Risk of depression following traumatic limb amputation-a general population-based cohort study.
Background: Individuals with traumatic limb amputation (TLA) may be at risk of depression, but evidence of increased depression after TLA from longitudinal studies has been limited. It is also unknown whether physical function, cognitive function, and employment prior to amputation affects depression risk. We aimed to examine longitudinal associations between TLA and depression in working age men, and to explore the role of pre-amputation occupational and individual characteristics. Methods: A Swedish national register-based cohort of 189,220 men born between 1952 and 1956, and who attended conscription assessments in adolescence, was followed from 1985 to 2009. Physical, cognitive, and psychological characteristics were measured at the conscription examination, and occupational information was obtained from the 1985 census. Main outcome measures were hospital inpatient and outpatient admissions for depression. Results: In total, 401 men underwent TLA; mean age at amputation was 42.5 years (SD 7.4). Cox regression produced an unadjusted hazard ratio (95% confidence interval) of 2.61 (1.62-4.21) for risk of subsequent depression associated with TLA compared with the general population. Adjustment for occupational, physical, cognitive, and psychological characteristics did not change the association much, producing a hazard ratio of 2.53 (1.57-4.08). Conclusions: TLA is associated with an increased risk of depression in men over more than two decades of follow up. Occupational and individual characteristics prior to amputation did not greatly change depression risk following amputation. We speculate that a coordinated combination of social support and medical management may help reduce persistent depression risk in men who experience amputation.


Electron-transfer pathways between redox enzymes and electrode surfaces: reagentless biosensors based on thiol-monolayer-bound and polypyrrole-entrapped enzymes.
Based on previous results which showed that quinohemo-protein alcohol dehydrogenase (QH-ADH) entrapped within polypyrrole is able to directly transfer electrons via the conducting polymer to the electrode surface, the electron-transfer properties of this multi-cofactor enzyme adsorbed and covalently-bound to self-assembled thiol monolayers and bare electrode surfaces has been investigated more closely. While the dissolved enzyme is able to transfer electrons to the electrode via heme c as well as via the more deeply buried PQQ (fast adsorption-chemical reaction-desorption mechanism), an orientation of adsorbed QH-ADH on hydrophobic electrode surfaces, as well as of adsorbed and covalently bound QH-ADH on negatively-charged thiol monolayers could be observed. In these cases the heme c units are pointing towards the electrode surfaces resulting in an optimised direct ET rate.


Symbiotic seed germination and protocorm development of Aa achalensis Schltr., a terrestrial orchid endemic from Argentina.
Aa achalensis is an endangered terrestrial orchid endemic from Argentina. In vitro symbiotic seed germination was evaluated for its propagation. Five different fungal strains were isolated from this species: two Rhizoctonia-like related to Thanatephorus cucumeris and three ascomicetaceous fungi belonging to Phialophora graminicola and one to an uncultured Pezizaceae. All five isolates promoted seed germination being one T. cucumeris strain the most effective. After 16 weeks of growth, 30% of A. achalensis protocorms developed until seedlings with two/four leaves in this treatment. These findings open an opportunity to the knowledge and preservation of this species.


Biological variability influencing lung dosimetry for inhaled 222Rn and 220Rn decay products.
Particle deposition, clearance and dosimetry of inhaled short-lived 222Rn and 220Rn decay products in the human respiratory tract have been modeled by a stochastic compartment system, based on an already existing deterministic model (Ho82a). In order to allow for statistical uncertainties and biological variabilities, input parameters and transfer coefficients were described by truncated lognormal frequency distributions. Applying Monte Carlo techniques, a set of parameter values is selected randomly in an iterative manner from these prespecified distributions, yielding finally basal-cell doses as probability density distributions instead of single mean doses. The resulting highly skewed dose distributions illustrate the fact that a small percentage of individuals in an exposed population receives considerably higher doses than indicated by a deterministic mean value.


Effects of FlrBC on flagellar biosynthesis of Shewanella oneidensis.
As a most conserved complex molecular machine made up of a large number of structural subunits, the flagellum is under tight regulation by hierarchical arrangements. Although variations in polar flagellar systems are found, most of them are restricted to multiple-copy components, such as flagellins and stators. Therefore, these features are regarded to be peripheral relative to the comprehensive conservation. In this study, however, we present evidence to show that the difference in highly conserved polar flagellar systems can be surprisingly profound, even at the heart of the classical regulatory hierarchy. In Gram-negative Shewanella oneidensis, two-component system FlrBC, whose counterpart is essential for flagellar biosynthesis and motility by directly controlling expression of class III genes in polarly flagellated bacteria such as Vibrio cholerae, is dispensable for the process. The system directly controls expression of the flaA gene, encoding a flagellin of weak motility. We further show that the ratio of two flagellins, FlaA and FlaB, determines motility of a flagellum. More strikingly, overproduction of FlrC results in a peritrichously multi-flagellated phenotype, and FlrC is likely to function as an activator in its unphosphorylated form for transcription of the flaA gene, contrasting the previously characterized counterpart.


[Comparison of methods for the large-scale culture of cells and viruses].
The paper presents the results of experiments on propagation of primary, secondary, and continuous diploid and heteroploid human and animal cells in 2 different systems for large-scale propagation: in perfusion tank with Rashig rings and in tanks with microcarriers. Both methods of large-scale cultivation produce higher cell yields than the traditional cultivation methods. A yield of tick-borne encephalitis virus per 1 cell in the perfusion cultivator was 12 times as high as in roller cultures. Poliomyelitis virus titres were practically equal with both methods of large-scale cultivation and with the conventional method (7.73 lg PFU/ml in GMKC on microcarriers). The method of cultivation on microcarriers is more acceptable and advantageous as it gives higher yields of cells necessary for growth of poliomyelitis virus.


[Cognitive potential P300 in dementia with Levy's bodies and Alzheimer's disease].
Neurodegenerative diseases accompanied by cognitive impairment (dementia with Levy's bodies--DLB and Alzheimer's disease--AD) and others) are becoming more widespread due to increasing population life expectancy. The paper addresses use of cognitive event-related potential P300 to verbal and nonverbal stimuli with an assessment of interhemispheric asymmetry and topographic distribution of P300 in patients with DLB and AD comparied to age-matched healthy controls. Specific alterations of P300 amplitude and latency suggest a use of cognitive event-related potential P300 in differential diagnosis of DLB and AD.


Experimental investigation of a diffraction tomography technique in fluid ultrasonics.
Qualitative ultrasonic imaging of cylindrical fluid targets is investigated by a diffraction tomography technique applied to experimental data. The principles of the image formulation are stated and an experimental setup is described. Experimental difficulties related to the short wavelength used and respective advantages in collecting the data, either with a mechanically scanned single transducer or with an electronically scanned array of transducers, are emphasized. Representative images of simply structured phantoms and of real biological bodies are obtained in spite of the small number of views available.


Ultrastructure and localization of Neorickettsia in adult digenean trematodes provides novel insights into helminth-endobacteria interaction.
Neorickettsia are a group of intracellular α proteobacteria transmitted by digeneans (Platyhelminthes, Trematoda). These endobacteria can also infect vertebrate hosts of the helminths and cause serious diseases in animals and humans. Neorickettsia have been isolated from infected animals and maintained in cell cultures, and their morphology in mammalian cells has been described. However, limited information is available on the morphology and localization of Neorickettsia in the trematode host. We used a Neorickettsia-infected strain of the model trematode Plagiorchis elegans to infect Syrian Golden hamsters to produce adult worms. Ultrastructure of Neorickettsia was assessed by transmission electron microscopy of high pressure freezing/freeze substitution fixed specimens. A Neorickettsia surface protein from P. elegans (PeNsp-3) was cloned and antibodies against the recombinant protein were used to localize Neorickettsia by immunohistochemistry. Ultrastructural analysis revealed moderate numbers of pleomorphic endobacteria with a median size of 600 × 400 nm and characteristic double membranes in various tissue types. Endobacteria showed tubular membrane invaginations and secretion of polymorphic vesicles. Endobacteria were unevenly localized as single cells, or less frequently as small morula-like clusters in the ovary, Mehlis' gland, vitelline follicles, uterus, intrauterine eggs, testis, cirrus-sac, tegument, intestine and the oral and ventral sucker. Examination of hamster small intestine infected with P. elegans showed many endobacteria at the host-parasite interface such as the oral and ventral sucker, the tegument and the excretory pore. We conclude that adult P. elegans trematodes carry Neorickettsia endobacteria in varying numbers in many tissue types that support vertical transmission, trematode to trematode transmission via seminal fluid, and possibly horizontal transmission from trematodes to vertebrate hosts. These means appear to be novel mechanisms of pathogen transmission by endoparasitic worms.


The Development and Initial Findings of A Study of a Prospective Adult Research Cohort with Inflammatory Bowel Disease (SPARC IBD).
Clinical and molecular subcategories of inflammatory bowel disease (IBD) are needed to discover mechanisms of disease and predictors of response and disease relapse. We aimed to develop a study of a prospective adult research cohort with IBD (SPARC IBD) including longitudinal clinical and patient-reported data and biosamples. We established a cohort of adults with IBD from a geographically diverse sample of patients across the United States with standardized data and biosample collection methods and sample processing techniques. At enrollment and at time of lower endoscopy, patient-reported outcomes (PRO), clinical data, and endoscopy scoring indices are captured. Patient-reported outcomes are collected quarterly. The quality of clinical data entry after the first year of the study was assessed. Through January 2020, 3029 patients were enrolled in SPARC, of whom 66.1% have Crohn's disease (CD), 32.2% have ulcerative colitis (UC), and 1.7% have IBD-unclassified. Among patients enrolled, 990 underwent colonoscopy. Remission rates were 63.9% in the CD group and 80.6% in the UC group. In the quality study of the cohort, there was 96% agreement on year of diagnosis and 97% agreement on IBD subtype. There was 91% overall agreement describing UC extent as left-sided vs extensive or pancolitis. The overall agreement for CD behavior was 83%. The SPARC IBD is an ongoing large prospective cohort with longitudinal standardized collection of clinical data, biosamples, and PROs representing a unique resource aimed to drive discovery of clinical and molecular markers that will meet the needs of precision medicine in IBD.


