actin not significantly different from other close-contact regions of the cell. The major analytical problem of TIRF microscopy is separation of the effects of proximity to substrate, refractive index, and fluorescent probe concentration on the local brightness of the TIRF image. From our results, it appears possible to use TIRF microscopy to measure the proximity of different components of substrate contact regions of cells.


Sites and homeostatic control of auxin biosynthesis in Arabidopsis during vegetative growth.
The distribution and biosynthesis of indole-3-acetic acid (IAA) was investigated during early plant development in Arabidopsis. The youngest leaves analysed, less than 0.5 mm in length, contained 250 pg mg(-1) of IAA and also exhibited the highest relative capacity to synthesize this hormone. A decrease of nearly one hundred-fold in IAA content occurred as the young leaves expanded to their full size, and this was accompanied by a clear shift in both pool size and IAA synthesis capacity. The correlation between high IAA content and intense cell division was further verified in tobacco leaves, where a detailed analysis revealed that dividing mesophyll tissue contained ten-fold higher IAA levels than tissue growing solely by elongation. We demonstrated that all parts of the young Arabidopsis plant can potentially contribute to the auxin needed for growth and development, as not only young leaves, but also all other parts of the plant such as cotyledons, expanding leaves and root tissues have the capacity to synthesize IAA de novo. We also observed that naphthylphthalamic acid (NPA) treatment induced tissue-dependent feedback inhibition of IAA biosynthesis in expanding leaves and cotyledons, but intriguingly not in young leaves or in the root system. This observation supports the hypothesis that there is a sophisticated tissue-specific regulatory mechanism for auxin biosynthesis. Finally, a strict requirement for maintaining the pool sizes of IAA was revealed as reductions in leaf expansion followed both decreases and increases in the IAA levels in developing leaves. This indicates that leaves are not only important sources for IAA synthesis, but that normal leaf expansion depends on rigorous control of IAA homeostasis.


Effectiveness of planning and management interventions for improving age-appropriate immunization in rural India.
To study the effectiveness of planning and management interventions for ensuring children in India are immunized at the appropriate age. The study involved children aged less than 18 months recruited from Haryana, India, in 2005-2006: 4336 in a pre-intervention cohort and 5213 in a post-intervention cohort. In addition, immunization of 814 hospitalized children from outside the study area was also assessed. Operational barriers to age-appropriate immunization with diphtheria, pertussis and tetanus (DPT) vaccine were investigated by monitoring vaccination coverage, observing immunization sessions and interviewing parents and health-care providers. An intervention package was developed, with community volunteers playing a pivotal role. Its effectiveness was assessed by monitoring the ages at which the three DPT doses were administered. The main reasons for delayed immunization were staff shortages, non-adherence to plans and vaccine being out of stock. In the post-intervention cohort, 70% received a third DPT dose before the age of 6 months, significantly more than in the pre-intervention cohort (62%; P = 0.002). In addition, the mean age at which the first, second and third DPT doses were administered decreased by 17, 21 and 34 days, respectively, in the study area over a period of 18 months (P for trend < 0.0001). No change was observed in hospitalized children from outside the study area. An intervention package involving community volunteers significantly improved age-appropriate DPT immunization in India. The Indian Government's intention to recruit village-based volunteers as part of a health sector reform aimed at decentralizing administration could help increase timely immunization.


Our experience with transoral incisionless plication of gastroesophageal reflux disease: NOTES procedure.
Natural Orifice Transluminal Surgery (NOTES) has been introduced in endoscopic surgery as a new system offering the advantage of a less invasive procedure. Gastroesophageal reflux disease (GERD) appears to be the most promising application of NOTES treatment. The aims of our study were to evaluate the safety and efficacy of this procedure and length of hospital stay. Patients indicated for surgery of GERD were randomly assigned (ratio 2:1) to transoral incisionless fundoplication (TIF group, n=34) and control group, where gold standard Nissen laparoscopic fundoplication was performed (NLF group, n=18). For TIF the Plicator® method was initially used for 18 patients, but the company terminated production in 2008 without a follower. During the last 2 years the EsophyX® method was used for 16 patients. After the evaluation of 34 TIF patients and 18 NLF patients we observed similar efficacy of TIF procedures compared with NLF after 3 and 12 months. The hospital stay was significantly shorter (p<0.0001) in TIF group (average, 2.9±0.8 days) than in NLF group (6.4±0.7). The TIF procedure was safe; we observed 1 serious adverse event in the TIF group and 3 in the NLF group. It can be summarized that both NOTES TIF procedures are, after the initial learning curve, safe and effective methods for treatment of GERD, allowing substantial shortening of hospital stay. The effect of both procedures was sustained over 12 months. Longer follow-up is necessary to verify efficacy for more years.


Political priority of oral health in India: analysis of the reasons for neglect.
To examine the political priority of oral health in India and to understand the underlying reasons for the political support oral health receives. The analysis is based on the political power framework developed by Shiffman and Smith and modified by Benzian et al. to examine the factors that contribute to the political priority of oral health in India. The framework comprises four main analysis categories, further subdivided into 11 dimensions. Based on the set of criteria, each dimension was analyzed and rated by assigning a score to assess to what extend the criteria were met. There is a good understanding on what defines an oral health problem, however, there is no consolidated and comprehensive approach to address oral diseases. Despite India's efforts to improve oral health-related research, its poor utilization in terms of public health and population-based approaches is apparent. The absence of a national surveillance system for oral health masks the severity and extent of the oral disease burden and limits the basis for advocacy on improving oral health to health decision makers. The fragmentation of actors and institutions and the absence of leaders uniting various actors in oral public health impede changes toward improving the oral health status of the population. Limited accessibility to oral health care, poor portrayals of the severity and extent of the burden, and inertia to address-related challenges are important factors contributing to the low political priority of oral health.


Mechanisms of action of peripherally administered cholecystokinin octapeptide on brain stem neurons in the rat.
We have investigated the pathway and the mechanism by which cholecystokinin octapeptide (CCK-8), given systemically, may influence the discharge of brain stem neurons that have an input from the stomach. Extracellular recordings were made from neurons in the nucleus of the solitary tract (NTS), where vagal afferents terminate, and from neighboring regions of the dorsal medial medulla. Gastric distension and CCK-8 injected intra-aortically close to the stomach evoked either excitatory or inhibitory responses that were abolished by cervical vagal section. In animals from which the celiac/superior mesenteric ganglia were removed, or the gastric antrum resected 2 weeks earlier, responses to gastric distension and CCK-8 were maintained. The effects of CCK-8 are unlikely to be secondary to changes in smooth muscle tone because CCK-8 decreased pressure in the body of the stomach, while distension increased it. Moreover, intravenous noradrenaline and vasoactive intestinal peptide had effects similar to CCK-8 on intragastric pressure, but evoked different patterns of responses from brain stem neurons. The results are consistent with the idea that CCK-8 acts directly on vagal mechanoreceptive endings in the gastric corpus wall. It is well known that peripheral administration of CCK-8 influences short-term regulation of food intake. The effects described here may reflect the pathway by which peripheral CCK influences CNS function.


Cytologic appearance of hibernoma in two dogs.
Hibernomas are rare benign tumors of brown fat (adipose tissue) that have been reported in several different species. The cytologic characterization of these tumors has not been described in dogs. In this case report, we describe two dogs with hibernomas, focusing on the cytologic appearance of these unique neoplasms. Both cytologic specimens were highly cellular and predominated by vacuolated neoplastic cells with no evidence of concurrent inflammation. The cells contained a moderate to large number of variably sized cytoplasmic vacuoles, with occasional, irregularly shaped pink granular material. Most cells contained a single nucleus; however, cells displayed moderate anisokaryosis. A biopsy with histologic examination was performed in both cases, confirming the cytologic suspicion of hibernoma. Immunohistochemistry revealed that both tumors were positive for UCP1 and vimentin, and negative for cytokeratin. Hibernoma is an important differential diagnosis in dogs with conjunctival and periocular swellings that exfoliate numerous, mildly atypical, vacuolated cells.


