In vitro and in vivo characterization of three major dadAX promoters in Escherichia coli that are regulated by cyclic AMP-CRP and Lrp.
We have shown previously that the dadAX operon of Escherichia coli expresses multiple transcripts, which are repressed by leucine-responsive regulatory protein (Lrp). Here we used site-directed mutagenesis and in vitro and in vivo transcription assays to show that each of the three major dad transcripts requires a specific promoter. These promoters, P1-P3, overlap and are positively regulated in vivo by cyclic AMP-CRP. DNase I footprinting experiments localized two CRP binding sites in this region: CRP1, which is positioned upstream of P1-P3, and CRP2, which is located within the promoters. Site-directed mutagenesis of each site provided evidence that CRP1 is necessary for the effects of cyclic AMP-CRP on dad expression in vivo and in vitro, and that CRP2 probably plays little or no role in this process.


Disease flare patterns and predictors of systemic lupus erythematosus in a monocentric cohort of 423 Japanese patients during a long-term follow-up: The JUDE study.
To clarify the clinical features of systemic lupus erythematosus (SLE) patients, factors associated with flares, and changes over time. Patients having SLE with a visiting history were entered into the Juntendo University Database of Erythematosus. We included 423 cases in the long-term follow-up analysis, and 383 cases were followed for 10 years after the initiation of any therapeutic intervention (comparative analysis: 1973-1982, 82 cases; 1983-1992, 141, and 1993-2002, 160). We assessed changes in the patients' background characteristics, disease symptoms, flare rates, etc. Among the 423 cases, the mean follow-up period was 25.9 years, and mean number of flares was 0.51. Of those, 31.9% had ≥1 flares. Thrombocytopenia at onset contributed to the flares. For disease symptoms at onset, a recent trend in increasing thrombocytopenia was observed. The combination rate of immunosuppressive agents for diseases other than lupus nephritis was slightly increased, and there was no improvement until the first flare or in the flare rate. Thrombocytopenia at onset is predictive factor for flares. Since SLE is a diverse disease with varying symptoms at recurrence, the treatment guidelines should be improved for thrombocytopenia from a long-term perspective.


Mechanistic Insights into the Chemo- and Regio-Selective B(C6F5)3 Catalyzed C-H Functionalization of Phenols with Diazoesters.
The Lewis acidic B(C6F5)3 was recently demonstrated to be effective for the C-H alkylation of phenols with diazoesters. The method avoids the general hydroxyl activation in transition-metal catalysis. Ortho-selective C-H alkylation occurs regardless of potential para-selective C-H alkylation and O-H alkylation. In the present study, a theoretical calculation was carried out to elucidate the reaction mechanism and the origin of chemo- and regio-selectivity. It is found that the previously proposed B(C6F5)3/N or B(C6F5)3/C bonding-involved mechanisms are not favorable, and a more favored one involves the B(C6F5)3/C═O bonding, rate-determining N2 elimination, selectivity-determining electrophilic attack, and proton transfer steps. Meanwhile, the new mechanism is consistent with KIE and competition experiments. The facility of the mechanism is attributed to two factors. First, the B(C6F5)3/C═O bonding reduces the steric hindrance during electrophilic attack. Second, the bonding forms the conjugated system by which the LUMO energy is reduced via the electron-withdrawing B(C6F5)3. The ortho-selectivity resulted from the greater ortho-C-C (than para-C-C) interaction and the O-H···O and O-H···F hydrogen-bond interaction during electrophilic attack. The greater C-C (than C-O) interaction and the π-π stacking between the benzene rings of phenol and diazoester concerted contribute to the chemo-selective C-H alkylation.


Transcellular transport of organic anions across a double-transfected Madin-Darby canine kidney II cell monolayer expressing both human organic anion-transporting polypeptide (OATP2/SLC21A6) and Multidrug resistance-associated protein 2 (MRP2/ABCC2).
Human organic anion transporting polypeptide 2 (OATP2/SLC21A6) and multidrug resistance-associated protein 2 (MRP2/ABCC2) play important roles in the vectorial transport of organic anions across hepatocytes. In the present study, we have established a double-transfected Madin-Darby canine kidney (MDCK II) cell monolayer, which expresses both OATP2 and MRP2 on basal and apical membranes, respectively. The basal-to-apical transport of 17 beta estradiol 17 beta-d-glucuronide (E(2)17 beta G), pravastatin, and leukotriene C(4) (LTC(4)), which are substrates of OATP2 and MRP2, was significantly higher than that in the opposite direction in the double-transfected cells. Such vectorial transport was also observed for taurolithocholate sulfate, which is transported by rat oatp1 and Mrp2. The K(m) values of E(2)17 beta G and pravastatin for the basal-to-apical flux were 27.9 and 24.3 microm, respectively, which were comparable with those reported for OATP2. Moreover, the MRP2-mediated export of E(2)17 beta G across the apical membrane was not saturated. In contrast, basal-to-apical transport of estrone-3-sulfate and dehydroepiandrosterone sulfate, which are significantly transported by OATP2, but not by MRP2, was not stimulated by MRP2 expression. The double-transfected MDCK II monolayer expressing both OATP2 and MRP2 may be used to analyze the hepatic vectorial transport of organic anions and to screen the transport profiles of new drug candidates.


Helix packing and subunit conformation in horse spleen apoferritin.
An electron density map of horse spleen apoferritin at 0.28-nm (2.8 A) resolution and its preliminary interpretation have been described previously. Rigorous examination of this and newer maps at the same nominal resolution but calculated from more extensive data sets, including model building in a Richards' comparator, now allows us to report on structural features in more detail. We list inter-helical angles within and between neighbouring subunits, and describe a new short region of inter-subunit anti-parallel pleated sheet. A short section of electron density not properly accounted for in the first model has also been found. We also report on two alternative ways of connecting helical regions which account almost equally well for the observed electron density, and we assess these two alternative conformations and compare them with the conformations of other known proteins.


Mapping correspondence between facial mimicry and emotion recognition in healthy subjects.
We aimed at verifying the hypothesis that facial mimicry is causally and selectively involved in emotion recognition. For this purpose, in Experiment 1, we explored the effect of tonic contraction of muscles in upper or lower half of participants' face on their ability to recognize emotional facial expressions. We found that the "lower" manipulation specifically impaired recognition of happiness and disgust, the "upper" manipulation impaired recognition of anger, while both manipulations affected recognition of fear; recognition of surprise and sadness were not affected by either blocking manipulations. In Experiment 2, we verified whether emotion recognition is hampered by stimuli in which an upper or lower half-face showing an emotional expression is combined with a neutral half-face. We found that the neutral lower half-face interfered with recognition of happiness and disgust, whereas the neutral upper half impaired recognition of anger; recognition of fear and sadness was impaired by both manipulations, whereas recognition of surprise was not affected by either manipulation. Taken together, the present findings support simulation models of emotion recognition and provide insight into the role of mimicry in comprehension of others' emotional facial expressions.


Interaction of human beta-endorphin with nonopiate binding sites on the terminal SC5b-9 complex of human complement. Significance of COOH-terminal beta H-endorphin fragments.
We have characterized the binding of 125I-labeled human beta-endorphin (125I-beta H-endorphin) to sites present on the terminal fluid-phase complex of human complement, consisting of complement components C5b, C6, C7, C8, C9, and the S-protein (SC5b-9 complex). Specific binding exhibited saturability, reversibility, structural specificity, temperature dependence, and absence of negative cooperative effects. Binding was maximal at 4 degrees C and pH 7.0; it was diminished by monovalent and divalent cations as well as by increasing concentrations of urea and Triton X-100 and apparently required intact disulfide groups. Binding was not inhibited by a number of opioid peptides sharing common sequences with the NH2 terminus of beta H-endorphin. In contrast, binding was inhibited by beta H-endorphin, N-acetyl-beta H-endorphin, and a series of COOH-terminal beta H-endorphin fragments, where of the COOH-terminal dipeptide Gly-Glu represented the minimal effective structure. Stepwise extension towards the NH2 terminus led to an increased binding affinity of the respective fragment. Computer resolution of competition curves yielded one binding component for several shorter COOH-terminal beta H-endorphin fragments and for beta H-endorphin (1-5) + (16-31), whereas two distinct binding components were obtained when beta H-endorphin (27-31), beta H-endorphin (6-31), N-acetyl-beta H-endorphin or beta H-endorphin were used as inhibitors. This study presents detailed data on the binding of COOH-terminal beta H-endorphin fragments to specific nonopiate binding sites present on the terminal SC5b-9 complex of human complement. We suggest that through this interaction, beta H-endorphin may modulate certain functions within the immune system.


Morphine stimulates vascular endothelial growth factor-like signaling in mouse retinal endothelial cells.
Go/Gi coupled G-protein receptor mediated transactivation is critical in the activation of receptor tyrosine kinases (RTK). Here we show that mu opioid receptor (MOR) transactivates Flk1 and platelet-derived growth factor-beta (PDGF-beta) receptors and its agonist morphine stimulates pro-angiogenic and survival-promoting signaling in mouse retinal endothelial cells (mREC). Morphine stimulates mREC proliferation in a dose dependent fashion and promotes survival to the same extent as vascular endothelial growth factor164 (VEGF164). Morphine stimulates mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and Akt phosphorylation in a time dependent manner like VEGF in mREC. Moreover, analogous to VEGF, morphine stimulates oncogenic signal transducer and activator of transcription 3 (STAT3) signaling. Morphine as well as VEGF-induced phospho-STAT3 and phospho-Flk1 immunoprecipitated with MOR-associated proteins. In addition morphine also stimulated MOR associated PDGF-beta receptor phosphorylation. Consistent with the relationship between VEGF and MOR we found that VEGF upregulates MOR protein and RNA expression in mREC. These data suggest that MOR associates and transactivates RTKs for Flk1 and PDGF-beta, which may have a compounding effect on angiogenic signaling in endothelium. Therefore, G-Protein coupled receptors including MOR provide novel targets to develop anti-angiogenic agents.


