Liquid chromatography/electrospray ionization tandem mass spectrometry assay for determination of nicotine and metabolites, caffeine and arecoline in breast milk.
A procedure based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) is described for the determination of nicotine and its principal metabolites cotinine, trans-3-hydroxycotinine and cotinine-N-oxide, caffeine and arecoline in breast milk, using N-ethylnorcotinine as internal standard. Liquid/liquid extraction with chloroform/isopropanol (95:5, v/v) was used for nicotine, cotinine, trans-3-hydroxycotinine, cotinine-N-oxide and caffeine under neutral conditions and for arecoline under basic conditions. Chromatography was performed on a C(8) reversed-phase column using a gradient of 50 mM ammonium formate, pH 5.0, and acetonitrile as a mobile phase at a flow rate of 0.5 mL/min. Separated analytes were determined by electrospray ionization tandem mass spectrometry in the positive ion mode using multiple reaction monitoring. Limits of quantification were 5 microg/L for nicotine, cotinine, trans-3-hydroxycotinine, cotinine-N-oxide and caffeine, and 50 microg/L for arecoline using 1 mL human milk per assay. Calibration curves were linear over the calibration ranges for all the substances under investigation, with a minimum r(2) > 0.998. At three concentrations spanning the linear dynamic range of the assay, mean recoveries from breast milk ranged between 71.8 and 77.4% for different analytes. This method was applied to the analysis of analytes in human milk to assess substance exposure in breast-fed infants in relation to eventual clinical outcomes. This LC/MS/MS assay provides adequate sensitivity and performance characteristics for the simultaneous quantification of biomarkers of three of the drugs most commonly used worldwide (tobacco, caffeine and areca nut).


Molecular analysis of FMRFamide- and FMRFamide-related peptides (FaRPS) in the cuttlefish Sepia officinalis.
The display of complex color patterns of the cuttlefish Sepia officinalis is under the regulation of the FMRFamide-related peptide (FaRP) family, but their exact identities are unknown. We report the isolation and characterization of a full-length FaRP cDNA from the brain of S. officinalis. This cDNA is 1850 base pairs long, including an open reading frame of 996 base pairs. The cDNA encodes a precursor protein containing four FaRPs: ALSGDAFLRF, FIRF, FLRF and FMRF. Each propeptide has a C-terminal glycine residue that is presumably converted post-translationally to an amide. Every FaRP propeptide is also flanked by basic amino acid residues at the amino and carboxy termini, indicative of putative cleavage sites during post-translational processing. Each of the four FaRPs encoded by this cDNA causes chromatophore expansion when assayed in an in vitro chromatophore bioassay. Thus, it is likely that one or more of the FaRPs identified in this study are involved in controlling chromatophore activity in cuttlefish.


Routine clinical evaluation of cerebrovascular reserve capacity using carbogen in patients with intracranial stenosis.
A promising method for identifying hemodynamic impairment that may serve as a biomarker for stroke risk in patients with intracranial stenosis is cerebrovascular reactivity (CVR) mapping using noninvasive MRI. Here, abilities to measure CVR safely in the clinic using hypercarbic hyperoxic (carbogen) gas challenges, which increase oxygen delivery to tissue, are investigated. In sequence with structural and angiographic imaging, blood oxygenation level-dependent carbogen-induced CVR scans were performed in patients with symptomatic intracranial stenosis (n=92) and control (n=10) volunteers, with a subgroup of patients (n=57) undergoing cerebral blood flow-weighted pseudocontinuous arterial spin labeling CVR. Subjects were stratified for 4 substudies to evaluate relationships between (1) carbogen and hypercarbic normoxic CVR in healthy tissue (n=10), (2) carbogen cerebral blood flow CVR and blood oxygenation level-dependent CVR in intracranial stenosis patients (n=57), (3) carbogen CVR and clinical measures of disease in patients with asymmetrical intracranial atherosclerotic (n=31) and moyamoya (n=29) disease, and (4) the CVR scan and immediate and longer-term complications (n=92). Noninvasive blood oxygenation level-dependent carbogen-induced CVR values correlate with (1) lobar hypercarbic normoxic gas stimuli in healthy tissue (R=0.92; P<0.001), (2) carbogen-induced cerebral blood flow CVR in patients with intracranial stenosis (R=0.30-0.33; P<0.012), and (3) angiographic measures of disease severity both in atherosclerotic and moyamoya patients after appropriate processing. No immediate stroke-related complications were reported in response to carbogen administration; longer-term neurological events fell within the range for expected events in this patient population. Carbogen-induced CVR elicited no added adverse events and provided a surrogate marker of cerebrovascular reserve consistent with intracranial vasculopathy.


SWADAPT1: assessment of an electric wheelchair-driving robotic module in standardized circuits: a prospective, controlled repeated measure design pilot study.
The objective of this study is to highlight the effect of a robotic driver assistance module of powered wheelchair (PWC), using infrared sensors and accessorizing a commercial wheelchair) on the reduction of the number of collisions in standardized circuit in a population with neurological disorders by comparing driving performance with and without assistance. This is a prospective, single-center, controlled, repeated measure design, single-blind pilot study including patients with neurological disabilities who are usual drivers of electric wheelchairs. The main criterion for evaluating the device is the number of collisions with and without the assistance of a prototype anti-collision system on three circuits of increasing complexity. Travel times, cognitive load, driving performance, and user satisfaction are also analyzed. 23 Patients, 11 women and 12 men with a mean age of 48 years old completed the study. There was a statistically significant reduction in the number of collisions on the most complex circuit: 61% experienced collisions without assistance versus 39% with assistance (p = 0.038). This study concludes that the PWC driving assistance module is efficient in terms of safety without reducing the speed of movement in a population of people with disabilities who are habitual wheelchair drivers. The prospects are therefore to conduct tests on a target population with driving failure or difficulty who could benefit from this device so as to allow them to travel independently and safely.


Disaggregating census data for population mapping using random forests with remotely-sensed and ancillary data.
High resolution, contemporary data on human population distributions are vital for measuring impacts of population growth, monitoring human-environment interactions and for planning and policy development. Many methods are used to disaggregate census data and predict population densities for finer scale, gridded population data sets. We present a new semi-automated dasymetric modeling approach that incorporates detailed census and ancillary data in a flexible, "Random Forest" estimation technique. We outline the combination of widely available, remotely-sensed and geospatial data that contribute to the modeled dasymetric weights and then use the Random Forest model to generate a gridded prediction of population density at ~100 m spatial resolution. This prediction layer is then used as the weighting surface to perform dasymetric redistribution of the census counts at a country level. As a case study we compare the new algorithm and its products for three countries (Vietnam, Cambodia, and Kenya) with other common gridded population data production methodologies. We discuss the advantages of the new method and increases over the accuracy and flexibility of those previous approaches. Finally, we outline how this algorithm will be extended to provide freely-available gridded population data sets for Africa, Asia and Latin America.


Common and distinctive features of verbal and pictorial stimuli as determinants of psychophysiological responsivity.
The present study focuses on the relation between stimulus features and psychophysiological responsivity by using a modified version of the information detection paradigm. Compound pictorial and verbal stimuli (schematic faces with beard, glasses, and hat, and descriptions of people in terms of occupation, city of residence, a hobby, and a personality trait) were used as the relevant stimuli that subjects were instructed to memorize. Skin conductance responses were measured during the subsequent presentation of a sequence of test stimuli. Each sequence included a critical stimulus that shared from one to four common components with the relevant stimulus in each of two pictorial and two verbal experiments. We hypothesized that the electrodermal responsivity to the critical stimulus would reflect the degree it matches the relevant one. The results indicated that when the critical stimulus was identical to the relevant stimulus, responsivity was maximal. Neutral stimuli (i.e., those that shared no components with the relevant stimulus) produced minimal responsivity, and critical stimuli that only partially matched the relevant one produced intermediate levels of responsivity (in spite of the subjects' awareness of the differences between the critical and the relevant stimuli). The monotonic relation between the degree of match and responsivity supports the proposed model, which assumes that each stimulus in the sequence is being compared with the relevant stimulus by a feature-matching process, and electrodermal responsivity is related to the outcome of this process. In a fifth experiment, we compared geometric and contrast models for similarity and found that the pattern of responsivity violated the minimality and symmetry assumptions of the geometric model. The relation between cognitive processes and psychophysiological responsivity is discussed, as are implications for the application of the guilty knowledge technique for detecting information.


[Experimental and clinical studies on cefmetazole in the field of internal medicine].
Susceptibilities in various types of clinical isolates to cefmetazole (CMZ) were determined by the disk method and the serial agar dilution method for MIC measurement. CMZ showed high antibacterial activity for all Gram-positive cocci except E. faecalis, and for H. influenzae, E. coli, Klebsiella sp. and Proteus sp.. CMZ was administered on 33 patients with infections, mainly biliary tract infections, in the field of internal medicine. The clinical efficacy of the drug was 86.7% for infections of the biliary tract, 80.0% for respiratory tract infections, 100% for urinary tract infections and 76.7% for all cases. There were no adverse reaction or changes in laboratory findings caused by CMZ in any of the patients.


Reliability of self-assessed reading of skin tests: a possible approach in research and clinical practice?
In the investigation and management of skin disease, various testing protocols are of importance. The extent to which clinical judgments and decisions on therapy are supported by the performance of such testing can be affected negatively by the lack of time and resources for the performance of tests. In the present study, the possibility of utilizing self-reporting by subjects is investigated. Determination of irritation threshold for sodium lauryl sulphate (SLS) and minimal erythema dose for ultraviolet B were chosen as suitable self-reading protocols. Test reading by 26 subjects instructed in "present" or "absent" reporting of test reactions were compared to trained observer reading. Absolute agreement was found in 76.9 percent of the SLS reactions and in 85 percent of the UVB reactions. Weighted Kappa for the agreement between observations showed values of 0.76 for the SLS reactions and 0.83 for UVB reactions. We conclude that use of the protocols here studied, and other test protocols modified to accommodate a binomial assessment outcome ("+" or "-"), could well lead to an increase in the performance of skin testing. This could be a qualitative advantage for diagnosis and management of skin diseases. Additionally, population studies and even prevention initiatives could be facilitated.