Estrogen in vascular smooth muscle cells: A friend or a foe?
Cardiovascular disease (CVD) continues to be the leading cause of death worldwide. The effect of estrogen on these diseases has been assessed in in vitro and in vivo models, as well as in observational studies. Collectively, these studies alluded to a cardiovasculo-protective effect of estrogen. However, comprehensive clinical investigation failed to produce concrete proof of a cardiovascular protective effect for hormone replacement therapy (HRT), let alone rule out potential harm. These seemingly paradoxical effects of estrogen were explained by the 'theory of timing and opportunity'. This theory states that the effect of estrogen, whether cardiovasculo-protective or pathological, significantly depends on the age of the individual when estrogen administration takes place. Here, we review the conflicting effects of estrogen on vascular smooth muscle cells, mainly proliferation and migration as two cellular capacities intimately related to physiology and pathophysiology of the cardiovascular system. Furthermore, we critically discuss the major parameters and signaling pathways that may account for the aforementioned paradoxical observations, as well as the key molecular players involved.


Transient kinetics of copper-containing lentil (Lens culinaris) seedling amine oxidase.
The reaction between lentil (Lens culinaris) seedling amine oxidase and its chromogenic substrate, p-dimethylaminomethylbenzylamine, has been studied by the stopped-flow technique. Upon being mixed with substrate in the absence of oxygen, the enzyme is bleached in a complex kinetic process. A yellow intermediate absorbing at 464 nm and the first product (aldehyde) are formed in subsequent steps. When oxygenated buffer is mixed with substrate-reduced amine oxidase, the 496 nm absorption of the oxidized enzyme is very rapidly restored in a second-order process (k = 2.5 X 10(7) M-1 X S-1). This reaction is appreciable even at very low oxygen concentration, in keeping with the fairly low Km for O2 measured by steady-state kinetics.


Effects of ventriculoperitoneal shunt removal on cerebral oxygenation and brain compliance in chronic obstructive hydrocephalus.
The pathophysiology of shunt malfunction has not been fully examined, probably because of the paucity of appropriate animal models. Using a canine model of chronic obstructive hydrocephalus, the effects of shunt placement and removal on physiological parameters were evaluated. Fifteen dogs, nine in which chronic hydrocephalus was induced and six controls, were used in the experiment. Thirteen weeks after the induction of hydrocephalus, intracranial pressure (ICP), tissue and cerebrospinal fluid O2 saturation, response to hyperventilation, and brain compliance at low (5-15 mm Hg) and high (15-25 mm Hg) pressures were measured (untreated stage). Following this procedure, ventriculoperitoneal shunts were implanted in the dogs suffering from hydrocephalus. Two weeks later, the same series of measurements were repeated (shunted stage), following which the shunt systems were removed. One week after shunt removal, the last measurements were obtained (shunt-removed stage). All dogs underwent magnetic resonance imaging four times: before induction of hydrocephalus and before each measurement. All dogs with hydrocephalus also had ventriculomegaly (1.42 +/- 0.89 ml before induction of hydrocephalus compared with 3.4 +/- 1.64 ml 13 weeks after induction, p = 0.0064). In dogs in the untreated hydrocephalus stage, ICP remained within the normal range (8.33 +/- 2.60 mm Hg)--although it was significantly higher than that in the control group (5 +/- 1.41 mm Hg, p = 0.014). Tissue O2 saturation in the dogs in the hydrocephalus group (26.1 +/- 5.33 mm Hg) was lower than that in the dogs in the control group (48.7 +/- 4.27 mm Hg, p < 0.0001). After the dogs underwent shunt placement, significant improvement was observed in their ICP (5.22 +/- 2.17 mm Hg, p = 0.012) and tissue O2 saturation (35.2 +/- 6.80 mm Hg, p = 0.0084). However, removal of the shunt reversed these improvements back to the preshunt status. Hyperventilation induced significant decreases in ICP and O2 saturation at every measurement time and induced a significant decrease in tissue O2 saturation during the shunted stage, but not during the untreated and shunt-removed stages. Brain compliance measured at high pressure demonstrated a significant gradual decrease at every measurement. In chronic obstructive hydrocephalus, shunt placement improves ICP and cerebral oxygenation as well as the response to hyperventilation in the tissue. Shunt removal reverses these improvements back to levels present during the untreated stage. The decrease in brain compliance may be one of the factors responsible for symptoms in shunt malfunction.


The Predictive Validity of the Berlin Definition of Acute Respiratory Distress Syndrome for Patients With COVID-19-Related Respiratory Failure Treated With High-Flow Nasal Oxygen: A Multicenter, Prospective Cohort Study.
The Berlin definition of acute respiratory distress syndrome (ARDS) was constructed for patients receiving invasive mechanical ventilation (IMV) with consideration given to issues related to reliability, feasibility, and validity. Notwithstanding, patients with acute respiratory failure (ARF) may be treated with high-flow nasal oxygen (HFNO) and may not fall within the scope of the original definition. We aimed to evaluate the predictive validity of the Berlin definition in HFNO-treated patients with COVID-19-related respiratory failure who otherwise met ARDS criteria. Multicenter, prospective cohort study. Five ICUs of five centers in Argentina from March 2020 to September 2021. We consecutively included HFNO-treated patients older than 18 years with confirmed COVID-19-related ARF, a Pa o2 /F io2 of less than 300 mm Hg, bilateral infiltrates on imaging, and worsening respiratory symptoms for less than 1 week. None. We evaluated the predictive validity of mortality at day 28 using the area under the receiver operating characteristics curve (AUC), compared the predictive validity across subgroups, and characterized relevant clinical outcomes. We screened 1,231 patients and included 696 ARDS patients [30 (4%) mild, 380 (55%) moderate, and 286 (41%) severe]. For the study cohort, the AUC for mortality at day 28 was 0.606 (95% CI, 0.561-0.651) with the AUC for subgroups being similar to that of the overall cohort. Two hundred fifty-six patients (37%) received IMV. By day 28, 142 patients (21%) had died, of whom 81 (57%) had severe ARDS. Mortality occurred primarily in patients who were transitioned to IMV. The predictive validity of the Berlin ARDS definition was similar for HFNO-treated patients as compared with the original population of invasively ventilated patients. Our findings support the extension of the Berlin definition to HFNO-treated patients with ARDS.


An evaluation of the Swiss staging model for hypothermia using hospital cases and case reports from the literature.
The Swiss staging model for hypothermia uses clinical indicators to stage hypothermia and guide the management of hypothermic patients. The proposed temperature range for clinical stage 1 is < 35-32 °C, for stage 2 is < 32-28 °C, for stage 3 is < 28-24 °C, and for stage 4 is below 24 °C. Our previous study using 183 case reports from the literature showed that the measured temperature only corresponded to the clinical stage in the Swiss staging model in approximately 50% of cases. This study, however, included few patients with moderate hypothermia. We aimed to expand this database by adding cases of hypothermic patients admitted to hospital to perform a more comprehensive evaluation of the staging model. We retrospectively included patients aged ≥18 y admitted to hospital between 1.1.1994 and 15.7.2016 with a core temperature below 35 °C. We added the cases identified through our previously published literature review to estimate the percentage of those patients who were correctly classified and compare the theoretical with the observed temperature ranges for each clinical stage. We included 305 cases (122 patients from the hospital sampling and the 183 previously published). Using the theoretically derived temperature ranges for clinical stages resulted in 185/305 (61%) patients being assigned to the correct temperature range. Temperature was overestimated using the clinical stage in 55/305 cases (18%) and underestimated in 65/305 cases (21%); important overlaps in temperature existed among the four stage groups. The optimal temperature thresholds for discriminating between the four stages (32.1 °C, 27.5 °C, and 24.1 °C) were close to those proposed historically (32 °C, 28 °C, and 24 °C). Our results provide further evidence of the relationship between the clinical state of patients and their temperature. The historical proposed temperature thresholds were almost optimal for discriminating between the different stages. Adding overlapping temperature ranges for each clinical stage might help clinicians to make appropriate decisions when using clinical signs to infer temperature. An update of the Swiss staging model for hypothermia including our methodology and findings could positively impact clinical care and future research.


PON2 Deficiency Leads to Increased Susceptibility to Diet-Induced Obesity.
(1) Background: Paraoxonase 2 (PON2) is a ubiquitously expressed protein localized to endoplasmic reticulum and mitochondria. Previous studies have shown that PON2 exhibits anti-oxidant and anti-inflammatory functions, and PON2-deficient (PON2-def) mice are more susceptible to atherosclerosis. Furthermore, PON2 deficiency leads to impaired mitochondrial function. (2) Methods: In this study, we examined the susceptibility of PON2-def mice to diet-induced obesity. (3) Results: After feeding of an obesifying diet, the PON2-def mice exhibited significantly increased body weight due to increased fat mass weight as compared to the wild-type (WT) mice. The increased adiposity was due, in part, to increased adipocyte hypertrophy. PON2-def mice had increased fasting insulin levels and impaired glucose tolerance after diet-induced obesity. PON2-def mice had decreased oxygen consumption and energy expenditure. Furthermore, the oxygen consumption rate of subcutaneous fat pads from PON2-def mice was lower compared to WT mice. Gene expression analysis of the subcutaneous fat pads revealed decreased expression levels of markers for beige adipocytes in PON2-def mice. (4) Conclusions: We concluded that altered systemic energy balance, perhaps due to decreased beige adipocytes and mitochondrial dysfunction in white adipose tissue of PON2-def mice, leads to increased obesity in these mice.


Effects of rolipram on U46619-induced contraction and cyclic nucleotide content in the porcine coronary artery.
The effects of various selective phosphodiesterase (PDE) inhibitors on muscle contractility and cyclic nucleotide content in the porcine coronary artery were investigated. Various selective PDE inhibitors, vinpocetine (type 1), erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA, type 2), milrinone (type 3), rolipram (type 4), Ro20-1724 (type 4), and zaprinast (type 5), inhibited U46619-induced contractions in a concentration-dependent manner. The rank order of potency for the porcine coronary artery was rolipram > Ro20-1724 >milrinone > vinpocetine > zaprinast > EHNA, which was different from that of both the porcine carotid artery and aorta. Rolipram inhibited the U46619-induced muscle tension with a decreased [Ca(2+)](i) level, but inhibited the high K(+)-induced contraction without a change in [Ca(2+)](i) level. Rolipram increased cAMP but not cGMP content. Iberiotoxin restored the inhibition of muscle tension and the [Ca(2+)](i) levels induced by rolipram. U46619 and caffeine induced a transient increase in the [Ca(2+)](i) levels in a Ca(2+)-free solution, but rolipram only inhibited the U46619-induced Ca(2+) transient. In conclusion, rolipram is the most potent inhibitor in the porcine coronary artery, but not in the carotid artery and aorta. Moreover it is suggested that the mechanism by which rolipram causes relaxation is due to a decrease in the [Ca(2+)](i) levels and of the Ca(2+) sensitivity of the contractile elements to cAMP.