How doctors learn and perform sustained inflations with a self-inflating bag: a manikin study with a newborn lung simulator.
Sustained inflations during initial resuscitation may help a depressed infant make a more efficient transition to air-filled lungs. This study examined whether doctors could perform sustained inflations with a self-inflating bag in high and low compliance settings and with an open or blocked pressure-relief valve. We asked 43 doctors to carry out sustained inflations for more than 5-sec in a manikin connected to a newborn lung simulator with randomised compliance settings. Tidal volume, inflation time, peak and mean inflating pressure were measured, and 34 were retested 3 months later. The majority of the doctors - 72% in the initial study and 62% in the retest - managed sustained inflations within three ventilation attempts, irrespective of lung compliance setting and years of work experience. Using a blocked pressure-relief valve produced higher tidal volume (27.8 versus 22.6 mL, p < 0.001), inflation time (8.9 versus 8.1 sec, p = 0.025), peak inflating pressure (34.0 versus 28.0 cmH2O; p = 0.012) and mean inflating pressure (28.1 versus 22.8 cmH2O; p < 0.001). The majority of doctors could deliver sustained inflation with a self-inflating bag in a newborn lung simulator for more than 5-sec. Using a blocked pressure-relief valve resulted in higher inflation time, tidal volume and inflation pressure.


Cytoplasmic origin of an intranuclear DNA virus (duck plague virus).
Electron microscopic and autoradiographic studies have revealed that the assembly and maturation of duck plague virus (DPV, an intranuclear DNA virus) occur both in the nucleus and in the cytoplasm. A dense cytoplasmic matrix (tentatively called viroplast) is the basic organelle in which the assembly of nucleocapsids occurs. At the early stage of virion maturation, dense particles, approximately 40 nm in size, appear and become assembled nucleocapsid structures later on. As capsids become mature, they obtain a diameter of 95-105 nm. The viroplast matrix gradually decreases in size, and a loose structure appears inside as the nucleocapsids increase in number. When viroplast has almost disappeared, nucleocapsids become abundant, regularly arranged around the cytoplasmic vacuoles. They bud into these vacuoles, thereby acquiring their envelopes. These nucleocapsids differ from those found in the nucleus both in morphology and in distribution. Electron microscopic autoradiography has shown that there is a huge amount of DNA in viroplast. DNA for the assembly of the nucleocapsids in the cytoplasm is derived from the very viroplast.


Deletion analysis of the mini-P1 plasmid origin of replication and the role of Escherichia coli DnaA protein.
The mini-P1 plasmid origin of replication is contained on a 246 base pair (bp) piece of DNA. At one end there are five 19-bp binding sites for the P1 initiator protein, RepA, and near the other end there are two 9-bp DnaA protein-binding sites. To further define the limits of the origin, we cloned the origin region in M13 and constructed deletions of either end. We sequenced the DNA and tested the replicative form I DNA of the deletion phages for their ability to support RepA-dependent DNA replication in an in vitro system. The origin that is functional in vitro could be reduced to 202 bp. It includes three intact and one incomplete RepA-binding sites at one end and the two DnaA-binding sites at the other end. When the two naturally occurring DnaA-binding sites were replaced with one or two synthetic sites, only the construction containing two sites was active in vitro. We found that the minimal origin that is functional in vivo contains all of the five RepA and the two DnaA-binding sites. Mini-P1 plasmid replication both in vivo and in vitro requires two initiator proteins, the Escherichia coli DnaA protein and the P1 RepA protein. We have found that the ADP form of DnaA is as active as the ATP form of the protein in the in vitro replication of mini-P1. In contrast, only the ATP form is active for in vitro replication of plasmids carrying the E. coli origin (Bramhill, D., and Kornberg, A. (1988) Cell 52, 743-755).


Inhibitory effect of oatmeal extract oligomer on vasoactive intestinal peptide-induced inflammation in surviving human skin.
The aim of this study was to evaluate the antiinflammatory effect of oatmeal extract oligomer on skin fragments stimulated by a neuromediator, vasoactive intestinal peptide (VIP). Skin fragments (from plastic surgery) were maintained in survival conditions for 6 h. To induce inflammation, VIP was placed in contact with dermis by culture medium. Histological analysis was then performed on hematoxylin- and eosin-stained slides. Edema was evaluated with semiquantitative scores. Vasodilation was studied by quantifying the percentage of dilated vessels according to scores and by measuring their surface by morphometrical image analysis. TNF-alpha dosage was made on culture supernatants. Vasodilation was significantly increased after application of VIP. After treatment with oatmeal extract oligomer, the mean surface of dilated vessels and edema were significantly decreased compared with VIP-treated skin. Moreover, treatment with this extract decreased TNF-alpha.


[Ambiguous polyuridylic acid translation by transport RNA from various eukaryotic substances].
Various eucaryotic tRNAs were compared from the standpoint of their ability to misread poly-U in the tRNA-dependent cell-free system of wheat germs. The highest level of 14C-leucine incorporation into the product of translation is observed for tRNA from differentiated (lactating) mammary gland under conditions of the increased Mg2+ concentration. Possible reasons may be connected with different tRNALeu/tRNAPhe ratios in these tissues and with the appearance of specific isoacceptory tRNA4Leu in the total set of tRNA from the lactating gland.


Proline and glycinebetaine induce antioxidant defense gene expression and suppress cell death in cultured tobacco cells under salt stress.
Salt stress causes oxidative damage and cell death in plants. Plants accumulate proline and glycinebetaine (betaine) to mitigate detrimental effects of salt stress. The aim of this study was to investigate the protective effects of proline and betaine on cell death in NaCl-unadapted tobacco (Nicotiana tabacum) Bright Yellow-2 suspension-cultured cells subjected to salt stress. Salt stress increased reactive oxygen species (ROS) accumulation, lipid peroxidation, nuclear deformation and degradation, chromatin condensation, apoptosis-like cell death and ATP contents. Neither proline nor betaine affected apoptosis-like cell death and G(1) phase population, and increased ATP contents in the 200mM NaCl-stressed cells. However, both of them effectively decreased ROS accumulation and lipid peroxidation, and suppressed nuclear deformation and chromatin condensation induced by severe salt stress. Evans Blue staining experiment showed that both proline and betaine significantly suppressed increment of membrane permeability induced by 200mM NaCl. Furthermore, among the ROS scavenging antioxidant defense genes studied here, mRNA levels of salicylic acid-binding (SAbind) catalase (CAT) and lignin-forming peroxidase (POX) were found to be increased by proline and betaine under salt stress. It is concluded that both proline and betaine provide a protection against NaCl-induced cell death via decreasing level of ROS accumulation and lipid peroxidation as well as improvement of membrane integrity.


Proteogenomics of the house dust mite, Dermatophagoides farinae: Allergen repertoire, accurate allergen identification, isoforms, and sex-biased proteome differences.
The allergen repertoire of the house dust mite, Dermatophagoides farinae, is incomplete despite most mite allergens having been described in this species. Using proteogenomics, we aimed to compare proteins and allergens between sexes and provide a foundation for the identification of novel allergens. Overall, 6297 protein hits were identified, and 2899 and 886 were male- and female-specific, respectively. Removal of trace results narrowed the dataset to 3478 hits, including 275 and 157 male- and female-specific hits, respectively. All 34 WHO/IUIS-approved D. farinae allergens (omitting Der f 17) were identified, and we also identified homologs of the yet undescribed Der f 9 and 38. Der f 27/serpin exhibited the largest sex-dependent difference and was dominant in females. Using official protein sequences, Der f 11, 14, 23, 28 and 30 were identified with low success. However, identification success of Der f 11 and 14 was greatly increased by using longer/complete sequences. Der f 30 is characterized by the same tryptic digests as the more abundant Der f 30 (isoform) identified here. Der f 23 appears to be of low abundance in mite bodies. Der f 28.0101 and Der f 28.0201 were detected at low abundance and in trace amounts, respectively. SIGNIFICANCE: In this work, we performed a proteogenomic annotation of the house dust mite, Dermatophagoides farinae, which is the most important source of house dust allergens. The proteogenomic analysis performed here provides a foundation for not only understanding the biology of the mite but also the identification of novel allergens. This study generated a robust proteomic dataset for D. farinae and reviewed existing and candidate allergens in this species. We stress some pitfalls of high-throughput analyses, especially that improper headers of allergen protein records provided in databases can lead to confusion. Using partial sequences in proteomic identification and quantification can lead to low identification success (low signal intensity or MS/MS counts). Thus, we individually curated the protein sequences for proper identification and quantification. The discovered sex differences can be one factor affecting allergen/immunogen variations in mite extracts. Overall, this work provides a benchmark for accurate identification of mite immunogenic proteins using proteomics.


[Determining the aortic and renal hemodynamic effects of propofol on rabbits using Doppler ultrasonography].
The hemodynamic effects of propofol-fentanyl anesthesia were known to reduce arterial pressure and heart rate. We aimed to determine the effects of propofol-fentanyl anesthesia on renal artery blood flow and aorta by using Doppler ultrasonography on an animal model. The study was conducted on 8 male New Zaeland White strain rabbits. The initial aortic and renal hemodynamic measurements were determined on animals using ketamin anesthesia. Later the right femoral arteries were cannulated for determining arterial blood pressure and pulse oximetry was followed from the ear. Propofol 5-7 mg/kg with fentanyl 1-2 microg/kg was injected via the ear vein. Doppler parameters (RI, RI, Vmax, Vmin) of the renal artery and aorta were determined after the anesthesia. According to the basal measurements, arterial blood pressure and breath rates declined significantly (27% and 53% respectively, p<0.05); however there were no statistical alteration of aortic and renal artery Doppler parameters. Anesthesia induced by propofol-fentanyl did not induce a statistically significant difference in Doppler parameters obtained from the renal artery and aorta. It is clinically important to know that determining the changes in RI and PI during the postoperative period is not related to the propofol-fentanyl anesthesia.


Therapeutic angiogenesis with recombinant fibroblast growth factor-2 for intermittent claudication (the TRAFFIC study): a randomised trial.
Recombinant fibroblast growth factor-2 (rFGF-2) improves perfusion in models of myocardial and hindlimb ischaemia. We investigated whether one or two doses of intra-arterial rFGF-2 improves exercise capacity in patients with moderate-to-severe intermittent claudication. 190 patients with intermittent claudication caused by infra-inguinal atherosclerosis were randomly assigned (1:1:1) bilateral intra-arterial infusions of placebo on days 1 and 30 (n=63); rFGF-2 (30 microg/kg) on day 1 and placebo on day 30 (single-dose, n=66); or rFGF-2 (30 microg/kg) on days 1 and 30 (double-dose, n=61). Primary outcome was 90-day change in peak walking time. Secondary outcomes included ankle-brachial pressure index and safety. The main analysis was per protocol. Before 90 days, six patients had undergone peripheral revascularisation and were excluded, and ten withdrew or had missing data. 174 were therefore assessed for primary outcome. Peak walking time at 90 days was increased by 0.60 min with placebo, by 1.77 min with single-dose, and by 1.54 min with double-dose. By ANOVA, the difference between groups was p=0.075. In a secondary intention-to-treat analysis, in which all 190 patients were included, the difference was p=0.034. Pairwise comparison showed a significant difference between placebo and single-dose (p=0.026) but placebo and double-dose did not differ by much (p=0.45). Serious adverse events were similar in all groups. Intra-arterial rFGF-2 resulted in a significant increase in peak walking time at 90 days; repeat infusion at 30 days was no better than one infusion. The findings of TRAFFIC provide evidence of clinical therapeutic angiogenesis by intra-arterial infusion of an angiogenic protein.


