Effect of recombinant human leukocyte, fibroblast, and immune interferons on expression of class I and II major histocompatibility complex and invariant chain in early passage human melanoma cells.
Twenty-five early-passage (less than or equal to 8) melanoma cell lines, isolated from ten patients with metastatic melanoma, were analyzed by a combination of serological, immunochemical, and molecular methods for mRNA levels, synthesis, and surface expression of MHC class I and class II antigens prior to and following exposure to recombinant human leukocyte (IFN-alpha A), fibroblast (IFN-beta), and immune (IFN-gamma) interferon. All the cell lines expressed variable levels of HLA class I gene products that were up-regulated to different extents upon exposure to specific interferons (IFNs). HLA class II antigens were expressed in 22 of the 25 melanoma lines and IFN-gamma increased the levels of class II mRNA, protein synthesis, and surface expression in all cultures displaying baseline expression. A significant up-regulation of class II antigen expression by IFN-alpha or -beta, associated with higher levels of class II transcripts and enhanced synthesis, was found only in two early-passage human melanoma cell lines. In three lesions from the same patient which did not constitutively express class II antigens, no expression of these glycoproteins could be induced with any of the IFNs. These results indicate that IFN-gamma does not act as a de novo inducer of class II antigen expression in early-passage human melanoma cell lines. This hypothesis is further supported by analysis of class II-associated invariant chain (Ii) expression, which is expressed and induced by IFNs in a manner similar to that of class II antigens. The present study also indicates that early-passage metastatic melanoma lesions from the same patient are heterogeneous in their de novo expression of major histocompatibility antigens and in their modulation by IFNs.


Acute amiodarone promotes drift and early termination of spiral wave re-entry.
Intravenous application of amiodarone is commonly used in the treatment of life-threatening arrhythmias, but the underlying mechanism is not fully understood. The purpose of the present study is to investigate the acute effects of amiodarone on spiral wave (SW) re-entry, the primary organization machinery of ventricular tachycardia/fibrillation (VT/VF), in comparison with lidocaine. A two-dimensional ventricular myocardial layer was obtained from 24 Langendorff-perfused rabbit hearts, and epicardial excitations were analyzed by high-resolution optical mapping. During basic stimulation, amiodarone (5 microM) caused prolongation of action potential duration (APD) by 5.6%-9.1%, whereas lidocaine (15 microM) caused APD shortening by 5.0%-6.4%. Amiodarone and lidocaine reduced conduction velocity similarly. Ventricular tachycardias induced by DC stimulation in the presence of amiodarone were of shorter duration (sustained-VTs >30 s/total VTs: 2/58, amiodarone vs 13/52, control), whereas those with lidocaine were of longer duration (22/73, lidocaine vs 14/58, control). Amiodarone caused prolongation of VT cycle length and destabilization of SW re-entry, which is characterized by marked prolongation of functional block lines, frequent wavefront-tail interactions near the rotation center, and considerable drift, leading to its early annihilation via collision with anatomical boundaries. Spiral wave re-entry in the presence of lidocaine was more stabilized than in control. In the anisotropic ventricular myocardium, amiodarone destabilizes SW re-entry facilitating its early termination. Lidocaine, in contrast, stabilizes SW re-entry resulting in its persistence.


Transcriptional profiling of Helicobacter pylori Fur- and iron-regulated gene expression.
Intracellular iron homeostasis is a necessity for almost all living organisms, since both iron restriction and iron overload can result in cell death. The ferric uptake regulator protein, Fur, controls iron homeostasis in most Gram-negative bacteria. In the human gastric pathogen Helicobacter pylori, Fur is thought to have acquired extra functions to compensate for the relative paucity of regulatory genes. To identify H. pylori genes regulated by iron and Fur, we used DNA array-based transcriptional profiling with RNA isolated from H. pylori 26695 wild-type and fur mutant cells grown in iron-restricted and iron-replete conditions. Sixteen genes encoding proteins involved in metal metabolism, nitrogen metabolism, motility, cell wall synthesis and cofactor synthesis displayed iron-dependent Fur-repressed expression. Conversely, 16 genes encoding proteins involved in iron storage, respiration, energy metabolism, chemotaxis, and oxygen scavenging displayed iron-induced Fur-dependent expression. Several Fur-regulated genes have been previously shown to be essential for acid resistance or gastric colonization in animal models, such as those encoding the hydrogenase and superoxide dismutase enzymes. Overall, there was a partial overlap between the sets of genes regulated by Fur and those previously identified as growth-phase, iron or acid regulated. Regulatory patterns were confirmed for five selected genes using Northern hybridization. In conclusion, H. pylori Fur is a versatile regulator involved in many pathways essential for gastric colonization. These findings further delineate the central role of Fur in regulating the unique capacity of H. pylori to colonize the human stomach.


Adaptive significance of synchronous chorusing in an acoustically signalling wolf spider.
Synchronous sexual signalling is a behavioural phenomenon that has received considerable theoretical interest, but surprisingly few empirical tests have been conducted. Here, we present a set of experiments designed to determine (i) whether the sexual signalling of the drumming wolf spider Hygrolycosa rubrofasciata is synchronous, and (ii) whether the synchrony may have evolved through female preference. Using controlled playback experiments, we found that males actively synchronized their drumming bouts with other males and females significantly preferred closely synchronized drumming clusters compared with loose clusters. In loose clusters, the first drumming signals attracted the most female responses, whereas in close clusters, the last drumming signals were the most heeded. We suggest that this female preference for the last drummer can maintain male synchronous signalling in H. rubrofasciata.


Neuraminidase and hemagglutinin matching patterns of a highly pathogenic avian and two pandemic H1N1 influenza A viruses.
Influenza A virus displays strong reassortment characteristics, which enable it to achieve adaptation in human infection. Surveying the reassortment and virulence of novel viruses is important in the prevention and control of an influenza pandemic. Meanwhile, studying the mechanism of reassortment may accelerate the development of anti-influenza strategies. The hemagglutinin (HA) and neuraminidase (NA) matching patterns of two pandemic H1N1 viruses (the 1918 and current 2009 strains) and a highly pathogenic avian influenza A virus (H5N1) were studied using a pseudotyped particle (pp) system. Our data showed that four of the six chimeric HA/NA combinations could produce infectious pps, and that some of the chimeric pps had greater infectivity than did their ancestors, raising the possibility of reassortment among these viruses. The NA of H5N1 (A/Anhui/1/2005) could hardly reassort with the HAs of the two H1N1 viruses. Many biological characteristics of HA and NA, including infectivity, hemagglutinating ability, and NA activity, are dependent on their matching pattern. Our data suggest the existence of an interaction between HA and NA, and the HA NA matching pattern is critical for valid viral reassortment.


Sodium hydroxide chemical matricectomy for the treatment of ingrown toenails: comparison of three different application periods.
Sodium hydroxide matricectomy is a successful method for the treatment of ingrown toenails. This study was designed to evaluate the optimal sodium hydroxide application period providing high success rates with minimal postoperative morbidity. Sixty-six patients with 225 ingrown nail edges were treated in three groups receiving 30-second, 1-minute, and 2-minute applications of sodium hydroxide. Each patient was reviewed postoperatively for pain, drainage, and tissue damage. The median long-term follow-up period was 14 months. The success rate of the therapy was 70.9% in the first group, 92.7% in the second group, and 94.4% in the third group. In all groups, about half of the patients experienced minimal pain within 48 hours following the operation, but only in the third group, 20% of the patients had minimal pain, which continued about 1 week. Drainage and tissue damage were minimal or mild in all groups and disappeared within 3 weeks in the first and second groups but were prolonged to 6 weeks in the third group. The success rate of 30-second application is significantly lower than 1-minute and 2-minute applications. Although the success rates of the latter two procedures are similar, the prolonged healing time is the disadvantage of the 2-minute application. We conclude that 1-minute application of 10% sodium hydroxide is simple, safe, and highly effective for the treatment of ingrown nails.


Role of basic fibroblast growth factor in the suppression of apoptotic caspase-3 during chronic gastric ulcer healing.
In the course of ulcer healing an array of factors compel mucosal cells to proliferate, differentiate, and migrate to the site of injury. The recognition of triggering cues requires close interaction between the regulatory proteins integrating the growth factor and cytokine- mediated signals that propel cells through the cycle events, or to signal apoptosis. In this study, we analyzed the interplay between mucosal expression of the receptor-bound basic fibroblast growth factor (bFGF-R) and cyclin-dependent kinase (Cdk2), and the activity of apoptotic protease, caspase-3, and constitutive nitric oxide synthase (cNOS) during chronic gastric ulcer healing. The experiments were conducted with gastric mucosa of rats at different stages of acetic acid-induced chronic gastric ulcer healing. The ulcer onset (2 days following injury) was characterized by a massive epithelial apoptosis associated with a 33-fold increase in caspase-3 activity and a 7.6-fold drop in cNOS, while the mucosal expression of Cdk2 fell by an 18% and that of bFGF-R by 12%. The ulcer healing was accompanied by a rapid elevation in bFGF-R and Cdk2, and a slow recovery in cNOS activity, while the caspase-3 activity and epithelial apoptosis showed a marked decline. The bFGF-R and Cdk2 reached their maximums of 2.2-2.3-fold at 4-6 day of healing, while the caspase-3 activity and the apoptotic DNA fragmentation showed a 3-fold decline by the 7th day of healing. However, the activity of cNOS remained about 50% lower than that of the controls. Taken together, these results provide strong indications that the initial phase of ulcer healing involves the inhibition of apoptotic caspase activities by a signaling events initiated by bFGF-receptor activation and propagated by the regulatory kinases that propel the cell cycle progression. Our findings also point towards participation of cNOS in the suppression of proapoptotic activities in gastric mucosa.