Role of axial base coordination in isonitrile binding and chalcogen atom transfer to vanadium(III) complexes.
The enthalpy of oxygen atom transfer (OAT) to V[(Me3SiNCH2CH2)3N], 1, forming OV[(Me3SiNCH2CH2)3N], 1-O, and the enthalpies of sulfur atom transfer (SAT) to 1 and V(N[t-Bu]Ar)3, 2 (Ar = 3,5-C6H3Me2), forming the corresponding sulfides SV[(Me3SiNCH2CH2)3N], 1-S, and SV(N[t-Bu]Ar)3, 2-S, have been measured by solution calorimetry in toluene solution using dbabhNO (dbabhNO = 7-nitroso-2,3:5,6-dibenzo-7-azabicyclo[2.2.1]hepta-2,5-diene) and Ph3SbS as chalcogen atom transfer reagents. The V-O BDE in 1-O is 6.3 ± 3.2 kcal·mol(-1) lower than the previously reported value for 2-O and the V-S BDE in 1-S is 3.3 ± 3.1 kcal·mol(-1) lower than that in 2-S. These differences are attributed primarily to a weakening of the V-Naxial bond present in complexes of 1 upon oxidation. The rate of reaction of 1 with dbabhNO has been studied by low temperature stopped-flow kinetics. Rate constants for OAT are over 20 times greater than those reported for 2. Adamantyl isonitrile (AdNC) binds rapidly and quantitatively to both 1 and 2 forming high spin adducts of V(III). The enthalpies of ligand addition to 1 and 2 in toluene solution are -19.9 ± 0.6 and -17.1 ± 0.7 kcal·mol(-1), respectively. The more exothermic ligand addition to 1 as compared to 2 is opposite to what was observed for OAT and SAT. This is attributed to less weakening of the V-Naxial bond in ligand binding as opposed to chalcogen atom transfer and is in keeping with structural data and computations. The structures of 1, 1-O, 1-S, 1-CNAd, and 2-CNAd have been determined by X-ray crystallography and are reported.


Novel scientific methods in court.
In recent decades the use of forensic science in investigations and therefore its subsequent presentation within the courts has increased exponentially, fuelled by an increase in scientific advances, development of databases and greater access to scientists and their expertise. This explosion in the use of forensic evidence has not been limited to one single scientific domain, as there are a broad range of scientific disciplines, encompassed by the general umbrella term' forensic science'. Many of these involve commonly applied methodologies and are accepted by the courts with limited scrutiny. Where tensions exist concerning the use of science in the courtroom is when novel or emerging sciences and scientific techniques are introduced. This may be particularly evident when the demands of the investigatory phase, where those working want to apply all possible tools at their disposal to gather as much evidence as possible and the needs of the courts, where the evidence must scientifically robust and admissible for it to be presented before a jury, come together. This paper examines the implications for the court for emerging or novel sciences and scientific techniques. In such cases, the potential rewards of implementing the scientific process and the information these may contribute to an investigation provides a temptation to investigators to push for their operational use, with the unintended consequence of posing an issue to the court when considering whether to admit the evidence into the judicial process.


Clinical and demographic characteristics of 13,911 medical emergency patients.
In order to provide better quality of care at Taipei Veterans General Hospital, 13,911 emergency patients coming into the medical emergency room were studied using the computer to key in all demographic data including registration time, time to be seen, desposition time, impression, triage category, discipline as well as daily dynamic status in the observation room from August through December 1989. The study showed that 8.6% were triage category 1 (life-threatening cases) and 22.08% were triage 4 (pseudo-emergency patients). Of the average 101 studied patients seen per day, 20 (20.27%) should have been theoretically admitted, but only 14.7% were admitted, and 18 (18.08%) were sent to the observation room. In general, the daily dynamic status of the patients in the observation room were: (1) Out of 45 overnight patients, 12 (27%) were waiting for admission; and (2) 9 (20.14%) were waiting for a transfer to other convalescent hospitals. We conclude that less than one-tenth of the emergency patients were really emergencies in such a large and busy emergency department, and there was enormous patients stasis in the observation room causing overcrowding of the emergency department, which is the main issue we have to resolve if the quality assurance of the emergency department is to be improved.


X-ray fluorescence analysis in biology.
Our efforts to understand the important biological roles of many "minor" elements are limited by the capabilities of the existing instruments. X-ray fluorescence analysis is not the most sensitive method for the assay of minor elements, but it is the instrumental technique of choice both for certain clinical studies and for certain research problems. The technique will be more widely used as it becomes more widely known (25).


Multiplexed quantification of bacterial 16S rRNA by solution hybridization with oligonucleotide probes and affinity capture.
Multiplexed and quantitative analysis of nucleic acid sequences in complex mixtures is essential in various applications of microbiological research. We have developed a method based on solution hybridization between biotinylated nucleic acid targets and multiple fluorophore-labeled oligonucleotide probes of distinct sizes. The biotin-nucleic acid-probe complexes are captured on magnetic streptavidin-coated microparticles and washed. The hybridized probes are eluted and their identity and quantity are determined by capillary electrophoresis. The signal intensities of the recorded probes correspond to the amount of target nucleic acid in the mixture, and the size indicates the target. Based on this principle and 16S rRNA-specific oligonucleotide probes, we set up an application for the relative quantification of different groups of clostridia and related organisms in a mixed bacterial population. The lower detection limit is 0.05 ng of total RNA and the linear range of measurement is 10(2). The method allowed accurate and highly repeatable quantification of the proportion of clostridia in human feces. Further, we discuss other applications of the method such as quantitative transcriptional analysis of eukaryotic microorganisms, which can be performed without conversion of mRNA to cDNA.


Harmonization of four biomarkers across nine nationally representative studies of older persons.
A growing number of international population surveys have included measurement of biomarkers, but differ in the type of specimens collected, sample processing procedures, shipment protocols, and laboratory assay platforms. The purpose of this study is to harmonize biomarker data from nine nationally representative studies of people 50 years of age and over by adjusting for assay platforms and type of specimens for total cholesterol (total-C), high-density lipoprotein cholesterol (HDL-C), glycosylated hemoglobin (HbA1c), and C-reactive protein (CRP). Sets of 24 identical serum, plasma, whole blood, and dried blood spot harmonization samples with known analyte levels were generated at a reference laboratory, shipped at -80°C to the respective study laboratories, and subsequently assayed following the study laboratory's protocol. Both original and harmonized study data were used to calculate mean values and at-risk prevalence. The correlation coefficients between the biomarker values of the harmonization samples obtained by the study laboratories and the reference laboratory were 0.99 or above for all analytes and laboratories, indicating the high quality of assays at all laboratories. However, using the harmonized data from each study, there were significant differences in the mean values and country ranking of the prevalence of at-risk levels of these four biomarkers. While the biomarker data from the different study laboratories were highly correlated, indicating very high correlation of rank order of specimens, absolute values did vary significantly. This can have a major impact on assessment of international differences in estimates of risks for chronic morbidity and mortality.


Feature Selection Based on Machine Learning in MRIs for Hippocampal Segmentation.
Neurodegenerative diseases are frequently associated with structural changes in the brain. Magnetic resonance imaging (MRI) scans can show these variations and therefore can be used as a supportive feature for a number of neurodegenerative diseases. The hippocampus has been known to be a biomarker for Alzheimer disease and other neurological and psychiatric diseases. However, it requires accurate, robust, and reproducible delineation of hippocampal structures. Fully automatic methods are usually the voxel based approach; for each voxel a number of local features were calculated. In this paper, we compared four different techniques for feature selection from a set of 315 features extracted for each voxel: (i) filter method based on the Kolmogorov-Smirnov test; two wrapper methods, respectively, (ii) sequential forward selection and (iii) sequential backward elimination; and (iv) embedded method based on the Random Forest Classifier on a set of 10 T1-weighted brain MRIs and tested on an independent set of 25 subjects. The resulting segmentations were compared with manual reference labelling. By using only 23 feature for each voxel (sequential backward elimination) we obtained comparable state-of-the-art performances with respect to the standard tool FreeSurfer.


Diagnosis and treatment of botulism in lions.
Six circus lions (Panthera leo) showed neurological and gastrointestinal signs after consuming casualty broiler chickens. Signs included ataxia, hindlimb paralysis and recumbency. Neurological examination of two affected males showed paralysis of extraocular muscles, fixed dilated pupils and inability to swallow. Replacement fluids and antibiotics were given and Clostridium botulinum type C antitoxin was found in serum samples. Type C antitoxin was not then available and therapy was started in one lioness with guanidine hydrochloride. Convulsions were controlled by diazepam but this animal died. One of the two males was given type C antitoxin; both were given anabolic steroids. All the remaining animals made slow recoveries over varying periods; one lion was recumbent for 41 days. No lion developed respiratory paralysis; other animals which had consumed the chickens remained healthy. Aspects of the treatment of botulism in animals are discussed.


Hydroxyl radical production in body fluids. Roles of metal ions, ascorbate and superoxide.
Hydroxyl radical production, detected by ethylene formation from methional, has been investigated in plasma, lymph and synovial fluid. In the presence of added iron--EDTA, addition of either H2O2 or xanthine and xanthine oxidase gave rise to hydroxyl radical formation that in most cases was not superoxide-dependent. The ascorbate already present in the fluid appeared to participate in the reaction. In the absence of added catalyst, the reaction was hardly detectable, the rate being less than 5% of that observed with 1 microM-iron--EDTA added. This implies that the fluids had little if any capacity to catalyse hydroxyl radical production via this mechanism.


Mutations in enterobacterial common antigen biosynthesis restore outer membrane barrier function in Escherichia coli tol-pal mutants.
The outer membrane (OM) is an essential component of the Gram-negative bacterial envelope that protects the cells against external threats. To maintain a functional OM, cells require distinct mechanisms to ensure balance of proteins and lipids in the membrane. Mutations in OM biogenesis and/or homeostasis pathways often result in permeability defects, but how molecular changes in the OM affect barrier function is unclear. Here, we seek potential mechanism(s) that can alleviate permeability defects in Escherichia coli cells lacking the Tol-Pal complex, which accumulate excess PLs in the OM. We identify mutations in enterobacterial common antigen (ECA) biosynthesis that re-establish OM barrier function against large hydrophilic molecules, yet did not restore lipid homeostasis. Furthermore, we demonstrate that build-up of biosynthetic intermediates, but not loss of ECA itself, contributes to the rescue. This suppression of OM phenotypes is unrelated to known effects that accumulation of ECA intermediates have on the cell wall. Finally, we reveal that an unusual diacylglycerol pyrophosphoryl-linked lipid species also accumulates in ECA mutants, and might play a role in the rescue phenotype. Our work provides insights into how OM barrier function can be restored independent of lipid homeostasis, and highlights previously unappreciated effects of ECA-related species in OM biology.