Brittle smiles: positive biases toward stigmatized and outgroup targets.
We examined individuals' tendencies to exaggerate their positive responses toward stigmatized others (i.e., overcorrect) and explored how overcorrection, because of its fragile nature, could be disrupted. The first 2 studies demonstrate overcorrection: White participants paired with Black partners (Experiment 1A) smiled, laughed, and showed more positive behavior than those paired with same-race partners. Experiment 1B replicated the general effect with a physically stigmatized sample (i.e., facial birthmarks) and then demonstrated that overcorrection is moderated by bias; participants who exhibited more positive behavior toward their partner showed the most physiological "threat" during a stressful task with their partner. We then examined the idea that if overcorrection requires cognitive resources and is effortful, then it may be fragile when resources are taxed. In Experiments 2 and 3, we observed that overcorrection was easily disrupted when resources were compromised (e.g., with stress or cognitive load). Taken together, these studies suggest that positive biases toward stigmatized and outgroup members are fragile and can be undermined when resources are taxed.


Multipole expansion of Bessel and Gaussian beams for Mie scattering calculations.
Multipole expansions of Bessel and Gaussian beams, suitable for use in Mie scattering calculations, are derived. These results allow Mie scattering calculations to be carried out considerably faster than existing methods, something that is of particular interest for time evolution simulations where large numbers of scattering calculations must be performed. An analytic result is derived for the Bessel beam that improves on a previously published expression requiring the evaluation of an integral. An analogous expression containing a single integral, similar to existing results quoted, but not derived, in literature, is derived for a Gaussian beam, valid from the paraxial limit all the way to arbitrarily high numerical apertures.


Salivary glands.
A review of the more common inflammatory and neoplastic conditions affecting salivary glands has been presented. The use of hydration, massage, antibiotics, and steroids is effective initial treatment for suppurative sialadenitis and usually negates the need for surgical drainage. Total excision of the salivary gland and its duct is necessary in procedures for recurrent infection. Our technique for closure of the floor of the mouth after excision of the submandibular gland and Wharton's duct is described. Salivary neoplasms involving the parotid gland, the submandibular gland, and the minor salivary glands are treated on the basis of their histologic and local findings. Stepwise illustrations of our technique of parotidectomy and surgical considerations, including the counseling of a patient with a parotid mass, are presented to assist surgeons who care for patients with salivary disorders.


A randomised controlled trial of the effectiveness of providing free fluoride toothpaste from the age of 12 months on reducing caries in 5-6 year old children.
To assess the impact of regularly supplying free fluoride toothpaste regularly to children, initially aged 12 months, and living in deprived areas of the north west of England on the level of caries in the deciduous dentition at 5-6 years of age. A further aim was to compare the effectiveness of a programme using a toothpaste containing 440 ppmF (Colgate 0-6 Gel) with one containing 1,450 ppmF (Colgate Great Regular Flavour) in reducing caries. Randomised controlled parallel group clinical trial. Clinical data were collected from test and control groups when the children were 5-6 years old. A programme of posting toothpaste with dental health messages to the homes of children initially aged 12 months. Clinical examinations took place in primary schools. 7,422 children born in 3-month birth cohorts living in high caries areas in nine health districts in north west England. Within each district children were randomly assigned to test or control groups. Toothpaste, containing either 440 ppmF or 1450 ppmF, and dental health literature posted at three monthly intervals to children in test groups until they were aged 5-6 years. The dmft index, missing teeth and the prevalence of caries experience. An analysis of 3,731 children who were examined and remained in the programme showed the mean dmft to be 2.15 for the group who had received 1,450 ppmF toothpaste and 2.49 for the 440 ppmF group. The mean dmft for the control group was 2.57. This 16% reduction between the 1,450 ppmF and control group was statistically significant (P<0.05). The difference between the 440 ppmF group and control was not significant. Further analyses to estimate the population effect of the programme also confirmed this relationship. This study demonstrates that a programme distributing free toothpaste containing 1,450 ppmF provides a significant clinical benefit for high caries risk children living in deprived, non-fluoridated districts.


Secondary ureteroscopy: results and management strategy at a referral center.
In an era when extracorporeal shock wave lithotripsy occupies a dominant place in the treatment of urolithiasis ureteroscopy retains an important role in certain circumstances. While often a definitive procedure, ureteroscopy can be associated with potential risks and complications. The treatment of patients who have undergone a failed attempt at ureteroscopic stone retrieval or have a complication may be complex. As a tertiary care stone referral center we review our experience with performing salvage ureteroscopy following a previous unsuccessful attempt at endoscopic stone removal. Between May 1990 and February 1996, 79 patients were referred following an unsuccessful attempt at retrograde endoscopic or basket manipulation for ureteral calculi. A retrospective review of the outcomes of these patients was conducted. Of the patients 11 presented with associated complications, which included ureteral perforation (4), intramural false passage (1) and fever or sepsis (6). Complications were managed by early establishment of urinary tract drainage by stenting or nephrostomy. Among patients without complications elective salvage ureteroscopy was performed. Ureteroscopy was used in 79 patients with a successful outcome (stone-free) in 75 (95%). Followup imaging with renal ultrasound or excretory urography at least 3 months after secondary ureteroscopy was available in 65 patients and showed no evidence of hydronephrosis or delayed stricture formation. Treating the patient who undergoes a failed attempt at ureteroscopy may be problematic and requires access to a wide array of endourological equipment. Each subsequent treatment should be individualized with consideration given to stone size, location and general health. In the presence of a ureteral injury establishment of early urinary tract drainage is essential. Following stabilization, secondary ureteroscopy can be performed yielding high stone-free rates with minimal complications.


Diurnal intraocular pressure. Correlation to automated perimetry.
Patients referred to a centralized glaucoma laboratory obtained intraocular pressure measurements every two hours from 5:00 am to 3:00 pm. Analysis revealed 400 eyes with visual field defects and 400 eyes without visual field defects as determined by Octopus perimetry of the central 20 degrees. The diurnal variation in intraocular pressure was 6.2 mmHg +/- 3.6 for those with visual field defects and 5.5 mmHg +/- 2.7 for those without visual field defects. There was no statistical significance in the mean diurnal variation between the two groups (P = 0.91). The highest intraocular pressure tended to occur at either 5:00 am to 7:00 am or 11:00 am to 1:00 pm in both groups. The lowest intraocular pressure tended to occur between 7:00 am to 9:00 am or 1:00 pm to 3:00 pm in both groups. No significant differences were noted in the distribution between the two groups with regard to the time of the highest or lowest intraocular pressure. In the group with visual field defects, 30% had an intraocular pressure of less than 23 mmHg and 23% had an intraocular pressure of greater than or equal to 23 mmHg at all five time periods.


[Conjugal plasmid transfer in Bacillus subtilis in soil microcosms].
Conjugal transfer of the small plasmid pUB110 between Bacillus subtilis strains was studied under conditions of microcosms with sterile and nonsterile soil. Plasmid transfer proved to be possible after soil inoculation with vegetative partner cells or with their spores. Plasmid transfer occurred at temperatures of 30 degrees C and 22-23 degrees C.


Comparison of success rates between two video laryngoscope systems used in a prehospital clinical trial.
The primary aims of this study were to compare paramedic success rates and complications of two different video laryngoscopes in a prehospital clinical study. This study was a multi-agency, prospective, non-randomized, cross over clinical trial involving paramedics from four different EMS agencies. Following completion of training sessions, six Storz CMAC™ video laryngoscopes and six King Vision™ (KV) video laryngoscopes were divided between agencies and placed into service for 6 months. Paramedics were instructed to use the video laryngoscope for all patients estimated to be ≥ 18 years old who required advanced airway management per standard operating procedure. After 6 months, the devices were crossed over for the final 6 months of the study period. Data collection was completed using a telephone data collection system with a member of the research team (available 24/7). First attempt success, overall success, and success by attempt, were compared between treatment groups using exact logistic regression adjusted for call type and user experience. Over a 12-month period, 107 patients (66 CMAC, 41 KV) were treated with a study device. The CMAC had a significantly higher likelihood of first attempt success (OR = 1.85; 95% CI 0.74, 4.62; p = 0.188), overall success (OR = 7.37; 95% CI 1.73, 11.1; p = 0.002), and success by attempt (OR = 3.38; 95% CI 1.67, 6.8; p = 0.007) compared to KV. Providers reverted to direct laryngoscopy in 80% (27/34) of the video laryngoscope failure cases, with the remaining patients having their airways successfully managed with a supraglottic airway in 3 cases and bag-valve mask in 4 cases. The provider-reported complications were similar and none were statistically different between treatment groups. Complication rates were not statistically different between devices. The CMAC had a higher likelihood of successful intubation compared to the King Vision. Complication rates were not statistically different between groups. Video laryngoscope placement success rates were not higher than our historical direct laryngoscopy success rates.


[Improvement of drug bioavailability using protease inhibitors].
Although many proteins and peptides were produced by gene-technology, their administration routes are limited to be i.v. route. To increase the clinical use of these products, oral dosage form is required. However, in the case of oral administration, proteins are degraded by digestive fluid having strong protease activity. To decrease the protease activity, protease inhibitors are administered concomitantly with proteins. Aprotinin, soybean trypsin inhibitors and gel-forming polymer such as Polycarbophil are representative protease inhibitors. Gel-forming polymers have both protease inhibiting activity and absorption enhancing effect on protein/peptides. The usefulness of these protease inhibitors have been suggested as an additive for the oral delivery of G-CSF and vasopressin derivatives.