Reactions to others' mistakes: an empirical test of fairness theory.
Drawing on fairness theory (R. Folger & R. Cropanzano, 1998, 2001), the authors examined undergraduates' reactions to advisor mistakes made in an academic advisement scenario. The authors hypothesized that perceived fairness, blame, and behavioral intentions to address wrongdoing would be influenced by outcome severity and the nature of the mistake (knowledgeable mistakes vs. ignorant mistakes; errors of commission vs. errors of omission). Results partially supported the hypotheses. Participants perceived scenarios as less fair and expressed higher intent to address the wrongdoing when the consequences were high in severity and when a knowledgeable person made the mistake. Participants attributed significantly more blame to the advisor in the high severity outcome conditions. Counterfactual thoughts mediated the effects of target knowledge but not outcome severity. The authors discuss theoretical and practical implications.


Sick leave in patients with obstructive lung disease is related to psychosocial and work variables rather than to FEV1.
The aim of this exploratory study was to investigate associations between sick leave in workers with asthma or COPD and disease-related variables, psychosocial variables, and work characteristics. Hundred and eighty-nine patients with physician-diagnosed asthma (N=118) or COPD (n=71) who had paid work in the past 12 months completed questionnaires on sick leave, health complaints, functional limitations, work characteristics and psychosocial issues, and underwent a pulmonary function test (FEV1 and FVC before and after bronchodilation). Logistic regression analyses were performed to investigate variables independently associated with high sick leave (i.e. more than twice a year and/or longer than 1 month per episode). Asthma patients, not having an emotionally difficult job, with low job satisfaction, who had changed employers, utilized job control, and who encountered pulmonary aggravating factors at work were found to have a higher incidence of sick leave. COPD patients, who had informed the employer or colleagues about the disease, who did not have difficult tasks at work, who did not hide dyspnea and limitations, and who reported high fatigue were showing higher sick leave. FEV1 and FVC were not associated with sick leave in either group. It was concluded that psychosocial variables, work characteristics, functional limitations, and complaints play a more important role in sick leave in workers with asthma and COPD than FEV1.


[Amino acids in human nutrition].
The article presents data on amino acids metabolism in human organism. The review described structure and function of amino acids, essentiality criteria, amino acids energy source role, amino acids specific functions, amino acids requirements.


Methods for multiplex template sampling in digital PCR assays.
The efficient use of digital PCR (dPCR) for precision copy number analysis requires high concentrations of target molecules that may be difficult or impossible to obtain from clinical samples. To solve this problem we present a strategy, called Multiplex Template Sampling (MTS), that effectively increases template concentrations by detecting multiple regions of fragmented target molecules. Three alternative assay approaches are presented for implementing MTS analysis of chromosome 21, providing a 10-fold concentration enhancement while preserving assay precision.


Concise approach toward tetrazolo[1,5-a][1,4]benzodiazepines via a novel multicomponent isocyanide-based condensation.
A novel and efficient method for the synthesis of heteroannulated [1,4]benzodiazepines via an isocyanide-based multicomponent reaction is reported. The tetrazolo[1,5-a][1,4]benzodiazepines were obtained by a facile azide Ugi five-center four-component reaction (U-5C-4CR) using ketones, sodium azide, ammonium chloride, and corresponding isocyanide. The aforementioned tetrazolodiazepines represent a notable class of compounds with proven platelet aggregation inhibitory and cholecystokinin agonist activities.


Identification of new molecular targets for PET imaging of the microglial anti-inflammatory activation state.
Microglia are potential targets for therapeutic intervention in neurological and neurodegenerative diseases affecting the central nervous system. In order to assess the efficacy of therapies aimed to reduce the tissue damaging activities of microglia and/or to promote the protective potential of these cells, suitable pre-clinical and clinical tools for the in vivo analysis of microglia activities and dynamics are required. The aim of this work was to identify new translational markers of the anti-inflammatory / protective state of microglia for the development of novel PET tracers. Methods: New translational markers of the anti-inflammatory/protective activation state of microglia were selected by bioinformatic approaches and were in vitro and ex vivo validated by qPCR and immunohistochemistry in rodent and human samples. Once a viable marker was identified, a novel PET tracer was developed. This tracer was subsequently confirmed by autoradiography experiments in murine and human brain tissues. Results: Here we provide evidence that P2RY12 expression increases in murine and human microglia following exposure to anti-inflammatory stimuli, and that its expression is modulated in the reparative phase of experimental and clinical stroke. We then synthesized a novel carbon-11 labeled tracer targeting P2RY12, showing increased binding in brain sections of mice treated with IL4, and low binding to brain sections of a murine stroke model and of a stroke patient. Conclusion: This study provides new translational targets for PET tracers for the anti-inflammatory/protective activation state of microglia and shows the potential of a rationale-based approach. It therefore paves the way for the development of novel non-invasive methodologies aimed to monitor the success of therapeutic approaches in various neurological diseases.


A novel Mcm1-dependent element in the SWI4, CLN3, CDC6, and CDC47 promoters activates M/G1-specific transcription.
We have identified a novel promoter element that confers M/G1-specific transcription in Saccharomyces cerevisiae. This element, which we call an ECB (early cell cycle box), was first identified in the SWI4 promoter, but it is also present in the promoter of a G1 cyclin CLN3, as well as in the promoters of three DNA replication genes: CDC6, CDC47, and CDC46. Transcripts from all five of these genes oscillate during the cell cycle and peak at the M/G1 boundary, as do isolated ECB elements in reporter constructs. The ECB element contains an Mcm1 binding site to which Mcm1 binds in vitro, and an Mcm1-VP16 fusion, which places a constitutive activator on Mcm1-binding sites in vivo, can deregulate ECB-containing promoters. Mcm1 is a transcription factor that is also required for minichromosome maintenance. We provide evidence that the replication defect of mcm1 mutants can be suppressed by ectopic CDC6 transcription. Periodic expression of SWI4 and CLN3 may be important for cell cycle progression, as we find that these genes are both haploinsufficient and rate limiting for G1 progression. We suggest that ECB-regulated gene products play critical roles in promoting the initiation of S-phase, both by regulating CLN1 and CLN2 transcription and as components of the initiation complex on origins of replication.


Parvalbumin Cell Ablation of NMDA-R1 Leads to Altered Phase, But Not Amplitude, of Gamma-Band Cross-Frequency Coupling.
Altered gamma-band electrophysiological activity in individuals with autism spectrum disorder (ASD) is well documented, and analogous gamma-band alterations are recapitulated in several preclinical murine models relevant to ASD. Such gamma-band activity is hypothesized to underlie local circuit processes. Gamma-band cross-frequency coupling (CFC), a related though distinct metric, interrogates local neural circuit signal integration. Several recent studies have observed perturbed gamma-band CFC in individuals with ASD, although the direction of change remains unresolved. It also remains unclear whether murine models relevant to ASD recapitulate this altered gamma-band CFC. As such, this study examined whether mice with parvalbumin (PV) cell-specific ablation of NMDA-R1 (PVcre/NR1fl/fl) demonstrated altered gamma-band CFC as compared with their control littermates (PVcre/NR1+/+-mice that do not have the PV cell-specific ablation of NMDA-R1). Ten mice of each genotype had 4 min of "resting" electroencephalography recorded and analyzed. First, resting electrophysiological power was parsed into the canonical frequency bands and genotype-related differences were subsequently explored so as to provide context for the subsequent CFC analyses. PVcre/NR1fl/fl mice exhibited an increase in resting power specific to the high gamma-band, but not other frequency bands, as compared with PVcre/NR1+/+. CFC analyses then examined both the standard magnitude (strength) of CFC and the novel metric PhaseMax-which denotes the phase of the lower frequency signal at which the peak higher frequency signal power occurred. PVcre/NR1fl/fl mice exhibited altered PhaseMax, but not strength, of gamma-band CFC as compared with PVcre/NR1+/+ mice. As such, this study suggests a potential novel metric to explore when studying neuropsychiatric disorders.


Atriopeptin III alters renal medullary hemodynamics and the pressure-diuresis response in rats.
The effects of atriopeptin III on the renal response to changes in renal perfusion pressure (RPP) were examined in uninephrectomized and adrenalectomized rats. Neural influences on the kidney were eliminated by renal denervation. Plasma levels of antidiuretic hormone (ADH), angiotensin II, aldosterone, corticosterone, and norepinephrine were controlled by intravenous infusion. Infusion of atriopeptin III at a dose of 30 or 100 ng X kg-1 X min-1 increased plasma levels of atrial natriuretic peptide from 101 +/- 8 pg/ml to 424 +/- 16 and 2,553 +/- 308 pg/ml, respectively. In control rats, increasing RPP in two steps from 100 to 125 to 150 mmHg produced sixfold increases in sodium and water excretion. The slopes of the relationships between fractional sodium and water excretion and RPP were enhanced by approximately 10, 20, and 40% in the rats infused with the 30, 100, and 500 ng X kg-1 X min-1 dose of atriopeptin III. Renal blood flow and glomerular filtration rate were not significantly different in vehicle- and atriopeptin III-infused rats at any RPP studied. Papillary blood flow measured with a laser-Doppler flowmeter increased 15% within the first 5 min of infusion of atriopeptin III at a dose of 100 ng X kg-1 X min-1. Cortical blood flow was not significantly altered. The rise in papillary blood flow during atriopeptin III infusion, however, may be a consequence and not the cause of the natriuresis and diuresis, since urine flow increased before significant changes in papillary blood flow were detected. These studies suggest that atrial natriuretic factor could influence the long-term control of arterial pressure by altering renal medullary hemodynamics and promoting the elimination of sodium and water. The plasma levels of atrial natriuretic peptide needed to alter the pressure-natriuresis relationship, however, may exceed levels that can be attained in vivo with endogenous release.


The two cytochromes c in the facultative methylotroph Pseudomonas am1.
It was previously suggested that there is only one soluble cytochrome c in Pseudomonas AM1, having a molecular weight of 20000, a redox midpoint potential of about +260mV and a low isoelectric pint [Anthony (1975) Biochem. J.146, 289-298; Widdowson & Anthony (1975) Biochem. J.152, 349-356]. A more thorough examination of the soluble fraction of methanol-grown Pseudomonas AM1 has now revealed the presence of two different cytochromes c. These were both purified to homogeneity by acid treatment, ion-exchange chromatography, gel filtration, chromatography on hydroxyapatite and preparative isoelectric focusing. Molecular weights were determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis; midpoint redox potentials were determined directly by using platinum and calomel electrodes; isoelectric points were estimated by electrophoresis and by the behaviour of the two cytochromes on ion-exchange celluloses. The more abundant cytochrome c(H) (lambda(max.) 550.5nm) had a low molecular weight (11000), a midpoint potential of about +294mV and a high isoelectric point, not being adsorbed on DEAE-cellulose in 20mm-Tris/HCl buffer, pH8.0. The less abundant cytochrome c(L) (lambda(max.) 549nm) was about 30% of the total; it had a high molecular weight (20900), a midpoint potential of about +256mV and a low isoelectric point, binding strongly to DEAE-cellulose in 20mm-Tris/HCl buffer, pH8.0. The pH-dependence of the midpoint redox potentials of the two cytochromes c were very similar. There were four ionizations affecting the redox potentials in the pH range studied (pH4.0-9.5), two in the oxidized form (pK values about 3.5 and 5.5) and two in the reduced form (pK values about 4.5 and 6.5), suggesting that the ionizing groups involved may be the two propionate side chains of the haem. Neither of the cytochromes c was present in mutant PCT76, which was unable to oxidize or grow on C(1) compounds, although still able to grow well on multicarbon compounds such as succinate. Whether or not these two cytochromes c have separate physiological functions is not yet certain.