A Chest Patch for Continuous Vital Sign Monitoring: Clinical Validation Study During Movement and Controlled Hypoxia.
The standard of care in general wards includes periodic manual measurements, with the data entered into track-and-trigger charts, either on paper or electronically. Wearable devices may support health care staff, improve patient safety, and promote early deterioration detection in the interval between periodic measurements. However, regulatory standards for ambulatory cardiac monitors estimating heart rate (HR) and respiratory rate (RR) do not specify performance criteria during patient movement or clinical conditions in which the patient's oxygen saturation varies. Therefore, further validation is required before clinical implementation and deployment of any wearable system that provides continuous vital sign measurements. The objective of this study is to determine the agreement between a chest-worn patch (VitalPatch) and a gold standard reference device for HR and RR measurements during movement and gradual desaturation (modeling a hypoxic episode) in a controlled environment. After the VitalPatch and gold standard devices (Philips MX450) were applied, participants performed different movements in seven consecutive stages: at rest, sit-to-stand, tapping, rubbing, drinking, turning pages, and using a tablet. Hypoxia was then induced, and the participants' oxygen saturation gradually reduced to 80% in a controlled environment. The primary outcome measure was accuracy, defined as the mean absolute error (MAE) of the VitalPatch estimates when compared with HR and RR gold standards (3-lead electrocardiography and capnography, respectively). We defined these as clinically acceptable if the rates were within 5 beats per minute for HR and 3 respirations per minute (rpm) for RR. Complete data sets were acquired for 29 participants. In the movement phase, the HR estimates were within prespecified limits for all movements. For RR, estimates were also within the acceptable range, with the exception of the sit-to-stand and turning page movements, showing an MAE of 3.05 (95% CI 2.48-3.58) rpm and 3.45 (95% CI 2.71-4.11) rpm, respectively. For the hypoxia phase, both HR and RR estimates were within limits, with an overall MAE of 0.72 (95% CI 0.66-0.78) beats per minute and 1.89 (95% CI 1.75-2.03) rpm, respectively. There were no significant differences in the accuracy of HR and RR estimations between normoxia (≥90%), mild (89.9%-85%), and severe hypoxia (<85%). The VitalPatch was highly accurate throughout both the movement and hypoxia phases of the study, except for RR estimation during the two types of movements. This study demonstrated that VitalPatch can be safely tested in clinical environments to support earlier detection of cardiorespiratory deterioration. ISRCTN Registry ISRCTN61535692; https://www.isrctn.com/ISRCTN61535692.


Altered psychobiological responsiveness in women with irritable bowel syndrome.
Although stress has been considered an important pathophysiological factor in irritable bowel syndrome (IBS), there is incomplete understanding of its physiological mechanisms. The current study was designed to compare diurnal hypothalamic-pituitary-adrenal (HPA) axis activity in IBS patients and controls and their psychobiological response to a psychosocial stressor. Basal and stimulated HPA axis activity was assessed in 57 women with IBS and 20 matched controls. Psychiatric comorbidity was assessed using a standardized clinical interview. Salivary morning cortisol and diurnal profile were obtained, and the Trier Social Stress Test (TSST) was administered. Levels of cortisol and adrenocorticotropic hormone (ACTH) were measured before and within 1 hour after the stressor. Overall stress experience and stress related to the TSST were assessed using standardized questionnaires. All subjects showed intact circadian variation of cortisol. However, IBS patients with predominant diarrhea exhibited substantially heightened cortisol levels at awakening (p < .03) and a blunted cortisol awakening response. In response to the TSST, patients exhibited significantly blunted cortisol (p < .05) and slightly attenuated ACTH secretion compared with controls. During the recovery period, ACTH levels were significantly lower (p < .04) in patients than those in healthy subjects. Women with IBS perceived higher stress susceptibility than control subjects did (p < .01). The enhanced morning cortisol levels in one subgroup of IBS patients may indicate an association between basal HPA axis activity and predominant bowel habit. The downregulated HPA axis reactivity in IBS after the TSST suggests a downregulated sensitivity of the endocrine system. On the contrary, all subjective stress ratings were increased in the IBS group, which may indicate increased stress susceptibility.


Parametric polytope reconstruction, an application to crystal shape estimation.
In situ imaging techniques are a promising direction for monitoring the distribution of crystal sizes and shapes during a crystallization process. Nevertheless, no tractable method yet exists for estimating complex crystal shapes. In this paper, an in situ imaging setup is presented and a novel algorithm for crystal shape estimation from a pair of images is presented. It is shown that such a shape estimation problem can be turned into parametric polytope reconstruction from projections. Based on results in polyhedral geometry, it is demonstrated that an accurate estimate of the crystal shape can be computed by solving a nonlinear least-squares problem built from samples in images and a prior model of the crystal. Effectiveness of the approach is proven on artificial and real images. Results show that very accurate estimations of crystal shapes can be obtained from well-chosen data points sampled on images.


The OmpU outer membrane protein, a potential adherence factor of Vibrio cholerae.
Expression of the OmpU outer membrane protein of Vibrio cholerae is positively regulated by toxR, which also regulates critical virulence factors such as cholera toxin and the toxin-coregulated pilus colonization factor. In this study, we have characterized the 38-kDa OmpU protein and investigated its role in the adhesion of V. cholerae to mammalian cells. The amino-terminal sequence of OmpU has similarity with the sequences of Haemophilus influenzae HMW1 and HMW2 adhesins, which, in turn, also have similarity with the sequence of Bordetella pertussis filamentous hemagglutinin. A monoclonal antibody directed against FHA recognized both V. cholerae OmpU and Escherichia coli OmpA, and polyclonal anti-OmpU antibodies recognized FHA and E. coli OmpA, suggesting the existence of common epitopes among these proteins. OmpU was strongly recognized by convalescent-phase serum from volunteers experimentally infected with virulent V. cholerae strains, indicating that OmpU is immunogenic and produced in vivo. OmpU selectively bound to fibronectin and to an arginine-glycine-asparagine (RGD) tripeptide but not to other matrix glycoproteins tested such as collagen or laminin. Antibodies directed against OmpU or their F(ab)2 fragments completely inhibited adhesion of several V. cholerae strains to HeLa, HEp-2, Caco-2, and Henle 407 epithelial cells and also inhibited intestinal colonization and conferred protection in newborn mice against both biotypes (El Tor and classical) of V. cholerae O1. Collectively, these data indicate that OmpU has adhesive properties which may play a role in the pathogenesis of cholera.


Chemical Proteomics Uncovers EPHA2 as a Mechanism of Acquired Resistance to Small Molecule EGFR Kinase Inhibition.
Tyrosine kinase inhibitors (TKIs) have become an important therapeutic option for treating several forms of cancer. Gefitinib, an inhibitor of the epidermal growth factor receptor (EGFR), is in clinical use for treating non-small cell lung cancer (NSCLC) harboring activating EGFR mutations. However, despite high initial response rates, many patients develop resistance to gefitinib. The molecular mechanisms of TKI resistance often remain unclear. Here, we describe a chemical proteomic approach comprising kinase affinity purification (kinobeads) and quantitative mass spectrometry for the identification of kinase inhibitor resistance mechanisms in cancer cells. We identified the previously described amplification of MET and found EPHA2 to be more than 10-fold overexpressed (p < 0.001) in gefitinib-resistant HCC827 cells suggesting a potential role in developing resistance. siRNA-mediated EPHA2 knock-down or treating cells with the multikinase inhibitor dasatinib restored sensitivity to gefitinib. Of all dasatinib targets, EPHA2 exhibited the most drastic effect (p < 0.001). In addition, EPHA2 knockdown or ephrin-A1 treatment of resistant cells decreased FAK phosphorylation and cell migration. These findings confirm EPHA2 as an actionable drug target, provide a rational basis for drug combination approaches, and indicate that chemical proteomics is broadly applicable for the discovery of kinase inhibitor resistance.


Effect of a two-day extensive continuing medical education course on participants' knowledge of clinical and operative urology.
Continuing Medical Education (CME) is an established method for facilitating the lifelong learning and developing knowledge, skills and attitudes to ensure delivery of a medical care which is up-to-date, evidence based, safe and patient-centered. An extensive 2 day urology course was conducted to meet the needs of learners. The purpose of the current study was to measure the effect of this CME activity on the knowledge of the participants. This quasi-experimental, single group pre-, and post-test study measured the gain in learning as a result of the two-day extensive CME course conducted by the section of Urology at Department of Surgery, the Aga Khan University, Karachi-Pakistan. Gain in knowledge, defined as the difference between the pre-test and the post-test scores, was taken as a measure of course effectiveness. The test comprised of 40 one-best type carefully constructed multiple choice questions (MCQs). Item analyses were also performed. Forty-five out of a total of 70 participants from within and outside the city completed both pre and post-tests and were included in the study. The mean age of the subjects was 33.3+6.7 years. Of these 45 participants, 68.9% (n=31) of them were trainees at different levels. Mean gain in knowledge was 12.7±6.8% (p<0.01; 95% CI: 4.17-5.79). Mean test scores improved significantly from 37.8±11.3% to 50.3±10.8%. Difference in pre and post scores due to age, gender, practice type or years since start of training was not significant. The reliability of the test using Cronbach's α was 0.634. CME sessions when designed and delivered carefully are effective means of increasing the knowledge significantly. Pre- and post-test is a reliable and valid strategy to measure gain in participants' knowledge.