Role of Na-K pump potassium regulation and IPSPs in seizures and spreading depression in immature rabbit hippocampal slices.
1. Using the immature (8-12 days postnatal) rabbit hippocampal slice preparation, we investigated regional extracellular potassium concentration [( K+]o) changes that occur during spontaneous and evoked spreading depression (SD) episodes. We report here a difference between the CA1 and CA3 cell populations in the immature hippocampus with regard to 1) resting [K+]o, 2) magnitude of the [K+]o change during seizurelike events and SDs, and 3) susceptibility to SD episodes. Experiments were also performed to elucidate the roles that the Na-K pump and synaptic inhibition play in controlling SD onset, duration, and recovery. We demonstrated a major role for potassium regulation by the Na-K pump and a lesser modulatory role for inhibitory postsynaptic potentials (IPSPs) in preventing SD in the CA3 region. 2. Simultaneous intra- and extracellular recordings were made in the CA1 and CA3 regions of the immature rabbit hippocampus during spontaneous or evoked SD, while potassium ion-sensitive microelectrodes (K-ISMs) monitored changes in [K+]o. The CA1 region had 1) a higher frequency of spontaneous SD episodes than CA3, 2) a lower threshold to potassium-triggered SD, 3) a longer duration SD episode, and 4) smaller post-SD membrane potential and [K+]o undershoots (below the original resting membrane potential and resting [K+]o). 3. During the onset of a SD episode in the CA1 region, the local [K+]o rose either before or at the same time as the membrane potential depolarization. 4. In the CA3 region, spontaneous ictallike events consisting of tonic cell depolarization with repetitive activity followed by clonic afterdischarges were more likely to occur than SD episodes. During these ictallike episodes, [K+]o rose above the 10- to 12-mM ceiling level reported for adult CNS tissue during seizures. Increases in [K+]o evoked by repetitive stimulation were regulated at a lower level in CA3 (average [K+]o rise to 11.4 mM) than in CA1 (average [K+]o rise to 18.3 mM). 5. In CA3, bath application of 10 microM bicuculline or 3.4 mM penicillin did not change the frequency of spontaneously occurring SDs or the SD response threshold to local pressure ejection of 2 M KCl. However, blockade of IPSPs did lead to lower thresholds for SD or seizurelike episodes elicited by stimulation of the mossy fibers. 6. A single application of ouabain (10 microM) to CA3 by local pressure ejection caused a slow rise in local [K+]o measured with K-ISMs. The ouabain treatment also increased the frequency of spontaneous postsynaptic potential activity and decreased the amplitude and duration of CA3 pyramidal cell afterhyperpolarizations (AHPs).(ABSTRACT TRUNCATED AT 400 WORDS)


Sleep-wake patterns in children with intrauterine growth retardation.
The purpose of this study was to characterize the sleep patterns of children with intrauterine growth retardation, known to be at risk for neurodevelopmental disorders, and seek a possible correlation with behavior, concentration, and attention problems. The sleep patterns of 26 children with intrauterine growth retardation aged 4 to 7 years were compared with those of 47 control children using activity monitors (actigraphs). In addition, data were collected from the parents regarding sleep habits, behavior, concentration, and attention. Children with intrauterine growth retardation aged 4 to 7 years were found to have a tendency toward poorer quality of sleep than their matched controls. This inclination was statistically significant only for one sleep measure, the true sleep time. A tendency toward increased fragmentation of sleep, prolonged waking, and decreased sleep efficiency, although not statistically significant in this study, was demonstrated. Our results showed that 58% of the children with intrauterine growth retardation, compared with 40% of the children in the control group, could be defined as "poor sleepers" (sleep efficiency lower than 90% or three or more waking episodes per night). This disturbed sleep profile is probably an integral part of the neurodevelopmental profile typical of these at-risk children. No significant correlations were found between sleep quality and behavior, concentration, and attention problems.


[Age-related features of the working capacity of rats during positive emotional reinforcement by hypothalamic self-stimulation].
Some age-related peculiarities found in rats striving for hypothalamic self-stimulation: (1) the capacity in old rats decreased more rapidly during fatique than in adult ones; (2) some old but not adult animals died after hard physical work; (3) some old rats "working" for a longer time have done greater work, than adult ones.


Long-term tumor control following stereotactic radiosurgery for jugular paraganglioma using 3D volumetric segmentation.
OBJECTIVEThe morbidity of gross-total resection of jugular paraganglioma (JP) is often unacceptable due to the potential for irreversible lower cranial neuropathy. Stereotactic radiosurgery (SRS) has been used at the authors' institution since 1990 for the treatment of JP and other benign intracranial tumors. Conventional means of assessing tumor progression using linear measurements or elliptical approximations are imprecise due to the irregular shape and insinuating growth pattern of JP. The objective of this study was to assess long-term tumor control in these patients by using slice-by-slice 3D volumetric segmentation of serial MRI data.METHODSRadiographic data and clinical records were reviewed retrospectively at a single, tertiary-care academic referral center for patients treated from 1990 to 2017. Volumetric analyses by integration of consecutive tumor cross-sectional areas (tumor segmentation) of serial MRI data were performed. Tumor progression was defined as volumetric growth of 15% or greater over the imaging interval. Primary outcomes analyzed included survival free of radiographic and clinical progression. Secondary outcomes included new or worsened cranial neuropathy.RESULTSA total of 85 patients were treated with Gamma Knife radiosurgery (GKRS) for JP at the authors' institution over the last 27 years. Sixty patients had pretreatment and serial posttreatment contrast-enhanced MRI follow-up suitable for volumetric analysis. A total of 214 MR images were analyzed to segment tumor images in a slice-by-slice fashion to calculate integral tumor volume. The median follow-up duration was 66 months (range 7-202 months). At 5 years the tumor progression-free survival rate was 98%. Three tumors exhibited progression more than 10 years after GKRS. Estimated survival free of radiographic progression rates (95% confidence interval [CI]; n = number still at risk) at 5, 10, and 15 years following radiosurgery were 98% (95% CI 94%-100%; n = 34), 94% (95% CI 85%-100%; n = 16), and 74% (95% CI 56%-98%; n = 6), respectively. One patient with tumor progression required treatment intervention using external beam radiation therapy, constituting the only case of clinical progression. Two patients (3%) without preexisting lower cranial nerve dysfunction developed new ipsilateral vocal fold paralysis following radiosurgery.CONCLUSIONSSRS achieves excellent long-term tumor control for JP without a high risk for new or worsened cranial neuropathy when used in primary, combined modality, or recurrent settings. Long-term follow-up is critical due to the potential for late radiographic progression (i.e., more than 10 years after SRS). As none of the patients with late progression have required salvage therapy, the clinical implications of this degree of tumor growth have yet to be determined.


[MRI-Based Ratio of Fetal Lung to Body Volume as New Prognostic Marker for Chronic Lung Disease in Patients with Congenital Diaphragmatic Hernia].
Background Our aim was to evaluate the prognostic value of magnetic resonance imaging (MRI)-based ratio of fetal lung volume (FLV) to fetal body volume (FBV) as a marker for development of chronic lung disease (CLD) in fetuses with congenital diaphragmatic hernia (CDH). Patients and Methods FLV and FBV were measured and the individual FLV/FBV ratio was calculated in 132 fetuses. Diagnosis of CLD was established following prespecified criteria and graded into mild/moderate/severe if present. Logistic regression analysis was used to calculate the probability of postnatal development of CLD in dependence of the FLV/FBV ratio. Receiver operating characteristic curves were analysed by calculating the area under the curve to evaluate the prognostic accuracy of this marker. Results 61 of 132 fetuses developed CLD (46.21%). The FLV/FBV ratio was significantly lower in fetuses with CLD (p=0.0008; AUC 0.743). Development of CLD was significantly associated with thoracic herniation of liver parenchyma (p<0.0001), requirement of extracorporal membrane oxygenation (ECMO) (p<0.0001) and gestational age at delivery (p=0.0052). Conclusion The MRI-based ratio of FLV to FBV is a highly valuable prenatal parameter for development of CLD. The ratio is helpful for early therapeutic decisions by estimating the probability to develop CLD. Perinatally, gestational age at delivery and ECMO requirement are useful additional parameters to further improve prediction of CLD.


A scanning electron-microscopic study of the muscle layer of the canine gallbladder.
A connective tissue digestion and removal technique utilizing collagenase and hyaluronidase incubation followed by treatment with 8 N hydrochloric acid successfully revealed the muscular components within the gallbladder wall of the dog, allowing three-dimensional visualization with the scanning electron microscope. The muscle adjacent to the serosa appeared as a homogeneous layer with few discontinuities. In contrast to this were the more superficial muscle layers which were arranged as a meshwork of interlacing smooth muscle bundles of various size, the interstices of which were filled with cellular connective tissue. These gaps in the muscle layer varied in size, becoming larger as the muscle bundles tapered out just below the mucosal aspect of the gallbladder wall.


Batrachochytrium salamandrivorans and the Risk of a Second Amphibian Pandemic.
Amphibians are experiencing devastating population declines globally. A major driver is chytridiomycosis, an emerging infectious disease caused by the fungal pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal). Bd was described in 1999 and has been linked with declines since the 1970s, while Bsal is a more recently discovered pathogen that was described in 2013. It is hypothesized that Bsal originated in Asia and spread via international trade to Europe, where it has been linked to salamander die-offs. Trade in live amphibians thus represents a significant threat to global biodiversity in amphibians. We review the current state of knowledge regarding Bsal and describe the risk of Bsal spread. We discuss regional responses to Bsal and barriers that impede a rapid, coordinated global effort. The discovery of a second deadly emerging chytrid fungal pathogen in amphibians poses an opportunity for scientists, conservationists, and governments to improve global biosecurity and further protect humans and wildlife from a growing number of emerging infectious diseases.