Modulation Of [35S]-tert-butylbicyclophosphorothionate binding by somatostatin in rat hypothalamus.
1. The present study was designed to assess the effect of the tetradecapeptide somatostatin on the GABA(A) receptor complex in the rat hypothalamus. 2. GABA(A) receptors were labelled with [35S]-tert-butylbicyclophosphorothionate (TBPS), which binds in or near the chloride channel, and binding as assessed by in vitro quantitative autoradiography using a computer-assisted image analysis system. 3. Somatostatin inhibited the binding of [35S]-TBPS to the convulsant site of the hypothalamic GABA(A) receptor complex of rat slide-mounted hypothalamic structures in a concentration-dependent manner with an affinity in the micromolar range (10(-6) to 3 x 10(-6) mol/L). Somatostatin appeared to mimic the effects of the neurosteroid 5alpha-pregnane-3alpha ol-one (5alpha3alphaP), GABA and picrotoxin on [35S]-TBPS binding in the rat hypothalamus in all structures examined. Furthermore, GABA or muscimol (a GABA(A) receptor agonist), when added to the incubation medium, enhanced the capacity of somatostatin to inhibit [35S]-TBPS binding, with an IC50 of 10(-7) mol/L. However, incubation with bicuculline (a GABA(A) receptor antagonist) led to the abolition of the inhibitory effect of somatostatin on [35S]-TBPS specific binding in rat hypothalamus. 4. The present results demonstrate the presence of a modulatory effect of somatostatin on the GABA(A) receptor complex in rat hypothalamic structures. Furthermore, the data suggest that somatostatin allosterically modifies [35S]-TBPS binding through a mechanism similar to that of GABA. Taken together, these results provide evidence for the presence of somatostatin- GABA interactions in rat hypothalamus.


Vitamin C alone does not improve treatment outcomes in mechanically ventilated patients with severe sepsis or septic shock: a retrospective cohort study.
Vitamin C has shown several beneficial effects on sepsis in preclinical studies. However, clinical data supporting these reports are scarce. This study aimed to evaluate whether adjunctive intravenous vitamin C therapy could reduce hospital mortality in patients with severe sepsis or septic shock requiring mechanical ventilation. For this retrospective cohort study, consecutive medical ICU patients with severe sepsis or septic shock requiring mechanical ventilation were included. The study patients were classified into the vitamin C or control groups depending on the administration of intravenous vitamin C (2 g every 8 hours). The primary outcome was hospital mortality. Thirty-five patients in the vitamin C group and 40 patients in the control group were included. The two groups were comparable in regards to the baseline characteristics at ICU admission. The hospital mortality was 46% (16 of 35 patients) in the vitamin C group and 40% (16 of 40 patients) in the control group, showing a statistically nonsignificant difference (P=0.62). The mortality at 90 days after ICU admission (60% vs. 48%) did not significantly differ between groups. The median time to shock reversal was 3 days [interquartile range (IQR), 2 to 5 days] in both groups. The changes in the Sepsis-related Organ Failure Assessment (SOFA) scores during the first 4 ICU days were -1.4±3.3 and -1.4±3.0 in the vitamin C and control groups, respectively. Adjunctive intravenous vitamin C therapy alone did not reduce hospital mortality in mechanically ventilated patients with severe sepsis or septic shock.


UBC-Nepal expedition: phenotypical evidence for evolutionary adaptation in the control of cerebral blood flow and oxygen delivery at high altitude.
Sherpa have lived in the Nepal Himalaya for 25-40 thousand years and display positive physiological adaptations to hypoxia. Sherpa have previously been demonstrated to suffer less negative cerebral side effects of ascent to extreme altitude, yet little is known as to whether or not they display differential regulation of oxygen delivery to the brain compared to lowland natives. We demonstrate that Sherpa have lower brain blood flow during ascent to and acclimatization at high altitude compared to lowlanders and that this difference in flow is not attributable to factors such as mean arterial pressure, blood viscosity and pH. The observed lower cerebral oxygen delivery in Sherpa likely represents a positive adaptation that may indicate a cerebral hypometabolic conservation of energy at altitude and/or decreased risk of other cerebral consequences such as vasogenic oedema. Debilitating side effects of hypoxia manifest within the central nervous system; however, high-altitude natives of the Tibetan plateau, the Sherpa, experience negligible cerebral effects compared to lowland natives at extreme altitude. Phenotypical optimization of the oxygen cascade has been demonstrated in the systemic circulation of Tibetans and Sherpa, likely underscoring their adapted capacity to thrive at altitude. Yet, little is known as to how the cerebral circulation of Sherpa may be adapted. To examine potential differences in cerebral oxygen delivery in Sherpa compared to lowlanders we measured arterial blood gases and global cerebral blood flow (duplex ultrasound) during a 9 day ascent to 5050 m. Although cerebral oxygen delivery was maintained during ascent in lowlanders, it was significantly reduced in the Sherpa at 3400 m (-30.3 ± 21.6%; P < 0.01) and 4371 m (-14.2 ± 10.7%; P = 0.03). Furthermore, linear mixed effects modelling indicated that independent of differences in mean arterial pressure, pH and blood viscosity, race accounts for an approximately 100 mL min-1 (∼17-34%) lower cerebral blood flow in Sherpa compared to lowlanders across ascent to altitude (P = 0.046). To ascertain the role of chronic hypoxia independent of the ascent, Sherpa who had not recently descended were also examined at 5050 m. In these Sherpa, cerebral oxygen delivery was also lower compared to lowlanders (∼22% lower; P < 0.01). We highlight new information about the influence of race and genetic adaptation in the regulation of cerebral oxygen delivery. The lower cerebral oxygen delivery in the Sherpa potentially represents a positive adaptation considering Sherpa endure less deleterious cerebral consequences than lowlanders at altitude.


On the variety of methods for calculating confidence intervals by bootstrapping.
Researchers often want to place a confidence interval around estimated parameter values calculated from a sample. This is commonly implemented by bootstrapping. There are several different frequently used bootstrapping methods for this purpose. Here we demonstrate that authors of recent papers frequently do not specify the method they have used and that different methods can produce markedly different confidence intervals for the same sample and parameter estimate. We encourage authors to be more explicit about the method they use (and number of bootstrap resamples used). We recommend the bias corrected and accelerated method as giving generally good performance; although researchers should be warned that coverage of bootstrap confidence intervals is characteristically less than the specified nominal level, and confidence interval evaluation by any method can be unreliable for small samples in some situations.


Current practice patterns in urologic management of upper-tract transitional-cell carcinoma.
To determine the current practice patterns in the management of upper-tract transitional-cell carcinoma (TCC) among a large group of urologists. A survey was sent to 220 practicing members of the Society of Urologic Oncology (SUO) and the Endourological Society (ES) and members of the American Urological Association who did not belong to either society. The survey consisted of 16 focused questions pertaining to the surveillance and management of upper-tract TCC. The responses were used to create a database, which was then analyzed to determine practice trends. Eighty-four of the urologists responded, for a response rate of 38%. Fourteen responses were excluded because of multiple answers to a given question, so 70 were included in the final analysis. Eighty percent of the respondents were in academic practice. A CT urogram was the favored initial procedure for diagnosis of upper-tract TCC and an intravenous urogram was the next commonest choice (53% and 40%, respectively). Ureterorenoscopy was the surveillance tool of choice (70%) after conservative treatment of upper- tract TCC. Laparoscopic nephroureterectomy was the preferred procedure (73%) for a high-grade, large renal-pelvic TCC. Twenty-one percent of the endourologists recommended ureteroscopic ablation for a high-grade, large distal ureteral tumor. This was in sharp contrast to 77% of the respondents who favored a distal ureterectomy for the same clinical scenario. This study confirms that most urologists treating upper-tract TCC follow the principles reported in the published literature regarding the management of these patients. Further, most urologists, regardless of society affiliations or years in practice, favor minimally invasive techniques for the management of upper-tract TCC. This information may be useful in formulating clear guidelines for the management of this disease.


Venezuelan equine encephalitis virus: comparison of infectivity and virulence of strains V-38 and P676 in donkeys.
Two strains of Venezuelan equine encephalitis (VEE) virus were examined for the ability to replicate in, as well as to produce death among donkeys. One, a low passage strain known as strain P676 was originally isolated from mosquitos in Venezuela. The other, strain V-38 was isolated from a horse brain in 1938 and had undergone an unknown number of laboratory passages; it is used extensively for the preparation of inactivated VEE vaccine. Both strains were found to be approximately equal in their ability to infect donkeys. However, a quantity as small as 50% hamster intraperitoneal infectious units of strain V-38 resulted in fatal infection. On the other hand, as much as 631 million infectious units of strain P676 were nonfatal in one of two donkeys. It appears that strain V-38 is approximately 100 million times more virulent than strain P676 in equine species. One donkey which received strain P676 demonstrated a biphasic pattern of clinical illness and viremia, and there is suggestive evidence that another animal experienced a second and fatal clinical response 3 weeks after virus inoculation.


A New Look at the Structural and Magnetic Properties of Potassium Neptunate K2NpO4 Combining XRD, XANES Spectroscopy, and Low-Temperature Heat Capacity.
The physicochemical properties of the potassium neptunate K2NpO4 have been investigated in this work using X-ray diffraction, X-ray absorption near edge structure (XANES) spectroscopy at the Np-L3 edge, and low-temperature heat capacity measurements. A Rietveld refinement of the crystal structure is reported for the first time. The Np(VI) valence state has been confirmed by the XANES data, and the absorption edge threshold of the XANES spectrum has been correlated to the Mössbauer isomer shift value reported in the literature. The standard entropy and heat capacity of K2NpO4 have been derived at 298.15 K from the low-temperature heat capacity data. The latter suggest the existence of a magnetic ordering transition around 25.9 K, most probably of the ferromagnetic type.


