Adrenal-dependent change in vascular reactivity in stroke-prone spontaneously hypertensive rats.
Tail arteries from stroke-prone spontaneously hypertensive rats (SHRSP) exhibit oscillatory contractions in response to norepinephrine. This type of oscillatory behavior does not occur in tail arteries from normotensive Wistar-Kyoto rats (WKY). We have shown that the traits of norepinephrine-induced oscillatory activity and high blood pressure are genetically associated in SHRSP, suggesting that oscillatory activity is a primary vascular abnormality that contributes to hypertension in this strain. In the present experiments, two approaches were used to test the hypothesis that adrenal mineralocorticoids modulate expression of this genetically determined vascular abnormality in SHRSP. First, the effect of adrenalectomy on blood pressure and oscillatory activity was determined in SHRSP that underwent bilateral adrenalectomy 3 weeks before experimentation. Second, the effect of deoxycorticosterone acetate (DOCA)-salt treatment on blood pressure and oscillatory activity was determined in 1) rats with no genetic background for oscillatory activity (WKY) and 2) progeny of SHRSP x WKY (F1 rats). Helically cut tail artery strips from all rats were mounted in isolated tissue baths for isometric force recording. Vessels were exposed to norepinephrine (6 x 10(-10) to 6 x 10(-6) M) for 20 minutes at each concentration. Oscillatory activity was defined as the sum of the phasic contractile amplitudes for all oscillations occurring during the final 10 minutes of norepinephrine incubation. Adrenalectomy markedly decreased blood pressure and oscillatory activity in SHRSP.(ABSTRACT TRUNCATED AT 250 WORDS)


A Natural Language Processing Framework for Assessing Hospital Readmissions for Patients With COPD.
With the passage of recent federal legislation, many medical institutions are now responsible for reaching target hospital readmission rates. Chronic diseases account for many hospital readmissions and chronic obstructive pulmonary disease has been recently added to the list of diseases for which the United States government penalizes hospitals incurring excessive readmissions. Though there have been efforts to statistically predict those most in danger of readmission, a few have focused primarily on unstructured clinical notes. We have proposed a framework, which uses natural language processing to analyze clinical notes and predict readmission. Many algorithms within the field of data mining and machine learning exist, so a framework for component selection is created to select the best components. Naïve Bayes using Chi-Squared feature selection offers an AUC of 0.690 while maintaining fast computational times.


Unilateral extrapulmonary airway bypass in advanced emphysema.
Gas trapping in emphysema results in resting and dynamic hyperinflation. We tested the hypothesis that a direct connection between the lung parenchyma and the atmosphere could increase expiratory flow and thereby potentially improve dyspnea through the relief of gas trapping. Ex vivo we studied 7 emphysematous lungs and 3 fibrotic lungs (as controls) and measured expiratory flow before and after airway bypass insertion during a forced maneuver in an artificial thorax. Pilot studies were conducted in vivo in 6 patients with advanced emphysema using a size 9 endotracheal tube as a bypass surgically placed through the chest wall into the upper lobe. In the ex vivo emphysematous lungs the volume expelled during a forced expiratory maneuver increased from 169 to 235 mL (p < 0.05). In the in vivo group 4 patients retained the bypass tube for 3 months or more; total lung capacity was reduced, and the forced expiratory volume in 1 second increased by 23% (mean percent predicted at baseline versus 3 months, 24.4% versus 29.5%). An extrapulmonary airway bypass increases expiratory flow in emphysema. This may be a useful approach in hyperinflated patients with homogeneous emphysema.


A physically transient form of silicon electronics.
A remarkable feature of modern silicon electronics is its ability to remain physically invariant, almost indefinitely for practical purposes. Although this characteristic is a hallmark of applications of integrated circuits that exist today, there might be opportunities for systems that offer the opposite behavior, such as implantable devices that function for medically useful time frames but then completely disappear via resorption by the body. We report a set of materials, manufacturing schemes, device components, and theoretical design tools for a silicon-based complementary metal oxide semiconductor (CMOS) technology that has this type of transient behavior, together with integrated sensors, actuators, power supply systems, and wireless control strategies. An implantable transient device that acts as a programmable nonantibiotic bacteriocide provides a system-level example.


Competitive inhibitors of Mycobacterium tuberculosis ribose-5-phosphate isomerase B reveal new information about the reaction mechanism.
Ribose-5-phosphate isomerase (Rpi), an important enzyme in the pentose phosphate pathway, catalyzes the interconversion of ribulose 5-phosphate and ribose 5-phosphate. Two unrelated isomerases have been identified, RpiA and RpiB, with different structures and active site residues. The reaction catalyzed by both enzymes is thought to proceed via a high energy enediolate intermediate, by analogy to other carbohydrate isomerases. Here we present studies of RpiB from Mycobacterium tuberculosis together with small molecules designed to resemble the enediolate intermediate. The relative affinities of these inhibitors for RpiB have a different pattern than that observed previously for the RpiA from spinach. X-ray structures of RpiB in complex with the inhibitors 4-phospho-d-erythronohydroxamic acid (K(m) 57 microm) and 4-phospho-d-erythronate (K(i) 1.7 mm) refined to resolutions of 2.1 and 2.2 A, respectively, allowed us to assign roles for most active site residues. These results, combined with docking of the substrates in the position of the most effective inhibitor, now allow us to outline the reaction mechanism for RpiBs. Both enzymes have residues that can catalyze opening of the furanose ring of the ribose 5-phosphate and so can improve the efficiency of the reaction. Both enzymes also have an acidic residue that acts as a base in the isomerization step. A lysine residue in RpiAs provides for more efficient stabilization of the intermediate than the corresponding uncharged groups of RpiBs; this same feature lies behind the more efficient binding of RpiA to 4-phospho-d-erythronate.