Total and added sugar intakes, sugar types, and cancer risk: results from the prospective NutriNet-Santé cohort.
Excessive sugar intake is now recognized as a key risk factor for obesity, type 2 diabetes, and cardiovascular diseases. In contrast, evidence on the sugar-cancer link is less consistent. Experimental data suggest that sugars could play a role in cancer etiology through obesity but also through inflammatory and oxidative mechanisms and insulin resistance, even in the absence of weight gain. The objective was to study the associations between total and added sugar intake and cancer risk (overall, breast, and prostate), taking into account sugar types and sources. In total, 101,279 participants aged >18 y (median age, 40.8 y) from the French NutriNet-Santé prospective cohort study (2009-2019) were included (median follow-up time, 5.9 y). Sugar intake was assessed using repeated and validated 24-h dietary records, designed to register participants' usual consumption for >3500 food and beverage items. Associations between sugar intake and cancer risk were assessed by Cox proportional hazard models adjusted for known risk factors (sociodemographic, anthropometric, lifestyle, medical history, and nutritional factors). Total sugar intake was associated with higher overall cancer risk (n = 2503 cases; HR for quartile 4 compared with quartile 1: 1.17; 95% CI: 1.00, 1.37; Ptrend = 0.02). Breast cancer risks were increased (n = 783 cases; HRQ4vs.Q1 = 1.51; 95% CI: 1.14, 2.00; Ptrend = 0.0007). Results remained significant when weight gain during follow-up was adjusted for. In addition, significant associations with cancer risk were also observed for added sugars, free sugars, sucrose, sugars from milk-based desserts, dairy products, and sugary drinks (Ptrend ≤ 0.01). These results suggest that sugars may represent a modifiable risk factor for cancer prevention (breast in particular), contributing to the current debate on the implementation of sugar taxation, marketing regulation, and other sugar-related policies. This trial was registered at clinicaltrials.gov as NCT03335644.


Translation elongation factor 2 is part of the target for a new family of antifungals.
Translation elongation factor 2 (EF2), which in Saccharomyces cerevisiae is expressed from the EFT1 and EFT2 genes, has been found to be targeted by a new family of highly specific antifungal compounds derived from the natural product sordarin. Two complementation groups of mutants resistant to the semisynthetic sordarin derivative GM193663 were found. The major one (21 members) consisted of isolates with mutations on EFT2. The minor one (four isolates) is currently being characterized but it is already known that resistance in this group is not due to mutations on EFT1, pointing to the complex structure of the functional target for these compounds. Mutations on EF2 clustered, forming a possible drug binding pocket on a three-dimensional model of EF2, and mutant cell extracts lost the capacity to bind to the inhibitors. This new family of antifungals holds the promise to be a much needed and potent addition to current antimicrobial treatments, as well as a useful tool for dissection of the elongation process in ribosomal protein synthesis.


The failing human heart is unable to use the Frank-Starling mechanism.
There is evidence that the failing human left ventricle in vivo subjected to additional preload is unable to use the Frank-Starling mechanism. The present study compared the force-tension relation in human nonfailing and terminally failing (heart transplants required because of dilated cardiomyopathy) myocardium. Isometric force of contraction of electrically driven left ventricular papillary muscle strips was studied under various preload conditions (2 to 20 mN). To investigate the influence of inotropic stimulation, the force-tension relation was studied in the presence of the cardiac glycoside ouabain. In skinned-fiber preparations of the left ventricle, developed tension was measured after stretching the preparations to 150% of the resting length. To evaluate the length-dependent activation of cardiac myofibrils by Ca2+ in failing and nonfailing myocardium, the tension-Ca2+ relations were also measured. After an increase of preload, the force of contraction gradually increased in nonfailing myocardium but was unchanged in failing myocardium. There were no differences in resting tension, muscle length, or cross-sectional area of the muscles between both groups. Pretreatment with ouabain (0.02 mumol/L) restored the force-tension relation in failing myocardium and preserved the force-tension relation in nonfailing tissue. In skinned-fiber preparations of the same hearts, developed tension increased significantly after stretching only in preparations from nonfailing but not from failing myocardium. The Ca2+ sensitivity of skinned fibers was significantly higher in failing myocardium (EC50, 1.0; 95% confidence limit, 0.88 to 1.21 mumol/L) compared with nonfailing myocardium (EC50, 1.7; 95% confidence limit, 1.55 to 1.86 mumol/L). After increasing the fiber length by stretching, a significant increase in the sensitivity of the myofibrils to Ca2+ was observed in nonfailing but not in failing myocardium. These experiments provide evidence for an impaired force-tension relation in failing human myocardium. On the subcellular level, this phenomenon might be explained by a failure of the myofibrils to increase the Ca2+ sensitivity after an increase of the sarcomere length.


Familial hypersecretion of adrenal androgens transmitted as a dominant, non-HLA linked trait.
Clinical evidence of adrenal androgen hyperfunction (premature pubarche, hirsutism, amenorrhea) occurred in the studied proband, her mother, maternal aunt (twin sisters), and maternal great-grandmother. The basal levels of androgen in the first three were variably elevated. In all the members of this family who were tested, the response of 17-hydroxyprogesterone and progesterone to adrenocorticotropic hormone stimulation was either normal or of the type seen in heterozygotes for congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Of particular importance is the fact that neither the proband nor her mother or maternal aunt had the type of response seen in homozygotes presenting the attenuated form of congenital adrenal hyperplasia. The disorder appears to be a familial condition resulting in excessive levels of adrenal androgens beginning during childhood years, causing hirsutism and amenorrhea and interfering with normal pubertal and adult ovarian function. Glucocorticoid therapy suppresses adrenal androgen levels; in two individuals, conception occurred twice in each during such treatment in otherwise amenorrheic individuals. The pattern of transmission of the disorder appears to be either autosomal or X-linked dominant, and not linked to the homologous leucocytic antibodies (HLA) region of the sixth chromosome.


Parameters affecting the exposure to furfuryl alcohol from coffee.
Recently, furfuryl alcohol (FFA) was labelled a human potential carcinogen (group 2B) by the International Agency for Research on Cancer. Its alimentary exposure is mostly from coffee since in any other foods the concentrations are significantly lower. The various storage conditions of roasted coffee, the different brewing techniques applied and the bioaccessibility after ingestion are potential parameters that might alter the exposure to FFA from coffee. An 8 weeks stability study at varying temperatures showed that FFA is stable in the ground coffee matrix. Moreover, different brewing techniques extracted different amounts of FFA and affected its final concentration. The evaluation of the relative exposure to four furans (FFA, 5-hydroxymethyl-furaldehyde, 2-furoic acid, and 5-hydroxymethyl-2-furoic acid) revealed that FFA amounts were at least 2-fold the amounts of other studied furans in the same brew. A 22-fold variation in the concentration of the four furans in brews prepared using different coffee grounds and brewing techniques could be observed. 90% of the four furans were extracted by the first 25-30% fraction of the filter brew. A significant decrease of FFA is observed after stressing with simulated gastric fluid. However, this decrease could not be reproduced when mimicking a regular coffee ingestion situation.


Effect of Short-Term Aging on the Flexural Strength of a Novel Experimental CAD/CAM Fiber-Reinforced Composite Material: A Pilot Study.
To investigate the three-point flexural strength of a novel CAD/CAM fiber-reinforced composite (FRC) material following different aging conditions. The specimens were randomly assigned to one of five groups based on aging condition: (1) control (no treatment); (2) short-term water storage; (3) thermal degradation with autoclaving; (4) chemical degradation with hydrochloric acid; and (5) chemical degradation with citric acid (n = 10 per group). The specimens in the control group received no treatment. Following each treatment protocol, the three-point bending test was used to calculate the flexural strength. Data were statistically analyzed (α = .05), and scanning electron microscopy (SEM) analysis of the specimens was conducted. No significant differences in flexural strength were observed among the groups (P = .199). In addition, no distinct morphologic differences were detected in the SEM images of the specimens. The flexural strength of this novel CAD/CAM FRC material was unaffected by different aging methods.


Scales to assess sleep impairment in Parkinson's disease: critique and recommendations.
There is a broad spectrum of sleep disturbances observed in Parkinson's disease (PD). A variety of scales have been applied to the evaluation of PD sleep and wakefulness, but only a small number have been assessed specifically for clinimetric properties in the PD population. The movement disorder society has commissioned this task force to examine these scales and to assess their use in PD. A systematic literature review was conducted to explore the use of sleep scales in PD and to determine which scales qualified for a detailed critique. The task force members, all of whom have extensive experience in assessing sleep in PD reviewed each of the scales using a structured proforma. Scales were categorized into recommended, suggested and listed according to predefined criteria. A total of 48 potential scales were identified from the search and reviewed. Twenty-nine were excluded because they did not meet review criteria or were variations of scales already included, leaving 19 scales that were critiqued and rated by the task force based on the rating criteria. Only six were found to meet criteria for recommendation or suggestion by the task force: the PD sleep scale (PDSS) and the Pittsburgh sleep quality index (PSQI) are recommended for rating overall sleep problems to screen and to measure severity, the SCOPA-sleep (SCOPA) is recommended for rating overall sleep problems both to screen and to measure severity, and for rating daytime sleepiness; the Epworth sleepiness scale (ESS) is recommended for rating daytime sleepiness to screen and to measure severity; the inappropriate sleep composite score (ISCS) is suggested for rating severe daytime sleepiness or sleep attacks to screen and to measure severity; and the Stanford sleepiness scale (SSS) is suggested for rating sleepiness and to measure severity at a specific moment. The task force does not recommend the development of new scales, but emphasizes the need for educational efforts to train physicians in sleep interview techniques and polysomnography.