Arsenic in drinking-water and risk for cancer in Denmark.
Arsenic is a well-known carcinogen, which is often found in drinking-water. Epidemiologic studies have shown increased cancer risks among individuals exposed to high concentrations of arsenic in drinking-water, whereas studies of the carcinogenic effect of low doses have had inconsistent results. Our aim was to determine if exposure to low levels of arsenic in drinking-water in Denmark is associated with an increased risk for cancer. The study was based on a prospective Danish cohort of 57,053 persons in the Copenhagen and Aarhus areas. Cancer cases were identified in the Danish Cancer Registry, and the Danish civil registration system was used to trace and geocode residential addresses of the cohort members. We used a geographic information system to link addresses with water supply areas, then estimated individual exposure to arsenic using residential addresses back to 1970. Average exposure for the cohort ranged between 0.05 and 25.3 microg/L (mean = 1.2 microg/L). Cox's regression models were used to analyze possible relationships between arsenic and cancer. We found no significant association between exposure to arsenic and risk for cancers of the lung, bladder, liver, kidney, prostate, or colorectum, or melanoma skin cancer; however, the risk for non-melanoma skin cancer decreased with increasing exposure (incidence rate ratio = 0.88/microg/L average exposure; 95% confidence interval, 0.84-0.94). Results adjusted for enrollment area showed no association with non-melanoma skin cancer. The results indicate that exposure to low doses of arsenic might be associated with a reduced risk for skin cancer.


What determines adherence to treatment in cardiovascular disease prevention? Protocol for a mixed methods preference study.
Background Significant gaps exist between guidelines-recommended therapies for cardiovascular disease prevention and current practice. Fixed-dose combination pills ('polypills') potentially improve adherence to therapy. This study is a preference study undertaken in conjunction with a clinical trial of a polypill and seeks to examine the underlying reasons for variations in treatment adherence to recommended therapy. Methods/design A preference study comprising: (1) Discrete Choice Experiment for patients; and (2) qualitative study of patients and providers. Both components will be conducted on participants in the trial. A joint model combining the observed adherence in the clinical trial (revealed preference) and the Discrete Choice Experiment data (stated preference) will be estimated. Estimates will be made of the marginal effect (importance) of each attribute on overall choice, the extent to which respondents are prepared to trade-off one attribute for another and predicted values of the level of adherence given a fixed set of attributes, and contextual and socio-demographic characteristics. For the qualitative study, a thematic analysis will be used as a means of exploring in depth the preferences and ultimately provide important narratives on the experiences and perspectives of individuals with regard to adherence behaviour. Ethics and dissemination Primary ethics approval was received from Sydney South West Area Health Service Human Research Ethics Committee (Royal Prince Alfred Hospital zone). In addition to usual scientific forums, the findings will be reported back to the communities involved in the studies through site-specific reports and oral presentations.


Benzyl Isothiocyanate, a Vegetable-Derived Compound, Induces Apoptosis via ROS Accumulation and DNA Damage in Canine Lymphoma and Leukemia Cells.
Treatment of neoplastic diseases in companion animals is one of the most important problems of modern veterinary medicine. Given the growing interest in substances of natural origin as potential anti-cancer drugs, our goal was to examine the effectiveness of benzyl isothiocyanate (BITC), a compound found in cruciferous vegetables, against canine lymphoma and leukemia. These are the one of the most common canine cancer types, and chemotherapy is the only treatment option. The study involved established cell lines originating from various hematopoietic malignancies: CLBL-1, GL-1, CLB70 and CNK-89, immortalized noncancerous cell lines: MDCK and NIH-3T3 and canine peripheral blood mononuclear cells (PBMCs). The cytotoxic activity of BITC, apoptosis induction, caspase activity and ROS generation were evaluated by flow cytometry. H2AX phosphorylation was assessed by western blot. The study showed that the compound was especially active against B lymphocyte-derived malignant cells. Their death resulted from caspase-dependent apoptosis. BITC induced ROS accumulation, and glutathione precursor N-acetyl-l-cysteine reversed the effect of the compound, thus proving the role of oxidative stress in BITC activity. In addition, exposure to the compound induced DNA damage in the tested cells. This is the first study that provides information on the activity of BITC in canine hematopoietic malignancies and suggests that the compound may be particularly useful in B-cell neoplasms treatment.


Prognostic factors and treatment outcomes for patients with Fournier's gangrene: a retrospective study.
Fournier's gangrene is a gas-forming, necrotising soft tissue infection affecting the perineum. It spreads rapidly along the deep fascial planes and is associated with a high mortality rate. With a growing elderly population with comorbidities, the frequency of severe cases of Fournier's gangrene is expected to increase. We retrospectively reviewed 20 patients diagnosed with Fournier's gangrene at our institution from 2003 to 2014 and analysed data. Thirteen patients had diabetes mellitus, two had been diagnosed with liver cirrhosis, and four were chronic alcoholics. Of 15 patients admitted to an intensive care unit, 11 underwent colostomy, and 4 required skin grafts for wound healing. The wide wounds of two patients were healed using vacuum-assisted closure (VAC® ) dressing without additional surgery. The mortality rate was 25%, and the patients whose Fournier's gangrene severity index (FGSI) score was higher than 9 points or whose blood urea nitrogen (BUN) level was higher than 50 mg/dl had a poor prognosis. In order to treat Fournier's gangrene, aggressive surgical treatment, including wide debridement and stoma creation, should be considered as soon as possible to improve survival rates. Additionally, VAC dressing is helpful in healing the wide debridement wound without additional reconstructive surgery.


Diurnal migration of Echinostoma caproni (Digenea: Echinostomatidae) in ICR mice.
Twenty-four female ICR mice, 12 acclimated to a 12 ∶ 12 light-dark cycle and 12 to a 12 ∶ 12 dark-light cycle for 7 days, were each infected with 10 metacercariae of Echinostoma caproni. Infected mice were maintained on their respective lighting regimes for 28 days. Six mice (3 from each group) were necropsied at 4-hr intervals beginning at 0700 hr. The small intestine was removed, opened, and the position of individual worms and worm clusters was measured to the nearest 0.1 cm. Each intestine was subsequently divided into 20 equal segments and individual worms and worm clusters were assigned to the appropriate segment based on the original measurements. All worms were found in the posterior 55% of the intestine (ileum). All posterior segments (10-20), with the exception of segment 18, harbored at least 1 worm at some time. A Monte Carlo simulation of worm abundance in segments 10-17 over all time periods indicated a random distribution, while the same analysis of segments 10-20 indicated a non-random distribution due to large numbers of worms in segment 20 and to the absence of worms in segment 18. To analyze temporal changes in worm distribution, mice were grouped by time of necropsy as follows: night (1900 and 2300 hr), morning (0300 and 0700 hr), and day (1100 and 1500 hr). During the night and morning, E. caproni was heavily concentrated in segments 10-17 and, during the day, worms were located more posteriorly, with a heavy concentration in the last segment (20).


Delirium in the intensive care setting dependent on the Richmond Agitation and Sedation Scale (RASS): Inattention and visuo-spatial impairment as potential screening domains.
In the intensive care setting, delirium is a common occurrence; however, the impact of the level of alertness has never been evaluated. Therefore, this study aimed to assess the delirium characteristics in the drowsy, as well as the alert and calm patient. In this prospective cohort study, 225 intensive care patients with Richmond Agitation and Sedation Scale (RASS) scores of -1 - drowsy and 0 - alert and calm were evaluated with the Delirium Rating Scale-Revised-1998 (DRS-R-98) and the Diagnostic and Statistical Manual 4th edition text revision (DSM-IV-TR)-determined diagnosis of delirium. In total, 85 drowsy and 140 alert and calm patients were included. Crucial items for the correct identification of delirium were sleep-wake cycle disturbances, language abnormalities, thought process alterations, psychomotor retardation, disorientation, inattention, short- and long-term memory, as well as visuo-spatial impairment, and the temporal onset. Conversely, perceptual disturbances, delusions, affective lability, psychomotor agitation, or fluctuations were items, which identified delirium less correctly. Further, the severities of inattentiveness and visuo-spatial impairment were indicative of delirium in both alert- or calmness and drowsiness. The impairment in the cognitive domain, psychomotor retardation, and sleep-wake cycle disturbances correctly identified delirium irrespective of the level alertness. Further, inattentiveness and - to a lesser degree - visuo-spatial impairment could represent a specific marker for delirium in the intensive care setting meriting further evaluation.


Tolerance of radiotherapy and chemotherapy in elderly patients with bladder cancer.
Bladder-sparing radiotherapy with concurrent chemotherapy may be a reasonable alternative to cystectomy in patients with invasive bladder cancer. The purpose of this study was to determine the tolerance of combined treatment in elderly patients. In this retrospective study, the records of patients 70 or more years of age with stage T2-T4a, N0, M0 disease who were treated with bladder-sparing regimens between 1985 and 2000 were examined for toxicity. Of 149 consecutive patients treated for cancer of the bladder, 14 patients met eligibility criteria. The median age was 79 years. At a median follow-up of 17 months, the median survival was 19 months. All patients had at least mild toxicity, with 6 of 14 patients having grade III to IV toxicity. Grade III to IV toxicities included one patient with grade IV neutropenia, three with grade III gastrointestinal toxicities, one patient with grade III urinary frequency, one patient with grade IV ureteral obstruction who required stent placement, and one episode of hydration-induced grade III heart failure. Two of 14 patients stopped chemotherapy and 5 patients required dose reductions for toxicity. The observed rates of toxicity compare favorably with studies of bladder-sparing therapy in patients with median ages less than 70 years. Our study shows that bladder-sparing radiotherapy with concurrent chemotherapy is feasible in patients 70 or more years of age, and should be considered for such patients if they are inoperable or strongly wish to avoid cystectomy.


[Experimental production of anencephalic monsters by chick embryo microsurgery. Initial observations].
42 hour-old chick embryos are operated by microsurgical means. The encephalon, which is normally closed at this stage, is reopened and the incision extended to the level of 2nd or 3rd somites. The experimental embryos are anencephalic. Their comparison with a spontaneous anencephalus shows a large morphological similarity. These first experiments confirm the hypothesis by which anencephaly is a direct consequence of the failure of neural tube closure.