Systematic Review and Meta-Analysis of Avascular Necrosis and Posttraumatic Arthritis After Traumatic Hip Dislocation.
To determine the incidence rate and associative factors for the development of avascular necrosis (AVN) and posttraumatic arthritis (PTA) after traumatic hip dislocation and time to reduction. A comprehensive search of databases including PubMed, Cochrane Database, and Embase through April 2014 for English articles reporting complications of AVN and PTA after hip dislocation was performed. Inclusion criteria were English-only studies, a patient population of adults, study outcomes of AVN and/or PTA reported, and articles reported at least type I dislocations. Two authors independently extracted data from the selected studies and the data collected were compared to verify agreement. Random-effects models were used for meta-analysis. The overall event rate of AVN and PTA was calculated and stratified based on Thompson-Epstein of the hip dislocation. Odds ratios were calculated for those articles that reported rates of AVN based on time to reduction. For anterior dislocations, the event rate for AVN ranged from 0.087 to 0.333, whereas the event rate for PTA ranged from 0.125 to 0.700. Analysis of posterior dislocations revealed that the event rate for AVN ranged from 0.106 to 0.430; additionally, the event rate for PTA ranged from 0.194 to 0.586. For posterior hip dislocations and type I and II anterior dislocations, the severity of the injury correlates with an increase in the development of AVN and PTA. The odds ratio of AVN for those hip dislocations reduced after 12 hours versus those reduced before 12 hours was 5.627. Prognostic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Sustained co-cultivation with human placenta-derived MSCs enhances ALK5/Smad3 signaling in human breast epithelial cells, leading to EMT and differentiation.
The interaction between mammary epithelial cells and their surrounding microenvironment are important in the development of the mammary gland. Thus, mesenchymal stem cells (MSCs), which retain pluripotency for various mesenchymal lineages, may provide a permissive environment for the morphologic alteration and differentiation of mammary epithelial cells. To this end, we investigated whether the interactions between mammary epithelial cells and human placenta-derived MSCs (hPMSC) affect the morphology, proliferation, and differentiation of epithelial cells in a co-culture system. We show that after co-culture with hPMSCs, human mammary epithelial cell lines (MCF-10F and HEMC) underwent significant morphologic alterations and a dramatic increase in ductal-alveolar branching, which was accompanied by a decrease or loss of the epithelial marker E-cadherin and a gain of the mesenchymal markers, alpha-SMA and vimentin. MCF-10F and HEMC proliferation was also inhibited in the presence of hPMSCs, and this retardation in growth was due to cell cycle arrest. Furthermore, in MCF-10F and HMEC cells, hPMSCs induced the production of lipid droplets, milk fat globule protein, and milk protein lactoferrin, which are markers of functional mammary differentiation. We also noticed an elevation in ALK5 and phosphorylated Smad3 protein levels upon hPMSC co-culture. Strikingly, the changes in morphology, proliferation, and differentiation were reversed by treatment with ALK5 or Smad3 knockdown in MCF-10F/hPMSC co-cultures. Collectively, our findings suggest that co-cultivation with hPMSCs leads to epithelial to mesenchymal transition (EMT) and differentiation of human breast epithelial cells through the ALK5/Smad3 signaling pathway.


Axons containing a prolactin-like peptide project into the perivascular layer of the median eminence: an immunocytochemical light and electron microscope study in adult and infant rats.
A light and electron microscopic immunocytochemical study was undertaken to explore the fine structural organization of prolactin-immunoreactive axons in the rat median eminence. In adult intact males and females and in hypophysectomized females, light microscopic immunocytochemical labeling of the mediobasal hypothalamus revealed a marked concentration of prolactin-like immunoreactive fibers in the perivascular layer throughout the median eminence and the hypophysial stalk. At the electron microscopic level, immunostaining was associated with typical neurosecretory axons located either in the palisade layer where they displayed numerous contacts with tanycyte processes, or in the perivascular layer where they frequently contacted the perivascular space. Within the labeled axonal profiles, immunostaining was essentially located on secretory granules, 90-120 nm in diameter, whereas the microvesicles accumulated in some perivascular profiles constantly remained unlabeled. These data strongly suggest that most prolactin-immunoreactive axons of the median eminence release their content into the hypophysial portal vessels. In 1-day-old infant rats, intensely prolactin-like immunoreactive fibers were similarly localized in the most external layer of the median eminence, in which, contrary to adult animals, very slight if any tyrosine-hydroxylase-immunoreactive fibers were detected. Since earlier studies have provided evidence for a nondopaminergic prolactin-release-inhibiting factor in the hypothalamus of infant rats, and for an inhibitory effect of prolactin on pituitary mammotrophs, we propose that hypothalamic prolactin may contribute, as an additional prolactin-release-inhibiting factor, to the multifactorial control of pituitary mammotrophs.


In vivo neuroimaging correlates of the efficacy of paroxetine in the treatment of mood and anxiety disorders.
The advent of neuroimaging technology has brought with it a deeper understanding of brain function and structure in health and psychiatric illness. This article overviews pertinent findings from neuroimaging studies in mood and anxiety disorders. Paroxetine is a particularly well-studied psychopharmacologic agent in this regard. The findings of neuroimaging studies of paroxetine will be placed into perspective for a better understanding of the interaction of this selective serotonin reuptake inhibitor (SSRI) with the serotonergic and noradrenergic systems in the brain that mediate clinical efficacy. When considered in the context of a burgeoning literature on neuroimaging research of the pathophysiology of mood and anxiety disorders, the findings of paroxetine studies suggest a neurobiological explanation for the mechanisms whereby chronic administration of paroxetine affects neural systems involved in the pathophysiology of major depression and several anxiety disorders such as obsessive-compulsive disorder, posttraumatic stress disorder, and social anxiety disorder.


Chronic intestinal pseudo-obstruction: manifestations, natural history and management.
Chronic intestinal pseudo-obstruction (CIPO) is a rare pathological condition characterized by a marked derangement of gut propulsive motility mimicking mechanical obstruction, in the absence of any lesion occluding the gut lumen. This disease is often associated with a disabling and potentially life-threatening complications and is still too often unrecognized even in referral centres. As a result, patients receive neither appropriate care nor recognition of their severe health condition. Medical and surgical therapies are often unsatisfactory and long-term outcome turns out to be poor in the vast majority of cases. This article focuses on the main clinical features, the management and long-term outcome of patients affected by CIPO, with particular emphasis on those aspects which remain a matter of debate.


Blue-light-emitting ZnSe random laser.
We observed blue-light-emitting random laser action with coherent feedback in ZnSe particles. Below the threshold excitation, we observed a broad spontaneous emission band peaking at approximately 470 nm. Above the threshold, several discrete lasing lines appear at the center of the spontaneous emission band (approximately 475 nm). The linewidth of each lasing emission is less than 0.4 nm, which is about 40 times narrower than that of the spontaneous emission band. Although the lasing intensity of each line varies from pulse to pulse, the peak wavelengths do not shift significantly. The lasing emission was also found to radiate in all directions.


Mechanism of antagonizing cyanide-induced lethality by alpha-ketoglutaric acid.
alpha-Ketoglutaric acid (alpha-KG) has been shown to be an effective antagonist for cyanide-induced lethality. The mechanism of this antagonism is hypothesized to result from alpha-KG binding with cyanide. Several investigative approaches were taken to determine the existence of this binding. First, mixtures of various molar ratios of alpha-KG:cyanide were injected into a high pressure liquid chromatograph. The addition of cyanide reduced the peak area of alpha-KG at a molar ratio of greater than 1:5. Second, blood from naive male ICR mice was spiked with alpha-KG and cyanide. Headspace above these blood samples was injected into a gas chromatograph and analyzed for released hydrogen cyanide. alpha-KG reduced the peak area of hydrogen cyanide released into the headspace at molar ratios of greater than 1:2.5. Third, the effect of cyanide on the ultraviolet spectrum of alpha-KG was determined as an indication of binding. In the presence of cyanide the absorption peak at 316 nm for alpha-KG was eliminated. Inhibition of cytochrome oxidase is an accepted target enzyme for cyanide-induced lethality. Fourth, further evidence of alpha-KG's mechanism was determined by the effect of alpha-KG on brain cytochrome oxidase (BRCYTOX) and its ability to antagonize cyanide-induced inhibition of BRCYTOX. BRCYTOX activity was determined in the presence of alpha-KG and was found to be unaffected between 0.01 and 0.06 M of alpha-KG. Greater concentrations of alpha-KG inhibited BRCYTOX activity. The complete inhibition of BRCYTOX activity by 10(-5) M cyanide was prevented with 0.05 and 0.06 M alpha-KG. Fifth, BRCYTOX activity of animals pretreated with saline and then an LD80 dose (8.5 mg/kg) of cyanide was 80% inhibited, while BRCYTOX activity of animals pretreated with 2 g alpha-KG/kg, i.p., and then an LD80 dose (7.75 mg/kg) of cyanide was not different from control values. Thus, these data suggest that alpha-KG does bind with cyanide, and this binding can account for the antagonism of cyanide-induced lethality.


Potency after permanent prostate brachytherapy for localized prostate cancer.
The evaluation of potency preservation after treatment of localized prostate cancer with transperineal permanent prostate brachytherapy (PPB) and the efficacy of sildenafil were studied. This study comprised 482 patients who were able to maintain an erection suitable for intercourse before treatment from a cohort of 1166 patients with clinically localized prostate cancer treated with PPB. All patients have been followed prospectively, and actuarial analysis was performed to assess potency preservation over time. Patients treated with sildenafil were evaluated as to its efficacy. The median follow-up of this cohort was 34 months (6--92), with a median age of 68 years (47--80). Potency was preserved in 311 of the 482 patients, with a 5-year actuarial potency rate of 52.7%. The 5-year actuarial potency rate for patients treated with PPB as monotherapy was 76%, and, for those treated with combination external beam radiotherapy (EBT) + PPB, 56% (p = 0.08). Patients treated with neoadjuvant androgen deprivation (NAAD) + PPB had a 5-year potency rate of 52%, whereas those with combination EBT + PPB + NAAD had a potency rate of 29% (p = 0.13). Cox regression analysis identified that pretreatment use of NAAD and patient age predicted for impotence (p = 0.0001 and 0.04, respectively). Of 84 patients treated with sildenafil, 52 had a successful outcome (62%). The response to sildenafil was significantly better in those patients not treated with NAAD (p = 0.04). The actuarial potency rates at 5 years for patients treated with PPB are lower than generally acknowledged, except for those patients treated with PPB as monotherapy. Patients who received sildenafil exhibited improved potency in a majority of cases.


Bone disease in patients with primary hypercalciuria and calcium nephrolithiasis.
In patients affected by calcium nephrolithiasis, primary hypercalciuria is frequently accompanied by bone demineralisation and increased susceptibility to fragility fractures. The relationship between bone loss and primary hypercalciuria is multifactorial. Organs and tissues which control calcium and phosphate metabolism - bone, intestine, and kidney - are actively involved in the pathogenesis of bone alterations which together form a multi-factorial metabolic disorder. We conducted a comprehensive evaluation of the published data concerning hypercalciuria and nephrolithiasis included in Medline from 1985 and 2005.


Phase II trial of 4'-epi-doxorubicin in metastatic colorectal carcinoma.
Twenty-three patients with metastatic colorectal carcinoma were treated with 4'-epi-doxorubicin (Epirubicin, 4'-epi-DX) 90 mg/m2 i.v. every 4 weeks. Out of 18 patients evaluable for response, 11 were previously untreated. One complete response of 21 + months was observed in a patient with rectosigmoid carcinoma. Furthermore there was one minor response in pretreated rectosigmoid series, and 6 patients had stabilization of disease. Median survival for all patients, including early deaths, was 4 months. For patients who were not pretreated and who received at least 2 cycles of therapy, median survival was 15 months. Further studies in patients with rectosigmoid carcinoma are warranted.