Dietary fiber intake and risk of colorectal cancer and incident and recurrent adenoma in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial.
Dietary fiber has been associated with a reduced risk of colorectal cancer. However, it remains unclear at which stage in the carcinogenic pathway fiber may act or which food sources of dietary fiber may be most beneficial against colorectal cancer development. The objective was to prospectively evaluate the association between dietary fiber intake and the risk of incident and recurrent colorectal adenoma and incident colorectal cancer. Study participants were identified from the intervention arm of the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. Participants received flexible sigmoidoscopy at baseline and 3 or 5 y after. Dietary fiber intake was measured by using a self-reported dietary questionnaire. The colorectal cancer, incident adenoma, and recurrent adenoma analyses were based on 57,774, 16,980, and 1667 participants, respectively. Unconditional logistic regression was used to assess the risk of incident and recurrent adenoma, and Cox proportional hazards models were used to assess the risk of colorectal cancer across categories of dietary fiber intake, with adjustment for potential confounders. Elevated total dietary fiber intake was associated with a significantly reduced risk of incident distal colorectal adenoma (ORhighest vs. lowest tertile of intake: 0.76; 95% CI: 0.63, 0.91; P-trend = 0.003) but not recurrent adenoma (P-trend = 0.67). Although the association was not statistically significant for colorectal cancer overall (HR: 0.85; 95% CI: 0.70, 1.03; P-trend = 0.10), a reduced risk of distal colon cancer was observed with increased total fiber intake (HR: 0.62; 95% CI: 0.41, 0.94; P-trend = 0.03). Protective associations were most notable for fiber originating from cereals or fruit. This large, prospective study within a population-based screening trial suggests that individuals consuming the highest intakes of dietary fiber have reduced risks of incident colorectal adenoma and distal colon cancer and that this effect of dietary fiber, particularly from cereals and fruit, may begin early in colorectal carcinogenesis. This trial was registered at clinicaltrials.gov as NCT01696981.


Photoelectrocatalytic production of active chlorine on nanocrystalline titanium dioxide thin-film electrodes.
The production of chlorine and hypochlorite is of great economical and technological interest due to their large-scale use in many kinds of commercial applications. Yet, the current processes are not without problems such as inevitable side reactions and the high cost of production. This work reports the photoelectrocatalytic oxidation of chloride ions to free chlorine as it has been investigated by using titanium dioxide (TiO2) and several metal-doped titanium dioxide (M-TiO2) material electrodes. An average concentration of 800 mg L(-1) of free chlorine was obtained in an open-air reactor using a TiO2 thin-film electrode biased at +1.0 V (SCE) and illuminated by UV light. The M-doped electrodes have performed poorly compared with the pure TiO2 counterpart. Test solutions containing 0.05 mol L(-1) NaCl pH 2.0-4.0 were found to be the best conditions for fast production of free chlorine. A complete investigation of all parameters that influence the global process of chlorine production by the photo electrocatalytic method such as applied potential, concentration of NaCl, pH solution, and time is presented in detail. In addition, photocurrent vs potential curves and the reaction order are also discussed.


Sexual Assault and HIV Postexposure Prophylaxis at an Urban African Hospital.
Antiretroviral postexposure prophylaxis (PEP) may be underutilized in sexual assault cases in Kenya. This study evaluated the characteristics of survivors of sexual violence attending the Gender-Based Violence Recovery Center (GBVRC) at the Kenyatta National Hospital and reviewed the uptake, adherence, and outcomes of those initiated on PEP. In a retrospective cohort, data from charts of the assaulted seen at the GBVRC from 2009 to 2012 were abstracted. Data were collected describing sociodemographic characteristics, nature of sexual assault, HIV serostatus, and aspects of the PEP care cascade. Characteristics of participants who received PEP were compared with those who did not receive PEP. We enrolled 385 assaulted persons; 331 (86%) were female; the median age of the assaulted persons was 21 (interquartile range 14-28) years; and 61 (15.8%) were children aged 10 years and younger. Of 379 assaults with descriptions, 330 (85.7%) were vaginal assaults and 40 (10.3%) were penile-anal assaults. Most perpetrators were unknown to the assaulted 220 of 384 (57.3%). All assaulted persons were offered HIV testing and 359 (93%) accepted testing; 346 (96.4%) of 359 assaulted persons tested HIV negative. In total, 207 (53.8%) of 385 sexual assault survivors initiated PEP. Only 70 (34%) completed 28 days of PEP, and only 21 (10.1%) returned for repeat HIV test at 3 months. In conclusion, PEP was only initiated in 54% of sexual assault cases. The care cascade showed that late presentation and poor adherence were the greatest gaps in PEP provision. Earlier presentation for PEP should be promoted among sexual assault in areas of high HIV prevalence.


Severe peri-ictal respiratory dysfunction is common in Dravet syndrome.
Dravet syndrome (DS) is a severe childhood-onset epilepsy commonly due to mutations of the sodium channel gene SCN1A. Patients with DS have a high risk of sudden unexplained death in epilepsy (SUDEP), widely believed to be due to cardiac mechanisms. Here we show that patients with DS commonly have peri-ictal respiratory dysfunction. One patient had severe and prolonged postictal hypoventilation during video EEG monitoring and died later of SUDEP. Mice with an Scn1aR1407X/+ loss-of-function mutation were monitored and died after spontaneous and heat-induced seizures due to central apnea followed by progressive bradycardia. Death could be prevented with mechanical ventilation after seizures were induced by hyperthermia or maximal electroshock. Muscarinic receptor antagonists did not prevent bradycardia or death when given at doses selective for peripheral parasympathetic blockade, whereas apnea, bradycardia, and death were prevented by the same drugs given at doses high enough to cross the blood-brain barrier. When given via intracerebroventricular infusion at a very low dose, a muscarinic receptor antagonist prevented apnea, bradycardia, and death. We conclude that SUDEP in patients with DS can result from primary central apnea, which can cause bradycardia, presumably via a direct effect of hypoxemia on cardiac muscle.


Work productivity and health of informal caregivers of persons with advanced cancer.
The purpose of this study was to describe health promotion behaviors and work productivity loss in informal caregivers of individuals with advanced stage cancer. Using a cross-sectional, correlational design, 70 caregivers completed measures of health behaviors, mood, social support, and burden. Absenteeism and presenteeism were evaluated in employed caregivers (n = 40). Caregivers reported low levels of physical activity. The mean percentage of work productivity loss due to caregiving was 22.9%. Greater work productivity loss was associated with greater number of caregiving hours, higher cancer stage, married status, and greater anxiety, depression, and burden related to financial problems, disrupted schedule, and health. Nurses should assess caregivers and provide health promotion interventions, which may ultimately reduce the economic impact of caregiving.


Zone of traction injury of the common peroneal nerve.
The history of functional motor recovery after reconstruction of traction injury to the common peroneal nerve is poor, regardless of technique and regardless of the experience of the surgeon doing the reconstruction. The hypothesis tested is that the failure of functional motor recovery after common peroneal nerve traction injury is because the zone of injury extends beyond the visible region of peroneal nerve in continuity injury and into the muscle entry zone of the motor nerve terminations. The opportunity arose to examine pathologically this suspected distal zone of injury in one patient. Histology from this patient was compared with that from a similar zone in a leg amputated for vascular indications. With a peroneal traction injury severe enough to cause disruption of the nerve, histology using Masson trichrome stain for collagen, neurofilament and S-100 stain for nerve fibers and Schwann cells demonstrated collagen deposition between the peroneal nerve and the muscle. This fibrosis was not seen in the in the same location from the amputation specimen. These findings were corroborated by electron microscopy of the myoneural junction in both specimens. It is concluded that stretch/traction injury zone extends into the myoneural junction, preventing otherwise successful neural regeneration through nerve grafts to reinnervate muscle. This suggests that with a distal zone of injury extending into the myoneural junction region, peroneal motor function may be better achieved by direct neurotization than with nerve grafting.


Joint effects of N-terminal pro-B-type-natriuretic peptide and C-reactive protein vs angiographic severity in predicting major adverse cardiovascular events and clinical restenosis after coronary angioplasty in patients with stable coronary artery disease.
This study was designed to evaluate the joint effects of plasma C-reactive protein (CRP) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) vs coronary angiographic severity on cardiovascular risk stratification. A total of 345 patients with stable coronary artery disease (CAD) were recruited after successful percutaneous coronary intervention (PCI). Endpoints were major adverse cardiovascular events (MACE) and cumulative clinical restenosis rate after 18-36-month follow-up. Plasma NT-proBNP and CRP levels were among the strongest predictors of MACE. Adjusted hazard ratios of MACE according to combined biomarkers were 2.4 (p=0.05) for elevated CRP only, 5.22 (p<0.001) for elevated NT-proBNP only, and 7.04 (p<0.001) for elevation of both. The differential capacity using both plasma CRP and NT-proBNP in a receiver-operating-characteristics curve analysis (area under curve, AUC: 0.82) was significantly higher than using either biomarker alone or conventional risk factors (AUC: 0.67). Significant predictors of clinical restenosis were plasma NT-proBNP and the Gensini score. The combination of NT-proBNP and the Gensini score was the strongest predictor (AUC: 0.77) for clinical restenosis. Plasma NT-proBNP, CRP, and the Gensini score are complementary in risk stratification. Combined use of these biomarkers has provided substantial extra information to conventional risk factors in stable CAD patients.