Handling test of eye drop dispenser--comparison of unit-dose pipettes with conventional eye drop bottles.
The aims of this study were to investigate how elderly people handle single-use eye drop dispensers (unit-dose pipettes) and to compare the performance with conventional eye drop bottles. In this open-label study, the handling of unit-dose pipettes and conventional eye drop bottles was compared in 41 elderly people who had little or no prior regular use of eye drop dispensers. The participants tested both types of dispenser once, and the following 7 variables were studied: ease/difficulty of opening the dispenser; influence of the size for handling of the dispenser; influence of the shape for handling of the dispenser; observation of the contents in the dispenser; the feeling of the dispenser in the hand; ease/difficulty of drop instillation on the eye from the dispenser; and overall performance of the eye drop dispenser. The dispensers contained isotonic saline, and a visual analog scale was used for assessment of each of the above variables. The mean age of the participants was 73 years. A statistically significant difference in favor of the unit-dose pipettes was found with respect to observation of the contents in the dispenser, ease of administration, and the overall performance. Women regarded the unit-dose pipettes generally better than the bottles, but such a difference was not seen in men. The study participants managed the unit-dose pipettes at least as well as the conventional eye drop bottles. If anything, the unit-dose pipettes appeared to be easier to use.


DNA Damage Tolerance by Eukaryotic DNA Polymerase and Primase PrimPol.
PrimPol is a human deoxyribonucleic acid (DNA) polymerase that also possesses primase activity and is involved in DNA damage tolerance, the prevention of genome instability and mitochondrial DNA maintenance. In this review, we focus on recent advances in biochemical and crystallographic studies of PrimPol, as well as in identification of new protein-protein interaction partners. Furthermore, we discuss the possible functions of PrimPol in both the nucleus and the mitochondria.


[Sister Mary Joseph's nodule. Its clinical significance and management].
The eponym Sister (Mary) Joseph's nodule is referred to the existence of umbilical metastasis. Historically it has been considered a sign of ominous vital prognosis and therapeutic abstention. The analysis of 407 cases published at the medical literature (MEDLINE 1966-1997) is against this idea, because recent series have showed survivals longer than a year after treatment. On the other hand 60% of umbilical nodules are benign. Nowadays this sign must be considered as an early indicator of inner neoplasms, mainly from the abdominal cavity (52% digestive origin, 28% gynecologic origin), and in 40% of cases they are the only expression of relapse of previously known neoplasms. When a umbilical nodule is found it is necessary to make an accurate histologic differential diagnosis between a primary (benign or neoplastic) and metastatic lesions. Fine needle aspiration has demonstrated to be an easy and reliable method for diagnosis. Once the metastatic origin of the nodule is confirmed treatment with curative intention must be tried.


Selection for fast and slow exploration affects baseline and stress-induced corticosterone excretion in Great tit nestlings, Parus major.
In nestlings, glucocorticoid (GC) secretion has short-term and long-term fitness consequences. For example, short-time elevations trigger begging activity, whereas chronically elevated GC levels impair body condition, growth and cognitive abilities. Despite a growing body of literature on personality traits, the effects of selection for fast and slow exploration on GC secretion have received little attention. We compared baseline and stress-induced hypothalamic-pituitary-adrenal (HPA) axis activity of hand-reared great tit nestlings of lines selected for fast and slow exploration. Nestling droppings were collected under three conditions: control, test (following handling stress, day 14 after hatching) and the following day. The concentrations of excreted immunoreactive corticosterone metabolites (CM) were determined via an enzyme immunoassay. We also observed nestlings' begging behaviour. CM differed significantly between the lines. Nestlings of the fast line excreted lower CM than slow-line birds. In response to handling stress, nestlings excreted significantly higher concentrations of CM than during the control and on the day after handling. Sex and begging activity were not related to CM levels. Under the control condition, but not after handling, males begged significantly more often than females. In both lines, adults excreted significantly less CM compared to nestlings. Both nestlings and adults of the slow line produced higher baseline CM values than fast-line birds. Fast-line nestlings excreted lower baseline CM than nestlings of a wild population not selected for fast or slow exploration. Slow-line nestlings did not. Our results show that selection on the basis of exploratory behaviour affected HPA axis reactivity.


SeMet alleviates LPS-induced eggshell gland necroptosis mediated inflammation by regulating the Keap1/Nrf2/HO-1 pathway.
Exposure to lipopolysaccharide (LPS) can lead to inflammation in a variety of tissues and organs. Selenium (Se) plays a crucial role in mitigating inflammatory damage. Compared with inorganic selenium, organic selenium, such as selenomethionine (SeMet), has the advantages of a higher absorption rate and lower toxicity in animals. This study examined the protective effects of SeMet on eggshell gland tissue damage caused by LPS. Hy-Line Brown laying hens were chosen as the experimental animals and were randomly assigned to four groups: control group (C), lipopolysaccharide group (LPS), SeMet group (Se), and SeMet + lipopolysaccharide group (Se + LPS). H&E staining and transmission electron microscope were performed to observe the pathological changes of eggshell glands, oxidative stress related indicators were measured using relevant kits, qRT‒PCR and western blotting were used to evaluate the mRNA and protein levels of the Nrf2 pathway, necroptosis, and inflammation related indicators. The results showed that LPS treatment increased the content of malondialdehyde (MDA), decreased the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX), and decreased the content of glutathione (GSH). LPS increased the levels of Keap1, RIPK1, RIPK3, MLKL, TNF-α, COX-2, and NF-κB, while decreasing the levels of HO-1, NQO1, Nrf2, and Caspase-8. However, SeMet treatment effectively reversed the changes of the above indicators, indicating that SeMet alleviates eggshell gland cell necroptosis-mediated inflammation induced by LPS via regulating the Keap1/Nrf2/HO-1 pathway. This study elucidated the mechanism by which SeMet alleviates LPS-induced eggshell gland tissue damage in Hy-Line Brown laying hens and provided a new direction for expanding the application of SeMet in the feeding and production of laying hens.


Viral Interactions and Pathogenesis during Multiple Viral Infections in Agaricus bisporus.
Viral interactions during multiple viral infections were examined in Agaricus bisporus cultures harboring 9 viruses (comprising 18 distinct viral RNAs) by statistically analyzing their relative abundance in fruitbodies. Four clusters of viral RNA were identified that suggested synergism and coreplication. Pairwise correlations revealed negative and positive correlations between clusters, indicating further synergisms and an antagonism involving a group containing a putative hypovirus and four nonhost ORFan RNAs (RNAs with no similarity to known sequences) possibly acting as defective interfering RNAs. The disease phenotype was observed in 10 to 15% of the fruitbodies apparently randomly located among asymptomatic fruitbodies. The degree of symptom expression consistently correlated with the levels of the multipartite virus AbV16. Diseased fruitbodies contained very high levels of AbV16 and AbV6 RNA2; these levels were orders of magnitude higher than those in asymptomatic tissues and were shown statistically to be discretely higher populations of abundance, indicating an exponential shift in the replicative capacity of the virus. High levels of AbV16 replication were specific to the fruitbody and not found in the underlying mycelium. There appeared to be a stochastic element occurring in these viral interactions, as observed in the distribution of diseased symptoms across a culture, differences in variance between experiments, and a number of additional viruses undergoing the step-jump in levels between experiments. Possible mechanisms for these multiple and simultaneous viral interactions in single culture are discussed in relation to known virus-host regulatory mechanisms for viral replication and whether additional factors could be considered to account for the 1,000-fold increase in AbV16 and AbV6 RNA2 levels.IMPORTANCE How viruses interact in a multiple-virus infection was examined by quantifying the levels of 18 viral RNAs in fruiting cultures of the agriculturally cultivated fungus Agaricus bisporus and statistically analyzing and modeling their abundance. Synergistic, antagonistic, and neutral interactions occurred simultaneously in cultures. The viral RNAs were grouped into four clusters, each displaying similar relative abundance, and between clusters, further interactions were found with positive, negative, or no correlations. Mushroom fruitbodies showing disease symptoms were distributed apparently randomly across the culture. These symptoms were associated with the presence of viral RNAs from two different clusters at very high levels, 1,000-fold higher than asymptomatic fruitbodies. The role of viral interaction together with stochastic factors and the regulation of host antiviral defenses in pathogenesis are discussed.


Guidelines for the surgical treatment of unruptured intracranial aneurysms: the first annual J. Lawrence pool memorial research symposium--controversies in the management of cerebral aneurysms.
The management of unruptured cerebral aneurysms remains one of the most controversial topics in neurosurgery. To this end, we discuss the diagnosis and estimated prevalence of these lesions as well as review the literature regarding the rate of rupture for cerebral aneurysms and risks of operative intervention. Our interpretation of the literature concludes that aneurysms are present in approximately 1% of the adult population, varying between less than 1% in young adults to 4% in the elderly. The yearly risk of subarachnoid hemorrhage for an unruptured intracranial aneurysm is approximately 1% for lesions 7 to 10 mm in diameter. Based on these assumptions, we recommend that 1) with rare exceptions, all symptomatic unruptured aneurysms should be treated; 2) small, incidental aneurysms less than 5 mm in diameter should be managed conservatively in virtually all cases; 3) aneurysms larger than 5 mm in patients younger than 60 years of age should be seriously considered for treatment; 4) large, incidental aneurysms larger than 10 mm should be treated in nearly all patients younger than 70 years of age; and 5) microsurgical clipping rather than endovascular coiling should be the first treatment choice in low-risk cases. Critical to our guidelines is collaboration by a highly experienced cerebrovascular team of microneurosurgeons and endovascular neurosurgeons working at a tertiary medical center with a high case volume and using a decision-making paradigm designed to offer only low-risk treatments. In certain patients for whom both treatment and natural history carry high risks, such as those with giant aneurysms, nonoperative management is typically elected.


The Aging Cardiovascular System: Understanding It at the Cellular and Clinical Levels.
Cardiovascular disease (CVD) presents a great burden for elderly patients, their caregivers, and health systems. Structural and functional alterations of vessels accumulate throughout life, culminating in increased risk of developing CVD. The growing elderly population worldwide highlights the need to understand how aging promotes CVD in order to develop new strategies to confront this challenge. This review provides examples of some major unresolved clinical problems encountered in daily cardiovascular practice as we care for elderly patients. Next, the authors summarize the current understanding of the mechanisms implicated in cardiovascular aging, and the potential for targeting novel pathways implicated in endothelial dysfunction, mitochondrial oxidative stress, chromatin remodeling, and genomic instability. Lastly, the authors consider critical aspects of vascular repair, including autologous transplantation of bone marrow-derived stem cells in elderly patients.