Health effects of hazardous waste.
Since 1995, the Agency for Toxic Substances and Disease Registry (ATSDR) has evaluated environmental contaminants and human health risks at nearly 3000 sites. Hazardous substances at these sites include newly emerging problems as well as historically identified threats. ATSDR classifies sites according to the degree of hazard they represent to the public. Less than 1% of the sites investigated are considered urgent public health hazards where chemical or physical hazards are at levels that could cause an immediate threat to life or health. Approximately 20% of sites have a potential for long-term human exposures above acceptable risk levels. At almost 40% of sites, hazardous substances do not represent a public health hazard. Completed exposure pathways for contaminants in air, water, and soil have been reported at approximately 30% of evaluated sites. The most common contaminants of concern at these sites include heavy metals, volatile organic compounds, and polychlorinated biphenyls. This article reviews ATSDR's ongoing work by examining the historic hazard of lead, the contemporary hazard of asbestos, and the emerging issue of perchlorate contamination.


Redox-Dependent Conformational Dynamics of Decameric 2-Cysteine Peroxiredoxin and its Interaction with Cyclophilin 20-3.
2-Cysteine peroxiredoxins (2-CysPrxs) switch between functions as a thiol peroxidase, chaperone, an interaction partner and possibly a proximity-based oxidase in a redox-dependent manner. In photosynthetic eukaryotes, 2-CysPrx localizes to the plastid, functions in the context of photosynthesis and enables an ascorbate peroxidase-independent water-water cycle for detoxifying H2O2 The high degree of evolutionary conservation of 2-CysPrx suggests that the switching is an essential characteristic and needed to transduce redox information to downstream pathways and regulation. The study aimed at exploring the dissociation behavior of 2-CysPrx and its interactions with cyclophilin depending on bulk phase conditions. Isothermal titration microcalorimetry (ITC), dynamic light scattering and size exclusion chromatography (SEC) proved the previously suggested model that reduced 2-CysPrx below a critical transition concentration (CTC) exists in its dimeric state, and above the CTC adopts the decameric state. The presence of cyclophilin 20-3 (Cyp20-3) affected the CTC of a 2-CysPrx decamer suggesting interaction which was further quantified by direct titration of 2-CysPrx with Cyp20-3, and in overlays. Finally catalytic inactivation assays showed the higher catalytic efficiency of 2-CysPrx at pH 8 compared with pH 7.2, but also revealed increased inactivation by hyperoxidation at pH 8. Interestingly, calculation of the average turnover number until inactivation gave rather similar values of 243 and 268 catalytic cycles at pH 8 and pH 7.2, respectively. These quantitative data support a model where 2-CysPrx and Cyp20-3, by interaction, form a redox-sensitive regulatory module in the chloroplast which is under control of the photosynthesis-linked stromal pH value, the redox state and additional stromal protein factor(s).


Emerging haemostatic agents and patient blood management.
The transfusion of allogeneic blood products has been considered as a life-saving procedure for patients suffering from major traumatic injury and those who are undergoing major surgery. The safety of blood products has improved in terms of infectious complications over the last three decades due to advanced donor screening procedures and tests. Nevertheless, non-infectious complications including a blood-type mismatch, volume overload and immunologic and non-immunologic reactions to blood products can adversely affect clinical outcomes. It is thus important to implement a patient-specific strategy in diagnosing bleeding cause(s) and optimising haemostatic therapy. This strategy is an integral part of patient blood management applicable to many perioperative patients. Recent advances in the haemostatic management and transfusion include better understanding of the pathomechanisms of coagulopathy, availability of point-of-care coagulation monitoring and introductions of pathogen-inactivated plasma and factor concentrates as well as recombinant coagulation factors. Understanding the indications and limitations of conventional haemostatic therapy, and potential indications and complications relating to emerging haemostatic agents, is important for perioperative physicians. In this article, we discuss current issues related to allogeneic plasma products and emerging biological haemostatic agents and techniques. Further, we review the mechanisms of action and available preclinical or clinical data for each therapeutic agent.


Indirect laryngoscopic assessment for the diagnosis of difficult intubation in patients undergoing microlaryngeal surgery.
The aim of this study is to investigate the feasibility of using indirect laryngoscopy for the diagnosis of difficult intubations in patients who are undergoing microlaryngeal surgery. In the pre-anesthetic examination the thyromental distance, sternomental distance, interincisor distance, neck circumference, modified Mallampati scores, and Wilson risk scores were measured. An otolaryngologist conducted an indirect laryngoscopic assessment. Direct laryngoscopic profiles were classified according to their Cormack-Lehane scores. Forty patients underwent microlaryngeal surgery. Their mean ages, neck circumferences, Cormack-Lehane scores of the patients who had intubation difficulties were statistically significantly higher than those who did not have intubation difficulties (p < 0.05). The significant differences between the indirect laryngoscopic assessment distributions of patients who either had or did not have intubation difficulty were assessed (p < 0.05). Indirect laryngoscopic assessment is a simple and valuable technique that can be used for the diagnosis of difficult intubation in patients who are undergoing microlaryngeal surgery.


[A case of post-poliomyelitis muscular atrophy with cranial nerve signs and widespread muscular atrophy of the extremities].
Here we report a case of a 56-year-old male with post-poliomyelitis muscular atrophy (PPMA), who presented with cranial nerve signs and widespread atrophy of the extremities. He had suffered from poliomyelitis at the age of 2 years. After recovery from the acute stage, the paralysis remained in his left arm. He noticed muscle weakness of the right upper and lower extremities at the age of 45 years and the muscle atrophy progressed to his arms, hip and thigh at the age of 55 years. Neurological examination revealed muscle atrophy of the neck and disturbance of left V, VIII, IX, X and bilateral XI cranial nerves. We diagnosed this case as PPMA from his history and electromyographic and muscle biopsy findings which suggested chronic denervation. Among the 21 PPMA cases in the past in which the acute poliomyelitis had resulted in paralysis of the only one limb, ours was the only case that had muscle atrophy of all the limbs. Cranial nerve involvement is known to occur in acute poliomyelitis; therefore, there is a possibility that the involvement of the cranial nerves in our case might be a delayed progressive symptoms.


Electrochemical analysis of Shewanella oneidensis engineered to bind gold electrodes.
Growth in three-electrode electrochemical cells allows quantitative analysis of mechanisms involved in electron flow from dissimilatory metal reducing bacteria to insoluble electron acceptors. In these systems, gold electrodes are a desirable surface to study the electrophysiology of extracellular respiration, yet previous research has shown that certain Shewanella species are unable to form productive biofilms on gold electrodes. To engineer attachment of Shewanella oneidensis to gold, five repeating units of a synthetic gold-binding peptide (5rGBP) were integrated within an Escherichia coli outer membrane protein, LamB, and displayed on the outer surface of S. oneidensis. Expression of LamB-5rGBP increased cellular attachment of S. oneidensis to unpoised gold surfaces but was also associated with the loss of certain outer membrane proteins required for extracellular respiration. Loss of these outer membrane proteins during expression of LamB-5rGBP decreased the rate at which S. oneidensis was able to reduce insoluble iron, riboflavin, and electrodes. Moreover, poising the gold electrode resulted in repulsion of the engineered cells. This study provides a strategy to specifically immobilize bacteria to electrodes while also outlining challenges involved in merging synthetic biology approaches with native cellular pathways and cell surface charge.


Mitogen-activated lymphocytes: a good model for characterising lung CYP1A1 inducibility.
The CYP1A1 hyperinducibility phenotype occurring in some 10% of the human population corresponds to a higher risk of developing lung cancer. This study was undertaken to assess whether the inducibility factor, generally evaluated on mitogen-activated lymphocytes after PAH induction, represents correctly the lung situation. Optimal experimental conditions were determined for evaluating, on both lymphocytes and lung tissue explants, the inducibility factor, defined as the ratio of EROD activity (CYP1A1-specific) to cytochrome c reductase activity (unaffected by PAH induction). Paired results for lymphocytes and lung tissue samples from 10 lung cancer patients were compared. A good correlation was observed between lymphocyte and lung tissue inducibilities (R = 0.809; p = 0.005). In conclusion, mitogen-activated lymphocyte inducibility is indicative of lung tissue inducibility and constitutes a good marker for evaluating individual PAH inducibilities.


Infants' responses to frustrating situations: continuity and change in reactivity and regulation.
We examined continuity, stability, and change in behaviors reflecting infant reactivity and regulation. Healthy infants were observed in laboratory situations designed to elicit frustration when they were 5 (N = 87) and 10 months of age (N = 82). Behaviors indicative of reactivity included objective ratings of average intensity cry, peak intensity cry, and latency to cry. In addition, durations of orienting, avoidant, and nonnegative communicative behaviors were assessed as measures of regulation at each age. Results showed that several behaviors changed in level over time. In addition, Confirmatory Factor Analysis revealed both structural continuity and discontinuity: Behaviors were organized into 2 similar factors at 5 and 10 months (Reactivity and Regulation), but the relation between reactivity and regulation became increasingly independent over time, such that reactivity and regulation were negatively correlated at 5 months but not at 10 months. Finally, model-fitting revealed cross-dimension but not within-dimension stability (5-month reactivity predicted 10-month regulation).


Prevalence of high level aminoglycoside and vancomycin resistance among enterococci in Turkey.
Enterococcus spp. have become the third most common cause of nosocomial infections. High-level aminoglycoside resistance (HLAR), an important clinical concern, has been associated with some species of enterococci. The synergistic effect obtained by the combination of aminoglycosides with penicillin or vancomycin disappears in strains that show high-level resistance (HLR) to the aminoglycosides. We evaluated the agar dilution and high content disk diffusion tests for the detection of HLAR for 60 isolated enterococci. In our study 4 strains of Enterococcus faecalis (6.6%) and 8 strains of Enterococcus faecium (13.3%), totally 20% were found to be highly resistant to gentamicin; 2 strains of E. faecalis (13.3%), 8 strains of E. faecium (3.3%), totally 16.6% showed HLR to streptomycin by agar dilution method and the same rates were found by high-content disk diffusion method. 2 strains of E. faecalis (3.3%), 4 strains of E. faecium (6.6%), totally 10% showed vancomycin resistance by agar dilution method. The ratio for beta lactamase activity was found to be 26.6% for E. faecalis and 3.3% for E. faecium. Our results demonstrate that high-content disk diffusion test should be performed by using both gentamicin and streptomycin in routine screening of HLAR among enterococci and the determination of beta lactamase activity at the same time is also useful for detecting resistance to a beta lactam agent.