Assessing Adherence to Antihypertensive Therapy in Primary Health Care in Namibia: Findings and Implications.
Namibia has the highest burden and incidence of hypertension in sub-Sahara Africa. Though non-adherence to antihypertensive therapy is an important cardiovascular risk factor, little is known about potential ways to improve adherence in Namibia following universal access. The objective of this study is to validate the Hill-Bone compliance scale and determine the level and predictors of adherence to antihypertensive treatment in primary health care settings in sub-urban townships of Windhoek, Namibia. Reliability was determined by Cronbach's alpha. Principal component analysis (PCA) was used to assess construct validity. The PCA was consistent with the three constructs for 12 items, explaining 24.1, 16.7 and 10.8% of the variance. Cronbach's alpha was 0.695. None of the 120 patients had perfect adherence to antihypertensive therapy, and less than half had acceptable levels of adherence (≥ 80%). The mean adherence level was 76.7 ± 8.1%. Three quarters of patients ever missed their scheduled clinic appointment. Having a family support system (OR = 5.4, 95% CI 1.687-27.6, p = 0.045) and attendance of follow-up visits (OR = 3.1, 95% CI 1.1-8.7, p = 0.03) were significant predictors of adherence. Having HIV/AIDs did not lower adherence. The modified Namibian version of the Hill-Bone scale is reliable and valid for assessing adherence to antihypertensives in Namibia. There is sub-optimal adherence to antihypertensive therapy among primary health cares in Namibia. This needs standardized systems to strengthen adherence monitoring as well as investigation of other factors including transport to take full advantage of universal access.


The effect of multidisciplinary care teams on intensive care unit mortality.
Critically ill patients are medically complex and may benefit from a multidisciplinary approach to care. We conducted a population-based retrospective cohort study of medical patients admitted to Pennsylvania acute care hospitals (N = 169) from July 1, 2004, to June 30, 2006, linking a statewide hospital organizational survey to hospital discharge data. Multivariate logistic regression was used to determine the independent relationship between daily multidisciplinary rounds and 30-day mortality. A total of 112 hospitals and 107 324 patients were included in the final analysis. Overall 30-day mortality was 18.3%. After adjusting for patient and hospital characteristics, multidisciplinary care was associated with significant reductions in the odds of death (odds ratio [OR], 0.84; 95% confidence interval [CI], 0.76-0.93 [P = .001]). When stratifying by intensivist physician staffing, the lowest odds of death were in intensive care units (ICUs) with high-intensity physician staffing and multidisciplinary care teams (OR, 0.78; 95% CI, 0.68-0.89 [P < .001]), followed by ICUs with low-intensity physician staffing and multidisciplinary care teams (OR, 0.88; 95% CI, 0.79-0.97 [P = .01]), compared with hospitals with low-intensity physician staffing but without multidisciplinary care teams. The effects of multidisciplinary care were consistent across key subgroups including patients with sepsis, patients requiring invasive mechanical ventilation, and patients in the highest quartile of severity of illness. Daily rounds by a multidisciplinary team are associated with lower mortality among medical ICU patients. The survival benefit of intensivist physician staffing is in part explained by the presence of multidisciplinary teams in high-intensity physician-staffed ICUs.


Prevalence and pathogenesis of dental and periodontal lesions in children with X-linked hypophosphatemic rickets.
To assess the prevalence and to investigate the pathogenetic mechanisms of dental and periodontal lesions in children with X-linked hypophosphatemic rickets (XLH) examined at diagnosis or during treatment. Nine children with XLH (age 7.2 +/- 3.3 years) were enrolled in the study (at diagnosis, n = 2; during treatment with oral inorganic phosphate salts combined with 1,25-dihydroxyvitamin D3, n = 7). Oral examination was performed according to the evidence of carious and gingival lesions. Decayed or filled teeth (dft) index for primary teeth, and the decayed, missing, or filled teeth (DMFT) index for permanent teeth was assessed. All patients with a history of spontaneous dental abscesses underwent orthopantomography examination. d/D ranged from 0 to 9 and f/F from 0 to 3. DMFT/dft index was 0 in the three youngest patients. One patient had enamel hypoplasia and two had enamel dyschromic alterations. Six out of nine patients (67%) had a history of spontaneous fistulae as a consequence of periapical abscesses occurring in the absence of dental decay or history of injury. In these patients, orthopantomographies showed enlarged pulp chambers associated with prominent pulp horns extending up to the dentino-enamel junction in both primary and permanent dentition. XLH patients show some peculiar dentinal abnormalities. Treatment prevents only in part dental and periodontal lesions. Genetic mechanisms have a main role in causing defective dentin mineralisation.


Influence of ventricular contractility on non-work-related myocardial oxygen consumption.
The relationship between myocardial oxygen consumption (MVO2) and the total pressure-volume area (PVA), which represents the total mechanical work performed during a cardiac cycle, has been shown to be linear and independent of loading conditions: MVO2 = aPVA + b. When inotropic state is enhanced, the MVO2-PVA relation shifts upward (increase in b), and when inotropic state is depressed the relation shifts downward (decrease in b). However, the quantitative relationship between contractility and b (the non-work-related myocardial oxygen consumption) determined over a wide range of contractilities is not known. In seven isolated blood perfused canine hearts, left ventricular (LV) contractility was increased by dobutamine and decreased with nifedipine or reduction of coronary blood flow. At each level of contractility, the end-systolic pressure-volume relationship (ESPVR) and the MVO2-PVA relation were determined. For each heart, the resulting values of b (ml O2/beat) were plotted as a function of Emax (mmHg/ml), an index of contractility defined as the slope of the ESPVR. There was a linear relation between Emax and b over a wide range of contractilities; on average, b (ml O2/beat) = 0.0036 Emax (mmHg/ml) + 0.0101 [r = 0.929-0.978 (95% confidence interval)], when Emax was varied over an average range of 2.8-9.6 mmHg/ml. These results suggest a common underlying determinant of contractility and non-work-related oxygen consumption.


Effect of antiplatelet antibody on platelet shape change, volume, and morphology.
We extended our studies of the effect of antibody on human platelets. Monomeric immunoglobulin G (IgG) (rabbit) antiplatelet antibody was interacted with human platelets in the absence of complement and the early morphological changes monitored. Platelet shape change, assessed by a rheooptical approach, was noted within 20 s and was proportional to antibody concentration. Antiplatelet antibody-induced shape change was also accompanied by a change in apparent platelet volume, as measured in the resistive particle counter. Scanning and transmission electron microscopy confirmed a disk-to-sphere transformation associated with the formation of bulbous, short surface projections or pseudopodia. In contrast, ADP-induced disk-to-sphere transformation was associated with the development of long, thin filopodia emanating from the platelet. These specific effects of antiplatelet antibody may result in altered platelet function.


The impact of covariance priors on arm-based Bayesian network meta-analyses with binary outcomes.
Bayesian analyses with the arm-based (AB) network meta-analysis (NMA) model require researchers to specify a prior distribution for the covariance matrix of the treatment-specific event rates in a transformed scale, for example, the treatment-specific log-odds when a logit transformation is used. The commonly used conjugate prior for the covariance matrix, the inverse-Wishart (IW) distribution, has several limitations. For example, although the IW distribution is often described as noninformative or weakly informative, it may in fact provide strong information when some variance components are small (eg, when the standard deviation of study-specific log-odds of a treatment is smaller than 1/2), as is common in NMAs with binary outcomes. In addition, the IW prior generally leads to underestimation of correlations between treatment-specific log-odds, which are critical for borrowing strength across treatment arms to estimate treatment effects efficiently and to reduce potential bias. Alternatively, several separation strategies (ie, separate priors on variances and correlations) can be considered. To study the IW prior's impact on NMA results and compare it with separation strategies, we did simulation studies under different missing-treatment mechanisms. A separation strategy with appropriate priors for the correlation matrix and variances performs better than the IW prior, and should be recommended as the default vague prior in the AB NMA approach. Finally, we reanalyzed three case studies and illustrated the importance, when performing AB-NMA, of sensitivity analyses with different prior specifications on variances.


Resistance exercise training does not affect postexercise hypotension and wave reflection in women with fibromyalgia.
The purpose of this study was to assess the effects of resistance exercise training (RET) on aortic wave reflection and hemodynamics during recovery from acute resistance exercise in women with fibromyalgia (FM) and healthy women (HW). Nine women with FM (aged 42 ± 5 years; mean ± SD) and 14 HW (aged 45 ± 5 years) completed testing at baseline and after 12 weeks of whole-body RET that consisted of 3 sets of 5 exercises. Heart rate (HR), digital blood pressure (BP, plethysmography), aortic BP, and wave reflection (radial tonometry) were assessed before and 20 min after acute leg resistance exercise. Aortic and digital diastolic blood pressure (DBP) were significantly decreased (p < 0.05) and aortic and digital pulse pressures (PP) were significantly increased (p < 0.05) after acute exercise before RET. Acute resistance exercise had no effect on HR, wave reflection (augmentation index and reflection time), digital, or aortic systolic BP. RET improved muscle strength without affecting acute DBP and PP responses. Acute resistance exercise produces postexercise diastolic hypotension without affecting systolic blood pressure, HR, and wave reflection responses in women with and without FM. RET does not alter resting and postexercise hemodynamics and aortic wave reflection in premenopausal women.


Immediate and delayed reaching in hemispatial neglect.
Milner and Goodale (The visual brain in action, Oxford: Oxford University Press, 1995; The visual brain in action, 2nd ed., Oxford: Oxford University Press, 2006) propose a model of vision that makes a distinction between vision for perception and vision for action. One strong claim of the model is that the dorsal stream's control of action is designed for dealing with target stimuli in the 'here and now', yet when time is allowed to pass and a reaction has to be made on the basis of a visual memory, the ventral stream is required for successful performance. Regarding the syndrome of hemispatial neglect, Milner and Goodale further claim that the visual dorsal stream is relatively spared in these patients. In the current study we tested whether neglect patients would indeed be unimpaired in immediate pointing, yet show inaccurate pointing in a condition where a delay is interposed between the presentation of the stimulus and the response signal (in particular in left space). We tested the ability of nine neglect patients (and healthy and right hemisphere no neglect control groups) to perform reaches towards immediate and delayed targets, placed in left, central and right locations. Neglect patients showed no accuracy impairments when asked to perform an immediate action. Conversely, when pointing towards remembered leftward locations, they markedly overshoot the target or failed to initiate a reach altogether. These results confirm that patients with neglect are not specifically impaired when performing 'here and now' actions, but rather present deficits when the visuomotor task taps into more perceptual 'off-line' representations thought to depend on ventral visual stream activation.