Recent developments in the specific dermatoses of pregnancy.
During pregnancy, the mother undergoes changes to sustain and enable normal growth and development of the fetus. Common physiological changes include linea nigra, fibroepithelial polyps, striae, spider angioma, palmar erythema and pruritis gravidarum. However, there are some changes that are purely pathological, and these are termed the pregnancy-specific dermatoses (PSDs). The PSDs occur during pregnancy or in the immediate postpartum period. They do not include the various benign conditions or pre-existing dermatoses and tumours that may present or worsen with pregnancy. They do include a number of distinct and identifiable conditions: atopic eruption of pregnancy (AEP), polymorphic eruption of pregnancy (PEP), intrahepatic cholestasis of pregnancy (ICP) and pemphigoid gestationis (PG). These are a heterogeneous group of skin conditions characterized by pruritis and inflammatory changes. In addition, pruritis gravidarum is sometimes considered pathophysiological and thus part of this group, rather than a physiological process. Each of these conditions has a distinct, but not fully understood, pathogenesis. The mechanisms leading to PSD may be a reflection on the hormonal and immunological changes associated with pregnancy. AEP and PEP are benign conditions, and although they can cause distress to the mother, they are otherwise minor. However, ICP and PG are more serious conditions, and both carry the potential for serious risks to both the mother and the fetus. Thus, the pathophysiology of these latter two conditions is considered in more detail in the following article.


Vector competence is strongly affected by a small deletion or point mutations in bluetongue virus.
Transmission of vector-borne virus by insects is a complex mechanism consisting of many different processes; viremia in the host, uptake, infection and dissemination in the vector, and delivery of virus during blood-feeding leading to infection of the susceptible host. Bluetongue virus (BTV) is the prototype vector-borne orbivirus (family Reoviridae). BTV serotypes 1-24 (typical BTVs) are transmitted by competent biting Culicoides midges and replicate in mammalian (BSR) and midge (KC) cells. Previously, we showed that genome segment 10 (S10) encoding NS3/NS3a protein is required for virus propagation in midges. BTV serotypes 25-27 (atypical BTVs) do not replicate in KC cells. Several distinct BTV26 genome segments cause this so-called 'differential virus replication' in vitro. Virus strains were generated using reverse genetics and their growth was examined in vitro. The midge feeding model has been developed to study infection, replication and disseminations of virus in vivo. A laboratory colony of C. sonorensis, a known competent BTV vector, was fed or injected with BTV variants and propagation in the midge was examined using PCR testing. Crossing of the midgut infection barrier was examined by separate testing of midge heads and bodies. A 100 nl blood meal containing ±105.3 TCID50/ml of BTV11 which corresponds to ±20 TCID50 infected 50% of fully engorged midges, and is named one Midge Alimentary Infective Dose (MAID50). BTV11 with a small in-frame deletion in S10 infected blood-fed midge midguts but virus release from the midgut into the haemolymph was blocked. BTV11 with S1[VP1] of BTV26 could be adapted to virus growth in KC cells, and contained mutations subdivided into 'corrections' of the chimeric genome constellation and mutations associated with adaptation to KC cells. In particular one amino acid mutation in outer shell protein VP2 overcomes differential virus replication in vitro and in vivo. Small changes in NS3/NS3a or in the outer shell protein VP2 strongly affect virus propagation in midges and thus vector competence. Therefore, spread of disease by competent Culicoides midges can strongly differ for very closely related viruses.


Mesenteric Panniculitis: An Unusual Presentation of Abdominal Pain.
Sclerosing mesenteritis is a rare autoimmune disease that eventually evolves into fibrotic changes that usually affect the adipose tissue around the mesenteric vessels. It can present through a myriad of gastroenterological as well as constitutional symptoms, including but not limited to abdominal pain, diarrhea, fever, nausea, or vomiting. Although the exact etiology of the disease is yet to be determined, there are several predisposing factors, the most common of which is a previous history of abdominal trauma and/or surgery. Several case series have reported the association of sclerosing mesenteritis with prior abdominal surgery ranging from as low as 24% to as high as 53%.


Characterization and biological activity of a Brazilian isolate of Bacillus sphaericus (Neide) highly toxic to mosquito larvae.
Primary powders of Bacillus sphaericus strain S2 isolated from soil samples in Brazil, and strain 2362 were produced in a 14 liter fermentor. Growth patterns and sporulation observed in three trials with strains S2 and 2362 in the fermentor were similar. Second-instar larvae of Culex quinquefasciatus, Anopheles albimanus, Anopheles quadrimaculatus, and Aedes aegypti exposed for 48 hr to strain S2 responded with LC50 values of 0.25, 5.95, 12.28 and 140.0 ppb of lyophilized primary powder, respectively. Under the same conditions, strain 2362 resulted in LC50 values of 0.39, 7.16, 16.93 and 307.0 ppb of lyophilized primary powder, respectively, in those mosquito larvae. Statistical analysis of the bioassay data did not show significant differences among LC50 values observed in B. sphaericus strains S2 and 2362, at the 0.05 level. Toxins of strains S2 and 2362 were extracted at pH 12 with NaOH. Electrophoresis of the extracts in polyacrylamide gel under denaturing conditions revealed the 51 and 42 kDa toxins in both S2 and 2362 B. sphaericus strains. The presence of the 42 kDa peptide in the extracts was confirmed by Western blot and Elisa, with anti-42 kDa IgG previously prepared from strain 2362.


Effect of selective serotonin reuptake inhibitor treatment following diagnosis of depression on suicidal behaviour risk: a target trial emulation.
There is concern regarding the impact of selective serotonin reuptake inhibitors (SSRIs) on suicidal behaviour. Using the target trial framework, we investigated the effect on suicidal behaviour of SSRI treatment following a depression diagnosis. We identified 162,267 individuals receiving a depression diagnosis aged 6-59 years during 2006-2018 in Stockholm County, Sweden, after at least 1 year without antidepressant dispensation. Individuals who initiated an SSRI within 28 days of the diagnosis were assigned as SSRI initiators, others as non-initiators. Intention-to-treat and per-protocol effects were estimated; for the latter, individuals were censored when they ceased adhering to their assigned treatment strategy. We applied inverse probability weighting (IPW) to account for baseline confounding in the intention-to-treat analysis, and additionally for treatment non-adherence and time-varying confounding in the per-protocol analysis. The suicidal behaviour risk difference (RD), and risk ratio (RR) between SSRI initiators and non-initiators were estimated at 12 weeks. In the overall cohort, we found an increased risk of suicidal behaviour among SSRI initiators (intention-to-treat RR = 1.50, 95% CI = 1.25, 1.80; per-protocol RR = 1.69, 95% CI = 1.20, 2.36). In age strata, we only found evidence of an increased risk among individuals under age 25, with the greatest risk among 6-17-year-olds (intention-to-treat RR = 2.90, 95% CI = 1.72, 4.91; per-protocol RR = 3.34, 95% CI = 1.59, 7.00). Our finding of an increased suicidal behaviour risk among individuals under age 25 reflects evidence from RCTs. We found no evidence of an effect in the high-risk group of individuals with past suicidal behaviour. Further studies with information on a wider array of confounders are called for.


Using word recognition tests to estimate premorbid IQ in early dementia: longitudinal data.
This study examined the utility of word recognition scores for estimating actual Verbal IQ scores obtained from 1-5 years earlier. Participants were 271 persons remaining normal and 24 initially normal persons who developed cognitive impairment over longitudinal follow-up. A previously published regression equation based on education and the American modification of the National Adult Reading Test (Nelson, 1982) was used to estimate Mayo Verbal IQ. In spite of correlating well with prior obtained scores (r = .7) the predicted score tended to overestimate the obtained Mayo Verbal IQ. A revised equation was developed in the normal sample, which improved accuracy of prediction. Among the 24 persons who developed cognitive impairment over a 5-year span, the revised predicted scores provided reasonable estimates of initial Mayo Verbal IQ. To aid in clinical interpretation, a table of the normal frequencies of predicted Mayo Verbal IQ minus contemporaneously obtained Mayo Verbal IQ is provided.


Distribution and pollination services of wild bees and hoverflies along an altitudinal gradient in mountain hay meadows.
Extensively managed and flower-rich mountain hay meadows, hotspots of Europe's biodiversity, are subject to environmental and climatic gradients linked to altitude. While the shift of pollinators from bee- to fly-dominated communities with increasing elevation across vegetation zones is well established, the effect of highland altitudinal gradients on the community structure of pollinators within a specific habitat is poorly understood. We assessed wild bee and hoverfly communities, and their pollination service to three plant species common in mountain hay meadows, in eighteen extensively managed yellow oat grasslands (Trisetum flavescens) with an altitudinal gradient spanning approx. 300 m. Species richness and abundance of pollinators increased with elevation, but no shift between hoverflies and wild bees (mainly bumblebees) occurred. Seedset of the woodland cranesbill (Geranium sylvaticum) increased with hoverfly abundance, and seedset of the marsh thistle (Cirsium palustre) increased with wild bee abundance. Black rampion (Phyteuma nigrum) showed no significant response. The assignment of specific pollinator communities, and their response to altitude in highlands, to different plant species underlines the importance of wild bees and hoverflies as pollinators in extensive grassland systems.


Aggressive posterior retinopathy of prematurity in two cohorts of patients in South India: implications for primary, secondary, and tertiary prevention.
Aggressive posterior retinopathy of prematurity (APROP), which has a poor visual prognosis, is common in low- and middle-income countries (LMICs) as a result of suboptimal oxygen monitoring (primary prevention). The purpose of this study was to compare outcomes in APROP eyes treated with laser to eyes treated with antivascular endothelial growth factor (anti-VEGF) therapy. The medical records of a cohort of APROP eyes treated with anti-VEGF (2010-2018) and another of eyes treated with laser photocoagulation (2002-2010) at the same institution in South India were reviewed retrospectively and compared. The main outcome was the proportion of eyes developing retinal detachment during resolution of acute ROP. A total of 398 eyes of 199 preterm babies with APROP were included: 168 eyes were treated with photocoagulation; 230, with anti-VEGF. From 2002 to 2010, compared to the more recent cohort, babies diagnosed with APROP tended to be heavier (P < 0.001), older (P < 0.001), and exposed to fewer days of oxygen (P = 0.02). In the laser-treated cohort, 17 of 168 eyes (10%) developed retinal detachment (7, stage 5; 12, stage 4), compared with 3 of 230 (1%) in the anti-VEGF cohort (all stage 4 [P = 0.002]). The incidence of retinal detachment was significantly lower in eyes treated with anti-VEGF compared with laser-.treated eyes In the absence of a randomized trial, these data suggest that anti-VEGF may lead to better anatomic outcomes, although questions remain concerning dosage, timing, and risks.