Animal growth during liberation from appetite suppression.
Superalimentation (SA) of swine at 120% (ad libitum = 100%) for 23 days elicited a 40% greater gain of body mass (P less than .005) than littermate control (C) pigs. The accretion of lean and fat tissues during this excess and rapid growth were about the same as if the animal were allowed to attain the same weight over a longer period by conventional feeding practices. Superalimentation is defined as feed intake distinctly in excess of normal voluntary intake on a grams-dry-matter-per-kilogram-body-weight basis. The substantial body mass response shows that growth is limited by a lack of nutrient intake (appetite) during normal ad libitum feeding. Either digestion-absorption processes are not saturated during normal feed intake or digestion and absorption capacities are enhanced during excess food intake; furthermore, either cellular hyperplasia or hypertrophy are not regulated by inherited neuroendocrine factors or cellular growth is somehow released from neuroendocrine regulation during periods of excess circulating nutrients. The overall gain efficiency (kg total gain per kg total alimentation dry matter) during superalimentation was slightly improved (7.3%) but was not significant. Efficiency of the differential gain (delta gain/delta dry matter intake; delta = SA-C) was 25% higher than the overall gain efficiency of the C pigs (0.374 vs 0.300 kg/kg) and suggests that the additional nutrients may have been utilized for growth with greater efficiency than nutrient consumed ad libitum.


Tissue-Negative Transient Ischemic Attack: Is There a Role for Perfusion MRI?
Approximately 60% of patients with a clinical transient ischemic attack (TIA) do not have DWI evidence of cerebral ischemia. The purpose of this study was to assess the added diagnostic value of perfusion MRI in the evaluation of patients with TIA who have normal DWI findings. The inclusion criteria for this retrospective study were clinical presentation of TIA at admission with a discharge diagnosis of TIA confirmed by a stroke neurologist, MRI including both DWI and perfusion-weighted imaging within 48 hours of symptom onset, and no DWI lesion. Cerebral blood flow (CBF) and time to maximum of the residue function (Tmax) maps were evaluated independently by two observers. Multivariate analysis was used to assess perfusion findings; clinical variables; age, blood pressure, clinical symptoms, diabetes (ABCD2) score; duration of TIA; and time between MRI and onset and resolution of symptoms. Fifty-two patients (33 women, 19 men; age range, 20-95 years) met the inclusion criteria. A regional perfusion abnormality was identified on either Tmax or CBF maps of 12 of 52 (23%) patients. Seven (58%) of the patients with perfusion abnormalities had hypoperfused lesions best detected on Tmax maps; the other five had hyperperfusion best detected on CBF maps. In 11 of 12 (92%) patients with abnormal perfusion MRI findings, the regional perfusion deficit correlated with the initial neurologic deficits. Multivariable analysis revealed no significant difference in demographics, ABCD2 scores, or presentation characteristics between patients with and those without perfusion abnormalities. Perfusion MRI that includes Tmax and CBF parametric maps adds diagnostic value by depicting regions with delayed perfusion or postischemic hyperperfusion in approximately one-fourth of TIA patients who have normal DWI findings.


Cancer mortality of granite workers.
A retrospective cohort study was undertaken to investigate the cancer mortality of granite workers. The study comprised 1,026 workers hired between 1940 and 1971. The number of person-years was 20,165, and the number of deaths 235. During the total follow-up 46 tumors were observed and 44.9 were expected. An excess mortality from tumors was observed for the workers followed for 20 years or more, the greatest excess occurring during the follow-up period of 25-29 years (observed 11, expected 5.2). Of the 46 tumors, 22 were lung cancers (expected 17.1) and 15 were gastrointestinal cancers (expected 9.7), nine of which were cancers of the stomach (expected 6.0). Mortality from lung cancer was excessive for workers with at least 15 years since entry into granite work (latency) (21 observed and 9.5 expected), being highest during the follow-up period of 25-29 years (observed 8, expected 2.1). The results indicate that granite exposure per se may be an etiologic factor in the initiation or promotion of malignant neoplasms.


The antrochoanal polyp.
The different varieties of choanal polyps are reviewed by the authors according to their implantation pedicle. Their histology, possible pathogenesis, the various symptoms, steps in the diagnostic procedure and differential diagnoses are also described. Finally, the various modes of treatment are discussed, focussing in particular on the surgical methods and the new possibilities offered by developments in the field of endoscopy.


Comparison of prebiotic effects of arabinoxylan oligosaccharides and inulin in a simulator of the human intestinal microbial ecosystem.
In this study, the prebiotic potential of arabinoxylan oligosaccharides (AXOS) was compared with inulin in two simulators of the human intestinal microbial ecosystem. Microbial breakdown of both oligosaccharides and short-chain fatty acid production was colon compartment specific, with ascending and transverse colon being the predominant site of inulin and AXOS degradation, respectively. Lactate levels (+5.5 mM) increased in the ascending colon during AXOS supplementation, while propionate levels (+5.1 mM) increased in the transverse colon. The concomitant decrease in lactate in the transverse colon suggests that propionate was partially formed over the acrylate pathway. Furthermore, AXOS supplementation strongly decreased butyrate in the ascending colon, this in parallel with a decrease in Roseburia spp. and Bacteroides/Prevotella/Porphyromonas (-1.4 and -2.0 log CFU) levels. Inulin treatment had moderate effects on lactate, propionate and butyrate levels. Denaturing gradient gel electrophoresis analysis revealed that inulin changed microbial metabolism by modulating the microbial community composition. In contrast, AXOS primarily affected microbial metabolism by 'switching on' AXOS-degrading enzymes (xylanase, arabinofuranosidase and xylosidase), without significantly affecting microbial community composition. Our results demonstrate that AXOS has a higher potency than inulin to shift part of the sugar fermentation toward the distal colon parts. Furthermore, due to its stronger propionate-stimulating effect, AXOS is a candidate prebiotic capable of lowering cholesterol and beneficially affecting fat metabolism of the host.


Water expansion dynamics after pulsed IR laser heating.
A nanosecond pulsed IR (1.9 microm) laser rapidly heated water, in an open vessel, to temperatures well below the boiling point. The subsequent dynamics of volume expansion were monitored using time-resolved interferometry in order to measure the increase in the water level in the heated area. The water expanded at less than the speed of sound, taking just less than 100 ns to increase its height by approximately 500 nm at surface temperature jumps of 20 K. The initial expansion was followed by an apparent contraction and then a re-expansion. The first expansion phase occurred more slowly than the timescale for bulk H-bond re-structuring of the water, as determined from vibrational bands in the Raman spectra, and represents the limit to the rate at which the overpressure caused by sudden heating can be released. The second phase of the expansion was caused by hydrodynamic effects and is accompanied by morphological changes resulting in light scattering as well as droplet spallation.


Evidence-based periodontal treatment. II. Predictable regeneration treatment.
Clinical experience, technical ability, and intuition are indispensable but are no longer sufficient as the sole clinical skills necessary to provide the best outcomes from periodontal and peri-implant regeneration treatment. This article describes a comprehensive and rigorous methodologic framework to assess the available evidence contained within the literature. This assessment tool is referred to as the evidence-based method. The methodology was used by a series of task forces convened to evaluate three common areas of clinical regenerative treatment. The major goals of these task forces were: (1) to increase the strength of the inference that practitioners can derive from the base of knowledge contained within the literature, (2) to develop algorithms to improve the predictability of regeneration treatment, and (3) to determine methods that can be used to predictably transfer the value of therapy to the patient.


Aortic, coronary, and myocardial lesions in relation to various factors.
The relative extent of raised and calcified coronary aortic lesions, prevalence of coronary stenosis and of fresh and old myocardial infarction, and mean heart weight were expressed on a continuous scale for 4 disease groups (coronary heart disease, cerebral infarction/haemorrhage, hypertension/diabetes, cancer). Within these groups the relative position for each of the lesions was calculated by subgroups of disease in order to show the elevating or depressing effect of hypertension, diabetes, obesity, and combinations of disease. The strength of association between 33 factors (town 5, disease 24, obesity, heart weight, age, and sex) and the lesions was calculated. An analysis of variance was carried out and the proportion of the variance of the different lesions accounted for by town, disease, etc., is shown. The extent of raised aortic lesions is strongly associated with age and hypertension. It is positively associated with coronary heart disease and inversely associated with cancer. Town factors have a small positive association which is larger than that due to obesity. 33 factors taken together account for 50% of the variance. Age alone accounts for 37% and sex for only 0.3%. The extent of calcified aortic lesion is strongly associated with age, town, hypertension, coronary heart disease and diabetes mellitus. It is inversely associated with cancer, more strongly in cancer of the bronchus and the liver than in other cancers. It is inversely associated with obesity. 33 factors together account for 30% of the variance and age alone accounts for about 13%. The extent of raised coronary lesions is strongly associated with age, coronary heart disease, sex, diabetes mellitus, hypertension and obesity. It is inversely associated with cancer, more strongly in cancer of the prostate and the liver than in other cancers. Town factors have a small association. 33 factors taken together account for about 43% of the variance. Age alone accounts for 25%. The extent of calcified coronary lesions is associated with coronary heart disease and age. There is a low association with hypertension and diabetes, an inverse association with cancer, and a low inverse association with obesity. 33 factors together account for 20% of the variance. Coronary heart disease alone accounts for 12% and age alone accounts for 8%. The prevalence of coronary stenosis is strongly associated with coronary heart disease, age, and sex. There is a small association with town factors, obesity, and hypertension, and an inverse association with cancer. All the factors together account for about 30% of the variance. Coronary heart disease alone accounts for 23% and age 10%.