Does Racial Isolation in School Lead to Long-Term Disadvantages? Labor Market Consequences of High School Racial Composition.
School racial composition has modest effects on test score gaps, but evidence of a longer-term impact is scarce. Perpetuation theory suggests that blacks who attend schools with higher proportions of white classmates may have better job outcomes. Multilevel analyses of two national longitudinal surveys reveal no effects of high school racial composition on occupational status, employment, or annual earnings for blacks or whites. For other minority groups, attending schools with more whites impedes occupational advancement. For all groups, however, school racial composition predicts workplace racial composition: Whites who attend high schools with higher proportions of white students have higher proportions of white coworkers, while nonwhites who attend schools with higher proportions of whites have fewer same-race coworkers. The findings are modest in size but robust to alternative specifications, and sensitivity analyses support a causal interpretation for same-race coworkers. These results support perpetuation theory for workplace composition but not for stratification outcomes.


Influence of Pleistocene glacial/interglacial cycles on the genetic structure of the mistletoe cactus Rhipsalis baccifera (Cactaceae) in Mesoamerica.
Phylogeographical work on cloud forest-adapted species provides inconsistent evidence on cloud forest dynamics during glacial cycles. A study of Rhipsalis baccifera (Cactaceae), a bird-dispersed epiphytic mistletoe cactus, was conducted to investigate genetic variation at sequence data from nuclear [internal transcribed spacer (ITS), 677 bp] and chloroplast (rpl32-trnL, 1092bp) DNA for 154 individuals across the species range in Mesoamerica to determine if such patterns are consistent with the expansion/contraction model of cloud forest during glacial cycles. We conducted population and spatial genetic analyses as well as gene flow and divergence time estimates between 24 populations comprising the distribution of R. baccifera in Mexico and Guatemala to gain insight of the evolutionary history of these populations, and a complementary species distribution modeling approach to frame information derived from the genetic analyses into an explicit paleoecological context. The results revealed a phylogeographical break at the Isthmus of Tehuantepec, and high levels of genetic diversity among populations and cloud forest areas. Despite the genetic differentiation of some R. baccifera populations, the widespread ITS ribotypes suggest effective nuclear gene flow via pollen and population differentiation shown by the rpl32-trnL suggests more restricted seed flow. Predictions of species distribution models under past last glacial maximum (LGM) climatic conditions and a significant signal of demographic expansion suggest that R. baccifera populations experienced a range expansion tracking the conditions of the cloud forest distribution and shifted to the lowlands with population connectivity during the LGM.


Early prediction of treatment failure in severe community-acquired pneumonia: The PRoFeSs score.
To identify a single/panel of biomarkers and to provide a point score that, after 48 h of treatment, could early predict treatment failure at fifth day of Intensive Care Unit (ICU) stay in severe community-acquired pneumonia (SCAP) patients. Single-center, prospective cohort study of 107 ICU patients with SCAP. Primary outcome included death or absence of improvement in Sequential Organ Failure Assessment score by ≥2 points within 5 days of treatment. Biomarkers were evaluated within 12 h of first antibiotic dose (D1) and 48 h after the first assessment (D3). A model based on Charlson's score and a panel of biomarkers (procalcitonin on D1 and D3, B-natriuretic peptide on D1, D-dimer and lactate on D3) had good discrimination for primary outcome in both derivation (AUC 0.82) and validation (AUC 0.76) samples and was well calibrated (X2 = 0.98; df = 1; p = .32). A point score system (PRoFeSs score) built on the estimates of regression coefficients presented good discrimination (AUC 0.81; 95% Confidence Interval 0.72-0.89) for primary outcome. In SCAP, a combination of biomarkers measured at admission and 48 h later may early predict treatment failure. PRoFeSs score may recognize patients with poor short-term prognosis.


Breviscapine confers a neuroprotective efficacy against transient focal cerebral ischemia by attenuating neuronal and astrocytic autophagy in the penumbra.
Breviscapine is a flavonoid derived from a traditional Chinese herb Erigerin breviscapus (Vant.) Hand-Mazz, and has been extensively used in clinical treatment for cerebral stroke in China, but the underlying pharmacological mechanisms are still unclear. In present study, we investigated whether breviscapine could confer a neuroprotection against cerebral ischemia injury by targeting autophagy mechanisms. A cerebral stroke model in Sprague-Dawley rats was prepared by middle cerebral artery occlusion (MCAO), rats were then randomly divided into 5 groups: MCAO+Bre group, rats were treated with breviscapine; MCAO+Tat-Beclin-1 group, animals were administrated with specific autophagy inducer Tat-Beclin-1; MCAO+Bre+Tat-Beclin-1 group, rats were treated with both breviscapine and Tat-Beclin-1, MCAO+saline group, rats received the same volume of physiological saline, and Sham surgery group. The autophagy levels in infarct penumbra were evaluated by western blotting, real-time PCR and immunofluorescence 7days after the insult. Meanwhile, infarct volume, brain water content and neurological deficit score were assessed. The results illustrated that the infarct volume, brain water content and neurofunctional deficiency were significantly reduced by 7days of breviscapine treatment in MCAO+Bre group, compared with those in MCAO+saline group. Meanwhile, the western blotting, quantitative PCR and immunofluorescence showed that the autophagy in both neurons and astrocytes at the penumbra were markedly attenuated by breviscapine admininstration. Moreover, these pharmacological effects of breviscapine could be counteracted by autophagy inducer Tat-Beclin-1. Our study suggests that breviscapine can provide a neuroprotection against transient focal cerebral ischemia, and this biological function is associated with attenuating autophagy in both neurons and astrocytes.


Galanin family peptides: Molecular structure, expression and roles in the neuroendocrine axis and in the spinal cord.
Galanin is a neurohormone as well as a neurotransmitter and plays versatile physiological roles for the neuroendocrine axis, such as regulating food intake, insulin level and somatostatin release. It is expressed in the central nervous system, including hypothalamus, pituitary, and the spinal cord, and colocalises with other neuronal peptides within neurons. Structural analyses reveal that the human galanin precursor is 104 amino acid (aa) residues in length, consisting of a mature galanin peptide (aa 33-62), and galanin message-associated peptide (GMAP; aa 63-104) at the C-terminus. GMAP appears to exhibit distinctive biological effects on anti-fungal activity and the spinal flexor reflex. Galanin-like peptide (GALP) has a similar structure to galanin and acts as a hypothalamic neuropeptide to mediate metabolism and reproduction, food intake, and body weight. Alarin, a differentially spliced variant of GALP, is specifically involved in vasoactive effect in the skin and ganglionic differentiation in neuroblastic tumors. Dysregulation of galanin, GALP and alarin has been implicated in various neuroendocrine conditions such as nociception, Alzheimer's disease, seizures, eating disorders, alcoholism, diabetes, and spinal cord conditions. Further delineation of the common and distinctive effects and mechanisms of various types of galanin family proteins could facilitate the design of therapeutic approaches for neuroendocrine diseases and spinal cord injury.


Synergistic effect of Bordetella pertussis lymphocytosis-promoting factor on protective activities of isolated Bordetella antigens in mice.
The effect of low levels of added lymphocytosis-promoting factor (LPF) on the ability of several antigenic preparations isolated from Bordetella pertussis and other bacteria to protect mice against intracerebral infection with B. pertussis was examined. LPF was found to enhance the protective activities of filamentous hemagglutinin, 22S antigen, and fimbriae isolated from B. pertussis. Outer membrane protein preparations from phase I B. pertussis which had LPF removed by haptoglobin affinity columns or inactivated by glutaraldehyde, sodium dodecyl sulfate, or Formalin had reduced protective activities but were made fully protective by the readdition of LPF. Similarly, outer membrane protein preparations from Bordetella bronchiseptica, Bordetella parapertussis, or phase IV B. pertussis lacking LPF were protective only when low levels of LPF were added to the preparations. Outer membrane protein preparations from Neisseria gonorrhoeae or Escherichia coli were nonprotective even in the presence of added LPF. The purified LPF by itself was nonprotective unless treated with glutaraldehyde. LPF that had been detoxified with glutaraldehyde was, however, ineffective at enhancing the protective activity of antigenic preparations. The synergistic effect of LPF is discussed in relation to its known biological properties.


Erythromycin stimulates gastric emptying after esophagectomy with gastric replacement: a randomized clinical trial.
Delayed gastric emptying after esophagogastrectomy can pose a significant early postoperative problem. Because erythromycin, which stimulates the gastric antral and duodenal motilin receptor, has been shown to significantly increase gastric emptying in patients with diabetic gastroparesis, we decided to evaluate its effect on gastric emptying after esophagogastrectomy. Twenty-four patients (18 men and six women, age range 41 to 79 years, median 66 years) were randomized to receive either erythromycin lactobionate (200 mg in 50 ml normal saline solution intravenously) (n = 13) or placebo (50 ml normal saline solution intravenously (n = 11) 11 days after esophagogastrectomy (with pyloric drainage procedure). After erythromycin or placebo had been infused over a 15-minute period, patients ingested a solid meal (scrambled egg with bread) labeled with technetium 99m sulfur colloid (500 microCi) over approximately 15 minutes. Dynamic images of the stomach were then acquired over 90 minutes in the supine position by gamma imaging. Results were expressed as percentage of counts retained in the stomach (percent gastric retention) over time. There were no side effects of erythromycin. In the placebo group, the mean percent of radiolabeled meal retained in the stomach after 90 minutes was 88%, which was significantly greater than in the erythromycin group, 37% (p < 0.001). In addition, analysis of covariance demonstrated that the rate of gastric emptying (slope of the line) was significantly greater in the erythromycin-treated group than in the placebo group (p < 0.0001). Early satiety after esophagogastrectomy may be due to delayed gastric emptying and not due to a decrease in the gastric reservoir. Intravenous erythromycin significantly improves gastric emptying in patients after esophagogastrectomy by stimulating gastric motility.


Kaposi's sarcoma with a non-Hodgkin's lymphoma. Its association in a male homosexual with human T-cell lymphotropic virus type III infection.
Combined tumor syndromes, specifically reticuloendothelial malignancies and Kaposi's sarcoma, have long been recognized. With the recognition of the acquired immunodeficiency syndrome (AIDS), several patients with concurrent non-Hodgkin's lymphoma and Kaposi's sarcoma have been reported at high risk for developing AIDS. The present Centers for Disease Control definition of AIDS excludes these patients on the assumption that one tumor is affecting the cellular immunity, allowing for the development of the second malignancy. In evaluating such a patient who had serologic evidence of human T-cell lymphotropic virus type III infection, the probable cause of AIDS, we have reviewed reports of patients with similar concurrent malignancies before and since the onset of the AIDS epidemic. We conclude that patients in high-risk groups for AIDS who develop similar combined tumor syndromes should be classified as having AIDS.