Efficacy of current molecular cytogenetic protocols for the diagnosis of chromosome aberrations in tumor specimens.
Molecular cytogenetics provides a powerful link between molecular genetic analysis and chromosome morphology, allowing one to pinpoint structurally aberrant chromosome regions on the molecular level. Fluorescence in situ hybridization with selected DNA probes allows the design of efficient and sensitive tools for the diagnosis of chromosomal aberrations present in tumor cells. Comparative genomic hybridization (CGH) allows the identification of chromosomal imbalances in a comprehensive manner, and is applied to solid tumors and hematological malignancies in order to (i) identify clonal differences within a specimen, (ii) contribute to tumor classifications, (iii) identify recurrent chromosomal gains and losses as starting points for the characterization and isolation of pathogenetically relevant genes, such as proto-oncogenes and tumor suppressor genes respectively, (iv) identify imbalances of prognostic relevance, (v) detect high-copy-number amplification and other markers of genetic instability, and (vi) analyze chromosomal imbalances during tumor progression.


Activating PRKACB somatic mutation in cortisol-producing adenomas.
Mutations in the gene encoding the protein kinase A (PKA) catalytic subunit α have been found to be responsible for cortisol-producing adenomas (CPAs). In this study, we identified by whole-exome sequencing the somatic mutation p.S54L in the PRKACB gene, encoding the catalytic subunit β (Cβ) of PKA, in a CPA from a patient with severe Cushing syndrome. Bioluminescence resonance energy transfer and surface plasmon resonance assays revealed that the mutation hampers formation of type I holoenzymes and that these holoenzymes were highly sensitive to cAMP. PKA activity, measured both in cell lysates and with recombinant proteins, based on phosphorylation of a synthetic substrate, was higher under basal conditions for the mutant enzyme compared with the WT, while maximal activity was lower. These data suggest that at baseline the PRKACB p.S54L mutant drove the adenoma cells to higher cAMP signaling activity, probably contributing to their autonomous growth. Although the role of PRKACB in tumorigenesis has been suggested, we demonstrated for the first time to our knowledge that a PRKACB mutation can lead to an adrenal tumor. Moreover, this observation describes another mechanism of PKA pathway activation in CPAs and highlights the particular role of residue Ser54 for the function of PKA.


Trophic upgrading and mobilization of wax esters in microzooplankton.
Heterotrophic protists play pivotal roles in aquatic ecosystems by transferring matter and energy, including lipids, from primary producers to higher trophic predators. Using Oxyrrhis marina as a model organism, changes to the non-saponifiable protist lipids were investigated under satiation and starvation conditions. During active feeding on the alga Cryptomonas sp., the O. marina hexane soluble non-saponifiable fraction lipid profile reflected its food source with the observed presence of long chain mono-unsaturated fatty alcohols up to C25:1. Evidence of trophic upgrading in O. marina was observed with long chain mono-unsaturated fatty alcohol accumulation of up to C35:1. To the best of our knowledge, this is the first evidence that heterotrophic dinoflagellates are capable of producing ester derived alcohols and that dinoflagellates like O. marina are capable of synthesizing fatty alcohols up to C35. Additionally, we show evidence of trophic upgrading of lipids. During a 20-day resource deprivation, the lipid profile remained constant. During starvation, the mobilization of wax esters as energy stores was observed with long chain fatty alcohols mobilized first. Changes in lipid class profile and utilization of wax esters in O. marina provides insight into the types of lipids available for energy demand, the transfer of lipids through the base of marine food webs, and the catabolic response induced by resource deprivation.


Amino acid supplements and recovery from high-intensity resistance training.
The purpose of this study was to investigate whether short-term amino acid supplementation could maintain a short-term net anabolic hormonal profile and decrease muscle cell damage during a period of high-intensity resistance training (overreaching), thereby enhancing recovery and decreasing the risk of injury and illness. Eight previously resistance trained males were randomly assigned to either a high branched chain amino acids (BCAA) or placebo group. Subjects consumed the supplement for 3 weeks before commencing a fourth week of supplementation with concomitant high-intensity total-body resistance training (overreaching) (3 x 6-8 repetitions maximum, 8 exercises). Blood was drawn prior to and after supplementation, then again after 2 and 4 days of training. Serum was analyzed for testosterone, cortisol, and creatine kinase. Serum testosterone levels were significantly higher (p < 0.001), and cortisol and creatine kinase levels were significantly lower (p < 0.001, and p = 0.004, respectively) in the BCAA group during and following resistance training. These findings suggest that short-term amino acid supplementation, which is high in BCAA, may produce a net anabolic hormonal profile while attenuating training-induced increases in muscle tissue damage. Athletes' nutrient intake, which periodically increases amino acid intake to reflect the increased need for recovery during periods of overreaching, may increase subsequent competitive performance while decreasing the risk of injury or illness.