Acute Nonhemorrhagic Adrenal Infarction in Pregnancy: 10-Year MRI Incidence and Patient Outcomes at a Single Institution.
The purpose of this study was to retrospectively investigate the MRI incidence of nonhemorrhagic adrenal infarction in pregnant women undergoing MRI evaluation of acute abdominal or flank pain, assess the MRI features quantitatively, and report patient outcomes. All abdominal MRI examinations of pregnant women with acute pain at one institution from May 2005 to April 2015 were reviewed. The adrenals were evaluated for abnormal morphologic and signal intensity characteristics described in the literature characterizing nonhemorrhagic adrenal infarction and were compared with the contralateral adrenal by paired t tests. The findings were correlated with clinical presentation. Patient demographics and outcomes were gathered from the medical record. Findings of nonhemorrhagic adrenal infarction were present in 5 of 379 (1.3%) examinations of four pregnant patients (mean age, 28 years; range, 20.8-33.9 years; mean gestational age, 26 weeks; range, 16-35 weeks). MRI features included lengthening (mean, 39.8 versus 21.2 mm) (p = 0.005) and increased T2 signal intensity (p = 0.001) of the infarcted adrenal with surrounding edema and without T1 signal intensity suggesting hemorrhage. No alternative diagnosis was identified. All patients presented with severe acute abdominal or flank pain on the same side as the MRI findings, tenderness to palpation, and mild leukocytosis. All women delivered healthy infants. Unilateral nonhemorrhagic adrenal infarction was identified in 1.3% of abdominal MRI examinations performed for pregnant women with acute abdominal or flank pain. Knowledge of the MRI characteristics of this entity is important for recognizing it and may prevent further potentially invasive tests, procedures, or missed diagnoses.


Misuse of fentanyl transdermal patch mixed with acute coronary syndrome.
Fentanyl transdermal patches have long been used in the palliative care of patients with chronic pain with a favorable safety profile. However, intoxications secondary to intentional and unintentional misuse have been widely reported. In this study, we report an otherwise healthy woman presented to emergency department who used three patches of fentanyl to alleviate her knee pain and with a picture mimicking acute coronary syndrome.


Reverse-biomineralization assembly of acid-sensitive biomimetic fibers for hard tissue engineering and drug delivery.
Biomimetic design and fabrication of tissue-engineered bone scaffolds that not only resemble natural bone in structure and performance, but also are endowed with specific functions, e.g., for drug delivery, are always an exciting research area. Herein, we report a kind of doxorubicin hydrochloride-loaded biomimetic ultrathin fiber, which is synthesized by preparing a kind of nanoporous bioactive glass fiber as a drug/protein carrier and bio-template and combining them in a reverse-biomineralization reaction. Protein adsorption experiments demonstrate that bovine serum albumin can be hosted in open large nanopores of bioactive glass fibers and the adsorption mechanism follows the intraparticle diffusion process. Biomineralization shows that proteins and drugs can be integrated at the nanoscale into minerals to form biomimetic and drug-loaded fibers, and the formation of such fibers depends on the functional ion (Ca, P, and Si) release of bioactive glass fibers and electrostatic interaction among bioactive glass fibers, proteins, and drugs. The drug-loaded composite fibers demonstrate bare homogeneous solid matrices in the fiber interior and surfaces upon which amorphous carbonated apatite resides. The drug release profiles show that the as-synthesized fibers are acid-sensitive and drugs can be released at pH 5, but not at neutral pH 7.4. Because of their structural advantages and the characteristics of acid-sensitive drug release, the biomimetic fibers have potential applications for repairing the bone defects resulting from tumour extirpation.


Genetic Manipulation of Homologous Recombination In Vivo Attenuates Intestinal Tumorigenesis.
Although disruption of DNA repair capacity is unquestionably associated with cancer susceptibility in humans and model organisms, it remains unclear if the inherent tumor phenotypes of DNA repair deficiency syndromes can be regulated by manipulating DNA repair pathways. Loss-of-function mutations in BLM, a member of the RecQ helicase family, cause Bloom's syndrome (BS), a rare, recessive genetic disorder that predisposes to many types of cancer. BLM functions in many aspects of DNA homeostasis, including the suppression of homologous recombination (HR) in somatic cells. We investigated whether BLM overexpression, in contrast with loss-of-function mutations, attenuated the intestinal tumor phenotypes of Apc(Min/+) and Apc(Min/+);Msh2(-/-) mice, animal models of familial adenomatous polyposis coli (FAP). We constructed a transgenic mouse line expressing human BLM (BLM-Tg) and crossed it onto both backgrounds. BLM-Tg decreased adenoma incidence in a dose-dependent manner in our Apc(Min/) (+) model of FAP, although levels of GIN were unaffected and concomitantly increased animal survival over 50%. It did not reduce intestinal tumorigenesis in Apc(Min/) (+);Msh2(-/-) mice. We used the pink-eyed unstable (p(un)) mouse model to demonstrate that increasing BLM dosage in vivo lowered endogenous levels of HR by 2-fold. Our data suggest that attenuation of the Min phenotype is achieved through a direct effect of BLM-Tg on the HR repair pathway. These findings demonstrate that HR can be manipulated in vivo to modulate tumor formation at the organismal level. Our data suggest that lowering HR frequencies may have positive therapeutic outcomes in the context of specific hereditary cancer predisposition syndromes, exemplified by FAP.


Renal potassium adaptation: Na-K-ATPase activity along the nephron after chronic potassium loading.
To identify nephron sites where renal potassium adaptation takes place, Na-K-ATPase was measured with a micromethod along the nephron of potassium-loaded mice. A possible role of aldosterone in this process was evaluated in K-loaded animals treated concurrently with pharmacologic doses of spironolactone. Animals fed a potassium-enriched diet for at least 2 wk excreted about 90% of ingested potassium in the urine, and fractional potassium clearance averaged 87 +/- 8%, compared with 13 +/- 2% in controls. Na-K-ATPase activity per millimeter tubule length increased by 225% in the cortical collecting tubule and by 177% in the medullary collecting tubule, but was not substantially affected in other nephron segments. Stimulation of Na-K-ATPase was identical in the cortical collecting tubule of K-loaded mice treated with spironolactone. These results indicate that chronic potassium loading in the mouse results in an adaptive increase potassium loading in the mouse results in an adaptive increase in Na-K-ATPase in the collecting tubule and suggest that this nephron segment is the major site of potassium adaptation in this species. This effect appears to be independent of aldosterone.


Body dysmorphia, self-mutilation and the reconstructive surgeon.
Body dysmorphic disorder (BDD) is a disabling preoccupation with a slight or an imagined defect in appearance. It is recognised in some patients who present to the plastic surgeon requesting multiple cosmetic procedures. Very rarely, BDD patients may wish for amputation of a healthy limb and may even mutilate themselves deliberately in order to necessitate amputation. These patients pose a diagnostic challenge as BDD is uncommon and they are often uncooperative whilst appearing mentally sound. Furthermore, they raise difficult ethical and legal issues for the surgeon. Although there is some guidance for the management of BDD patients seeking elective amputation, there is currently none for the management of those who present in the emergency setting. Illustrated by the case of a man who, having failed to find a complicit surgeon, attempted self-amputation of the hand, we review the relevant ethical, legal and management issues with advice by the British Medical Association and General Medical Council.


Exploration of a Au(i)-mediated three-component reaction for the synthesis of DNA-tagged highly substituted spiroheterocycles.
We demonstrate a Au(i)-mediated three-component reaction to DNA-tagged highly substituted 6-oxa-1,2-diazaspiro[4.4]nonanes from either DNA-coupled aldehydes, hydrazides, or alkynols. The choice of the starting material coupled to the DNA tag was critial for the purity of the product as the DNA-aldehyde conjugate yielded the purest products, whereas the alkynol- and hydrazide conjugates returned complex product mixtures. The reaction was compatible with thymine-, cytosine-, and, surprisingly, with adenine-DNA, while guanine-containing DNA strands were degraded under the reaction conditions.


An introduction to kernel-based learning algorithms.
This paper provides an introduction to support vector machines, kernel Fisher discriminant analysis, and kernel principal component analysis, as examples for successful kernel-based learning methods. We first give a short background about Vapnik-Chervonenkis theory and kernel feature spaces and then proceed to kernel based learning in supervised and unsupervised scenarios including practical and algorithmic considerations. We illustrate the usefulness of kernel algorithms by discussing applications such as optical character recognition and DNA analysis.


In vitro sensing of Cu(+) through a green fluorescence rise of pyranine.
Pyranine as a new class of fluorescent chemosensor for the Cu(+) ion is reported. The probe is capable of discriminating ranges of cations from the Cu(+) ion, even in competing environment. The dye displayed a rapid fluorescence response (t1/2 = 1.66 min) towards the Cu(+) ion, and the micromolar detection limit enabled the detection of the ion in environmental samples. The observed stoichiometry of complexation between pyranine and Cu(+) was 2 : 1. Interestingly, the sensing characteristic was specific to only neutral pH. A metal-to-ligand charge-transfer (MLCT)-based mechanism of sensing was proposed based on electron spin resonance (EPR), Raman spectroscopic and cyclic voltammetric studies.


Short-Term Grape Consumption Diminishes UV-Induced Skin Erythema.
Over three million Americans are affected by skin cancer each year, largely as a result of exposure to sunlight. The purpose of this study was to determine the potential of grape consumption to modulate UV-induced skin erythema. With 29 human volunteers, we report that nine demonstrated greater resistance to UV irradiation of the skin after consuming the equivalent of three servings of grapes per day for two weeks. We further explored any potential relationship to the gut-skin axis. Alpha- and beta-diversity of the gut microbiome were not altered, but grape consumption modulated microbiota abundance, enzyme levels, and KEGG pathways. Striking differences in the microbiome and metabolome were discerned when comparing the nine individuals showing greater UV resistance with the 20 non-responders. Notably, three urinary metabolites, 2'-deoxyribonic acid, 3-hydroxyphenyl acetic and scyllo-inositol, were depressed in the UV-resistant group. A ROC curve revealed a 71.8% probability that measurement of urinary 2'-deoxyribonic acid identifies a UV skin non-responder. 2'-Deoxyribonic acid is cleaved from the DNA backbone by reactive oxygen species. Three of the nine subjects acquiring UV resistance following grape consumption showed a durable response, and these three demonstrated unique microbiomic and metabolomic profiles. Variable UV skin sensitivity was likely due to glutathione S-transferase polymorphisms. We conclude that a segment of the population is capable of demonstrating greater resistance to a dermal response elicited by UV irradiation as a result of grape consumption. It is uncertain if modulation of the gut-skin axis leads to enhanced UV resistance, but there is correlation. More broadly, it is reasonable to expect that these mechanisms relate to other health outcomes anticipated to result from grape consumption.