Moderate-intensity statin plus ezetimibe vs high-intensity statin according to baseline LDL-C in the treatment of atherosclerotic cardiovascular disease: A post-hoc analysis of the RACING randomized trial.
Whether the effect of a combination strategy rather than increasing doses of one drug to lower low-density lipoprotein cholesterol (LDL-C) levels is consistent across baseline LDL-C levels remains uncertain. In the RACING trial, which showed a non-inferiority of moderate-intensity statin with ezetimibe (rosuvastatin 10 mg with ezetimibe 10 mg) to high-intensity statin (rosuvastatin 20 mg) for the primary outcome (3-year composite of cardiovascular death, major cardiovascular event, or stroke), the heterogeneity in treatment effect according to baseline LDL-C levels was assessed for the primary and secondary outcomes (clinical efficacy and safety). Of 3780 participants, 2817 participants (74.5%) had LDL-C <100 mg/dL, and 963 participants (25.5%) had LDL-C ≥100 mg/dL. The treatment effect of combination therapy versus high-intensity statin monotherapy was similar among the lower LDL-C subset (8.8% vs. 10.2%; hazard ratio [HR] 0.85, 95% confidence interval [CI] 0.67 to 1.08, p = 0.19) and the higher LDL-C subset (10.8% vs. 9.6 %; HR 1.14, 95% CI 0.76 to 1.7, p = 0.53) without a significant interaction (interaction p = 0.22). Of the secondary outcomes, the 1-, 2-, and 3-year achievement of LDL-C <70 mg/dL was greater in the combination therapy group regardless of baseline LDL-C levels. Among ASCVD patients, there was no heterogeneity in the effect of moderate-intensity statin plus ezetimibe combination therapy in the higher and lower baseline LDL-C levels for the 3-year composite of cardiovascular outcomes.


Loss of lactose metabolism in lactic streptococci.
Lactose-negative mutants occurred spontaneously in broth cultures of Streptococcus lactis C(2)F. Instability of lactose metabolism was noted in other strains of S. lactis, in strains of S. cremoris, and in S. diacetilactis. Colonies of S. lactis C(2)F grown with lactose as the carbohydrate source also possessed lac(-) cells. Treatment of lactic streptococci with the mutagen acriflavine (AF) increased the number of non-lactose-fermenting variants. The effect of AF on growth and on loss of lactose-fermenting ability in S. lactis C(2)F was consequently further examined. The presence of AF appears to favor competitively the growth of spontaneously occurring lactose-negative cells and appears to act in the conversion of lactose-positive to non-lactose-fermenting cells. The lactose-negative mutants partially revert to lactose-positive variants which remain defective in lactose metabolism and remain unable to coagulate milk. The lactose-negative cells become dominant in continuous culture growth and provide evidence that alterations in the characteristics of starter strains can be produced by continuous culture, in this case, the complete loss in ability to ferment lactose.


Talking With white Clients About Race.
Most white people do not believe that race is an important feature of their lives, and this belief continues into the therapy room where race is rarely a topic of conversation, especially for all-white dyads. However, research shows that race and racism are highly salient for white people's mental health, and this gap in understanding has negative effects on the well-being of both white people and people of color. This paper argues to embrace the ethical and moral call to actively address race and racism in therapy between white therapists and white clients. This embrace can be particularly challenging for white therapists who believe themselves to be social justice-oriented people, but who nevertheless contribute to racism in both conscious and unconscious ways. A model is offered for how psychotherapists can bring up and work with the topics of race and racism during the course of therapy. The model includes ways for white therapists to engage in the long-term process of self-critique, ways to introduce the salience of race in the white client's life, and how to connect race and racism to the client's explicit goals for therapy. Finally, a case example is explored using a well-meaning, self-defined liberal white client.


Medical diagnostic radiation exposures and risk of gliomas.
High-dose ionizing radiation is an established risk factor for glioma, but it remains unknown whether moderate- and low-dose radiation increase glioma risk. In this analysis, we assessed the evidence that self-reported exposures to diagnostic ionizing radiation, including computerized tomography (CT) scans, is associated with increased risk of adult glioma. While no independent association was observed for CT scans alone (3+ scans compared to none P = 0.08 and 1-2 scans compared to none P = 0.68), our findings suggest an increased risk of adult gliomas with cumulative exposure to three or more CT scans to the head and neck region (OR = 1.97, 95% CI: 0.92-4.23) limited to those who reported a family history of cancer: the P value for the interaction between having three or more CT scans and family history of cancer was 0.08. The stratum-specific adjusted OR for those with family history of cancer was more than three times that for the sub-group without family history of cancer. While there is some potential for symptom-related bias, one might expect this to be present for all diagnostic procedures rather than specific to one procedure. The interaction between CT scans and glioma with family history of cancer supports the biological plausibility of our findings, because similar results have been found for breast cancer and radiation. This observational data will increase awareness about potential risks associated with CT scans and the need to minimize the use of unnecessary examinations.


In situ hybridization detection and subtyping of rotaviruses in swine samples.
Rotavirus groups A, B, and C (RVA, RVB, and RVC, respectively) have been the most prevalent and pathogenic in pigs. To date, immunohistochemistry is only available for RVA because of the lack of commercial antibodies for RVB and RVC. We developed a novel in situ hybridization RNA-based chromogenic technique (ISH-RNA) to detect and subtype RVA, RVB, and RVC. We evaluated 33 samples that were reverse-transcription PCR positive for RVA, RVB, and/or RVC. ISH-RNA was able to detect as few as 103 RV RNA copies/mL. The new ISH-RNA test can be useful for routine investigation of rotavirus enteritis in order to guide strategies for control of the infection in pigs, but a full validation study needs to be completed. Pathogenesis studies may be conducted using ISH-RNA based on the identification of replicating virus.


Changes in expression and activity of the secretory pathway Ca2+ ATPase 1 (SPCA1) in A7r5 vascular smooth muscle cells cultured at different glucose concentrations.
Diabetes mellitus-related vascular disease is often associated with both a dysregulation of Ca2+ homoeostasis and enhanced secretory activity in VSMCs (vascular smooth muscle cells). Here, we employ a commonly used rat cell line for VSMCs (A7r5 cells) to investigate the effects of glucose on the expression and activity of the SPCA1 (secretory pathway Ca2+-ATPase 1; also known as ATP2C1), which is a P-type Ca2+ pump located in the Golgi apparatus that plays a key role in the secretory pathway. Our results show that mRNA expression levels of SPCA1 are significantly increased in A7r5 cells cultured in high glucose (25.0 mM)-supplemented medium compared with normal glucose (5.55 mM)-supplemented medium. SPCA1 protein expression levels and thapsigargin-insensitive Ca2+-dependent ATPase activity were also consistent with a higher than normal expression level of SPCA1 in high-glucose-cultured A7r5 cells. Analysis of AVP (arginine-vasopressin)-induced cytosolic Ca2+ transients in A7r5 cells (after pre-treatment with thapsigargin) showed faster rise and decay phases in cells grown in high glucose medium compared with cells grown in normal glucose medium, supporting the observation of increased SPCA expression/activity. The significant levels of both Ca2+-ATPase activity and AVP-induced Ca2+ transients, in the presence of thapsigargin, indicate that SPCA must play a significant role in Ca2+ uptake within VSMCs. We therefore propose that, if such increases in SPCA expression and activity also occur in primary VSMCs, this may play a substantial role in the aetiology of diabetes mellitus-associated vascular disease, due to alterations in Ca2+ homoeostasis within the Golgi apparatus.


Efficient Estimation of the Human Circadian Phase via Kalman Filtering.
Circadian rhythms play a vital role in maintaining a person's well-being but remain difficult to quantify accurately. Numerous approaches exist to measure these rhythms, but they often suffer from performance issues on the individual level. This work implements a Steady-State Kalman Filter as a method for estimating the circadian phase shifts from biometric signals. Our framework can automatically fit the filter's parameters to biometric data obtained for each individual, and we were able to consistently estimate the phase shift within 1 hour of melatonin estimates on 100% of all subjects in this study. The estimation method opens up the possibility of real-time control and assessment of the circadian system, as well as chronotherapeutic intervention.Clinical relevance- This establishes a near real-time alternative to melatonin measurements for the estimation of circadian phase shifts, with potential applications in feedback circadian control and chronotherapeutics.


Behavioral problems and emotional stress in children with bruxism.
Bruxism has a multifactorial etiology, and psychosocial factors have been considered to increase the risk of occurrence of this parafunction. The aim of this study was to evaluate the behavior profile of a group of children diagnosed with bruxism. Eighty 7-11-year-old children of both genders (mean age 8.8 years) first recruited as eligible participants. Twenty-nine children (18 males and 11 females) whose parents/guardians reported to present frequent episodes of tooth grinding/clenching while awake or during sleep (at least 3 nights a week) in the previous 3 months were enrolled in the study. The diagnosis of bruxism was established based on the parents/guardians' report about the children's behavior, habits and possible discomforts in the components of the stomatognathic system allied to the presence of signs and symptoms such as pain on the masticatory muscles, masseter muscle hypertrophy, wear facets, fractures of restorations, dental impressions on the cheek mucosa and tongue. As part of the psychological evaluation, the Rutter's Child Behavior Scale-A2 was applied to the parents/caregivers (one for each child) and the Child Stress Scale was applied to the children. Data were analyzed descriptively based on the frequency of each studied variable. Twenty-four (82.76%) children needed psychological or psychiatric intervention; 17 of them presented neurotic disorders and 7 children presented antisocial disorders. Six (20.70%) children presented significant physical and psychological manifestations of stress. The findings of the present study suggest that behavioral problems and potential emotional problems can be risk factors to bruxism in children.


Molecular basis of mast cell disease.
Mastocytosis is an incurable and sometimes fatal haematological disorder grossly described as the accumulation of abnormal mast cells in the bone marrow and other organs causing tissue and organ damage. The clinical manifestations of this disease are extremely variable; disease phenotypes range from indolent to aggressive, and often present with associated non-mast cell haematological disorders (AHNMD), mainly myeloproliferative neoplasm and myelodysplastic syndromes. Recent efforts to genetically dissect the mechanisms that define aggressive and non-aggressive mastocytosis have generated a list of recurrent somatic mutations in mastocytosis patients that are associated with and may predict the evolution towards aggressive disease phenotypes. Here we review these mutations and discuss the molecular mechanisms associated with these mutations in an effort to better understand the biology of this disease and to predict its onset and evolution, with the ultimate goal of devising new and improved treatment strategies.