[Embryonic histogenesis of the atrioventricular segment of the heart conduction system of swine (Sus porcus L.)].
Histological, histochemical and neurochemical methods were used in order to study the features of the histogenesis of structures of the atrioventricular area of the conducting system of the heart and the formation of their cholinergic innervation in the postnatal period of the hog. Under study were 175 hearts of embryos of the hog at the age of 3, 4, 5, 6, 8, 10, 12, 14 and 15 weeks and 22 hearts of adult hogs. The formation of different areas of this system was shown to be asynchronous. The atrioventricular fascicle is formed during the 4th week, the atrioventricular node -- during the 6th week and the conducting muscle fibres--during the 8th week of embryogenesis. The fascicle and the node have a complicated structure and different cellular composition. In the process of prenatal ontogenesis the increased amount of glycogen and increased activity of phosphorylase were noted in the cytoplasm of myocytes of the atrioventricular area of the conducting system of the heart. The cholinergic nerve fibres grow up to the structures of the atrioventricular area of the conducting system of the heart during the 6th week of embryogenesis, and by the end of the prenatal development they form a thick network in all its structures.


Recognition of ischaemia and infection in diabetic foot ulcers: Dataset and techniques.
Recognition and analysis of Diabetic Foot Ulcers (DFU) using computerized methods is an emerging research area with the evolution of image-based machine learning algorithms. Existing research using visual computerized methods mainly focuses on recognition, detection, and segmentation of the visual appearance of the DFU as well as tissue classification. According to DFU medical classification systems, the presence of infection (bacteria in the wound) and ischaemia (inadequate blood supply) has important clinical implications for DFU assessment, which are used to predict the risk of amputation. In this work, we propose a new dataset and computer vision techniques to identify the presence of infection and ischaemia in DFU. This is the first time a DFU dataset with ground truth labels of ischaemia and infection cases is introduced for research purposes. For the handcrafted machine learning approach, we propose a new feature descriptor, namely the Superpixel Colour Descriptor. Then we use the Ensemble Convolutional Neural Network (CNN) model for more effective recognition of ischaemia and infection. We propose to use a natural data-augmentation method, which identifies the region of interest on foot images and focuses on finding the salient features existing in this area. Finally, we evaluate the performance of our proposed techniques on binary classification, i.e. ischaemia versus non-ischaemia and infection versus non-infection. Overall, our method performed better in the classification of ischaemia than infection. We found that our proposed Ensemble CNN deep learning algorithms performed better for both classification tasks as compared to handcrafted machine learning algorithms, with 90% accuracy in ischaemia classification and 73% in infection classification.


Effects of PEEP on the intracranial system of patients with head injury and subarachnoid hemorrhage: the role of respiratory system compliance.
Positive end-expiratory pressure (PEEP) can be effective in improving oxygenation, but it may worsen or induce intracranial hypertension. The authors hypothesized that the intracranial effects of PEEP could be related to the changes in respiratory system compliance (Crs). A prospective study investigated 21 comatose patients with severe head injury or subarachnoid hemorrhage receiving intracranial pressure (ICP) monitoring who required mechanical ventilation and PEEP. The 13 patients with normal Crs were analyzed as group A and the 8 patients with low Crs as group B. During the study, 0, 5, 8, and 12 cm H2O of PEEP were applied in a random sequence. Jugular pressure, central venous pressure (CVP), cerebral perfusion pressure (CPP), intracranial pressure (ICP), cerebral compliance, mean velocity of the middle cerebral arteries, and jugular oxygen saturation were evaluated simultaneously. In the group A patients, the PEEP increase from 0 to 12 cm H2O significantly increased CVP (from 10.6 +/- 3.3 to 13.8 +/- 3.3 mm Hg; p < 0.001) and jugular pressure (from 16.6 +/- 3.1 to 18.8 +/- 3.2 mm Hg; p < 0.001), but reduced mean arterial pressure (from 96.3 +/- 6.7 to 91.3 +/- 6.5 mm Hg; p < 0.01), CPP (from 82.2 +/- 6.9 to 77.0 +/- 6.2 mm Hg; p < 0.01), and mean velocity of the middle cerebral arteries (from 73.1 +/- 27.9 to 67.4 +/- 27.1 cm/sec; F = 7.15; p < 0.001). No significant variation in these parameters was observed in group B patients. After the PEEP increase, ICP and cerebral compliance did not change in either group. Although jugular oxygen saturation decreased slightly, it in no case dropped below 50%. In patients with low Crs, PEEP has no significant effect on cerebral and systemic hemodynamics. Monitoring of Crs may be useful for avoiding deleterious effects of PEEP on the intracranial system of patients with normal Crs.


Hemostatic effect of Vivostat patient-derived fibrin sealant on split-thickness skin graft donor sites.
Topical hemostatic agents are used frequently to control bleeding of skin graft donor sites. In this study, the hemostatic properties of Vivostat (Vivolution A/S, Birkerød, Denmark) patient-derived fibrin sealant were compared with a control group of spray thrombin solution, which is considered an industry standard for topical hemostasis. Treatments were applied simultaneously to two randomly chosen halves of a single split-thickness single donor site in patients in five United States surgical centers. The time to achieve satisfactory hemostasis (< or =10 min) was estimated on each half of the wound as the time at which active bleeding had stopped and the wound was suitable for application of a surgical dressing. The time to hemostasis of wounds treated with Vivostat (Vivolution A/S) patient-derived sealant was significantly shorter in comparison with wounds treated with thrombin solution (medians: Vivostat, 31 seconds; thrombin, 58 seconds; p=0.0012). No abnormalities in wound healing were reported for either treatment site 1 week after the operation. Vivostat (Vivolution A/S) sealant is a more rapidly effective topical hemostatic agent than thrombin on split-thickness skin graft donor sites.


The role of Th17/IL-17 in the pathogenesis of primary nephrotic syndrome in children.
This work aims to explore the role of Th17 and IL-17 signaling in the pathogenesis of primary nephrotic syndrome (PNS) in children and podocyte injury, children with PNS were divided into minimal change nephrotic syndrome (MCNS) and non-minimal change nephrotic syndrome [NMCNS, including mesangial proliferative glomerulonephritis (MsPGN) and focal segmental glomerulosclerosis (FSGS)]. Flow cytometry (FCM) was used to observe the circulating frequency of Th17 cells and the apoptosis of podocytes by annexinV-FITC/PI. Serum IL-1β and IL-6 levels were measured using enzyme-linked immunosorbent assay. The Fas and FasL expressions in podocytes were examined by FCM analysis using a direct immunofluorescence method. Reverse transcription polymerase chain reaction was applied to measure the mRNA expressions of RORc, IL-23p19, Nephrin, WT1, Synaptopodin, Podocalyxin, Fas, and FasL. The IL-17 and IL-1β expression in renal biopsy tissue was detected by immunohistochemistry. The expressions of WT1, Caspase 8, and Caspase 3 in podocyte cell culture were also measured using immunocytochemistry. Circulating frequencies of Th17 cells, mRNA levels of RORc and IL-23p19, and serum levels of IL-6 and IL-1β were higher in the MCNS and NMCNS groups than in the control group (all P < 0.05), and were higher in the NMCNS group than in the MCNS group (all P < 0.05). The expressions of IL-17 and IL-1β in renal biopsy tissue were higher in the MCNS, MsPGN, and FSGS groups than in the control group (all P < 0.05). Recombinant murine IL-17 (rmIL-17) had no effect on the expressions of Nephrin, Synaptopodin, and WT1 of mouse podocytes, but caused an decrease in the expression of podocalyxin as well as promoted apoptosis in a dose- and time-dependent fashion. Moreover, rmIL-17 increased the expression of Fas, Casepase-8, and Casepase-3, but had no effect on that of FasL. Th17/IL-17 may contribute to the pathogenesis of PNS by decreasing the podocalyxin level and inducing podocyte apoptosis.


Review of photon interaction cross section data in the medical and biological context.
The probability of a photon (x-ray, gamma-ray, bremsstrahlung, etc) of a given energy E undergoing absorption or scattering when traversing a layer of material Z can be expressed quantitatively in terms of a linear attenuation coefficient mu (cm(-1)). Since mu is dependent on the material's density, rho (g cm(-3)), which can be variable, the quantity usually tabulated is the mass attenuation coefficient mu/rho (cm2 g(-1)) in which the dependence on the density has been removed. Mu/rho, in turn, can be obtained as the sum of the different types of possible interactions of photons with atoms of the material. For photon energies below 1 MeV the major interaction processes to be considered are incoherent (Compton) scattering, coherent (Rayleigh) scattering and atomic photoeffect absorption. Above 1 MeV one must also include nuclear-field pair production and atomic-field (triplet) production, and above 5 MeV one in principle should include photonuclear absorption, although the latter is neglected in data tabulations up to the present time. This review includes a selective history of measurements and theory relating to mu/rho from the turn of the century up to the present time, and is intended to provide a basis for further calculations and critical tabulations of photon cross section data, particularly as required by users in radiation medicine and biology. The mass energy-absorption coefficient mu(en)/rho is also briefly discussed.


The role of the tumour microenvironment in immunotherapy.
Recent success in immunomodulating strategies in lung cancer and melanoma has prompted much enthusiasm in their potential to treat other advanced solid malignancies. However, their applications have shown variable success and are even ineffective against some tumours. The efficiency of immunotherapies relies on an immunogenic tumour microenvironment. The current field of cancer immunology has focused on understanding the interaction of cancer and host immune cells to break the state of immune tolerance and explain how molecular patterns of cytokines and chemokines affect tumour progression. Here, we review our current knowledge of how inherent properties of tumours and their different tumour microenvironments affect therapeutic outcome. We also discuss insights into recent multimodal therapeutic approaches that target tumour immune evasion and suppression to restore anti-tumour immunity.