The Inspired Sinewave Technique: A Comparison Study With Body Plethysmography in Healthy Volunteers.
The inspired sinewave technique is a noninvasive method to measure airway dead space, functional residual capacity, pulmonary blood flow, and lung inhomogeneity simultaneously. The purpose of this paper was to assess the repeatability and accuracy of the current device prototype in measuring functional residual capacity, and also participant comfort when using such a device. To assess within-session repeatability, six sinewave measurements were taken over two-hour period in 17 healthy volunteers. To assess day-to-day repeatability, measurements were taken over 16 days in 3 volunteers. To assess accuracy, sinewave measurements were compared to body plethysmography in 44 healthy volunteers. Finally, 18 volunteers who experienced the inspired sinewave device, body plethysmography and spirometry were asked to rate the comfort of each technique on a scale of 1-10. The repeatability coefficients for dead space, functional residual capacity, and blood flow were 48.7 ml, 0.48L, and 2.4L/min respectively. Bland-Altman analyses showed a mean BIAS(SD) of -0.68(0.42)L for functional residual capacity when compared with body plethysmography. 14 out of 18 volunteers rated the inspired sinewave device as their preferred technique. The repeatability and accuracy of functional residual capacity measurements were found to be as good as other techniques in the literature. The high level of comfort and the non-requirement of patient effort meant that, if further refined, the inspired sinewave technique could be an attractive solution for difficult patient groups such as very young children, elderly, and ventilated patients.


Ba2+ influx measures the duration of membrane excitation in Paramecium.
We have developed an assay for the duration of membrane excitation in Paramecium tetraurelia by tracing the influx of 133Ba2+. This assay is performed at physiological temperatures and in physiological solutions. Ba2+ enters the paramecia through the Ca channels, since mutants defective in Ca channels show no significant Ba2+ influx. Ba2+ enters only when the Ca channels are opened during excitation which can be triggered by Na+ or Ba2+ itself. The ionic species and relative concentrations determine the duration of the action potentials and hence the duration of spinning or backward swimming. The longer the average period of excitation, the larger the Ba2+ influx. In a Ba-Ca solution the cells spend 30% of their time in the excited state. The rate of Ba2+ entry into the paramecia in that state is 1.3 mM/min. Ba2+ influx occurs over a 50-fold range of Ba2+ concentration. There is very little Ba2+ efflux. The fate of the entered Ba2+, the consequences of the large and rapid influx, the advantages and drawbacks of the Ba2+ influx assay, and the possible use of the assay for Ca channel function in cell-free preparations are discussed.


Truncation of the C-terminal acidic transcriptional activation domain of herpes simplex virus VP16 renders expression of the immediate-early genes almost entirely dependent on ICP0.
The herpes simplex virus (HSV) proteins VP16 and ICP0 play key roles in stimulating the onset of the viral lytic cycle. We sought to explore the regulatory links between these proteins by studying the phenotypes of viral mutants in which the activation functions of both were simultaneously inactivated. This analysis unexpectedly revealed that truncation of the C-terminal transcriptional activation domain of VP16 (allele V422) in an ICP0-deficient background almost completely eliminated immediate-early gene expression and virus replication in Vero and HEL cells. The doubly mutant viral genome persisted in a quiescent state for at least 10 days in HEL cells infected at high multiplicity and could be reactivated by superinfection with wild-type HSV. In contrast, the in1814 VP16 mutation produced a markedly less severe phenotype in the same ICP0-deficient background. These data demonstrate that expression of the immediate-early genes requires ICP0 when the C-terminal activation domain of VP16 is deleted and raise the possibility that the in1814 form of VP16 retains a residual ability to stimulate gene expression during virus infection.


Longitudinal phenotypes for alcoholism: Heterogeneity of course, early identifiers, and life course correlates.
Alcoholism is a heterogeneous disorder; however, characterization of life-course variations in symptomatology is almost nonexistent, and developmentally early predictors of variations are very poorly characterized. In this study, the course of alcoholic symptomatology over 32 years is differentiated, and predictors and covariates of trajectory class membership are identified. A community sample of alcoholic and neighborhood matched control families, 332 men and 336 women, was recruited based on alcoholism in the men. Symptoms were assessed retrospectively at baseline (mean age = 32) back to age 15 and prospectively from baseline every 3 years for 15 years. Trajectory classes were established using growth mixture modeling. Men and women had very similarly shaped trajectory classes: developmentally limited (men: 29%, women: 42%), developmentally cumulative (men: 26%, women: 38%), young adult onset (men: 31%, women: 21%), and early onset severe (men: 13%). Three factors at age 15 predicted class membership: family history of alcoholism, age 15 symptoms, and level of childhood antisocial behavior. Numerous measures of drinking and other psychopathology were also associated with class membership. The findings suggest that clinical assessments can be crafted where the profile of current and historical information can predict not only severity of prognosis but also future moderation of symptoms and/or remission over intervals as long as decades.


Lymphovascular and marginal invasion as useful prognostic indicators and the role of c-erbB-2 in patients with male extramammary Paget's disease: a study of 31 patients.
Extramammary Paget's disease (EMPD) is a rare intraepithelial neoplasm arising in skin that bears apocrine glands. We analyzed clinical, pathological and immunohistochemical staining patterns in 31 cases of EMPD to assess possible predictors of outcome, including patient age, sex, involved organs, surgical margin status, invasion depth, lymphovascular invasion, adnexal invasion, and immunoreactivity for cytokeratin 7 and 20, carcinoembryonic androgen, androgen receptor, p53, c-erbB-2 (Dako, Carpinteria, California) and Ki67. A total of 31 cases of EMPD were retrieved from the surgical pathology files of the department of pathology along with the medical records of the department of urology. Dermal invasion depth, tumor margin status, adnexal invasion and lymphovascular invasion were determined from the hematoxylin and eosin stained slides of each case. Eight of the 31 cases (25.8%) experienced local recurrence. Two of 3 patients with and 6 of 28 without lymphovascular invasion experienced recurrence (p = 0.029). Dermal invasion significantly correlated with Her-2 over expression (Wilcoxon 2-sample test p = 0.0007). When the criteria of 1 and 5 mm invasion were applied, each validly correlated with Her-2 over expression. Her-2 over expression (grades 2 and 3) significantly correlated with p53 over expression (Wilcoxon rank test p = 0.0015). C-erbB-2 was positive in 51.6% of EMPD cases, in which it was strongly associated with dermal invasion and p53 over expression. The most valid prognostic factor in male EMPD for predicting late recurrence is lymphovascular permeation and marginal status.


The Role of Corticostriatal Systems in Speech Category Learning.
One of the most difficult category learning problems for humans is learning nonnative speech categories. While feedback-based category training can enhance speech learning, the mechanisms underlying these benefits are unclear. In this functional magnetic resonance imaging study, we investigated neural and computational mechanisms underlying feedback-dependent speech category learning in adults. Positive feedback activated a large corticostriatal network including the dorsolateral prefrontal cortex, inferior parietal lobule, middle temporal gyrus, caudate, putamen, and the ventral striatum. Successful learning was contingent upon the activity of domain-general category learning systems: the fast-learning reflective system, involving the dorsolateral prefrontal cortex that develops and tests explicit rules based on the feedback content, and the slow-learning reflexive system, involving the putamen in which the stimuli are implicitly associated with category responses based on the reward value in feedback. Computational modeling of response strategies revealed significant use of reflective strategies early in training and greater use of reflexive strategies later in training. Reflexive strategy use was associated with increased activation in the putamen. Our results demonstrate a critical role for the reflexive corticostriatal learning system as a function of response strategy and proficiency during speech category learning.