[Intensive therapy for multiple organ dysfunction in neonatal infants after cardiosurgical interventions].
This study was undertaken to evaluate the efficiency of complex intensive therapy for multiple organ dysfunction syndrome (MODS) after cardiosurgical interventions at the resuscitative and intensive care unit of the A. N. Bakulev Research Center of Cardiovascular Surgery, Russian Academy of Medical Sciences. In 2003-2004, MODS developed in 70 (37%) of the neonatal infants operated on the heart and vessels. The babies' age ranged from 6 hours of life to 1 month (8.3 +/- 2.1 days of life, their body weight was from 1.7 to 4.1 kg (3.0 +/- 0.49 kg). All the patients were found to have significant renal and respiratory failures. There were more than 4 (4.1 +/- 0.5) failing vital viscera. The use of phosphodiesterase (III) inhibitors in therapy for acute left ventricular insufficiency significantly improved the performance of the left heart whereas nitric oxide inhalation significantly lowered pulmonary pressure in babies with acute right ventricular insufficiency and improved oxygenation in patients with MODS. The efficiency of nitric oxide inhalation in MODS significantly increased when it was used in combination with endotracheal administration of a surfactant and high-frequency oscillatory ventilation. Peritoneal dialysis effectively replaced renal function when acute renal failure (ARF) developed. Nevertheless, the development of ARF in the pattern of MODS is a marker of high mortality (89% in ARF versus 46% in MODS without ARF).


Direct inhibition of arginase attenuated airway allergic reactions and inflammation in a Dermatophagoides farinae-induced NC/Nga mouse model.
The expression of arginase I has been a focus of research into the pathogenesis of experimental asthma, because arginase deprives nitric oxide synthase (NOS) of arginine and therefore participates in the attenuation of bronchodilators such as nitric oxide (NO). The present study used an intranasal mite-induced NC/Nga mouse model of asthma to investigate the contribution of arginase to the asthma pathogenesis, using an arginase inhibitor, N(omega)-hydroxy-nor-l-arginine (nor-NOHA). The treatment with nor-NOHA inhibited the increase in airway hyperresponsiveness (AHR) and the number of eosinophils in bronchoalveolar lavage fluid. NOx levels in the lung were elevated despite suppressed NOS2 mRNA expression. Accompanied by the attenuated activity of arginase, the expression of arginase I at both the mRNA and protein level was downregulated. The levels of mRNA for T helper 2 cytokines such as IL-4, IL-5, and IL-13, and for chemotactants such as eotaxin-1 and eotaxin-2, were reduced. Moreover, the accumulation of inflammatory cells and the ratio of goblet cells in the bronchiole were decreased. The study concluded that the depletion of NO caused by arginase contributes to AHR and inflammation, and direct administration of an arginase inhibitor to the airway may be beneficial and could be of use in treating asthma due to its anti-inflammatory and airway-relaxing effects, although it is not clear whether the anti-inflammatory effect is direct or indirect.


Evaluating the use of pathology in improving diagnosis in rural Malawi.
Limited data exists on histologically confirmed cancers and tuberculosis in rural Malawi, despite the high burden of both conditions. One of the main reasons for the limited data is the lack of access to pathology services for diagnosis. We reviewed histopathology results of patients in Neno District, one of the poorest rural districts in Malawi, from May 2011 to July 2017, with an emphasis on cancers and tuberculosis. This is a retrospective descriptive study reviewing pathology results of samples collected at Neno health facilities and processed at Kamiza Pathology Laboratory. Data was entered into Microsoft Excel and cleaned and analysed using Stata 14. A total of 532 specimens were collected, of which 87% (465) were tissue biopsies (incision or core biopsies), and 13% (67) were cytology samples. Of all specimens, 7% (n=40) of the samples had non-diagnostic results. Among the results that were diagnostic (n=492), 37% (183) were malignancies, 33% (112) were infections and inflammatory conditions other than tuberculosis, 20% (97) were benign tumours, 7% (34) were tuberculosis, 4% (21) were pre-malignant lesions, 5% (23) were normal samples, and 4% (22) were other miscellaneous conditions. Among the malignancies (n=183), 62% (114) were from females and 38% (69) from males. Among females, almost half of the cancers were cervical (43%, n= 49), followed by Kaposi sarcoma (14%, n=16), skin cancers (9%, n=10), and breast cancer (8%, n=9). In males, Kaposi sarcoma was the most common cancer (35%, n=24), followed by skin cancers (17%, n=12). About 75% (n=137) of the cancers occurred in persons aged 15 to 60 years. Histopathology services at a rural hospital in Malawi provides useful diagnostic information on malignancies, tuberculosis and other diagnoses, and can inform management at the district level.


Identification of mouse CPX-1, a novel member of the metallocarboxypeptidase gene family with highest similarity to CPX-2.
The recent finding that Cpe(fat)/Cpe(fat) mice, which lack carboxypeptidase E (CPE) activity because of a point mutation, are still capable of a reduced amount of neuroendocrine peptide processing suggested that additional carboxypeptidases (CPs) participate in this processing reaction. Searches for novel members of the CPE gene family led to the discovery of CPD, CPZ, AEBP1, and CPX-2. In the present report, we describe mouse CPX-1, another novel member of this gene family. Like AEBP1 and CPX-2, CPX-1 contains an N-terminal region of 160 amino acids with sequence similarity to the discoidin domain of a variety of proteins. The 410-residue CP-like domain of CPX-1 has 54% to 62% amino acid sequence identity with AEBP1 and CPX-2 and 33% to 49% amino acid identity with other members of the CPE subfamily. However, several active-site residues that are important for catalytic activity of other CPs are not conserved in CPX-1. Furthermore, CPX-1 expressed in either the baculovirus system or the mouse AtT-20 cell line does not cleave standard CP substrates. Northern blot analysis showed the highest levels of CPX-1 mRNA in testis and spleen and lower levels in salivary gland, brain, heart, lung, and kidney. In situ hybridization of CPX-1 mRNA in embryonic and fetal mouse tissue showed expression throughout the head and thorax, with abundance in primordial cartilage and skeletal structures. In the head, high levels of CPX-1 mRNA were associated with the nasal mesenchyme, primordial cartilage structures in the ear, and the meninges. In the thorax, CPX-1 mRNA was expressed in multiple developing skeletal structures, including chondrocytes and perichondrial cells of the rib, vertebral, and long-bone primordia. Taken together, these findings suggest that it is unlikely that CPX-1 functions in the processing of neuroendocrine peptides. Instead, CPX-1 may have a role in development, possibly mediating cell interactions via its discoidin domain.


Effects of Anodal tDCS on Arithmetic Performance and Electrophysiological Activity.
Arithmetic abilities are among the most important school-taught skills and form the basis for higher mathematical competencies. At the same time, their acquisition and application can be challenging. Hence, there is broad interest in methods to improve arithmetic abilities. One promising method is transcranial direct current stimulation (tDCS). In the present study, we compared two anodal tDCS protocols in their efficacy to improve arithmetic performance and working memory. In addition, we investigated stimulation-related electrophysiological changes. Three groups of participants solved arithmetic problems (additions and subtractions) and an n-back task before, during, and after receiving either frontal or parietal anodal tDCS (25 min; 1 mA) or sham stimulation. EEG was simultaneously recorded to assess stimulation effects on event-related (de-) synchronisation (ERS/ERD) in theta and alpha bands. Persons receiving frontal stimulation showed an acceleration of calculation speed in large subtractions from before to during and after stimulation. However, a comparable, but delayed (apparent only after stimulation) increase was also found in the sham stimulation group, while it was absent in the group receiving parietal stimulation. In additions and small subtractions as well as the working memory task, analyses showed no effects of stimulation. Results of ERS/ERD during large subtractions indicate changes in ERS/ERD patterns over time. In the left hemisphere there was a change from theta band ERD to ERS in all three groups, whereas a similar change in the right hemisphere was restricted to the sham group. Taken together, tDCS did not lead to a general improvement of arithmetic performance. However, results indicate that frontal stimulation accelerated training gains, while parietal stimulation halted them. The absence of general performance improvements, but acceleration of training effects might be a further indicator of the advantages of using tDCS as training or learning support over tDCS as a sole performance enhancer.


Transition of Individuals With Autism to Adulthood: A Review of Qualitative Studies.
Many young adults with autism spectrum disorder experience poor transition outcomes in key areas, including postsecondary employment, higher education, health care, social connectedness, and independent living, yet we lack a clear understanding of the specific factors that impact these outcomes. We reviewed qualitative research in which the perspectives of youth and young adults with autism spectrum disorder, parents, services providers, and other stakeholders were gathered to identify barriers and facilitators to optimal outcomes. Findings revealed that poor transition outcomes are influenced by several factors, including poor person-environment fit, uncertainty about the roles of parents, and the lack of comprehensive or integrated services. These findings also revealed the aspects of familial, organizational, and policy contexts that may be targeted for interventions. Finally, stakeholders emphasized that supports should be individualized and focused on the changing aspects of the young adult's social and physical environment rather than behavior change. We discuss implications for policy and practice and provide recommendations for further research.


Long-Term Outcomes of Laparoscopic Distal Gastrectomy for Locally Advanced Gastric Cancer: The KLASS-02-RCT Randomized Clinical Trial.
It is unclear whether laparoscopic distal gastrectomy for locally advanced gastric cancer is oncologically equivalent to open distal gastrectomy. The noninferiority of laparoscopic subtotal gastrectomy with D2 lymphadenectomy for locally advanced gastric cancer compared with open surgery in terms of 3-year relapse-free survival rate was evaluated. A phase III, open-label, randomized controlled trial was conducted for patients with histologically proven locally advanced gastric adenocarcinoma suitable for distal subtotal gastrectomy. The primary end point was the 3-year relapse-free survival rate; the upper limit of the hazard ratio (HR) for noninferiority was 1.43 between the laparoscopic and open distal gastrectomy groups. From November 2011 to April 2015, 1,050 patients were randomly assigned to laparoscopy (n = 524) or open surgery (n = 526). After exclusions, 492 patients underwent laparoscopic surgery and 482 underwent open surgery and were included in the analysis. The laparoscopy group, compared with the open surgery group, suffered fewer early complications (15.7% v 23.4%, respectively; P = .0027) and late complications (4.7% v 9.5%, respectively; P = .0038), particularly intestinal obstruction (2.0% v 4.4%, respectively; P = .0447). The 3-year relapse-free survival rate was 80.3% (95% CI, 76.0% to 85.0%) for the laparoscopy group and 81.3% (95% CI, 77.0% to 85.0%; log-rank P = .726) for the open group. Cox regression analysis after stratification by the surgeon revealed an HR of 1.035 (95% CI, 0.762 to 1.406; log-rank P = .827; P for noninferiority = .039). When stratified by pathologic stage, the HR was 1.020 (95% CI, 0.751 to 1.385; log-rank P = .900; P for noninferiority = .030). Laparoscopic distal gastrectomy with D2 lymphadenectomy was comparable to open surgery in terms of relapse-free survival for patients with locally advanced gastric cancer. Laparoscopic distal gastrectomy with D2 lymphadenectomy could be a potential standard treatment option for locally advanced gastric cancer.