Persistent emotional distress after a first-time myocardial infarction and its association to late cardiovascular and non-cardiovascular mortality.
Patients with symptoms of depression and/or anxiety - emotional distress - after a myocardial infarction (MI) have been shown to have worse prognosis and increased healthcare costs. However, whether specific subgroups of patients with emotional distress are more vulnerable is less well established. The purpose of this study was to identify the association between different patterns of emotional distress over time with late cardiovascular and non-cardiovascular mortality among first-MI patients aged <75 years in Sweden. We utilized data on 57,602 consecutive patients with a first-time MI from the national SWEDEHEART registers. Emotional distress was assessed using the anxiety/depression dimension of the European Quality of Life Five Dimensions questionnaire two and 12 months after the MI, combined into persistent (emotional distress at both time-points), remittent (emotional distress at the first follow-up only), new (emotional distress at the second-follow up only) or no distress. Data on cardiovascular and non-cardiovascular mortality were obtained until the study end-time. We used multiple imputation to create complete datasets and adjusted Cox proportional hazards models to estimate hazard ratios. Patients with persistent emotional distress were more likely to die from cardiovascular (hazard ratio: 1.46, 95% confidence interval: 1.16, 1.84) and non-cardiovascular causes (hazard ratio: 1.54, 95% confidence interval: 1.30, 1.82) than those with no distress. Those with remittent emotional distress were not statistically significantly more likely to die from any cause than those without emotional distress. Among patients who survive 12 months, persistent, but not remittent, emotional distress was associated with increased cardiovascular and non-cardiovascular mortality. This indicates a need to identify subgroups of individuals with emotional distress who may benefit from further assessment and specific treatment.


To milliseconds and beyond: challenges in the simulation of protein folding.
Quantitatively accurate all-atom molecular dynamics (MD) simulations of protein folding have long been considered a holy grail of computational biology. Due to the large system sizes and long timescales involved, such a pursuit was for many years computationally intractable. Further, sufficiently accurate forcefields needed to be developed in order to realistically model folding. This decade, however, saw the first reports of folding simulations describing kinetics on the order of milliseconds, placing many proteins firmly within reach of these methods. Progress in sampling and forcefield accuracy, however, presents a new challenge: how to turn huge MD datasets into scientific understanding. Here, we review recent progress in MD simulation techniques and show how the vast datasets generated by such techniques present new challenges for analysis. We critically discuss the state of the art, including reaction coordinate and Markov state model (MSM) methods, and provide a perspective for the future.


Hyperosmolarity-induced cornification of human corneal epithelial cells is regulated by JNK MAPK.
To evaluate the effects of hyperosmolar stress on expression of cornified envelope (CE) precursors and transglutaminases (TGs) by primary cultured human corneal epithelial (PCHCE) cells and the regulatory effects of JNK MAPK on this process. Expression of CE precursors and TGs were evaluated in PCHCE cells exposed to media of increasing osmolarity (350-450 mOsM) for 24, 48, and 72 hours. JNK1 and -2 MAPKs were inhibited by addition of short interfering (si)RNA. Relative levels of mRNA transcripts and proteins were evaluated. TG activity, cell viability, and apoptosis were detected in PCHCE cells, with or without siRNA-JNKs. Exposure of PCHCE cells to hyperosmolar medium increased TG activity at 3 hours, levels of the CE precursors SPRR1b and -2a and membrane-associated TG1 mRNA at 6 hours, and tissue-type TG2 mRNA at 24 hours. Osmotic stress decreased corneal epithelial cell viability, which was due in part to stimulation of apoptosis and cornification death. Inhibiting JNK2 production by siRNA in osmotically stressed PCHCE cells prevented the stimulation of SPRR and membrane-associated TG1 production and TG activity, and improved cell viability, whereas inhibition of JNK1 prevented early apoptosis. Osmotic stress promotes production of certain CE proteins and cross-linking membrane-associated TG1 and decreases cell viability via JNK MAPK-mediated pathways. Strategies that inhibit JNK production downregulate the cornification response of PCHCE cells to osmotic stress. These findings have potential therapeutic implications for preventing cornification of the corneal epithelium in response to the hyperosmolar tear film in dry eye disease.


Influencing organisational culture to improve hospital performance in care of patients with acute myocardial infarction: a mixed-methods intervention study.
Hospital organisational culture affects patient outcomes including mortality rates for patients with acute myocardial infarction; however, little is known about whether and how culture can be positively influenced. This is a 2-year, mixed-methods interventional study in 10 US hospitals to foster improvements in five domains of organisational culture: (1) learning environment, (2) senior management support, (3) psychological safety, (4) commitment to the organisation and (5) time for improvement. Outcomes were change in culture, uptake of five strategies associated with lower risk-standardised mortality rates (RSMR) and RSMR. Measures included a validated survey at baseline and at 12 and 24 months (n=223; average response rate 88%); in-depth interviews (n=393 interviews with 197 staff); and RSMR data from the Centers for Medicare and Medicaid Services. We observed significant changes (p<0.05) in culture between baseline and 24 months in the full sample, particularly in learning environment (p<0.001) and senior management support (p<0.001). Qualitative data indicated substantial shifts in these domains as well as psychological safety. Six of the 10 hospitals achieved substantial improvements in culture, and four made less progress. The use of evidence-based strategies also increased significantly (per hospital average of 2.4 strategies at baseline to 3.9 strategies at 24 months; p<0.05). The six hospitals that demonstrated substantial shifts in culture also experienced significantly greater reductions in RSMR than the four hospitals that did not shift culture (reduced RSMR by 1.07 percentage points vs 0.23 percentage points; p=0.03) between 2011-2014 and 2012-2015. Investing in strategies to foster an organisational culture that supports high performance may help hospitals in their efforts to improve clinical outcomes.


Urinary and plasma magnesium and risk of ischemic heart disease.
Previous studies on dietary magnesium and risk of ischemic heart disease (IHD) have yielded inconsistent results, in part because of a lack of direct measures of actual magnesium uptake. Urinary excretion of magnesium, an indicator of dietary magnesium uptake, might provide more consistent results. The objective was to investigate whether urinary magnesium excretion and plasma magnesium are associated with IHD risk. We examined 7664 adult participants free of known cardiovascular disease in the Prevention of Renal and Vascular End-Stage Disease (PREVEND) study-a prospective population-based cohort study. Urinary magnesium excretion was measured in 2 baseline 24-h urine collections. Mean ± SD urinary magnesium excretion was 4.24 ± 1.65 mmol/24 h for men and 3.54 ± 1.40 mmol/24 h for women. During a median follow-up of 10.5 y (IQR: 9.9-10.8 y), 462 fatal and nonfatal IHD events occurred. After multivariable adjustment, urinary magnesium excretion had a nonlinear relation with IHD risk (P-curvature = 0.01). The lowest sex-specific quintile (men: <2.93 mmol/24 h; women: <2.45 mmol/24 h) had an increased risk of fatal and nonfatal IHD (multivariable HR: 1.60; 95% CI: 1.28, 2.00) compared with the upper 4 quintiles of urinary magnesium excretion. A similar increase in risk of the lowest quintile was observed for mortality related to IHD (HR: 1.70; 95% CI: 1.10, 2.61). No associations were observed between circulating magnesium and risk of IHD. Low urinary magnesium excretion was independently associated with a higher risk of IHD incidence. An increased dietary intake of magnesium, particularly in those with the lowest urinary magnesium, could reduce the risk of IHD.


Perception of family medicine as the career option among young Indian graduates.
1. To estimate the number of graduates who had exposure to family medicine specialty. 2. To estimate the number of graduates who consider family medicine as the carrier option. 3. To compare the perception of family medicine as career option among the young graduates who had exposure to family medicine during their training or practice with no exposure. It is a cross-sectional, questionnaire-based study. The doctors graduated 2018 onwards were contacted through SIMSA (South Indian Medical Students Association) and WhatsApp groups (students doing the rural health service and intern WhatsApp). Subjects who consented for the study filled the google form. Filled Google forms were evaluated. First choice of among the graduates is general medicine followed by general surgery and paediatrics. Family medicine is 4th in the order. There is overall a very positive perception about the family medicine among the respondents irrespective of exposure to family medicine. Majority of them feel that it gives good financial gain (55.55%), interesting (67.03%), work and family balance (75.55%), which contributes to individual and community health (84.07%) and essential part of healthcare system (83.33%). Graduates feel that career growth (26.66%) and academic growth (33.70%) are less and it is not popular (38.88%). If there is an opportunity, majority would choose family medicine (54.44%). Around 5.9% are open to choose family medicine as the career option. Medical graduates perceive family medicine subject to give them good work and family balance and the subject is essential part of healthcare system and contributes to the individual and community health, and at the same time, it may not give them great financial gain, academic and career growth.


Generalized additive model demonstrates fluoroquinolone use/resistance relationships for Staphylococcus aureus.
Mathematical models currently used to study the relationship between the prevalence of the resistance to an antibiotic and the amount of drug may not be adequate because they do not integrate temporal and area analyses simultaneously. Furthermore, the forms of such relationship are unknown. We applied the Generalized Additive Model (GAM) to study fluoroquinolone use and the incidence of fluoroquinolone-resistance in Staphylococcus aureus in our institution over a 3-year period. Overall institution and individual hospital unitrestricted analyses were performed. Relative risk (RR) observed for increasing fluoroquinolone use with a 4-month lag from the 25th percentile to the 75th percentile was 1.27 (95% CI: 1.13-1.42). In the surgery units, RR was 1.23 (95% CI: 1.01-1.50) for fluoroquinolone use with a 2-months lag. GAM enabled us to observe for the first time a significant temporal relationship between fluoroquinolone use and the incidence of fluoroquinolone-resistant nosocomial S. aureus isolates.


Telling stories: news media, health literacy and public policy in Canada.
Mass media are very influential in shaping discourses about health but few studies have examined the extent to which newspaper coverage of such stories reflect issues embedded in health policy documents. We estimate the relative distribution of health stories using content analysis. Nine meta-topics are used to sort stories across a range of major influences shaping the health status of populations adapted from the document Toward a Healthy Future (Second Report on the Health of Canadians (1999)) (TAHF). A total of 4732 stories were analyzed from 13 Canadian daily newspapers (10 English, 3 French language) using a constructed week per quarter method. Stories were sampled from each chosen newspaper for the years 1993, 1995, 1997 and 2001. 72% (n=3405) of stories in this analysis were from English-language papers, 28% (n=1327) were from French-language papers. Topics related to health care (dealing either with issues of service provision and delivery or management and regulation) dominated newspaper stories, accounting for 65% of all stories. Physical environment topics accounted for about 13% of all stories, the socio-economic environment about 6% of stories, personal health practices about 5% of stories, and scientific advances in health research about 4% of stories. Other influences upon health identified in TAHF were rarely mentioned. The overall prominence of topics in newspapers is not consistent with the relative importance assigned to health influences in TAHF. Canadian newspapers rarely report on socio-economic influences frequently cited in the research literature (and reflected in TAHF) as being most influential in shaping population health outcomes.