EMG activity during whole body vibration: motion artifacts or stretch reflexes?
The validity of electromyographic (EMG) data recorded during whole body vibration (WBV) is controversial. Some authors ascribed a major part of the EMG signal to vibration-induced motion artifacts while others have interpreted the EMG signals as muscular activity caused at least partly by stretch reflexes. The aim of this study was to explore the origin of the EMG signal during WBV using several independent approaches. In ten participants, the latencies and spectrograms of stretch reflex responses evoked by passive dorsiflexions in an ankle ergometer were compared to those of the EMG activity of four leg muscles during WBV. Pressure application to the muscles was used to selectively reduce the stretch reflex, thus permitting to distinguish stretch reflexes from other signals. To monitor motion artifacts, dummy electrodes were placed close to the normal electrodes. Strong evidence for stretch reflexes was found: the latencies of the stretch reflex responses evoked by dorsiflexions were almost identical to the supposed stretch reflex responses during vibration (differences of less than 1 ms). Pressure application significantly reduced the amplitude of both the supposed stretch reflexes during vibration (by 61 +/- 17%, p < 0.001) and the stretch reflexes in the ankle ergometer (by 56 +/- 13%, p < 0.01). The dummy electrodes showed almost no activity during WBV (7 +/- 4% of the corresponding muscle's iEMG signal). The frequency analyses revealed no evidence of motion artifacts. The present results support the hypothesis of WBV-induced stretch reflexes. Contribution of motion artifacts to the overall EMG activity seems to be insignificant.


Community water fluoridation and caries prevention: a critical review.
The aim of this paper was to critically review the current role of community water fluoridation in preventing dental caries. Original articles and reviews published in English language from January 2001 to June 2006 were selected through MEDLINE database. Other sources were taken from the references of the selected papers. For the past 50 years community water fluoridation has been considered the milestone of caries prevention and as one of the major public health measures of the 20th century. However, it is now accepted that the primary cariostatic action of fluoride occurs after tooth eruption. Moreover, the caries reduction directly attributable to water fluoridation have declined in the last decades as the use of topical fluoride had become more widespread, whereas enamel fluorosis has been reported as an emerging problem in fluoridated areas. Several studies conducted in fluoridated and nonfluoridated communities suggested that this method of delivering fluoride may be unnecessary for caries prevention, particularly in the industrialized countries where the caries level has became low. Although water fluoridation may still be a relevant public health measure in poor and disadvantaged populations, the use of topical fluoride offers an optimal opportunity to prevent caries among people living in both industrialized and developing countries.


Bonding and magnetism of Fe(6)-(C(6)H(6))(m), m = 1, 2.
The interactions of one and two benzene molecules with the superparamagnetic Fe(6) cluster were studied by means of gradient-corrected density functional theory. The ground state, GS, of bare Fe(6) presents a distorted octahedral structure with 2S = 20; S is the total spin. For the calculated 2S = 16 GS of the neutral Fe(6)-C(6)H(6) complex, as well as in the positive and negative ions both with 2S = 15, the benzene unit is adsorbed on one axial Fe(a) atom. The 2S = 14 GS for Fe(6)-(C(6)H(6))(2) resembles a sandwich structure, with the metal Fe(6) cluster separating the benzene rings that are bonded symmetrically on the two axial sites of Fe(6). The binding is accounted for by electrostatic interactions and by 3d-pi bonds, as revealed by the molecular orbitals. Though each C-Fe bond is weak, eta(6) coordinations were indicated by the topology of the electronic density. The 3d-pi bonding is reflected by the adiabatic ionization energies and electron affinities, which are smaller than those of bare Fe(6). The computed IR spectra show vibrational bands near those of bare benzene; some forbidden IR modes in benzene and in Fe(6) become IR active in Fe(6)-(C(6)H(6))(1,2). The results show a strong perturbation of the electronic structure of Fe(6). The decrease of its magnetic moment implies that the magnetic effects play an important role in the adsorption of benzene.


The effect and evolution of patient selection on outcomes in endoscopic third ventriculostomy for hydrocephalus: A large-scale review of the literature.
Endoscopic third ventriculostomy (ETV) has become a popular technique for the treatment of hydrocephalus, but small sample size has limited the generalizability of prior studies. We performed a large-scale review of all available studies to help eliminate bias and determine how outcomes have changed and been influenced by patient selection over time. A systematic literature search was performed for studies of ETV that contained original, extractable patient data, and a meta-analytic model was generated for correlative and predictive analysis. A total of 130 studies were identified, which included 11,952 cases. Brain tumor or cyst was the most common hydrocephalus etiology, but high-risk etiologies, post-infectious or post-hemorrhagic hydrocephalus, accounted for 18.4%. Post-operative mortality was very low (0.2%) and morbidity was only slightly higher in developing than in industrialized countries. The rate of ETV failure was 34.7% and was higher in the first months and plateaued around 20months. As anticipated, ETV is less successful in high-risk etiologies of hydrocephalus and younger patients. Younger patient age and high-risk etiologies predicted failure. ETVs were performed more often in high-risk etiologies over time, but, surprisingly, there was no overall change in ETV success rate over time. This study should help to influence optimal patient selection and offer guidance in predicting outcomes.


Relationships between central and autonomic measures of arousal at age 15 years and criminality at age 24 years.
Previous studies have indicated that criminality is in part genetically determined, but it is not clear how this predisposition manifests itself at a biological level. This prospective study tests the hypothesis that a psychophysiological predisposition to criminality partly manifests itself through autonomic and central nervous system underarousal. Psychophysiological measures, taken at the age of 15 years, were related to criminality status that was assessed at the age of 24 years. Criminals had a significantly lower resting heart rate, skin conductance activity, and more slow-frequency electroencephalographic activity than noncriminals. Differences were not mediated by social, demographic, and academic factors. These results constitute the first clear evidence that implicates underarousal in all three response systems (electrodermal, cardiovascular, and cortical) in the development of criminality. Although arousal variables correctly classified 74.7% of all subjects, psychophysiological factors alone cannot fully account for criminal behavior and do not negate the potential role of social variables in predicting criminal behavior.


Dual vaccination against IL-4 and IL-13 protects against chronic allergic asthma in mice.
Allergic asthma is characterized by elevated levels of IgE antibodies, type 2 cytokines such as interleukin-4 (IL-4) and IL-13, airway hyperresponsiveness (AHR), mucus hypersecretion and eosinophilia. Approved therapeutic monoclonal antibodies targeting IgE or IL-4/IL-13 reduce asthma symptoms but require costly lifelong administrations. Here, we develop conjugate vaccines against mouse IL-4 and IL-13, and demonstrate their prophylactic and therapeutic efficacy in reducing IgE levels, AHR, eosinophilia and mucus production in mouse models of asthma analyzed up to 15 weeks after initial vaccination. More importantly, we also test similar vaccines specific for human IL-4/IL-13 in mice expressing human IL-4/IL-13 and the related receptor, IL-4Rα, to find efficient neutralization of both cytokines and reduced IgE levels for at least 11 weeks post-vaccination. Our results imply that dual IL-4/IL-13 vaccination may represent a cost-effective, long-term therapeutic strategy for the treatment of allergic asthma as demonstrated in mouse models, although additional studies are warranted to assess its safety and feasibility.


Rethinking fast and slow based on a critique of reaction-time reverse inference.
Do people intuitively favour certain actions over others? In some dual-process research, reaction-time (RT) data have been used to infer that certain choices are intuitive. However, the use of behavioural or biological measures to infer mental function, popularly known as 'reverse inference', is problematic because it does not take into account other sources of variability in the data, such as discriminability of the choice options. Here we use two example data sets obtained from value-based choice experiments to demonstrate that, after controlling for discriminability (that is, strength-of-preference), there is no evidence that one type of choice is systematically faster than the other. Moreover, using specific variations of a prominent value-based choice experiment, we are able to predictably replicate, eliminate or reverse previously reported correlations between RT and selfishness. Thus, our findings shed crucial light on the use of RT in inferring mental processes and strongly caution against using RT differences as evidence favouring dual-process accounts.


In-hospital day-by-day systolic blood pressure variability during rehabilitation: a marker of adverse outcome in secondary prevention after myocardial revascularization.
Although it is known that increased visit-to-visit or home day-by-day variability of blood pressure (BP), independently of its average value, results in an increased risk of cardiovascular events, the prognostic value of in-hospital day-by-day BP variability in secondary cardiovascular prevention has not yet been established. We studied 1440 consecutive cardiac patients during a cardiovascular rehabilitation program of about 12 days after coronary artery bypass graft (CABG) and/or valve surgery. We measured auscultatory BP at the patient bed in each rehabilitation day twice, in the morning and the afternoon. We correlated SBP variability assessed as standard deviation (SBP-SD) and coefficient of variation (SBP-CoV) of the daily measures with overall mortality, cardiovascular mortality and major adverse cardiocerebrovascular events (MACCEs) after a mean follow-up of 49 months by Cox hazard analysis. In our patients (age 68 ± 11years, 61% hypertensive patients) the ranges of SBP-SD tertiles were: 4.1-9.1, 9.2-11.5 and 11.6-24.5 mmHg. Fifty-five percent of the patients underwent CABG, 33% underwent valve surgery, 12% both CABG and valve surgery. In CABG patients, the highest SBP-SD tertile showed the highest overall mortality, cardiovascular mortality and MACCEs (P < 0.01). Results remained significant after multivariate analysis adjusting for age, sex, mean SBP, BMI, hypertension, hyperlipidaemia, and diabetes. No association between SBP-SD and mortality or MACCEs was found in valve surgery patients. In-hospital day-by-day SBP variability predicts mortality and MACCEs in CABG patients, possibly representing a target during rehabilitation and treatment in secondary cardiovascular prevention.