The α1-adrenergic receptor antagonist, doxazosin, reduces alcohol drinking in alcohol-preferring (P) Rats.
Evidence supports a role for the noradrenergic system in alcohol drinking in animals and humans. Our previous studies demonstrated the efficacy of prazosin, an α1-adrenergic antagonist, in decreasing alcohol drinking in rat models of alcohol dependence. Prazosin has also been shown to decrease alcohol drinking in treatment-seeking alcohol-dependent men. Clinically, the use of prazosin is limited by the requirement for multiple daily administrations, whereas doxazosin, a structurally similar α1-adrenergic antagonist, requires only once-daily dosing. In this study, we tested the hypothesis that doxazosin, like prazosin, would decrease alcohol drinking in rats selectively bred for alcohol preference (P line). Adult male P rats were given 2 h/d scheduled access to a 2-bottle choice (15% v/v alcohol vs. water) session 5 d/wk (M-F), with food and water available ad libitum 24 h/d. Rats were injected with doxazosin (0 to 10 mg/kg, IP) 40 minutes prior to initiation of the alcohol access session in 3 trials (of 3, 5, and 5 consecutive days) each separated by 5 to 8 weeks. The third trial included 1 day without alcohol access (for locomotor testing), and 1 day of a single hour of alcohol access (for plasma alcohol determination). Doxazosin significantly reduced alcohol intake in all 3 trials. The 5 mg/kg dose consistently reduced alcohol intake, increased water drinking, did not affect locomotor activity, and resulted in lower plasma alcohol concentrations, suggesting that the doxazosin-induced reduction in alcohol drinking was not dependent on a motor impairment or an alteration in alcohol clearance. Doxazosin decreases voluntary alcohol consumption by male alcohol-preferring (P) rats, supporting a role for the noradrenergic system in alcohol drinking in P rats and suggesting that doxazosin could potentially be an effective once-daily pharmacotherapeutic agent for the treatment of alcohol use disorders.


Gain-of-function mutant of angiotensin II receptor, type 1A, causes hypertension and cardiovascular fibrosis in mice.
The role of the renin-angiotensin system has been investigated by overexpression or inactivation of its different genes in animals. However, there is no data concerning the effect of the constitutive activation of any component of the system. A knockin mouse model has been constructed with a gain-of-function mutant of the Ang II receptor, type 1A (AT(1A)), associating a constitutively activating mutation (N111S) with a C-terminal deletion, which impairs receptor internalization and desensitization. In vivo consequences of this mutant receptor expression in homozygous mice recapitulate its in vitro characteristics: the pressor response is more sensitive to Ang II and longer lasting. These mice present with a moderate (~20 mmHg) and stable increase in BP. They also develop early and progressive renal fibrosis and cardiac fibrosis and diastolic dysfunction. However, there was no overt cardiac hypertrophy. The hormonal parameters (low-renin and inappropriately normal aldosterone productions) mimic those of low-renin human hypertension. This new model reveals that a constitutive activation of AT(1A) leads to cardiac and renal fibrosis in spite of a modest effect on BP and will be useful for investigating the role of Ang II in target organs in a model similar to some forms of human hypertension.


Development and psychometric testing of the Canine Owner-Reported Quality of Life questionnaire, an instrument designed to measure quality of life in dogs with cancer.
OBJECTIVE To describe development and initial psychometric testing of an owner-reported questionnaire designed to standardize measurement of general quality of life (QOL) in dogs with cancer. DESIGN Key-informant interviews, questionnaire development, and field trial. SAMPLE Owners of 25 dogs with cancer for item development and pretesting and owners of 90 dogs with cancer for reliability and validity testing. PROCEDURES Standard methods for development and testing of questionnaire instruments intended to measure subjective states were used. Items were generated, selected, scaled, and pretested for content, meaning, and readability. Response items were evaluated with exploratory factor analysis and by assessing internal consistency (Cronbach α) and convergence with global QOL as determined with a visual analog scale. Preliminary tests of stability and responsiveness were performed. RESULTS The final questionnaire-which was named the Canine Owner-Reported Quality of Life (CORQ) questionnaire-contained 17 items related to observable behaviors commonly used by owners to evaluate QOL in their dogs. Several items pertaining to physical symptoms performed poorly and were omitted. The 17 items were assigned to 4 factors-vitality, companionship, pain, and mobility-on the basis of the items they contained. The CORQ questionnaire and its factors had high internal consistency (Cronbach α = 0.68 to 0.90) and moderate to strong correlations (r = 0.49 to 0.71) with global QOL as measured on a visual analog scale. Preliminary testing indicated good test-retest reliability and responsiveness to improvements in overall QOL. CONCLUSIONS AND CLINICAL RELEVANCE The CORQ questionnaire was a valid, reliable owner-reported questionnaire that measured general QOL in dogs with cancer and showed promise as a clinical trial outcome measure for quantifying changes in individual dog QOL occurring in response to cancer treatment and progression.


Preferences for access to the GP: a discrete choice experiment.
Access to primary care services is one of the key components of the NHS Plan which states that patients should be able to see a health professional within 24 hours and a GP within 48 hours. However, it is not clear how patients value speed of access in comparison with other aspects of primary care. To investigate patient preferences when making an routine appointment for a GP, and to describe the trade-offs and relationships between speed of access, choice of time and choice of doctor in different patient groups. Discrete choice experiment. Adults consulting a GP in six general practices in Sunderland. Choice sets based on three attributes (time to appointment, choice of time, choice of doctor) were presented in a self-completion questionnaire. We obtained 6985 observations from 1153 patients. We found that the waiting time to make an appointment was only important if the appointment is for a child or when attending for a new health problem. Other responders would trade-off a shorter waiting time and be willing to wait in order to either see their own choice of doctor or attend an appointment at their own choice of time. For responders who work, choice of time is six times more important than a shorter waiting time and they are willing to wait up to 1 day extra for this. Those with a long-standing illness value seeing their own GP more than seven times as much as having a shorter waiting time for an appointment and will wait an extra 1 day for an appointment with the GP of their choice, women will wait an extra 2 days, and older patients an extra 2.5 days. Speed of access is of limited importance to patients accessing their GP, and for many is outweighed by choice of GP or convenience of appointment.