DNA motif representation with nucleotide dependency.
The problem of discovering novel motifs of binding sites is important to the understanding of gene regulatory networks. Motifs are generally represented by matrices (position weight matrix (PWM) or position specific scoring matrix (PSSM) or strings. However, these representations cannot model biological binding sites well because they fail to capture nucleotide interdependence. It has been pointed out by many researchers that the nucleotides of the DNA binding site cannot be treated independently, e.g. the binding sites of zinc finger in proteins. In this paper, a new representation called Scored Position Specific Pattern (SPSP), which is a generalization of the matrix and string representations, is introduced which takes into consideration the dependent occurrences of neighboring nucleotides. Even though the problem of discovering the optimal motif in SPSP representation is proved to be NP-hard, we introduce a heuristic algorithm called SPSP-Finder, which can effectively find optimal motifs in most simulated cases and some real cases for which existing popular motif finding software, such as Weeder, MEME and AlignACE, fail.


DAX-1A (NR0B1A) expression levels are extremely low compared to DAX-1 (NR0B1) in human steroidogenic tissues.
The orphan nuclear receptor DAX-1 (dosage-sensitive sex reversal-AHC critical region on the X chromosome gene 1; NR0B1) is known for its role in human development, specifically sex determination and steroidogenesis. Several recent publications have described an alternatively spliced form of DAX-1 called DAX-1A ( NR0B1A). DAX-1A is encoded by exons 1 and 2A of DAX-1, with exon 2A located within the DAX-1 intron 1. DAX-1A expression has been observed in several tissues, including adrenal gland, ovary, and testis. Transfection studies have further shown that DAX-1A has an inhibitory effect on DAX-1, suggesting a role for DAX-1A in the regulation of adrenal and gonadal differentiation/function. However, the relative level of DAX-1 versus DAX-1A transcripts still remains unclear. Herein, we developed and performed quantitative real-time RT-PCR to measure DAX-1 and DAX-1A mRNA expression levels in H295R human adrenal carcinoma cell lines, human adult and fetal adrenal glands, corpus luteum, testis, whole pre- and postmenopausal ovaries, ovarian follicles, placenta, liver, and kidney. These mRNA expression levels were quantified using DAX-1 and DAX-1A standard curves. In addition, Western blotting analysis was performed to examine both DAX-1 and DAX-1A protein levels in H295R cells, adrenal glands, corpus luteum, and liver. Both DAX-1 and DAX-1A mRNA were detected in all samples of H295R cells, human fetal and adult adrenals, testis, ovary, ovarian follicles, and corpus luteum. However, DAX-1 mRNA levels were significantly higher (> 37-fold) than that seen for DAX-1A (p<0.01). DAX-1A mRNA expression levels were undetectable in human liver, placenta, and kidney. Western blotting analysis results demonstrated that DAX-1 protein was predominantly expressed in H295R cells, human adult adrenal, and corpus luteum. These results suggest that in comparison to DAX-1A, DAX-1 is, by far, the predominant mRNA isoform found in human adrenal glands and gonads.


Reduction of claudin-5 and aquaporin-4 in the rat hippocampal CA-1 and CA-3 regions of a learned helplessness model of depression.
Although findings from both animal and clinical research indicate that the blood-brain barrier (BBB) contributes to the pathogenesis of various psychiatric disorders (including depression), the underlying mechanisms are unknown. We investigated the levels of the tight-junction proteins claudin-5 and aquaporin-4 (AQP-4) in astrocytes of learned helplessness (LH) rats (an animal model of depression) and non-LH rats (a model of resilience). We administered inescapable mild electric shock to rats and then identified the LH and non-LH rats by a post-shock test. The expressions of claudin-5 and AQP-4 in several brain regions of the LH and non-LH rats were then evaluated by a western blot analysis. The levels of both claudin-5 and AQP-4 in the CA-1 and CA-3 hippocampal areas of the LH group were significantly lower than those of the control group, whereas those of the non-LH rats were not significantly different from those of the control and LH rats. These results suggest that LH rats but not non-LH rats experienced down-regulations of claudin-5 and AQP-4 in the CA-1 and CA-3. It is possible that a region-specific modulation of claudin-5 and AQP-4 is involved in the mechanisms of vulnerability but not resilience in depression.


Effects of training on timbre recognition and appraisal by postlingually deafened cochlear implant recipients.
The purpose of this study was to compare the effect of structured training on recognition and appraisal of the timbre (tone quality) of musical instruments by postlingually deafened cochlear implant recipients. Twenty-four implant users (Nucleus CI24M) were randomly assigned to a control or a training group. The control group experienced only incidental exposure to music in their usual daily routine. The training group participated in 12 weeks of training delivered via a laptop computer in which they were introduced to excerpts of musical instruments representing three frequency ranges and four instrumental families. Those implant recipients assigned to the training group showed significant improvement in timbre recognition (p < .0001) and timbre appraisal (p < .02) compared to the control group. Correlations between timbre measures and speech perception measures are discussed.


Ambiguity in knowledge transfer: The role of theory-practice gap.
In spite of much literature written about the theory-practice gap in the international nursing journals, there is evidence that indicates this subject has not been probed comprehensively since nursing education was transferred to universities in Iran. In the recent years, the public and the government have criticized Iranian nurses because of poor quality of patient care. Although this subject has been lamented by some researchers, there is no comprehensive work on how this gap resulted. In the process of a larger study on "nursing knowledge translation to practice", of one PhD thesis, this process was explored. Using grounded theory analysis, indepth interviews were undertaken with a purposive sample of 29 nurses, with different levels of experience, from the school of nursing in Tehran University of Medical Sciences in 2006 from January to August. Data were analyzed using the constant comparative method. Three main themes emerging from this study included clinical behavior structure, paradoxical knowledge and practice, and divergent nursing organization. It seems that nursing education with some praxis and paradoxes in the realm of nursing knowledge and practice, along with divergent organizational structure have decreased nurses' ability in applying their professional knowledge and skills in order to bridge the gap between theory and practice. Moreover, in spite of increased academic input into nursing education, clinical behaviors of both education and practice settings was perceived as "traditional routine-based".


Liquor licences issued to Australian schools.
Children's positive socialisation to alcohol is associated with early initiation of drinking and alcohol-related harm in adult life. Internationally, there have been reports of adults' alcohol consumption at school events in the presence of children. The aim of this research was to identify the conditions under which Australian schools are required to apply for a liquor licence and the associated prevalence of liquor licences for these events where children were likely to be present. A document review was conducted to examine temporary liquor licensing legislation. Quantitative analysis was used to examine relevant licensing data. Coding criteria was developed to determine school type, student year levels and the likely presence of children. Four jurisdictions provided data on 1817 relevant licences. The average annual licences/100 schools was highest amongst Independent schools followed by Catholic and public (government) schools. The rates were highest in Queensland and Victoria where children were present at 61% and 32% of events respectively. While there are legislative differences across jurisdictions, the prevalence of adults' alcohol use at school events in the presence of children may reflect the various education department policies and principals' and school communities' beliefs and attitudes. Licences are not required for all events where liquor is consumed so the prevalence of adults' use of alcohol at school events is likely to be higher than our analyses imply. Such practices may undermine teaching about alcohol use in the school curriculum and health promotion efforts to develop alcohol-free events when children are present.


A method for calculation of dose per unit concentration values for aquatic biota.
A dose per unit concentration database has been generated for application to ecosystem assessments within the FASSET framework. Organisms are represented by ellipsoids of appropriate dimensions, and the proportion of radiation absorbed within the organisms is calculated using a numerical method implemented in a series of spreadsheet-based programs. Energy-dependent absorbed fraction functions have been derived for calculating the total dose per unit concentration of radionuclides present in biota or in the media they inhabit. All radionuclides and reference organism dimensions defined within FASSET for marine and freshwater ecosystems are included. The methodology has been validated against more complex dosimetric models and compared with human dosimetry based on ICRP 72. Ecosystem assessments for aquatic biota within the FASSET framework can now be performed simply, once radionuclide concentrations in target organisms are known, either directly or indirectly by deduction from radionuclide concentrations in the surrounding medium.


[Peroxidation of lecithin in the presence of dihydroquercetin and its complex with ferrous iron ions].
The ability of the flavonoid dihydroquercetin to prevent or accelerate the accumulation of reactive oxygen species and the metabolites of oxidative stress, carbonyl compounds has been studied. It has been shown on a model of oxidation of lecithin that dihydroquercetin exhibits a prooxidant effect in the alkaline region of pH, whereas at neutral and acidic pH values dihydroquercetin is an effective antioxidant. In the presence of ferrous iron ions, which catalyze the Fenton's reaction, dihydroquercetin forms a complex with metal that shows the antioxidant activity in the region of high pH values. It has been found that the oxidation of lecithin in the presence of 20-200 microM ferrous iron is inhibited by dihydroquercetin to a concentration of 3.2 mM. At higher concentration of dihydroquercetin in the presence of ferrous iron, the accumulation of malonic dialdehyde occurs, indicating the presence of the prooxidant activity of dihydroquercetin.