Use of perfluorocarbon nanoparticles for non-invasive multimodal cell tracking of human pancreatic islets.
In vivo imaging of engraftment and immunorejection of transplanted islets is critical for further clinical development, with (1)H MR imaging of superparamagnetic iron oxide-labeled cells being the current premier modality. Using perfluorocarbon nanoparticles, we present here a strategy for non-invasive imaging of cells using other modalities. To this end, human cadaveric islets were labeled with rhodamine-perfluorooctylbromide (PFOB) nanoparticles, rhodamine-perfluoropolyether (PFPE) nanoparticles or Feridex as control and tested in vitro for cell viability and c-peptide secretion for 1 week. (19)F MRI, computed tomography (CT) and ultrasound (US) imaging was performed on labeled cell phantoms and on cells following transplantation beneath the kidney capsule of mice and rabbits. PFOB and PFPE-labeling did not reduce human islet viability or glucose responsiveness as compared with unlabeled cells or SPIO-labeled cells. PFOB- and PFPE-labeled islets were effectively fluorinated for visualization by (19)F MRI. PFOB-labeled islets were acoustically reflective for detection by US imaging and became sufficiently brominated to become radiopaque allowing visualization with CT. Thus, perfluorocarbon nanoparticles are multimodal cellular contrast agents that may find applications in real-time targeted delivery and imaging of transplanted human islets or other cells in a clinically applicable manner using MRI, US or CT imaging.


Using economics alongside medical audit. A case study of the management of endometriosis.
During 1993, the diagnosis and initial management of endometriosis in women was identified as an important topic to be addressed within the Gynaecology Audit Project in Scotland (GAPS). This paper documents the experience of using the medical audit data collected by this exercise to estimate the costs and outcomes associated with alternative treatments for endometriosis associated infertility. Its aim is not to draw firm policy conclusions but rather to demonstrate the economic methods and to highlight difficulties in the application of economics to audit. The average cost of medical management was found to be significantly higher than the average cost of expectant management because of the high costs of the drugs involved. There was little difference in clinical and health outcomes observed between those women managed expectantly and those women managed medically. The results of the economic assessment add cautious support to the growing volume of clinical evidence indicating expectant management as the first line treatment of choice although further research is required to substantiate these results in a larger sample of women presenting with this condition.


[Current problems in aviation physiology].
Soviet aviation has developed an efficient system of medical support based on physiological knowledge and aimed at the solution of various problems associated with pilot selection and training, his professional activity, arrangement of his working place, etc. It is understood that the following lines of research hold great promise in protecting the human body against adverse flight effects: methods of protection against high altitude effects which also help maintain high operational efficiency; conditioning of transport, regulation and energy mechanisms of oxygen supply; methods of predicting man's tolerance to motion sickness; study of mechanisms of photoreception of blinding brightnesses; development of photoprotectors, etc. With respect to professional selection, it is important to use EEG data for prognostic purposes, to investigate lateral functions of paired organs, and to explore their relationship with body tolerance to flight factors and efficiency of training. The results of physiological investigations of the functional state of the pilot form the basis to standardize his workload, to develop rational work-rest cycles, and to prevent performance deterioration. Physiological recommendations can be used to formulate requirements for the configuration of the pilot working place.


Retroviral transduction of human dendritic cells with a tumor-associated antigen gene.
Dendritic cells (DCs) are potent antigen-presenting cells that can activate quiescent T lymphocytes. When pulsed with tumor-associated antigen (TAA) peptide or protein, murine DCs can provide antitumor immunity. We reasoned that DCs retrovirally transduced with TAA genes might have important advantages over peptide- or protein-pulsed DCs, including long-term TAA presentation in vivo, and presentation of important but undefined epitopes. Therefore, we attempted to retrovirally transduce human DCs with a melanoma TAA gene (MART-1) and determine whether these transduced DCs could raise a specific antitumor response from quiescent autologous T lymphocytes. After retroviral transduction, human CD34+ cells were differentiated into DCs in vitro using granulocyte macrophage colony-stimulating factor, tumor necrosis factor alpha, and stem cell factor. This method consistently yielded a population of DCs as analyzed by morphology, phenotype, and MLR. Flow cytometric analysis revealed that 22-28% of cells expressing the DC phenotype also expressed a transduced marker gene. When DCs were transduced with the gene encoding MART-1, they stimulated much higher levels of cytokine release by MART-1-specific tumor-infiltrating lymphocytes than control DCs transduced with an irrelevant gene. In vitro stimulation using MART-1-transduced DCs but not control-transduced DCs raised specific antitumor CTLs from autologous quiescent T cells. These results provide evidence that human DCs can be retrovirally transduced with a TAA gene and that these transduced cells can raise a specific antitumor immune response in vitro. Transduced DCs may be useful for in vivo immunization against TAA.


Hearing rehabilitation in children with malformed ears: The endoscopic-assisted approach for cochlear implantation.
Cochlear implantation (CI) in children with malformed ears can be challenging through the standard surgical technique. Several alternative approaches have been described. The endoscopic-assisted approach can be chosen as an effective and safe surgical technique, overcoming the drawbacks of the traditional approach. We further describe a combined technique based on a limited mastoidectomy with no posterior tympanotomy and an endoscopic transmeatal approach to the round window (RW): the electrode is driven from the mastoid to the middle ear through the attic. The concomitant endoscopic assistance allows for improved surgical vision, reducing the risk of major complications. The main advantages of this technique are related to better visualization of the RW for safe insertion of the electrode; avoidance of damage to the facial nerve (FN), due to direct visualization, and sparing the posterior tympanotomy; avoidance of subtotal petrosectomy, if not necessary. The purpose of this article, supported with a video file, is to describe step by step this endoscopic-assisted procedure in a patient with middle ear malformation.


Expression of BMI-1 and Mel-18 in breast tissue--a diagnostic marker in patients with breast cancer.
Polycomb Group (PcG) proteins are epigenetic silencers involved in maintaining cellular identity, and their deregulation can result in cancer. Expression of Mel-18 and Bmi-1 has been studied in tumor tissue, but not in adjacent non-cancerous breast epithelium. Our study compares the expression of the two genes in normal breast epithelium of cancer patients and relates it to the level of expression in the corresponding tumors as well as in breast epithelium of healthy women. A total of 79 tumors, of which 71 malignant tumors of the breast, 6 fibroadenomas, and 2 DCIS were studied and compared to the reduction mammoplastic specimens of 11 healthy women. In addition there was available adjacent cancer free tissue for 23 of the malignant tumors. The tissue samples were stored in RNAlater, RNA was isolated to create expression microarray profile. These two genes were then studied more closely first on mRNA transcription level by microarrays (Agilent 44 K) and quantitative RT-PCR (TaqMan) and then on protein expression level using immunohistochemistry. Bmi-1 mRNA is significantly up-regulated in adjacent normal breast tissue in breast cancer patients compared to normal breast tissue from noncancerous patients. Conversely, mRNA transcription level of Mel-18 is lower in normal breast from patients operated for breast cancer compared to breast tissue from mammoplasty. When protein expression of these two genes was evaluated, we observed that most of the epithelial cells were positive for Bmi-1 in both groups of tissue samples, although the expression intensity was stronger in normal tissue from cancer patients compared to mammoplasty tissue samples. Protein expression of Mel-18 showed inversely stronger intensity in tissue samples from mammoplasty compared to normal breast tissue from patients operated for breast cancer. Bmi-1 mRNA level is consistently increased and Mel-18 mRNA level is consistently decreased in adjacent normal breast tissue of cancer patients as compared to normal breast tissue in women having had reduction mammoplasties. Bmi-1/Mel-18 ratio can be potentially used as a tool for stratifying women at risk of developing malignancy.


Intracranial pressure monitoring using a newly developed transducer-tipped ventricular drainage catheter.
A transducer-tipped ventricular drainage catheter (TVDC) was developed to allow continuous measurement of intracranial pressure (i.c.p.) without interrupting cerebrospinal fluid (CSF) drainage. The 8.5 Fr catheter with a double lumen has 16 side holes for CSF drainage and a small silicone strain-gauge transducer on the side of the catheter tip to measure the ICP directly. The transducer is composed of a 2 x 1 mm2 silicone plate. Calibration must be performed before insertion. The calibrated catheter is inserted into the ventricles, usually via the anterior horn of the lateral ventricles to the foramen of Monro. Recalibration is not necessary even if the patient's head is tilted. The TVDC was applied to patients in whom ICP was suspected to be high and in whom the ventricles were large enough for tapping. The ICP wave form and values measured by the TVDC and the manometer method were equivalent. The drift phenomenon did not occur throughout the entire measurement period. The TVDC is very useful for both controlling and monitoring ICP.


Estimating Lymphodynamic Conditions and Lymphovenous Anastomosis Efficacy Using (99m)Tc-phytate Lymphoscintigraphy with SPECT-CT in Patients with Lower-limb Lymphedema.
Diagnostic and therapeutic strategies for lower-limb lymphedema have not yet been established. The purpose of this study was to estimate the lymphodynamic condition and therapeutic efficacy of lymphovenous anastomosis (LVA) in lower-limb lymphedema patients using 2-phase (99m)Tc-phytate lymphoscintigraphy with single-photon emission computed tomography-computed tomography (SPECT-CT). In this study, consecutive patients with lower-limb lymphedema who underwent 2-phase lymphoscintigraphy using (99m)Tc-phytate were enrolled between June 2013 and June 2014. SPECT-CT was also performed to clarify the relationships between functional and morphological information. In both the early and delayed images, inguinal lymph node accumulation, dermal backflow, and their sequential alternations were evaluated, and liver-to-blood ratio and inguinal lymph node-to-blood ratio were calculated. All participants were classified into 6 types of lymphodynamic conditions based on the image findings. Patients with both dermal backflow and associated normal lymphatic vessel accumulation proceeded to LVA and underwent a second lymphoscintigraphy after the operation. Of all 30 participants, the largest population was categorized as type 4, which had consistent inguinal lymph node accumulation defect with dermal backflow. In 12 operated cases, dermal backflow was degraded in 10 cases by LVA. Liver-to-blood ratio in both early and delayed images and inguinal lymph node-to-blood ratio in delayed image significantly increased after LVA. Lymphoscintigraphy with SPECT-CT can provide both functional and morphological information simultaneously in patients with lower-limb lymphedema. Using these procedures, a type categorization for the patients was devised, which reflects their lymphodynamic conditions. The therapeutic efficacy of LVA could also be estimated quantitatively by the derived findings.