Microhealth insurance and the risk coping strategies for the management of illness in Karnataka: a case study.
Risk coping strategies adopted by the households in the event of illness depends on the accessibility to healthcare financing mechanisms including health insurance. The empirical evidence on the effect of microhealth insurance (MHI) on the risk coping strategies of the households is scarce. This paper evaluates the impact of Sampoorna Suraksha Program, a nongovernmental organization-initiated MHI scheme and the risk coping strategies of households faced with medical illness in Karnataka state, India. In this descriptive cross-sectional study, we collected data from 416 insured households, 366 newly insured households and 364 uninsured households in randomly selected 10 taluks in three districts of Karnataka state, India. We hypothesized that insured individuals rely less on ex post risk coping strategies (borrowing, use of savings and sale of assets) compared with uninsured and newly insured individuals. Our hypothesis was tested using logistic and linear regression analysis. A significant difference among insured, uninsured and newly insured individuals was found for borrowing but not in the use of savings or sale of assets. A positive impact of MHI on illness-induced borrowing (both incidence and amount) was evident. The evidence from this study reinforces the role of MHI as a pivotal financing alternative to out-of-pocket expenditure in India.


Dinitrotoluene exposure in the copper mining industry and renal cancer: a case-cohort study.
To evaluate the association between dinitrotoluene (DNT) exposure and renal cancer in a case-cohort study. This case-cohort study was conducted among men born between 1920 and 1974 (n=16 441) who were gainfully employed between 1953 and 1990 in one of two copper mines in Mansfeld, Saxony-Anhalt, former German Democratic Republic, and followed up till 31 December 2006. The study included 109 cases with renal cancer identified by record linkage with the Common Cancer Registry of the New Federal States of Germany (GKR) or by a network of pathology institutes. A comparison subcohort of 999 cohort members was selected at random from the total cohort. Duration and intensity of inhalation and dermal exposure to DNT were assessed on the basis of a job exposure matrix. A time-dependent Cox proportional hazards model modified for case-cohort design was used to assess the relationship between cumulative inhalation and dermal DNT exposure and renal cancer. Elevated risks were found for medium (HR=2.73; 95% CI 1.00 to 7.42) and high (HR=1.81; 95% CI 0.75 to 4.33) dermal exposure to DNT. Relative risks for medium inhalation exposure to DNT were not increased (HR=0.93; 95% CI 0.48 to 1.79) while relative risks for high inhalation exposure to DNT were elevated to 1.36 (95% CI 0.84 to 2.21). We found a statistically significant HR of 2.12 (95% CI 1.03 to 4.37) for combined medium or high inhalation and medium or high dermal exposure to DNT. According to our case-cohort study, dermal and inhalation exposure to DNT is associated with increased renal cancer risk.


Dendritic cells limit fibroinflammatory injury in nonalcoholic steatohepatitis in mice.
Nonalcoholic steatohepatitis (NASH) is the most common etiology of chronic liver dysfunction in the United States and can progress to cirrhosis and liver failure. Inflammatory insult resulting from fatty infiltration of the liver is central to disease pathogenesis. Dendritic cells (DCs) are antigen-presenting cells with an emerging role in hepatic inflammation. We postulated that DCs are important in the progression of NASH. We found that intrahepatic DCs expand and mature in NASH liver and assume an activated immune phenotype. However, rather than mitigating the severity of NASH, DC depletion markedly exacerbated intrahepatic fibroinflammation. Our mechanistic studies support a regulatory role for DCs in NASH by limiting sterile inflammation through their role in the clearance of apoptotic cells and necrotic debris. We found that DCs limit CD8(+) T-cell expansion and restrict Toll-like receptor expression and cytokine production in innate immune effector cells in NASH, including Kupffer cells, neutrophils, and inflammatory monocytes. Consistent with their regulatory role in NASH, during the recovery phase of disease, ablation of DC populations results in delayed resolution of intrahepatic inflammation and fibroplasia. Our findings support a role for DCs in modulating NASH. Targeting DC functional properties may hold promise for therapeutic intervention in NASH.


Purification and characterization of the extracellular aspartyl proteinase of Candida albicans: removal of extraneous proteins and cell wall mannoprotein and evidence for lack of glycosylation.
Aspartyl proteinase (AP) is an extracellular enzyme of Candida albicans implicated as a pathogenic factor. Previous reports on the purification and characterization of AP suggested that a single DEAE-Sephadex chromatographic step was sufficient for the removal of extraneous proteins and that the final product was glycosylated. We purified AP using a chromatographic series consisting of DEAE-Sephadex A25, Sephadex G75 and rechromatography on DEAE-Sephadex A25. Use of DEAE-Sephadex alone did not remove extraneous proteins and removed little contaminating mannoprotein (MP). The addition of a Sephadex G75 column to the purification scheme removed the majority of contaminating MP and proteins. The final DEAE-Sephadex A25 chromatographic step resulted in (a) removal of detectable extraneous proteins, (b) removal of immunologically detectable MP by dot blot and Western blot enzyme immunoassay, (c) loss of periodic acid-silver stain positivity, and (d) a high AP yield (1295 U l-1) and specific activity (1749 U mg-1). We conclude that a single DEAE-Sephadex A25 purification step is insufficient to remove extraneous proteins and MP, which could interfere with the production of AP-specific antibodies and the dissection of moieties responsible for immune reactivity. Reports of periodic acid-Schiff or anthrone positivity of AP preparations may reflect the presence of extraneous MP, which can be removed by the chromatographic series we describe.