Metformin treatment of polycystic ovary syndrome improves health-related quality-of-life, emotional distress and sexuality.
In polycystic ovary syndrome (PCOS), changes in physical appearance, menstrual disturbances and infertility result in psychological distress and reduced quality-of-life. Metformin improves biochemical, clinical and reproductive parameters in PCOS women. In a prospective, observational study, we analysed the effects of metformin treatment on health-related quality-of-life (HRQL), emotional well-being and sexuality in PCOS. No placebo-treated control group was included. Before, during and after 6 months of treatment, changes in clinical and endocrine parameters, quality-of-life, psychological disturbances and sexuality were assessed in 64 PCOS patients using validated questionnaires (SF-36, SCL-90-R) and visual analogue scales. Patients were also compared with published normative data for the validated questionnaires. During treatment, HRQL, particularly the psychosocial aspects (indicated by significant increases in SF-36 scales Vitality, Social Function, Emotional Role Function, Mental Health, Psychological Sum scale) and emotional well-being (reflected by significant lowering of SCL-90-R scales) improved. These improvements in HRQL were significantly correlated with a reduction in body weight and significantly more pronounced in patients with normalized menstrual cycles. In addition, PCOS women were significantly more satisfied with their sex life and reported higher frequencies of sexual intercourse following treatment. Treatment can improve the psychosocial, emotional and psychosexual situation of PCOS patients. Although at least some of these effects may be related to the reduction of individual clinical symptoms (i.e. weight loss, normalization of menstrual disturbances, improvement of acne), this observational study does not allow us to clearly discern the role of symptom constellation and does not preclude non-specific and/or placebo effects. Nevertheless, emotional distress and reduced quality-of-life are clearly not an inevitable consequence of PCOS and should be considered as adjunct treatment goals in future studies.


Involvement of the Reck tumor suppressor protein in maternal and embryonic vascular remodeling in mice.
Developmental angiogenesis proceeds through multiple morphogenetic events including sprouting, intussusception, and pruning. Mice lacking the membrane-anchored metalloproteinase regulator Reck die in utero around embryonic day 10.5 with halted vascular development; however, the mechanisms by which this phenotype arises remain unclear. We found that Reck is abundantly expressed in the cells associated with blood vessels undergoing angiogenesis or remodelling in the uteri of pregnant female mice. Some of the Reck-positive vessels show morphological features consistent with non-sprouting angiogenesis. Treatment with a vector expressing a small hairpin RNA against Reck severely disrupts the formation of blood vessels with a compact, round lumen. Similar defects were found in the vasculature of Reck-deficient or Reck conditional knockout embryos. Our findings implicate Reck in vascular remodeling, possibly through non-sprouting angiogenesis, in both maternal and embyonic tissues.


Effects of state-level public spending on health on the mortality probability in India.
This study uses the second National Family Health Survey of India to estimate the effect of state-level public health spending on mortality across all age groups, controlling for individual, household, and state-level covariates. We use a state's gross fiscal deficit as an instrument for its health spending. Our study shows a 10% increase in public spending on health in India decreases the average probability of death by about 2%, with effects mainly on the young, the elderly, and women. Other major factors affecting mortality are rural residence, household poverty, and access to toilet facilities.


Quantity and quality of physical activity are influenced by outdoor temperature in people with knee osteoarthritis.
Environmental factors are known to influence physical activity (PA) levels of healthy people, but little work has explored these factors in people with osteoarthritis (OA). The purpose of this study was to explore the influence of outdoor temperature on PA in people with knee OA. A total of 38 people with knee OA (10 women; mean age 54 y) wore an accelerometer around their waist for 7 consecutive days. Dependent variables from the accelerometers were three PA measures: (1) activity counts/day and time spent at or above moderate levels of PA with (2) a cut-point of 1,041 activity counts/minute (MVPA1041) and (3) a cut-point of 1,952 activity counts/minute (MVPA1952). Independent variables were age, sex, body mass index (BMI), and maximum daily outdoor temperature. Three linear regression analyses were conducted using the three PA dependent variables and independent variables. After controlling for age, sex, and BMI, maximum daily outdoor temperature explained 9% of the variance in activity counts/day (p=0.042), 10% of variance in MVPA1041 (p=0.032), and 14% of variance in MVPA1952 (p=0.016). Participants who engaged in more PA were younger and were exposed to warmer temperatures. Outdoor temperature and age influence the PA levels of people with knee OA. These factors should be considered when designing PA programmes.


Ruptured Distal Accessory Anterior Cerebral Artery Aneurysm: A case report and literature review.
Aneurysms of the distal anterior cerebral artery (dACA) are rare, consisting only 2-6% of all intracranial aneurysms. dACA aneurysms are often associated with anomalies such as azygous, bihemispheric and triplication of ACA. Among these anomalies accessory anterior cerebral artery (accACA) is an anatomical variant found in 3.3% to 15% of the population. Here we report a case of a 60-year-old female who presented with a Hunt and Hess grade II, Fisher grade III subarachnoid hemorrhage secondary to a ruptured saccular aneurysm originating from the pericallosal-callosomarginal artery bifurcation at the A3 segment of the accessory ACA and underwent an uncomplicated microsurgical clipping.


Effect of oxygen limitations on monoclonal antibody production by immobilized hybridoma cells.
The productivity of an immobilized cell biocatalyst is often limited by the amount of oxygen that reaches cells located at interior regions of the biocatalyst. These diffusive limitations depend on a multitude of factors including the oxygen supply, the cellular uptake kinetics, and the cell density of the material. Large cell densities, which are desired for high productivity, are also likely to reduce the percentage of cells that receive an adequate supply of oxygen. To develop a better understanding of how different conditions affect biocatalyst behavior, a computational model of immobilized hybridoma cells was developed. The model accounts for oxygen diffusion and consumption, cell proliferation and death, and monoclonal antibody production. This model assumes that cellular productivity is limited only by the supply of oxygen and that the growth media is continually replenished so that nutrient levels remain high and wastes are eliminated. Biocatalyst performance is evaluated by monitoring the amount of monoclonal antibody produced by the cells. Model predictions agree with experimental measurements reported in the literature and indicate that for long operation time the supply of oxygen, biocatalyst size, and cell kinetics have a significant effect on biocatalyst performance, whereas the initial cell loading has only a relatively small effect. Under typical culture conditions, we find that oxygen penetrates to a maximum depth of about 0.4 mm. Accordingly, cells immobilized farther than this threshold distance receive an insufficient supply of oxygen.


Fetuin-A serum levels are not correlated to kidney function in long-lived subjects.
Serum Fetuin A has been identified as an inhibitor of ectopic calcification. It is reduced in subjects with chronic kidney disease (CKD) and it has been proposed as a potential link between CKD and the higher prevalence of arterial calcification observed in these patients. During aging both the stiffening of arterial wall due to calcification and a decline in kidney function are frequent. The aim of the study is to investigate if Fetuin A serum levels are associated with aging and with AHSG T256S polymorphism. Moreover, we aim at investigate whether serum Fetuin A is correlated to kidney function in this setting of senescence. 256 health long-lived subjects (age 92 [81-100]) were recruited for the study. Serum Fetuin A was evaluated by ELISA, Cystatin C by immune-nephelometry. AHSG T256S was determinated by PCR-RFLP. Serum Fetuin A shows a significant correlation with age (r=0.20; P=0.0048). AHSG TS and SS genotypes are associated to lower levels of serum protein (0.27 [0.19-0.29] g/L vs 0.42 [0.32-0.49] g/L; P<0.027 and 0.34 [0.25-0.41] g/L vs 0.42 [0.32-0.49] g/L; P<0.001, respectively). No significant correlation between Fetuin A and Cystatin C was observed. Serum Fetuin A increases with age in elder individuals and subjects with the TS or SS AHSG polymorphism have lower levels of the circulating protein. No correlation with kidney function decline was observed. Other mechanisms should be investigated to explain the increase of Fetuin A with age.


Poly(ester amide)s from Poly(ethylene terephthalate) Waste for Enhancing Bone Regeneration and Controlled Release.
The present study elucidates the facile synthesis and exceptional properties of a family of novel poly(ester amide)s (PEAs) based on bis(2-hydroxy ethylene) terephthalamide that was obtained from the poly(ethylene terephthalate) waste. Fourier transform infrared and 1H NMR were used to verify the presence of ester and amide in the polymer backbone. Differential scanning calorimetry data showed that the glass transition temperature decreased with as the chain length of dicarboxylic acids increased. Dynamic mechanical analysis and contact angle studies proved that the modulus values and hydrophobicity increased with as the chain lengths of dicarboxylic acids increased. In vitro hydrolytic degradation and dye release studies demonstrated that the degradation and release decreased with as the chain lengths of dicarboxylic acids increased. Modeling these data illustrated that degradation and release follow first-order degradation and zero-order release, respectively. The in vitro cytocompatibility studies confirmed the minimal toxicity characteristic of these polymers. Osteogenic studies proved that these polymers can be highly influential in diverting the cells toward osteogenic lineage. Alizarin red staining evinced the presence of twice the amount of calcium phosphate deposits by the cells on these polymers when compared to the control. The observed result was also corroborated by the increased expression of alkaline phosphatase. These findings were further validated by the markedly higher mRNA expressions for known osteogenic markers using real time polymerase chain reaction. Therefore, these polymers efficiently promoted osteogenesis. This study demonstrates that the physical properties, degradation, and release kinetics can be altered to meet the specific requirements in organ regeneration as well as facilitate simultaneous polymer resorption through control of the chain length of the monomers. The findings of this study have significant implications for designing cost-effective biodegradable polymers for tissue engineering.


Controlling the angiogenic switch in developing atherosclerotic plaques: possible targets for therapeutic intervention.
Plaque angiogenesis may have an important role in the development of atherosclerosis. Vasa vasorum angiogenesis and medial infiltration provides nutrients to the developing and expanding intima and therefore, may prevent cellular death and contribute to plaque growth and stabilization in early lesions. However in more advanced plaques, inflammatory cell infiltration, and concomitant production of numerous pro-angiogenic cytokines may be responsible for induction of uncontrolled neointimal microvessel proliferation resulting in production of immature and fragile neovessels similar to that seen in tumour development. These could contribute to development of an unstable haemorrhagic rupture-prone environment. Increasing evidence has suggested that the expression of intimal neovessels is directly related to the stage of plaque development, the risk of plaque rupture, and subsequently, the presence of symptomatic disease, the timing of ischemic neurological events and myocardial/cerebral infarction. Despite this, there is conflicting evidence regarding the causal relationship between neovessel expression and plaque thrombosis with some in vivo experimental models suggesting the contrary and as yet, few direct mediators of angiogenesis have been identified and associated with plaque instability in vivo.In recent years, an increasing number of angiogenic therapeutic targets have been proposed in order to facilitate modulation of neovascularization and its consequences in diseases such as cancer and macular degeneration. A complete knowledge of the mechanisms responsible for initiation of adventitial vessel proliferation, their extension into the intimal regions and possible de-novo synthesis of neovessels following differentiation of bone-marrow-derived stem cells is required in order to contemplate potential single or combinational anti-angiogenic therapies. In this review, we will examine the importance of angiogenesis in complicated plaque development, describe the current knowledge of molecular mechanisms of its initiation and maintenance, and discuss possible future anti-angiogenic therapies to control plaque stability.