Inhibition of lymphocyte mitogenesis by immobilized antigen-antibody complexes.
Mouse spleen cells, cultured on surfaces coated with antigen-antibody complexes, are inhibited from responding to the B-cell mitogens, lipopolysaccharide, lipid A, Pneumococcal polysaccharide SIII, and poly I:C. The response to the T-cell mitogen, concanavalin A, is also substantially inhibited by immobilized antigen-antibody complexes, but specific inhibition of the response to phytohemagglutinin is minimal. Control experiments showed that immobilized complexes prepared from IgG F(ab')2 fragments and IgA antibodies (both of which fail to bind to Fc receptors when complexed to antigen) did not show significant inhibitory activity when compared with the inhibition observed with complexes prepared from whole IgG. Suspensions of antigen-antibody complexes prepared from the same antigen and intact IgG antibody did not inhibit mitogenesis. None of the mitogens used could be demonstrated to compete with the binding of aggregated immunoglobulin to the B-cell Fc receptor. It appears that the interaction of Fc receptor-bearing lymphocytes and/or macrophages with immobilized complexes prevents lymphocyte activation by mitogens. It is suggested that the mechanism(s) involved may be relevant to antibody feedback control of the humoral immune response.


Extending Delivery Coverage to Include Prenatal Care for Low-Income, Immigrant Women Is a Cost-Effective Strategy.
To compare the outcomes and cost effectiveness of two alternate policy strategies for prenatal care among low-income, immigrant women: coverage for delivery only (the federal standard) and prenatal care with delivery coverage (state option under the Children's Health Insurance Program). A decision-analytic model was developed to determine the cost effectiveness of two alternate policies for pregnancy coverage. All states currently provide coverage for delivery, and 19 states also provide coverage for prenatal care. An estimated 84,000 unauthorized immigrant women have pregnancies where no prenatal care is covered. Our outcomes were costs, quality-adjusted life-years, and cases of cerebral palsy and infant death before age 1. Model inputs were obtained from a database of Oregon Medicaid claims and the literature. Univariate and bivariate sensitivity analyses, as well as a Monte Carlo simulation, were performed. Extending prenatal coverage is a cost-effective strategy. Providing prenatal care for the 84,000 women annually who are currently uninsured could prevent 117 infant deaths and 34 cases of cerebral palsy. Prenatal care coverage costs $380 more per woman than covering the delivery only. For every 865 additional women receiving prenatal care, one infant death would be averted, at an average cost of $328,700. Cost-effectiveness acceptability curve analyses suggest a 99% probability that providing prenatal care is more cost effective at a willingness-to-pay threshold of $100,000 per quality-adjusted life-year. Extending prenatal care to low-income, immigrant women, regardless of citizenship status, is a cost-effective strategy.


Disease-associated XMRV sequences are consistent with laboratory contamination.
Xenotropic murine leukaemia viruses (MLV-X) are endogenous gammaretroviruses that infect cells from many species, including humans. Xenotropic murine leukaemia virus-related virus (XMRV) is a retrovirus that has been the subject of intense debate since its detection in samples from humans with prostate cancer (PC) and chronic fatigue syndrome (CFS). Controversy has arisen from the failure of some studies to detect XMRV in PC or CFS patients and from inconsistent detection of XMRV in healthy controls. Here we demonstrate that Taqman PCR primers previously described as XMRV-specific can amplify common murine endogenous viral sequences from mouse suggesting that mouse DNA can contaminate patient samples and confound specific XMRV detection. To consider the provenance of XMRV we sequenced XMRV from the cell line 22Rv1, which is infected with an MLV-X that is indistinguishable from patient derived XMRV. Bayesian phylogenies clearly show that XMRV sequences reportedly derived from unlinked patients form a monophyletic clade with interspersed 22Rv1 clones (posterior probability >0.99). The cell line-derived sequences are ancestral to the patient-derived sequences (posterior probability >0.99). Furthermore, pol sequences apparently amplified from PC patient material (VP29 and VP184) are recombinants of XMRV and Moloney MLV (MoMLV) a virus with an envelope that lacks tropism for human cells. Considering the diversity of XMRV we show that the mean pairwise genetic distance among env and pol 22Rv1-derived sequences exceeds that of patient-associated sequences (Wilcoxon rank sum test: p = 0.005 and p < 0.001 for pol and env, respectively). Thus XMRV sequences acquire diversity in a cell line but not in patient samples. These observations are difficult to reconcile with the hypothesis that published XMRV sequences are related by a process of infectious transmission. We provide several independent lines of evidence that XMRV detected by sensitive PCR methods in patient samples is the likely result of PCR contamination with mouse DNA and that the described clones of XMRV arose from the tumour cell line 22Rv1, which was probably infected with XMRV during xenografting in mice. We propose that XMRV might not be a genuine human pathogen.


Association of trough vedolizumab levels with clinical, biological and endoscopic outcomes during maintenance therapy in inflammatory bowel disease.
To establish the relationship between trough vedolizumab levels and outcomes during maintenance therapy. Cross-sectional service evaluation was performed on patients with inflammatory bowel disease (IBD) receiving maintenance vedolizumab therapy (minimum of 12 weeks following induction). Prior to infusion, data on clinical activity (Harvey-Bradshaw Index or partial Mayo score), trough C-reactive protein (CRP)/vedolizumab levels and faecal calprotectin were collected. Endoscopic data (±8 weeks from vedolizumab level testing) were obtained by review of medical records. Vedolizumab levels were processed using the Immundiagnostik monitor ELISA. The Edinburgh IBD Unit, Western General Hospital (tertiary IBD referral centre). Seventy-three patients (30 ulcerative colitis and 43 Crohn's disease) were identified who fulfilled inclusion criteria and had vedolizumab levels matched with clinical activity scores, CRP and faecal calprotectin. Of these, 40 patients also had matched endoscopic data. The association of trough vedolizumab levels with clinical remission (Harvey-Bradshaw Index <5 or partial Mayo <2), biologic remission (faecal calprotectin <250 µg/g+CRP <5 mg/L) and endoscopic remission (Mayo score 0/no inflammation and ulceration on colonoscopy). The median trough vedolizumab levels were similar between patients in and not in clinical remission (10.6 vs 9.9 µg/mL, p=0.54); biologic remission (10.6 vs 9.8 µg/mL, p=0.35) and endoscopic remission (8.1 vs 10.2 µg/mL, p=0.21). Quartile analysis revealed no significant increase in the proportion of patients in clinical remission, biologic remission or endoscopic remission with increasing trough vedolizumab levels (p<0.05). In this cohort, trough vedolizumab levels were not associated with clinical, biological or endoscopic outcomes during maintenance therapy.


ISL1 Is Necessary for Maximal Thyrotrope Response to Hypothyroidism.
ISLET1 is a homeodomain transcription factor necessary for development of the pituitary, retina, motor neurons, heart, and pancreas. Isl1-deficient mice (Isl1(-/-)) die early during embryogenesis at embryonic day 10.5 due to heart defects, and at that time, they have an undersized pituitary primordium. ISL1 is expressed in differentiating pituitary cells in early embryogenesis. Here, we report the cell-specific expression of ISL1 and assessment of its role in gonadotropes and thyrotropes. Isl1 expression is elevated in pituitaries of Cga(-/-) mice, a model of hypothyroidism with thyrotrope hypertrophy and hyperplasia. Thyrotrope-specific disruption of Isl1 with Tshb-cre is permissive for normal serum TSH, but T4 levels are decreased, suggesting decreased thyrotrope function. Inducing hypothyroidism in normal mice causes a reduction in T4 levels and dramatically elevated TSH response, but mice with thyrotrope-specific disruption of Isl1 have a blunted TSH response. In contrast, deletion of Isl1 in gonadotropes with an Lhb-cre transgene has no obvious effect on gonadotrope function or fertility. These results show that ISL1 is necessary for maximal thyrotrope response to hypothyroidism, in addition to its role in development of Rathke's pouch.