Who stays in addiction treatment groups? Anxiety and avoidant attachment styles predict treatment retention and relapse.
Attachment styles have been shown to be an important predictor of relationship quality and well-being. They have also been linked with ability to function well in groups. Insecure attachment styles are thought to be an underlying cause of addiction and represent a target for change in one-to-one therapy. How attachment styles themselves affect group therapy for addiction is understudied. The current study addresses this gap by examining the effects of attachment styles on relapse and treatment retention amongst a population of people attending addiction therapy groups. Fifty-eight individuals in such groups completed measures of attachment anxiety and avoidance. Participants were followed up 4 weeks later, and their continued treatment attendance and relapse status were recorded. In terms of treatment retention, high anxiety/high avoidant participants had the highest retention. In terms of relapse, low anxiety/high avoidant participants had the lowest rates. These results are discussed in terms of the potentially protective effects of avoidant attachment styles during group therapy and the role of anxiety attachment in the continued maintenance of both protective and risky personal relationships.


Structural and mechanistic insights into 5-lipoxygenase inhibition by natural products.
Leukotrienes (LT) are lipid mediators of the inflammatory response that are linked to asthma and atherosclerosis. LT biosynthesis is initiated by 5-lipoxygenase (5-LOX) with the assistance of the substrate-binding 5-LOX-activating protein at the nuclear membrane. Here, we contrast the structural and functional consequences of the binding of two natural product inhibitors of 5-LOX. The redox-type inhibitor nordihydroguaiaretic acid (NDGA) is lodged in the 5-LOX active site, now fully exposed by disordering of the helix that caps it in the apo-enzyme. In contrast, the allosteric inhibitor 3-acetyl-11-keto-beta-boswellic acid (AKBA) from frankincense wedges between the membrane-binding and catalytic domains of 5-LOX, some 30 Å from the catalytic iron. While enzyme inhibition by NDGA is robust, AKBA promotes a shift in the regiospecificity, evident in human embryonic kidney 293 cells and in primary immune cells expressing 5-LOX. Our results suggest a new approach to isoform-specific 5-LOX inhibitor development through exploitation of an allosteric site in 5-LOX.


Continuous cerebral and myocardial perfusion during aortic arch repair in neonates and infants.
To minimize myocardial ischemia, we repaired aortic arch obstruction with ventricular septal defect, using two different techniques of cerebral and myocardial perfusion. Seventy-one infants, ages 3 to 137 days, underwent primary repair of coarctation of the aorta (n = 49)/interruption of the aortic arch (n = 22) with ventricular septal defect. In 65 patients, an end-to-end arch anastomosis was performed with cerebral and myocardial perfusion through the innominate or the ascending arterial cannula (non-working beating heart: NWBH). In the remaining 6 patients, an arterial cannula was placed into the innominate artery. With partial cardiopulmonary bypass, the innominate artery was snared proximal to the cannulation site and the ascending aorta was cross-clamped. An extended arch anastomosis was carried out with cerebral perfusion and a working beating heart (WBH). Ten patients (15%) undergoing aortic arch repair with the NWBH technique required cardioplegic arrest to complete a proximal anastomosis, whereas in all 6 repairs with the WBH technique, the extended anastomoses were completed without myocardial ischemia. One hospital death and late death occurred, with an overall survival of 98%. End-to-end arch reconstruction is feasible without myocardial ischemia, using the NWBH technique in patients without hypoplastic arch and using the WBH technique in patients with hypoplastic arch.


An anterior approach to spinal pathology of the upper thoracic spine through a partial manubriotomy.
Surgical pathology in the region of the upper thoracic spine (T1-4) is uncommon compared with other regions of the spine. Often times posterior and posterolateral approaches can be used, but formal anterior decompression often requires a low anterior cervical approach combined with a sternotomy, which yields significant perioperative morbidity. The authors describe a modified low anterior cervical dissection combined with a partial manubriotomy that they have used to successfully access and decompress anterior pathology of the upper thoracic spine. Their modified approach spares the sternoclavicular joints and leaves the sternum intact, decreasing the morbidity associated with these added procedures.


The current and future applications of microorganism in the bioremediation of cyanide contamination.
Inorganic cyanide and nitrile compounds are distributed widely in the environment, chiefly as a result of anthropogenic activity but also through cyanide synthesis by a range of organisms including higher plants, fungi and bacteria. The major source of cyanide in soil and water is through the discharge of effluents containing a variety of inorganic cyanide and nitriles. Here the fate of cyanide compounds in soil and water is reviewed, identifying those factors that affect their persistence and which determine whether they are amenable to biological degradation. The exploitation of cyanides by a variety of taxa, as a mechanism to avoid predation or to inhibit competitors has led to the evolution in many organisms of enzymes that catalyse degradation of a range of cyanide compounds. Microorganisms expressing pathways involved in cyanide degradation are briefly reviewed and the current applications of bacteria and fungi in the biodegradation of cyanide contamination in the field are discussed. Finally, recent advances that offer an insight into the potential of microbial systems for the bioremediation of cyanide compounds under a range of environmental conditions are identified, and the future potential of these technologies for the treatment of cyanide pollution is discussed.