The use of unproven remedies for rheumatoid arthritis in Australia.
It is important for a medical practitioner to be aware of his or her patient's use of non-prescribed, unproven remedies. This is especially so in a chronic relapsing disease of unknown cause such as rheumatoid arthritis. We selected 90 consecutive patients with classic or definite rheumatoid arthritis who attended the rheumatology clinic of a teaching hospital in 1982. The patients were asked about their previous or current use of an unproven remedy; 82% had used more than one unproven remedy since the diagnosis was made and 52% were currently using an unproven remedy. In all, 352 separate unproven remedies were used, with a mean of 4 +/- 0.3 remedies per patient. Avoidance of a particular food substance or use of a copper bracelet were the most common of such remedies. Fourteen per cent of remedies were deemed to be useful and 3% were felt to have resulted in an adverse effect.


Ring 4 chromosome with terminal p and q deletions.
A ring 4 chromosome was found in a boy with low birth weight, microcephaly, micrognathia, rounded broad nose, malformed ears, cleft soft palate, and retardation in growth and development. The ring 4 is formed by union of the ends after breakage and loss of the terminal parts of the short (p) arm and the long (q) arm. G-banded chromosome studies showed our patient to have a union at p16q35. Clinical findings in our patient were compared with those in others with the union at p16q35, p15q35, and p16q33. The patients with p15q35 had a deletion of more p-arm genetic material than those with the p16q35 union, and the patient with p16q33 had a greater loss of q-arm material. The loss of only a small part of the p16 band is associated with low birth weight, microcephaly, and retardation in growth and development.


Detection of activities that interfere with the enzymatic assay of deoxyribonucleoside 5'-triphosphates.
Several enzymes that interfere with the enzymatic assay of deoxyribonucleoside 5'-triphosphates (dNTP's) are present as contaminants when nucleotides are extracted from HeLa cells with 60% methanol. These activities include a nuclease, nucleoside diphosphokinase, and deoxyribonucleoside monophosphokinases which phosphorylate dAMP, dGMP, and dCMP. Collectively, these enzymes are able to degrade and reutilize the DNA template which is used together with DNA polymerase for dNTP assays. This process introduces large errors when dNTP assays are performed in this manner. Attempts to block the enzymatic conversion of deoxyribonucleoside diphosphates to triphosphates by inhibition of nucleoside diphosphokinase were unsuccessful because of the inability to block completely the kinase activity. Acid extraction of nucleotides also results in the presence of an activity that interferes with the enzymatic dNTP assay. The error introduced by this interfering activity is much smaller than that arising from the enzymes present in methanol extracts. All of these interfering activities are removed when cells are first extracted with 60% methanol and the resulting extract is subsequently treated with perchloric acid.


[Abnormal systemic artery originating from descending aorta to normal basal segments of left lung. Report 2 cases].
Two patients with a large abnormal systemic artery originating from the descending aorta several centimeters above the diaphragm to the four basal segments of normal left lower lobe (without cystic change like that found in the bronchopulmonary sequestration) were treated. No pulmonary artery was found to supply the basal segments. The patients suffered from repeated hemoptysis. In one patient the abnormal artery was incidentally found during bronchial arteriography. Lower lobectomy was performed in the two patients. Microscopic examination of specimens revealed some dilated small blood vessels with extremely thin walls and their rupture may be the cause of hemoptysis. Abnormal systemic artery must be suspected if hemoptysis or local murmur during chest examination cannot be explained by other conditions, and care must be taken not to injure the artery in isolating pleural adhesion and pulmonary ligament.


Proportion of APOBEC3-induced defective HIV DNA after 1 year of dolutegravir + lamivudine simplification in the ANRS 167 LAMIDOL trial.
Hypermutated viruses induced by APOBEC3 (apolipoprotein B mRNA-editing, enzyme-catalytic, polypeptide-like 3) proteins comprise some of the defective viruses in the HIV reservoir. Here, we assessed the proportion of APOBEC3-induced defective proviruses in HIV-positive patients before and after receiving dolutegravir + lamivudine dual therapy. PBMCs of virologically suppressed patients enrolled in the ANRS 167 LAMIDOL trial, evaluating a switch from triple therapy to dolutegravir + lamivudine, were collected 8 weeks before (W-8) and 48 weeks after (W48) dual-therapy initiation. The Vif and RT regions were subject to next-generation sequencing. Bioinformatic algorithms were developed to identify APOBEC3-defective sequences and APOBEC3-related drug resistance mutations (APOMuts). All hypermutated sequences and those containing at least one stop codon were considered as defective. One hundred and four patients were enrolled (median virological suppression duration: 4.2 years; IQR: 2.0-9.1). Proviral defective reads at W-8 and W48 were detected in Vif in 22% and 29% of patients, respectively, and in RT in 38% and 42% of patients, respectively. At least one APOMut was present in proviruses of 27% and 38% of patients at W-8 and W48, respectively. The ratio of APOMuts/number of potential APOMut sites was significantly higher at W48 (16.5%) than at W-8 (9.8%, P = 0.007). The presence of APOBEC3-defective viruses at W-8 was not associated with HIV total DNA level, nor with the third drug class received prior to switching to dolutegravir + lamivudine, nor with the duration of virological suppression. Whereas no significant change in the proportion of patients with APOBEC3-defective proviruses was evidenced after 1 year of dolutegravir + lamivudine maintenance, enrichment in APOMuts was observed. Further longer-term studies are needed to assess the other forms of defective viruses with dual-therapy.


A food policy package for healthy diets and the prevention of obesity and diet-related non-communicable diseases: the NOURISHING framework.
This paper presents the NOURISHING framework of food policies to promote healthy diets, and uses the framework to summarize the policy actions taken by the Bellagio meeting countries. NOURISHING was developed by WCRF International to formalize a comprehensive policy package that brings together the key domains of action and policy areas. It aims to provide global level recommendations for a comprehensive response, within which policymakers have the flexibility to select specific policy options suitable for their national/local contexts and target populations. It also aims to provide a framework for reporting, categorizing and monitoring policy actions taken around the world, and for systematically categorizing, updating, interpreting and communicating the evidence for policy to policymakers. In this paper we explain the structure for NOURISHING and the rationale behind it. We also use the framework to report on and categorize the policy actions implemented in the Bellagio countries.


Stage-specific effects of Notch activation during skeletal myogenesis.
Skeletal myogenesis involves sequential activation, proliferation, self-renewal/differentiation and fusion of myogenic stem cells (satellite cells). Notch signaling is known to be essential for the maintenance of satellite cells, but its function in late-stage myogenesis, i.e. post-differentiation myocytes and post-fusion myotubes, is unknown. Using stage-specific Cre alleles, we uncovered distinct roles of Notch1 in mononucleated myocytes and multinucleated myotubes. Specifically, constitutive Notch1 activation dedifferentiates myocytes into Pax7 quiescent satellite cells, leading to severe defects in muscle growth and regeneration, and postnatal lethality. By contrast, myotube-specific Notch1 activation improves the regeneration and exercise performance of aged and dystrophic muscles. Mechanistically, Notch1 activation in myotubes upregulates the expression of Notch ligands, which modulate Notch signaling in the adjacent satellite cells to enhance their regenerative capacity. These results highlight context-dependent effects of Notch activation during myogenesis, and demonstrate that Notch1 activity improves myotube's function as a stem cell niche.


Transvenous pacemaker leads do not worsen tricuspid regurgitation: a prospective echocardiographic study.
Limited prospective data are available regarding the influence of pacemaker leads on tricuspid valve function. To examine the true incidence of these complications, 35 patients were prospectively examined by two-dimensional and Doppler echocardiography before and after implantation of either a permanent pacemaker or an automatic implantable cardioverter-defibrillator. Of the 35 patients imaged preoperatively, the amount of tricuspid regurgitation (TR) was judged as normal or trivial in 15 (43%), mild in 10 (29%), moderate in 8 (23%), and severe in 2 (6%). Following electrode implantation, TR was noted to be normal or trivial in 13 (38%), mild in 15 (48%), moderate in 6 (17%) and severe in 1 (3%). We conclude that implantation of permanent right ventricular electrodes is not usually associated with an acute worsening of tricuspid regurgitation in most patients.


Slow sodium channel inactivation in rat fast-twitch muscle.
1. Voltage-clamp Na+ currents (INa) were measured in rat fast-twitch fibres using the loose-patch-clamp technique. Changes in the conditioning membrane potential produced slow changes in the peak INa elicited by short test depolarizations, due to a slow inactivation process. 2. Inactivation was increased by application of steady depolarizing potentials and was reversed by steady hyperpolarizations. These changes in peak INa could be well fitted by single-exponential functions with time constants in the range of 1-4 min. 3. The steady-state values of the maximum peak INa at any potential could be well fitted by a function identical to the one describing the fast inactivation process. This gave a potential of -108 mV at which 50% of the channels were closed due to slow inactivation. 4. The maximum peak current densities obtained with the slow inactivation fully removed were as large as 20 mA cm-2.


Single cell genome sequencing of laboratory mouse microbiota improves taxonomic and functional resolution of this model microbial community.
Laboratory mice are widely studied as models of mammalian biology, including the microbiota. However, much of the taxonomic and functional diversity of the mouse gut microbiome is missed in current metagenomic studies, because genome databases have not achieved a balanced representation of the diverse members of this ecosystem. Towards solving this problem, we used flow cytometry and low-coverage sequencing to capture the genomes of 764 single cells from the stool of three laboratory mice. From these, we generated 298 high-coverage microbial genome assemblies, which we annotated for open reading frames and phylogenetic placement. These genomes increase the gene catalog and phylogenetic breadth of the mouse microbiota, adding 135 novel species with the greatest increase in diversity to the Muribaculaceae and Bacteroidaceae families. This new diversity also improves the read mapping rate, taxonomic classifier performance, and gene detection rate of mouse stool metagenomes. The novel microbial functions revealed through our single-cell genomes highlight previously invisible pathways that may be important for life in the murine gastrointestinal tract.