The acute toxicity of thallium to freshwater organisms: Implications for risk assessment.
The acute toxicity of Tl(I) to the microalga, Pseudokirchneriella subcapitata, the planktonic crustaceans, Daphnia magna and Daphnia pulex, and early-life stage of the zebrafish, Danio rerio, has been studied according to OECD protocols. Toxicological end-point concentrations for the microalga ranged from 17 μg l(-1) for a 72 h EyC25 (yield inhibition) to 80 μg l(-1) for a 72 h ErC50 (growth inhibition). Daphnia were less sensitive to Tl, with 48 h EC50s of about 1000 μg l(-1) and 1200 μg l(-1) for D. magna and D. pulex, respectively; however, end-point concentrations were reduced considerably (to about 510 μg l(-1) and 730 μg l(-1), respectively) when experiments were repeated in dechlorinated Plymouth tap water (rather than OECD medium). The 96 h LC50 for D. rerio was 870 μg l(-1) but a variety of sub-lethal effects, including enlargement of yolk sac and reduction in heart beat rate, were observed when larvae were exposed to lower concentrations. Based on these results, a predicted no effect concentration (PNEC) for Tl in freshwaters of 0.087 μg l(-1) is proposed. The PNEC is an order of magnitude lower than the only (Canadian) water quality guideline for Tl that appears to exist, and is lower than Tl concentrations reported in freshwaters impacted by historical or contemporary metal mining. Our results are also consistent with previous studies that employ different organisms and end-points in that Tl toxicity is dependent on the concentration of K+, the biogeochemical analogue of Tl+. Accordingly, regulation of Tl in the freshwater environment should factor in the relative abundance of K.


Effect of autogenic training on general improvement in patients with irritable bowel syndrome: a randomized controlled trial.
Autogenic training (AT) is a useful and comprehensive relaxation technique. However, no studies have investigated the effects of AT on irritable bowel syndrome (IBS). In this study we tested the hypothesis that AT improves symptoms of IBS. Twenty-one patients with IBS were randomly assigned to AT (n = 11, 5 male, 6 female) or control therapy (n = 10, 5 male, 5 female). AT patients were trained intensively, while the control therapy consisted of discussions about patients' meal habits and life styles. All patients answered a question related to adequate relief (AR) of IBS symptoms and four questionnaires: Self-induced IBS Questionnaire (SIBSQ), Self-reported Depression Scale (SDS), State-Trait Anxiety Inventory (STAI), and Medical Outcome Short Form 36 Health Survey (SF-36). The proportion of AR in the last AT session in the AT group (9/11, 81.8%) was significantly higher than that in the controls (3/10, 30.0%, Chi-square test, p = 0.048). Two subscales of the SF-36, i.e., social functioning and bodily pain, were significantly improved in the AT group (p < 0.05) as compared to the control group. Role emotional (p = 0.051) and general health (p = 0.068) showed a tendency for improvement in the AT group. AT may be useful in the treatment of IBS by enhancing self-control.


Colorectal Cancer Patient Characteristics, Treatment and Survival in Oman--a Single Center Study.
Colorectal cancer is the most common gastrointestinal cancer in Oman with an increasing incidence. We here report the presenting features, treatment outcomes and survival in a University hospital in Oman and compare our data with regional and international studies. Medical records of patients with colorectal cancer were reviewed retrospectively between June 2000 and December 2013 and were followed until June 2014. A total of 162 patients were diagnosed with colorectal cancer. The majority were males (58.6%), with a median age of 56 years. Rectum was involved in 29.6% of patients, followed by ascending and sigmoid colon. The majority of patients had stage III (42.6%) and stage IV (32.7%) disease at presentation. K-Ras status was checked for 79 patients, and 41 (51.9%) featured the wild type. Median relapse free survival was 22 months. Median overall survival for all patients was 43 months. Observed 5 year overall survival (OS) for stages I, II and III was 100%, 60% and 60% respectively. On Log rank univariate analysis, age, BMI, diabetes, hypertension, metformin use, stage, clinical nodal status for rectal cancer, pathological T and nodal status, site of metastasis, surgical intervention, chemotherapy, radiotherapy, chemotherapy regimen, no of cycles of chemotherapy, response, RFS, site of recurrence and administration of 2nd line chemotherapy were significant factors affecting OS. On Cox regression multivariate analysis none of the factors independently affected the OS. The majority of patients present with advanced disease and at young age. The survival rates are comparable to the published regional and international literature.


Epimedium polysaccharide and propolis flavone can synergistically stimulate lymphocyte proliferation in vitro and enhance the immune responses to ND vaccine in chickens.
Four prescriptions, epimedium flavone plus propolis flavone (EF-PF), epimedium flavone plus propolis extracts (EF-PE), epimedium polysaccharide plus propolis flavone (EP-PF) and epimedium polysaccharide plus propolis extracts (EP-PE), were prepared and their immune-enhancing effects were compared. In test in vitro, the effects of them on chicken peripheral lymphocyte proliferation were determined by MTT method. The results showed that EP-PF group presented the highest stimulating index at most concentrations. In immune test, 300 14-day-old chickens were randomly divided into six groups and vaccinated with ND vaccine except for blank control (BC) group, re-challenged at 28 days of age. At the same time of the first vaccination, the chickens in four experimental groups were injected, respectively, with four prescriptions. The changes of the lymphocyte proliferation and antibody titer were determined. On day 28 after the first vaccination, the chickens except for BC group were challenged with NDV, the immune protective effect was observed. The results displayed that in EP-PF group, the antibody titers, lymphocyte proliferation and protective rate were the highest, the morbidity and mortality were the lowest. In dose test, 14-day-old chickens were randomly divided into five groups. The treatment and determinations were the same as the immune test except that the chickens in experimental groups were injected, respectively, with high, medium and low doses of EP-PF. The results revealed that in medium dose group, the antibody titers, lymphocyte proliferation and protective rate were the highest, the morbidity and mortality were the lowest. These results indicated that EP and PF possessed synergistically immune enhancement, EP-PF had the best efficacy, especially at medium dose, and would be expected to exploit into a new-type immunopotentiator.


Abnormal vitamin D metabolism in Paget's disease of bone.
We have studied several biochemical indices of bone turnover and vitamin D metabolism in 32 untreated patients with Paget's disease and in 32 age-matched control subjects. patients with Paget's disease, as expected, were characterized by high bone turnover, as judged by alkaline phosphatase and urinary excretion of hydorxyproline. Serum values of 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) and the ratio of 24,25(OH)2D3 to 25-OHD were significantly lower in patients than in control subjects. Serum concentrations of 1,25(OH)2D3 were normal in Paget's disease. The distribution of values for 24,25(OH)2D3 was log normal. On the basis of the normal range computed from control subjects, patients were divided into those with low or normal values for 24,25(OH)2D3. Disease activity, as judged by biochemical indices was significantly higher in the patients with the lower values of 24,25(OH)2D3. We conclude that Paget's disease is characterized by low circulating concentrations of 24,25(OH)2D3, particularly in patients with more extensive or severe disease.


Effect of depth of flooding on the rice water weevil, Lissorhoptrus oryzophilus, and yield of rice.
The rice water weevil, Lissorhoptrus oryzophilus (Kuschel) (Coleoptera: Curculionidae), is a semi-aquatic pest of rice and is the most destructive insect pest of rice in the United States. Adults oviposit after floods are established, and greenhouse studies have shown that plants exposed to deep floods have more eggs oviposited in leaf sheaths than plants exposed to a shallow flood. Experiments were conducted in three mid-southern states in the USA to determine if the depth of flooding would impact numbers of L. oryzophilus on rice plants under field conditions. Rice was flooded at depths of approximately 5 or 10 cm in Arkansas in 2007 and 2008 and Louisiana in 2008, and at depths between 0-20 cm in Missouri in 2008. Plants were sampled three and four weeks after floods were established in all locations, and also two weeks after flood in Missouri. On all sampling dates in four experiments over two years and at three field sites, fewer L. oryzophilus larvae were collected from rice in shallow-flooded plots than from deep-flooded plots. The number of L. oryzophilus was reduced by as much as 27% in shallow-flooded plots. However, the reduction in insect numbers did not translate to a significant increase in rice yield. We discuss how shallow floods could be used as a component of an integrated pest management program for L. oryzophilus.


Improving the Recovery of Patients with Subacromial Pain Syndrome with the DAid Smart Textile Shirt.
Wearable technologies provide many possibilities for applications in medicine, and especially in physiotherapy, where tracking and evaluation of body motion are of utmost importance. Despite the existence of multiple smart garments produced for applications in physiotherapy, there is limited information available on the actual impact of these technologies on the clinical outcomes. The objective of this paper is to evaluate the impact of the Double Aid (DAid) smart shirt, a purely textile-based system, on the training process of patients with subacromial pain syndrome. A randomized controlled trial was performed where patients with subacromial pain syndrome had to perform the assigned training exercises while employing the DAid smart shirt system. The core point of each exercise was to perform a movement while holding the shoulders stationary. The smart shirt was designed to sense even slight shoulder motion thus providing the patient with feedback on the accuracy of the motion, and allowing the patient to adjust the movement. The appropriate muscles should be strengthened through an increased effort to control the shoulder motion. The recovery of patients using the feedback system at the end of the treatment was compared to that of a reference group through standardized tests-the Disabilities of the Arm, Shoulder, and Hand score (DASH score), Closed Kinetic Chain Upper Extremity Stability test (CKCUES test), and internal/external rotation ratio. The test group that used the DAid system demonstrated significantly better results of the performed tests for all applied outcome measures compared to the reference group (p < 0.001). An overall positive impact on the patient recovery was observed from the DAid smart shirt system when applied for rehabilitation training of patients with subacromial pain syndrome.