Phase variation of slime production in Staphylococcus aureus: implications in colonization and virulence.
Two methods commonly used for slime detection in coagulase-negative staphylococci (tube biofilm formation and colony morphology in Congo red agar) were used to study 144 ruminant mastitis Staphylococcus aureus strains. Slime production was detected in 21 strains. A majority of cells (85%) in slime-producing (SP) strains and a minority of cells (5%) in non-slime-producing (NSP) strains showed a condensed exopolysaccharide matrix (slime) surrounding the bacterial cell wall, as revealed by electron microscopy and immunofluorescence. In vivo slime production was also detected immunohistochemically after experimental infection of the mammary gland in sheep. Upon repeated subcultures in Congo red agar, NSP variants were obtained from four ovine and four bovine SP strains at a frequency ranging from 0.5 x 10(-4) to 10(-4). Because SP variants could not be obtained from NSP strains within this range or at a higher frequency, they were obtained by the tube biofilm formation (requiring repeated subculturing of NSP strains in tryptic soy broth containing 2% glucose for subsequent recovery of colonies adherent to the walls of the culture tubes). In experimental challenge, the SP variant showed a significantly higher colonization capacity than did the NSP variant of the same strain used (P < 0.001). However, the NSP variant had a higher virulence than did the SP variant (P < 0.001). These results may help to explain the different roles of S. aureus slime production cell types (SP and NSP) coexisting in disease.


Tu Salud, ¡Si Cuenta!: Exposure to a community-wide campaign and its associations with physical activity and fruit and vegetable consumption among individuals of Mexican descent.
Mexican Americans along the US-Mexico border have been found to be disproportionately affected by chronic diseases particularly related to lack of physical activity and healthful food choices. A community-wide campaign (CWC) is an evidence-based strategy to address these behaviors but with few examples of implementation in Mexican descent populations facing profound health disparities. We examined exposure to a CWC, titled Tu Salud ¡Sí Cuenta!, and its association with meeting the recommended minutes of moderate and vigorous physical activity weekly and consuming more portions of fruits and vegetables daily. A cross-sectional sample of 1438 Mexican descent participants was drawn from a city-wide, randomly-selected cohort interviewed between the years 2008 and 2012. Multivariable comparisons of participants exposed and not exposed to the CWC and meeting physical activity guidelines or their fruit and vegetable consumptions using mixed effects models were conducted. The community-wide campaign components included different forms of mass media and individually-focused components such as community health worker (CHW) home visits. After adjusting for gender, age, marital status, educational attainment, language preference, health insurance, and diabetes diagnosis, the strongest association was found between meeting physical activity guidelines and exposure to both CHW discussions and radio messages (adjusted OR = 3.83; 95% CI = [1.28, 6.21]; p = 0.0099). Participants who reported exposure to both radio and TV messages consumed more portions of fruits and vegetables than those who reported no exposure (adjusted RR = 1.30; 95% CI = [1.02, 1.66]; p = 0.0338). This study provides insights into the implementation and behavioral outcomes associated with exposure to a community-wide campaign, a potential model for addressing lifestyle modifications in populations affected by health disparities.


Genetic diagnosis identifies occult lymph node metastases undetectable by the histopathological method.
The mutant allele-specific amplification (MASA) method is capable of detecting one tumor cell containing genetic changes in a sample containing thousands of normal cells. To investigate whether MASA can be applied to sensitive detection of lymph node metastasis, we screened 22 colorectal cancers for K-ras and p53 mutations and examined corresponding regional lymph node at the genetic level by the MASA method. Six of the primary tumors were found to certain K-ras mutations, and nine exhibited mutations of the p53 gene. In seven of the 14 cases in which genetic alterations were identified (mutations in both genes were found in one tumor), we found discrepancies between the genetic and the histopathological diagnoses with respect to the presence or absence of cancer cells in lymph nodes, in that these patients were histologically diagnosed lymph node negative, hn(-) but genetically diagnosed lymph node positive, gn(+). Because disease recurs in 20-30% of cancer patients whose lymph nodes are histopathologically negative after surgery, genetic evaluation of lymph nodes for metastasis may become a useful prognostic indicator.


Asbestos Induces Oxidative Stress and Activation of Nrf2 Signaling in Murine Macrophages: Chemopreventive Role of the Synthetic Lignan Secoisolariciresinol Diglucoside (LGM2605).
The interaction of asbestos fibers with macrophages generates harmful reactive oxygen species (ROS) and subsequent oxidative cell damage that are key processes linked to malignancy. Secoisolariciresinol diglucoside (SDG) is a non-toxic, flaxseed-derived pluripotent compound that has antioxidant properties and may thus function as a chemopreventive agent for asbestos-induced mesothelioma. We thus evaluated synthetic SDG (LGM2605) in asbestos-exposed, elicited murine peritoneal macrophages as an in vitro model of tissue phagocytic response to the presence of asbestos in the pleural space. Murine peritoneal macrophages (MFs) were exposed to crocidolite asbestos fibers (20 µg/cm²) and evaluated at various times post exposure for cytotoxicity, ROS generation, malondialdehyde (MDA), and levels of 8-iso Prostaglandin F2α (8-isoP). We then evaluated the ability of LGM2605 to mitigate asbestos-induced oxidative stress by administering LGM2605 (50 µM) 4-h prior to asbestos exposure. We observed a significant (p < 0.0001), time-dependent increase in asbestos-induced cytotoxicity, ROS generation, and the release of MDA and 8-iso Prostaglandin F2α, markers of lipid peroxidation, which increased linearly over time. LGM2605 treatment significantly (p < 0.0001) reduced asbestos-induced cytotoxicity and ROS generation, while decreasing levels of MDA and 8-isoP by 71%-88% and 41%-73%, respectively. Importantly, exposure to asbestos fibers induced cell protective defenses, such as cellular Nrf2 activation and the expression of phase II antioxidant enzymes, HO-1 and Nqo1 that were further enhanced by LGM2605 treatment. LGM2605 boosted antioxidant defenses, as well as reduced asbestos-induced ROS generation and markers of oxidative stress in murine peritoneal macrophages, supporting its possible use as a chemoprevention agent in the development of asbestos-induced malignant mesothelioma.