Somatotype tendency in Chilean adolescents from Valparaíso: review from 1979 to 2011.
A good tool to determine the biotype of growing and developing subjects that has been observed is the somatotype (ST). Objetive: This research study aims to describe the ST tendency in Chilean adolescents along the years. The bibliographic review was carried out using data from Medline, ISI Web of Knowledge, and SciELO. The key words used for the search were: somatotype, somatotyping, adolescent (in English and Spanish). The selected articles must describe ST in Chilean adolescents between 15 and 18 years of age. The graphic representation was made through a somato-chart and the series ST. To analyze the trajectory of the ST, the moving average method was used (SM), and the tendency line (lineal; TL), which expresses the data direction and projection. Eight articles complied with the inclusion criteria. Samples of students between 1979 and 2011 from the Regions of Valparaíso, Araucanía, and the Metropolitan region of Chile were reported. All studies correspond to transversal samples. The authors cite and/or describe the Heath-Carter anthropometric methods to assess the ST. The results showed positive TL for the endomorph component in all ages and genders. The mesomorph component maintained in male, unlike the female's component, which was negative except for the one at 18 years old. SM trajectory describes a regularity of the mesomorphic endomorph profile in female, yet with an increase in the endomorphy. In male, a change was observed from an ectomorphic mesomorph to endomorphic mesomorph. This progressive rise of the adipose component in both genders increases the risk of suffering non-communicable diseases.


The mechanism for the enhanced piezoelectricity in multi-elements doped (K,Na)NbO3 ceramics.
(K,Na)NbO3 based ceramics are considered to be one of the most promising lead-free ferroelectrics replacing Pb(Zr,Ti)O3. Despite extensive studies over the last two decades, the mechanism for the enhanced piezoelectricity in multi-elements doped (K,Na)NbO3 ceramics has not been fully understood. Here, we combine temperature-dependent synchrotron x-ray diffraction and property measurements, atomic-scale scanning transmission electron microscopy, and first-principle and phase-field calculations to establish the dopant-structure-property relationship for multi-elements doped (K,Na)NbO3 ceramics. Our results indicate that the dopants induced tetragonal phase and the accompanying high-density nanoscale heterostructures with low-angle polar vectors are responsible for the high dielectric and piezoelectric properties. This work explains the mechanism of the high piezoelectricity recently achieved in (K,Na)NbO3 ceramics and provides guidance for the design of high-performance ferroelectric ceramics, which is expected to benefit numerous functional materials.


Women Veterans' Experiences of Intimate Partner Violence and Non-Partner Sexual Assault in the Context of Military Service: Implications for Supporting Women's Health and Well-Being.
Women who have served in the military in the United States experience high rates of intimate partner violence (IPV) and non-partner sexual assault (SA). The military setting presents challenges and opportunities not experienced in other employment contexts that may compound the negative impacts of IPV/SA on women's lives. The purpose of this study was to explore the intersection of women's experiences of IPV/SA and military service through analysis of women veterans' narrative accounts. We conducted in-depth face-to-face qualitative interviews with 25 women veterans receiving primary care at a U.S. Veterans Affairs Medical Center. We draw upon Adler and Castro's (2013) Military Occupational Mental Health Model to frame our understanding of the impact of IPV/SA as a stressor in the military cultural context and to inform efforts to prevent, and support women service members who have experienced, these forms of violence. Our findings highlight the impact of IPV/SA on women's military careers, including options for entering and leaving military service, job performance, and opportunities for advancement. Women's narratives also reveal ways in which the military context constrains their options for responding to and coping with experiences of IPV/SA. These findings have implications for prevention of, and response to, intimate partner or sexual violence experienced by women serving in the military and underscore the need for both military and civilian communities to recognize and address the negative impact of such violence on women service members before, during, and after military service.


A randomized controlled trial of guided imagery in bulimia nervosa.
The objective of this study was to test a guided imagery therapy designed to enhance self-comforting in bulimia nervosa. A randomized controlled trial compared 6 weeks of individual guided imagery therapy with a control group. Fifty participants who met DSM-III-R criteria for bulimia nervosa completed the study. Measures of eating disorder symptoms, psychological functioning and the experience of guided imagery therapy were administered. The guided imagery treatment had substantial effects on the reduction of bingeing and purging episodes; the imagery group had a mean reduction of binges of 74% and of vomiting of 73%. The imagery treatment also demonstrated improvement on measures of attitudes concerning eating, dieting and body weight in comparison to the control group. In addition, the guided imagery demonstrated improvement on psychological measures of aloneness and the ability for self-comforting. Evidence from this study suggests that guided imagery was an effective treatment for bulimia nervosa, at least in the short-term.


Life and mental health of medical students after the Great East Japan Earthquake.
Students of the Tohoku University School of Medicine experienced the Great East Japan Earthquake on March 11, 2011. We conducted a series of surveys to examine the relationships among their experiences and activities on the day of the earthquake, their physical, mental, and economic problems following the disaster, and how their problems changed over time. The initial survey was performed in April 2011, with three follow-up surveys in July 2011, February 2012, and April 2013. The initial survey focused on students' experiences and living conditions during the disaster, which contained questions on their locations and circumstances, family circumstances, lives after the earthquake, voluntary works, physical or mental health problems, and desire for counseling. The follow-up surveys included new items regarding their circumstances, changes in their health problems, and their desire for economic assistance. Students who answered the first survey to the 4th one, with response rates in the following bracket, were as follows: 472 (28.0%), 640 (29.9%), 681 (36.0%), and 678 (39.0%), respectively. Six months after the earthquake, about 20% having experienced physical and/or mental problems. Although there was a trend toward a reduction in suffering and health problems over time, some students' conditions remained unchanged or worsened. It is notable that students who had participated in voluntary activities, despite their own suffering of harm and distress, were identified as the group that required the closest attention. Our present results can be applied to appropriate supports for students in future large-scale disasters.


Radiologist detection of microcalcifications with and without computer-aided detection: a comparative study.
To compare the sensitivity and specificity of microcalcification detection by radiologists alone and assisted by a computer-aided detection (CAD) system. Films of 106 patients were masked, randomized, digitized and analysed by the CAD-system. Five readers interpreted the original mammograms and were blinded to demographics, medical history and earlier films. Forty-two mammograms with malignant microcalcifications, 40 with benign microcalcifications and 24 normal mammograms were included. Results were recorded on a standardized image interpretation form. The mammograms with suspicious areas flagged by the CAD-system were displayed on mini-monitors and immediately re-reviewed. The interpretation was again recorded on a new copy of the standard form and classified according to six groups. Forty-one out of 42 (98%) malignant microcalcifications and 32 of 40 (80%) benign microcalcifications were flagged by the CAD-system. There was an average of 1.2 markers per image. The sensitivity for malignant microcalcifications detection by mammographers without and with the CAD-system ranged from 81% to 98% and from 88% to 98%, respectively. The mean difference without and with CAD-system was 2.2% (range 0-7%). No statistically significant changes in sensitivity were found when experienced mammographers were assisted by the CAD-system, with no significant compromise in specificity.


ABO haemolytic disease of the newborn: Improved prediction by novel integration of causative and protective factors in newborn and mother.
Prediction of haemolytic disease of the foetus and newborn (HDFN) caused by maternal anti-A/-B enables timely therapy, thereby preventing the development of kernicterus spectrum disorder. However, previous efforts to establish accurate prediction methods have been only modestly successful. In a case-control study, we examined 76 samples from mothers and 76 samples from their newborns; 38 with and 38 without haemolysis. The IgG subclass profile of maternal anti-A and anti-B was determined by flow cytometry. Samples from newborns were genetically analysed for the A2 subgroup, secretor and FcγRIIa receptor alleles. Surprisingly, we found a correlation between the newborn secretor allele and haemolysis (p = 0.034). No correlation was found for FcγRIIa alleles. The A2 subgroup was found only in newborns without haemolysis. Unexpectedly, different reaction patterns were found for maternal anti-A and anti-B; consequently, the results were treated separately. For the prediction of haemolysis in A-newborns, the maternal IgG1 subclass determination resulted in an accuracy of 83% at birth. For B-newborns, an accuracy of 91% was achieved by the maternal IgG2 subclass determination. We improved the prediction of ABO-HDFN by characterizing maternal anti-A and anti-B by flow cytometry and we presented genetic traits in newborns with correlation to haemolysis. We propose a new understanding of A- and B-substances as immunogens that enhance the maternal immune response and protect the newborn, and we suggest that the development of ABO-HDFN is different when caused by maternal anti-A compared to maternal anti-B.


Prospective phase II trial of iriontecan, 5-fluorouracil, and leucovorin in combination as salvage therapy for advanced colorectal cancer.
Irinotecan (CPT11) has established activity in the treatment of advanced colorectal cancer without cross-resistance with established 5-fluorouracil/folinic acid-based therapy. This phase II study was conducted to establish the efficacy and tolerance of combination treatment with irinotecan and 5-fluorouracil as salvage treatment for this disease. Open phase II trial of CPT11 180 mg/m2 on day 1, leucovorin 200 mg/m2 on days 1 and 2, and 5-fluorouracil 400 mg/m2 loading dose followed by 600 mg/m2 infusion on days 1 and 2. Treatment was continued until progression or limiting toxicity. Responders could proceed to surgical resection of residual disease. Thirty-nine patients from 2 institutions received a total of 287 cycles of therapy (median 7 cycles/patient). Eight patients achieved an objective response (7 for liver metastasis and 1 for lung metastasis), and an additional 12 obtained stabilization of disease or minor responses (MR); of these patients, 8 with liver metastasis (7 partial response and 1 MR) underwent hepatic resection of metastases and all them obtained a complete response. The median duration of response was 14 months, and the median survival was 11 months. Hematologic toxicity (neutropenia) was the most common serious side effect (29% of patients in 2% of cycles), but significant fever developed in only 4 patients. Grade III diarrhea was experienced in at least 1 cycle by 10% of patients. The results of this schedule compare favorably with previously reported experience of a phase I study designed to establish the dose of CPT11. Efficacy in this poor prognosis group of patients is very encouraging, and the schedule is well tolerated by even previously treated patients.