Activation of extracellular signal-regulated kinase1/2 in the medial prefrontal cortex contributes to stress-induced hyperalgesia.
Stressful stimuli can exacerbate persistent pain disorder. However, the underlying mechanism is still unknown. Here, to reveal the underlying mechanism for stressful stimuli-induced hyperalgesia in chronic pain, we investigated the effect of extracellular signal-regulated kinase1/2 (ERK1/2) activation on pain hypersensitivity using single-prolonged stress (SPS) model, complete Freund's adjuvant (CFA) model and SPS + CFA model. The experimental results revealed significantly reduced paw withdrawal threshold in the SPS, CFA, and SPS + CFA group compared with the control group. However, the increased phosphorylation of ERK1/2 in the medial prefrontal cortex (mPFC) was observed in the SPS- or SPS + CFA-exposed group but not the CFA group compared with control group. There was also a significant increase in mPFC ERK1/2 phosphorylation and mechanical allodynia after SPS + CFA treatment compared to SPS or CFA treatment alone. Furthermore, inhibiting ERK1/2 phosphorylation by microinjection of U0126, a MAPK kinase (MEK) inhibitor, into the mPFC attenuated SPS + CFA- and SPS- but not CFA-induced mechanical allodynia, anxiety-like behavior, and cognitive impairments. These results suggest that the activation of ERK1/2 in the mPFC may contribute to the process of stress-induced cognitive and emotional disorders, leading to an increase in pain sensitivity.


Gene expression in closely related species mirrors local adaptation: consequences for responses to a warming world.
Local adaptation of populations could preclude or slow range expansions in response to changing climate, particularly when dispersal is limited. To investigate the differential responses of populations to changing climatic conditions, we exposed poleward peripheral and central populations of two Lepidoptera to reciprocal, common-garden climatic conditions and compared their whole-transcriptome expression. We found evidence of simple population differentiation in both species, and in the species with previously identified population structure and phenotypic local adaptation, we found several hundred genes that responded in a synchronized and localized fashion. These genes were primarily involved in energy metabolism and oxidative stress, and expression levels were most divergent between populations in the same environment in which we previously detected divergence for metabolism. We found no localized genes in the species with less population structure and for which no local adaptation was previously detected. These results challenge the assumption that species are functionally similar across their ranges and poleward peripheral populations are preadapted to warmer conditions. Rather, some taxa deserve population-level consideration when predicting the effects of climate change because they respond in genetically based, distinctive ways to changing conditions.


Granulomas of the penis: a rare complication of intravesical therapy with Bacillus Calmette-Guerin.
Immunotherapy with Bacillus Calmette-Guérin is widely used for treatment and prophylaxis of superficial urothelial cancer. Complications associated with Bacillus Calmette-Guérin treatment are common. The authors describe a case of granulomatous inflammation of the penis associated with intravesical Bacillus Calmette-Guérin therapy, presenting with multiple erythematous and painless nodules located on the glans. A review of the literature is also performed. Granulomatous balanoposthitis is a rare complication of Bacillus Calmette-Guérin immunotherapy, with heterogeneous clinical presentation, which can make the diagnosis difficult. Its clinical recognition is essential for early start of therapy with antitubercular agents and interruption of Bacillus Calmette-Guérin.


Specialized proresolving lipid mediators in humans with the metabolic syndrome after n-3 fatty acids and aspirin.
The metabolic syndrome (MetS) is associated with a chronic low-grade inflammatory state and may be affected by the ability to resolve inflammation, which is an active process that involves specialized proresolving lipid mediators (SPMs) derived from n-3 (ω-3) fatty acids. We compared plasma concentrations of SPMs in men and women with features of the MetS and in healthy matched control subjects in response to intakes of n-3 fatty acids and aspirin. MetS volunteers (n = 22) and healthy, matched controls (n = 21) were studied in parallel for 4 wk. Both groups took n-3 fatty acids (2.4 g/d) for 4 wk with the addition of aspirin (300 mg/d) during the last 7 d. Blood was collected at baseline and at 3 and 4 wk. Plasma SPMs were measured with the use of liquid chromatography-tandem mass spectrometry and included 18-hydroxyeicosapentaenoic acid (18-HEPE), E-series resolvins, 17-hydroxydocosahexaenoic acid (17-HDHA), D-series resolvins, 14-hydroxydocosahexaenoic acid (14-HDHA), and maresin-1. Baseline SPMs did not differ between groups. There was an increase in the SPM precursors 18-HEPE, 17-HDHA, and 14-HDHA after n-3 fatty acid supplementation that was significantly attenuated in the MetS (P < 0.05). However, the E-series resolvins increased to a similar extent in the groups after n-3 fatty acid supplementation, and the D-series resolvins were not different from those at baseline. The addition of aspirin to n-3 fatty acids did not alter any SPMs in either group. Volunteers with MetS had reduced plasma concentrations of the precursors of the E- and D- series resolvins as well as of 14-HDHA in response to n-3 fatty acid supplementation. However, plasma E-series resolvins were increased to a similar extent after n-3 fatty acid supplementation in both groups, and the addition of aspirin to n-3 fatty acid supplementation did not alter any of the plasma SPMs in MetS and control subjects. Additional studies in the MetS are required to determine whether SPMs affect the ability to mount an appropriate response to infection. This trial was registered at the Australian New Zealand Clinical Trials Registry as ACTRN12610000708055.


Decreased single-photon emission computed tomographic [123I]beta-CIT striatal uptake correlates with symptom severity in Parkinson's disease.
Previous studies have utilized single-photon emission computed tomography (SPECT) to demonstrate decreased [123I]beta-CIT striatal uptake in idiopathic Parkinson disease (PD) patients. The present study extends this work by examining SPECT outcome measures in a larger group of PD patients with varying disease severity. Twenty-eight L-dopa-responsive PD patients (Hoehn-Yahr stages 1-4) and 27 healthy controls had SPECT scans at 18 to 24 hours after injection of [123I]beta-CIT. Specific to nondisplaceable striatal uptake ratios (designated V3") were correlated with Hoehn-Yahr stage and Unified Parkinson's Disease Rating Scale (UPDRS) subscores. Linear discriminant function analyses utilizing striatal uptakes, putamen-to-caudate ratios, and ipsilateral-contralateral asymmetry indices were performed. Decreased striatal tracer uptake (V3") was correlated with total UPDRS score for both contralateral and ipsilateral striatum. Putamen uptake was relatively more reduced than caudate with mean putamen:caudate ratios of 0.50 +/- 0.17 and 0.82 +/- 0.09 for PD patients and controls, respectively. Ipsilateral:contralateral asymmetry was significantly greater in PD patients than controls. Discriminant function analysis utilizing V3" for ipsilateral and contralateral caudate and putamen correctly classified all 55 cases. These data demonstrate marked differences in [123I]beta-CIT SPECT measures in healthy controls and PD patients. The significant correlation of SPECT measures with motor severity suggests [123I]beta-CIT may be a useful marker of disease severity in PD.


Umbilical cord blood hematopoietic stem and repopulating cells in human clinical transplantation.
This paper reviews our recent laboratory and clinical studies demonstrating the efficacious use of human umbilical cord blood for HLA-matched allogeneic sibling stem/progenitor cell transplantation in cases of Fanconi's anemia. Future implications and potential problems are discussed with regards to (a) the possibility of maternal cell contamination, (b) the broadness of applicability with regards to other diseases that might be transplanted, and whether such transplants are feasible in adults, as well as in children, and (c) the immunological reactivity of cord blood cells, and whether these cells can be used to cross histocompatibility barriers more easily than that of bone marrow from adults.


Apheresis for MPO-ANCA-associated RPGN-indications and efficacy: lessons learned from Japan nationwide survey of RPGN.
A national survey concerning rapidly progressive glomerulonephritis (RPGN) was conducted in Japan between 1989 and 2000 and resulted in the registration of 715 patients with RPGN. Among the documented patients, the most frequent primary disease was primary pauci-immune crescentic glomerulonephritis (n = 283), and the second most frequent was microscopic polyangitis (n = 127). Overall, 370 patients had MPO-ANCA, and 23 patients had PR3-ANCA. We found that both renal and patient survivals were significantly worse in patients with MPO-ANCA-associated RPGN than patients with PR3-ANCA. Fifty-three patients received apheresis therapy with various combinations of immunosuppressive regimens. They had higher serum creatinine, higher CRP, and a higher frequency of complicated pulmonary involvements as compared to the controls without apheresis therapy. In dialysis-dependent patients, no additional benefit from apheresis therapy was observed. Only pulmonary renal syndrome patients with CRP > 6 mg/dl at presentation showed a slightly better prognosis (patient survival with apheresis; 66.7%, without apheresis; 56.7%). Furthermore, a rapid MPO-ANCA titer reduction was observed in patients treated with apheresis. Patients with MPO-ANCA-associated RPGN were older, and had more chronic and sclerotic lesions than patients with PR3-ANCA-associated RPGN. Based on these findings, we suggest that a lower dose of immunosuppressant should be considered in order to avoid opportunistic infection. In this situation, cytapheresis is the treatment of choice. Nevertheless, in patients with an aggressive form of RPGN with rapid deterioration of renal function like the PR3-ANCA-associated RPGN, or pulmonary renal syndrome complicated severe inflammation, or relapses with high MPO-ANCA titer, we conclude that apheresis therapy should be considered.