The induction of microsomal electron transport enzymes.
Liver endoplasmic reticulum contains as NADPH-dependent electron transport complex where the family of hemeproteins, termed cytochrome P-450, serve as catalysts for the oxidation of a variety of different organic chemicals. The content and inventory of the types of cytochrome P-450 is readily modified following in vivo treatment of animals with 'inducing agents' such as barbiturates, steroids and polycyclic hydrocarbons. Recent studies have applied the methods of molecular biology to evaluate changes in the transcription and translation of genomic information occurring concomitant with the initiation of synthesis of various types of cytochrome P-450. The ability to isolate unique cytochrome P-450 proteins and to prepare specific antibodies now permits the study of in vitro translation of mRNA and the preparation of specific cDNAs. The present review summarizes the historic background leading to current concepts of cytochrome P-450 induction and describes recent advances in our knowledge of the regulation of cytochrome P-450 synthesis in the liver.


Social class mortality differentials: artefact, selection or life circumstances?
Data from 10 years follow up of mortality in the OPCS Longitudinal Study are used to relate deaths of men in 1976-81 to their social class as recorded by the 1971 census. Explanations of social class mortality differentials are critically reviewed in the light of these new data. The similarity between the class differentials observed for men aged 15-64 years in this study and those reported in the 1970-2 Decennial Supplement on Occupational Mortality indicate that the published gradients were not in fact grossly distorted by numerator denominator biases. Distortions to gradients observed in the early years of the longitudinal study and ascribed to selective health related mobility out of employment from the principal social classes to the permanently sick had largely worn off after five years of follow up. Sharp gradients at ages over 75 years, similar to those at younger ages, suggest that, for men aged over 50 years, selective health related mobility between social classes does not contribute to differentials in mortality.


Patient-derived anti-β2GP1 antibodies recognize a peptide motif pattern and not a specific sequence of residues.
Antiphospholipid antibody syndrome is an autoimmune disease characterized by the presence of so-called antiphospholipid antibodies and clinical manifestations such as recurrent thromboembolic or pregnancy complications. Although the main antigenic determinant for antiphospholipid antibodies has been identified as the β-2-glycoprotein 1 (β2GP1), the precise epitope recognized by antiphospholipid antibodies still remains largely unknown. In the study herein, we wanted to identify a sequence in domain I of β2GP1 able to induce the proliferation of CD4+ T cells isolated from antiphospholipid antibody syndrome patients, but not from healthy donors, and to interact with antiphospholipid antibodies. We have characterized a sequence in domain I of β2GP1 that triggers CD4+ T-cell proliferation. A comparison of this sequence with the previously reported binding of antiphospholipid antibodies to discontinuous epitope R39-R43 reveals the presence of an indeterminate motif in β2GP1, in which the polarity determines the characteristics and specificity of antiphospholipid antibodies-interacting motifs. Using point mutations, we characterized the main antiphospholipid antibodies-interacting motif as ϕϕϕζζFxC, but also established ϕϕϕζζFxϕ-related motifs as potential antiphospholipid antibodies epitopes, in which ϕ represents nonpolar residues and ζ polar residues, with charges of the residues not being involved. Of specific importance, these different motifs are present at least once in all antiphospholipid antibodies-related receptors described so far. We have further demonstrated, in vitro, that peptides and domains of β2GP1 containing these motifs were able to interact with antiphospholipid antibodies and inhibit their monocyte activating activity. These results established that the antiphospholipid antibodies-interacting motifs are determined by the polarity, but not by the sequence or charge, of amino acids. These data could also contribute to the future development of more sensitive and specific diagnostic tools for antiphospholipid antibody syndrome determination and potential peptide- or β2GP1 domain-based clinical therapies.


Is it all about sex? Acupuncture for the treatment of pain from a biological and gender perspective.
Pain is a unique personal experience showing variability where gender and sex related effects might contribute. The mechanisms underlying the differences between women and men are currently unknown but are likely to be complex and involving interactions between biological, sociocultural and psychological aspects. In women, painful experimental stimuli are generally reported to produce a greater intensity of pain than in men. Clinical pain is often reported with higher severity and frequency, longer duration, and present in a greater number of body regions in women than in men. Women are also more likely to experience a number of painful conditions such as fibromyalgia, temporomandibular dysfunction, migraine, rheumatoid arthritis and irritable bowel syndrome. With regard to biological factors, quantitative as well as qualitative differences in the endogenous pain inhibitory systems have been implicated, as well as an influence of gonadal hormones. Psychosocial factors like sex role beliefs, pain coping strategies, and pain related expectancies may also contribute to the differences. Being exposed to repeated painful visceral events (eg menses, labour) during life may contribute to an increased sensitivity to, and greater prevalence of, pain among women. When assessing the outcome of pharmacological and non-pharmacological therapies in pain treatment, the factors of gender and sex should be taken into account as the response to an intervention may differ. Preferably, treatment recommendations should be based on studies using both women and men as the norm. Due to variability in results, findings from animal studies and experiments in healthy subjects should be interpreted with care.