Nucleation of diamond films on heterogeneous substrates: a review.
Diamond thin films are known to have properties similar to bulk diamond and have applications in both industry and fundamental studies in academia. The high surface energy of diamond makes it extremely difficult to grow diamond films on foreign substrates. Hence, to grow diamond films on non-diamond substrates, a nucleation step is needed. In this review various techniques used for diamond nucleation/seeding will be discussed. At present electrostatic seeding by diamond nanoparticles is the most commonly used seeding technique for nanocrystalline growth. In this technique the substrate is dipped in a nanodiamond solution to form a mono layer of diamond seeds. These seeds when exposed to appropriate conditions grow to form diamond layers. This technique is suitable for most substrates. For heteroepitaxial growth, bias enhanced nucleation is the primary technique. In this technique the substrate is biased to form diamond nuclei in the initial stages of growth. This technique can be used for any conducting flat surface. For growth on ceramics, polishing by diamond grit or electrostatic seeding can be used. Polishing the ceramics with diamond powder leaves small diamond particles embedded in the substrate. These small particles then act as seeds for subsequent diamond growth. Apart from these techniques, chemical nucleation, interlayer driven nucleation and mixed techniques have been discussed. The advantages and disadvantages of individual techniques have also been discussed.


Binaural interference with simulated electric acoustic stimulation.
Preserved low-frequency acoustic hearing in cochlear implant (CI) recipients affords combined electric-acoustic stimulation (EAS) that could improve access to low-frequency acoustic binaural cues and enhance spatial hearing. Such benefits, however, could be undermined by interactions between electrical and acoustical inputs to adjacent (spectral overlap) or distant (binaural interference) cochlear places in EAS. This study simulated EAS in normal-hearing listeners, measuring interaural time difference (ITD) and interaural level difference (ILD) discrimination thresholds for a low-frequency noise (simulated acoustic target) in the presence or absence of a pulsatile high-frequency complex presented monotically or diotically (simulated unilateral or bilateral electric distractor). Unilateral distractors impaired thresholds for both cue types, suggesting influences of both binaural interference (which appeared more consistently for ITD than ILD) and physical spectral overlap (for both cue types). Reducing spectral overlap with an EAS gap between 1 and 3 kHz consistently improved binaural sensitivity. Finally, listeners displayed significantly lower thresholds with simulated bilateral versus unilateral electric stimulation. The combined effects revealed similar or better thresholds in bilateral full spectral overlap than in unilateral EAS gap conditions, suggesting that bilateral implantation with bilateral acoustic hearing preservation could allow for higher tolerance of spectral overlap in CI users and improved binaural sensitivity over unilateral EAS.


Stimulation of progesterone production in human granulosa-lutein cells by lipoproteins: evidence for cholesterol-independent actions of high-density lipoproteins.
Low-density lipoproteins (LDL) have been consistently reported to stimulate ovarian steroidogenesis, apparently by the provision of cholesterol as a steroidogenic substrate. Recent studies suggest that high-density lipoproteins (HDL) can also deliver cholesterol to support progesterone synthesis in human granulosa-lutein cells. Therefore, this study investigated the contributions of (i) cholesterol delivery, (ii) cyclic AMP and (iii) protein kinase C (PKC) in the steroidogenic responses of human granulosa-lutein cells to HDL and LDL. Over a 24-h treatment incubation, HDL stimulated a larger increase in progesterone output than did LDL at equivalent cholesterol concentrations. Moreover, at equal protein concentrations (100 microg protein/ml), HDL doubled progesterone production by cells co-treated with a maximally effective concentration of 22R-hydroxycholesterol, whereas LDL had no effect on the progesterone response to this membrane-permeable sterol. These observations indicate that the progesterone response to HDL is not solely due to the delivery of cholesterol as a steroidogenic substrate. Over 24 h, the stimulation of progesterone synthesis by HDL was additive with the response to a maximally effective concentration of dibutyryl-cAMP, but was unaffected by the down-regulation of PKC activity (by chronic pre-treatment with a tumour-promoting phorbol ester). We have concluded that HDL appears to stimulate progesterone production in human granulosa-lutein cells by a mechanism not solely reliant on cholesterol delivery.


Two-step phosphorylation of Ana2 by Plk4 is required for the sequential loading of Ana2 and Sas6 to initiate procentriole formation.
The conserved process of centriole duplication requires Plk4 kinase to recruit and promote interactions between Sas6 and Sas5/Ana2/STIL (respective nomenclature of worms/flies/humans). Plk4-mediated phosphorylation of Ana2/STIL in its conserved STAN motif has been shown to promote its interaction with Sas6. However, STAN motif phosphorylation is not required for recruitment of Ana2 to the centriole. Here we show that in Drosophila, Ana2 loads onto the site of procentriole formation ahead of Sas6 in a process that also requires Plk4. However, whereas Plk4 is first recruited to multiple sites around the ring of zone II at the periphery of the centriole, Ana2 is recruited to a single site in telophase before Plk4 becomes finally restricted to this same single site. When we over-ride the auto-destruction of Plk4, it remains localized to multiple sites in the outer ring of the centriole and, if catalytically active, recruits Ana2 to these sites. Thus, it is the active form of Plk4 that promotes Ana2's recruitment to the centriole. We now show that Plk4 phosphorylates Ana2 at a site other than the STAN motif, which lies in a conserved region we term the ANST (ANa2-STil) motif. Mutation of this site, S38, to a non-phosphorylatable residue prevents the procentriole loading of Ana2 and blocks centriole duplication. Thus the initiation of procentriole formation requires Plk4 to first phosphorylate a single serine residue in the ANST motif to promote Ana2's recruitment and, secondly, to phosphorylate four residues in the STAN motif enabling Ana2 to recruit Sas6. We discuss these findings in light of the multiple Plk4 phosphorylation sites on Ana2.


Presence of fetal DNA in maternal plasma decades after pregnancy.
Cells of fetal origin and cell-free fetal DNA can be detected in the maternal circulation during pregnancy, and it has recently been shown that fetal cells can persist long after delivery. Given the various biological and clinical implications of this fact, we tested the hypothesis that cell-free fetal DNA can be present in maternal plasma decades after pregnancy. We extracted DNA from plasma samples and nucleated blood cells of 160 healthy women with male offspring at different time intervals after delivery (range 1-60 years). All of the samples were tested by means of a real-time quantitative PCR assay for a specific Y chromosome sequence (the SRY gene). Y chromosome-specific DNA was detected in 16 peripheral blood cell samples (10%) and 35 plasma samples (22%). The women with male sequences in the cell fraction had significantly greater total parity ( P=0.018). The proportion of women with detectable Y sequences in the plasma or cell samples was not related to the time since delivery. The fetal DNA concentrations in the genomic material extracted from plasma samples were significantly higher than those extracted from the Y-positive cell samples (149+/-140 vs 20+/-13 genome-equivalents/ml; P<0.001). There was no relationship between the concentration of fetal DNA and the time since delivery. Not only fetal cells, but also fragments of fetal DNA can be present in the maternal circulation indefinitely after pregnancy. This finding has practical implications for non-invasive prenatal diagnoses based on maternal blood, and may be considered for possible pathophysiological correlations.


Total synthesis and revision of stereochemistry of the marine metabolite trunkamide A.
The isolation of the cytotoxic Lissoclinum sp. metabolite trunkamide A was reported in 1996. After completion of a total synthesis in 1999, it became clear that the structure of this marine natural product had to be revised. We now report the first preparation of actual trunkamide A in a total synthesis that serves as an unambiguous structural and stereochemical proof. Highlights of our synthetic strategy are a Lewis acid assisted aziridine opening that was used for the preparation of the novel reverse-prenylated serine and threonine side chains as well as an efficient oxazoline-thiazoline interconversion on the macrocyclic skeleton. In addition, several stereoisomers prepared by complementary synthetic protocols serve to illustrate the general scope of our methodology and confirm the configurational assignment.


A Resazurin Reduction-Based Assay for Rapid Detection of Polymyxin Resistance in Acinetobacter baumannii and Pseudomonas aeruginosa.
A rapid test was developed for identification of polymyxin resistance in nonfermenting bacteria. This test detects viable cells after growth in a medium containing a defined concentration of colistin. The principle of this test is based on the visual detection of the reduction of the resazurin reagent, a viability colorant, as observed by its color change (blue to purple or pink). Its evaluation was performed by using 92 colistin-resistant and colistin-susceptible Acinetobacter baumannii and Pseudomonas aeruginosa isolates. Sensitivity and specificity were found to be 100% and 95%, respectively, by comparison with the standard broth microdilution method. The Rapid ResaPolymyxin Acinetobacter/Pseudomonas NP test is inexpensive, easy to perform, highly sensitive and specific, and can be completed in 4 hours. It could be useful in countries facing endemic spread of colistin-resistant nonfermenters.


Interplay between Polo kinase, LKB1-activated NUAK1 kinase, PP1βMYPT1 phosphatase complex and the SCFβTrCP E3 ubiquitin ligase.
NUAK1 (NUAK family SnF1-like kinase-1) and NUAK2 protein kinases are activated by the LKB1 tumour suppressor and have been implicated in regulating multiple processes such as cell survival, senescence, adhesion and polarity. In the present paper we present evidence that expression of NUAK1 is controlled by CDK (cyclin-dependent kinase), PLK (Polo kinase) and the SCFβTrCP (Skp, Cullin and F-boxβTrCP) E3 ubiquitin ligase complex. Our data indicate that CDK phosphorylates NUAK1 at Ser445, triggering binding to PLK, which subsequently phosphorylates NUAK1 at two conserved non-catalytic serine residues (Ser476 and Ser480). This induces binding of NUAK1 to βTrCP, the substrate-recognition subunit of the SCFβTrCP E3 ligase, resulting in NUAK1 becoming ubiquitylated and degraded. We also show that NUAK1 and PLK1 are reciprocally controlled in the cell cycle. In G2-M-phase, when PLK1 is most active, NUAK1 levels are low and vice versa in S-phase, when PLK1 expression is low, NUAK1 is more highly expressed. Moreover, NUAK1 inhibitors (WZ4003 or HTH-01-015) suppress proliferation by reducing the population of cells in S-phase and mitosis, an effect that can be rescued by overexpression of a NUAK1 mutant in which Ser476 and Ser480 are mutated to alanine. Finally, previous work has suggested that NUAK1 phosphorylates and inhibits PP1βMYPT1 (where PP1 is protein phosphatase 1) and that a major role for the PP1βMYPT1 complex is to inhibit PLK1 by dephosphorylating its T-loop (Thr210). We demonstrate that activation of NUAK1 leads to a striking increase in phosphorylation of PLK1 at Thr210, an effect that is suppressed by NUAK1 inhibitors. Our data link NUAK1 to important cell-cycle signalling components (CDK, PLK and SCFβTrCP) and suggest that NUAK1 plays a role in stimulating S-phase, as well as PLK1 activity via its ability to regulate the PP1βMYPT1 phosphatase.