A multidimensional functional trait analysis of resource exploitation in European ants.
The major factors explaining ecological variation in plants have been widely discussed over the last decade thanks to numerous studies that have examined the covariation that exists between pairs of traits. However, multivariate relationships among traits remain poorly characterized in animals. In this study, we aimed to identify the main multivariate trait dimensions that explain variance in important functional traits related to resource exploitation in ants. To this end, we created a large ant trait database. This database includes information on 11 traits that are important in ant resource exploitation; data were obtained for 150 European species found in different biomes. First, we examined the pairwise correlations between the traits included in the database. Second, we used multivariate analyses to identify potential trait dimensions. Our study shows that, to a great extent, resource exploitation strategies align along two main trait dimensions. The first dimension emerged in both the overall and group-specific analyses, where it accounted for the same pairwise trait correlations. The second dimension was more variable, as species were grouped by levels of taxonomy, habitat, and climate. These two dimensions included most of the significant pairwise trait correlations, thus highlighting that complementarity, but also redundancy, exists among different pairs of traits. The first dimension was associated with behavioral dominance: dominance was associated with large colony size, presence of multiple nests per colony, worker polymorphism, and a collective foraging 'strategy. The second dimension was associated with resource partitioning along dietary and microhabitat lines: it ranged from species that consume liquid foods, engage in group foraging, and mainly nest in the vegetation to species that consume insects and seeds, engage in individual foraging, and demonstrate strictly diurnal activity. Our findings establish a proficient ecological trait-based animal research that minimizes the number of traits to be measured while maximizing the number of relevant trait dimensions. Overall, resource exploitation in animals might be framed by behavioral dominance, foraging strategy, diet, and nesting habitat; the position of animal species within this trait space could provide relevant information about their distribution and abundance, for today as well as under future global change scenarios.


Activation of hemolysin toxin: relationship between two internal protein sites of acylation.
HlyC, hemolysin-activating lysine acyltransferase, catalyzes the acylation (from acyl-ACP) of Escherichia coli prohemolysin (proHlyA) on the epsilon-amino groups of specific lysine residues, Lys564 and Lys690 of the 1024-amino acid primary structure, to form hemolysin (HlyA). The amino acid sequences flanking the two acylation sites are not homologous except that each has a glycine residue immediately preceding the lysine which is acylated; there are, however, numerous GK sequences throughout proHlyA that are not acylation sites. The substrate specificity of acylation was examined. ProHlyA-derived structures, altered by substantial deletions and separation of the acylation sites into two different peptides and site-directed mutation analyses of acylation sites, often served as internal protein acylation substrates, and the kinetics of the acylations were measured. The two sites of acylation of proHlyA functioned independently of one another with HlyC; there did not appear to be a common HlyC binding site or processivity of the enzyme between the sites. Acyl-HlyC was likely the enzyme form that interacted with the final acylation substrate. In a variety of constructs, the two acylation sites had similar K(m) values, but their V(max) values and catalytic efficiencies as substrates differed. Internal protein acylation was inhibited by specific small peptides mimicking the primary structure of each acylation site except that the crucial lysines were replaced with arginines; similar small peptides containing the crucial lysine, however, were not acylated.


Disorder and nonconservation in a driven diffusive system.
We consider a disordered asymmetric exclusion process in which randomly chosen sites do not conserve particle number. The model is motivated by features of many interacting molecular motors such as RNA polymerases. We solve the steady state exactly in the two limits of infinite and vanishing nonconserving rates. The first limit is used as an approximation to large but finite rates and allows the study of Griffiths singularities in a nonequilibrium steady state despite the absence of any transition in the pure model. The disorder is also shown to induce a stretched exponential decay of system density with stretching exponent phi=2/5 .


Cataract in long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency (LCHADD).
Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency (LCHADD) is a rare metabolic disorder that can lead to acute encephalopathy, liver disease, cardiomyopathy, rhabdomyolysis, and long-term complications involving the eye and peripheral nerves. LCHADD is a peroxisome biogenesis disorder (PBD). Except for the series presented by Tyni and colleagues (Ophthalmology 1998;105:810-824), which described visually insignificant lens opacities in association with LCHADD, previous ophthalmic papers have only reported retinal complications. We report on one case with progressive asymmetrical cataract. The more mildly affected eye had a similar morphology to that previously reported and the more severely affected eye had an unusual morphology we believe is unique to LCHADD. We discuss the range of ophthalmic presentations in our cases and in the literature. The variability of the severity of ocular complications, even between eyes in one individual, makes it difficult to test the effectiveness of therapeutic options upon the ophthalmic complications.


Trace determination of anthracyclines in urine: a new high-performance liquid chromatography/tandem mass spectrometry method for assessing exposure of hospital personnel.
Health-care workers handling antineoplastic agents may be exposed to extremely low doses of these drugs. Very sensitive and specific analytical methods are therefore needed for biological monitoring. The aim of this study was to develop and validate a method for trace level determination of doxorubicin, epirubicin, daunorubicin and idarubicin in human urine, using epi-daunorubicin as an internal standard. Solid-phase extraction (SPE) was used for sample preparation. Urine samples were loaded onto Bond Elut C18 cartridges. The analytes were eluted in methylene chloride/2-propanol (1:1, v/v) and then evaporated to dryness. The residue was reconstituted with the mobile phase prior to high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) analysis. Quantitation of each analyte was performed using the multiple reaction monitoring (MRM) method. The urine assay was linear over the range 0.1-2.0 microg/L, with a lower limit of quantification (LLOQ) of 0.10 microg/L for doxorubicin and epirubicin, and 0.03 microg/L for daunorubicin and idarubicin. The respective limits of detection (LODs) were 0.04 and 0.01 microg/L. The precision and accuracy of the assay were determined on three different days. The within-series precision was found to be always less than 13.9% for all the analytes. The overall precision expressed as relative standard deviation (RSD) was always less than 10.6%. The recovery of anthracyclines was assessed at two concentrations of the range tested (0.1 and 2.0 microg/L) and it ranged from 87.7% (daunorubicin) to 102.0% (doxorubicin) and from 79.1% (daunorubicin) to 90.7% (idarubicin) for the lower and the higher level, respectively, with a RSD always less than 9.1%. The uncertainty of the present assay was also evaluated and the combined uncertainty was always less than 20% over all the days of the validation study. This is the first method that makes use of LC/MS/MS for the biological monitoring of occupational exposure to anthracyclines.


Assessment of obesity in a community sample of prepubertal children.
The aims of this study were to quantify the effect of obesity definition on estimates of prevalence, and to determine the sensitivity and specificity of three commonly used definitions of overweight/obesity in children: body mass index (BMI) s.d. score > 2.00; BMI s.d. score > 1.04; weight > 120% ideal. A representative community sample of children in Edinburgh, Scotland (n=240, 124 boys and 116 girls; mean age 8.5 s.d. 0.4y) was recruited. Obesity was defined by a criterion method based on % body fat: > 25% fat in boys; > 32% fat in girls. Sensitivity of BMI s.d. score > 2.00 was relatively poor in both sexes (60% in girls; 36% in boys) but had high specificity (98%). Sensitivity of the other two clinical definitions was higher, and was better in girls than boys, but with lower specificity. Choice of definition had a profound effect on prevalence estimates. In conclusion, sensitivity of the definitions of obesity currently recommended for children, when tested in this sample, was heavily dependent on the definition used and differed between boys and girls. This should be considered when choosing a definition of obesity in clinical practice and epidemiology.


Retrospective and prospective data collection compared in the Dutch End Of Life in Dementia (DEOLD) study.
Studying end of life in dementia patients is challenging because of ill-defined prognoses and frequent inability to self-report. We aim to quantify and compare (1) feasibility and (2) sampling issues between prospective and retrospective data collection specific to end-of-life research in long-term care settings. The observational Dutch End of Life in Dementia study (DEOLD; 2007 to 2011) used both prospective data collection (28 facilities; 17 nursing home organizations/physician teams; questionnaires between January 2007 and July 2010, survival until July 2011) and retrospective data collection (exclusively after death; 6 facilities; 2 teams, questionnaires between November 2007 and March 2010). Prospective collection extended from the time of admission to the time after death or conclusion of the study. Prospectively, we recruited 372 families: 218 residents died (59%) and 184 (49%) had complete physician and family after-death assessments. Retrospectively, 119 decedents were enrolled, with 64 (54%) complete assessments. Cumulative data collection over all homes lasted 80 and 8 years, respectively. Per complete after-death assessments in a year, the prospective data collection involved 37.9 beds, whereas this was 7.9 for the retrospective data collection. Although age at death, sex, and survival curves were similar, prospectively, decedents' length of stay was shorter (10.3 vs. 31.4 mo), and fewer residents had advanced dementia (39% vs. 54%). Regarding feasibility, we conclude that prospective data collection is many fold more intensive and complex per complete after-death assessment. Regarding sampling, if not all are followed until death, it results in right censoring and in different, nonrepresentative samples of decedents compared with retrospective data collection. Future work may adjust or stratify for dementia severity and length of stay as key issues to promote comparability between studies.


Social class and access to specialist palliative care services.
To determine any social class differences in place of death of cancer patients in South Bristol; to explore the experience of carers; and to identify inequalities in access to palliative care. Two-part study: (1) A cross-sectional survey of all 960 cancer deaths in South Bristol between September 1999 and December 2002. (2) A qualitative in-depth interview study of 18 carers of patients who died of cancer during the same period in South Bristol. Fourteen of those who died were from social class IIIM (manual), IV and V (i.e. lower social classes). Place of death of patients according to social class and geographical distance from the hospice. Carers' accounts of the way in which illness and death were conducted, and their response to the management of death and dying. The cross sectional survey showed that patients from social class V were less likely to die in the hospice. This finding was independent of geographical proximity. In the qualitative study, no class specific beliefs about death and dying were identified. Attitudes to the way dying should be conducted were common across the classes. Families expected to be present and centre stage at the time of death and for it to be conducted in a dignified and personal manner. Health care staff in all settings supported them in this aim. No one in this study died without a family member present. Some elderly carers were less open than younger carers in the way they talked about death and did not wish to be present at the death. Some carers from social class IV and V were less active in seeking information or asking for hospice admission than carers from other classes. Unrealistic expectations about the availability of hospice beds were common to all carers. Anxiety was common among carers. It was reduced by the provision of reliable and consistent healthcare support, by information provided in a timely and sensitive way, and by open and shared decision making between carer and patient. Most important in reducing anxiety was the support of a second carer who lived locally and was reliable. Carers from social classes IIIM (manual), IV and V were more likely to have this kind of support than carers from social classes I, II and IIIN (non-manual). Although cancer patients from social class V were less likely than others to die in the hospice, social inequality in access to or utilisation of healthcare in terminal illness was not prominent in carers' accounts. When it did arise, it was associated with passivity in seeking information and support on the part of some carers from social classes IV and V. Carers from social classes IIIM-V received more regular and reliable support from their families than those from social classes I-IIIN.


Drug tolerance and biomembranes.
Our understanding of the structure and function of biomembranes is now at a stage where we can begin to examine drug actions on the membrane bilayer. Cells need a certain optimal degree of fluidity in their membranes and some organisms can adapt their membrane composition to maintain the appropriate fluidity under changing environmental conditions. Chronic administration of membrane-disrupting drugs may evoke such an adaptive process, which would show itself in vivo as drug tolerance.