Multitasking versus multiplexing: Toward a normative account of limitations in the simultaneous execution of control-demanding behaviors.
Why is it that behaviors that rely on control, so striking in their diversity and flexibility, are also subject to such striking limitations? Typically, people cannot engage in more than a few-and usually only a single-control-demanding task at a time. This limitation was a defining element in the earliest conceptualizations of controlled processing; it remains one of the most widely accepted axioms of cognitive psychology, and is even the basis for some laws (e.g., against the use of mobile devices while driving). Remarkably, however, the source of this limitation is still not understood. Here, we examine one potential source of this limitation, in terms of a trade-off between the flexibility and efficiency of representation ("multiplexing") and the simultaneous engagement of different processing pathways ("multitasking"). We show that even a modest amount of multiplexing rapidly introduces cross-talk among processing pathways, thereby constraining the number that can be productively engaged at once. We propose that, given the large number of advantages of efficient coding, the human brain has favored this over the capacity for multitasking of control-demanding processes.


Potency and stability of C terminal truncated human epidermal growth factor.
Epidermal growth factor (EGF) is normally present as EGF(1-53). A variety of C terminal truncated forms have been used in preliminary trials for treating gastrointestinal injury but their relative potency and stability when used in a clinical setting are unclear. Therefore, we compared the biological activity of recombinant EGF(1-53), EGF(1-52), EGF(1-51), and the C terminal peptides EGF(44-53) and EGF(49-53). Purity of forms was confirmed by mass spectrometry. Bioactivity of the different EGF forms was determined using [methyl-(3)H] thymidine incorporation into primary rat hepatocytes and their ability to reduce indomethacin (20 mg/kg subcutaneously)/restraint induced gastric injury in rats. Stability of EGF peptides was determined by serial sampling from a syringe driver system containing EGF/4% albumin in saline. Biological activity assays of EGF(1-53), EGF(1-52), and EGF(1-51) gave almost identical thymidine uptake dose-response curves (maximal responses increasing baseline uptake from 4400 (600) cpm (mean (SEM)) to about 22 000 (2000) cpm when EGF was added at 1. 6 nM). EGF(44-53) and EGF(49-53) did not stimulate (3)H thymidine uptake. Control rats had 47 (4) mm(2) damage/stomach, EGF(1-51), EGF(1-52), and EGF(1-53) at 0.16 and 0.80 nmol/kg/h each reduced gastric injury by about 50% and 80%, respectively (both doses p<0.01 compared with control but no significant difference between the different forms). EGF was stable at room temperature for seven days but biological activity decreased by 35% and 40% at two and three weeks, respectively (both p<0.01). Exposure to light did not affect bioactivity. EGF(1-51) and EGF(1-52) are as biologically active as full length EGF(1-53) but the C terminal penta- and decapeptides are ineffective. Clinical trials of EGF can probably use infusion systems for at least 48 hours at room temperature and with exposure to light, without reducing biological efficacy.


Retroviral vector insertion sites associated with dominant hematopoietic clones mark "stemness" pathways.
Evidence from model organisms and clinical trials reveals that the random insertion of retrovirus-based vectors in the genome of long-term repopulating hematopoietic cells may increase self-renewal or initiate malignant transformation. Clonal dominance of nonmalignant cells is a particularly interesting phenotype as it may be caused by the dysregulation of genes that affect self-renewal and competitive fitness. We have accumulated 280 retrovirus vector insertion sites (RVISs) from murine long-term studies resulting in benign or malignant clonal dominance. RVISs (22.5%) are located in or near (up to 100 kb [kilobase]) to known proto-oncogenes, 49.6% in signaling genes, and 27.9% in other or unknown genes. The resulting insertional dominance database (IDDb) shows substantial overlaps with the transcriptome of hematopoietic stem/progenitor cells and the retrovirus-tagged cancer gene database (RTCGD). RVISs preferentially marked genes with high expression in hematopoietic stem/progenitor cells, and Gene Ontology revealed an overrepresentation of genes associated with cell-cycle control, apoptosis signaling, and transcriptional regulation, including major "stemness" pathways. The IDDb forms a powerful resource for the identification of genes that stimulate or transform hematopoietic stem/progenitor cells and is an important reference for vector biosafety studies in human gene therapy.


Atheroprotective effects of Alum are associated with capture of oxidized LDL antigens and activation of regulatory T cells.
The immune system represents a promising novel target for prevention of atherosclerosis. Several pilot vaccines that reduce atherosclerosis in experimental animals have been developed. The aluminum hydroxide adjuvant Alum has been shown to have antiatherogenic properties in itself, suggesting that it may be a suitable adjuvant in possible future atherosclerosis vaccines. To characterize the immune pathways mediating this protection, we treated wild-type C57BL/6 and Apoe(-)(/)(-) mice with Alum or PBS. Analyses of splenocytes isolated from 12-week-old mice demonstrated that Alum increased the presence of CD4(+)CD25(+)FoxP3(+) regulatory T cells and downregulated the expression of T cell activation markers CD28 and ICOS in Apoe(-)(/)(-) mice but not in C57BL/6 wild-type mice. A similar immunosuppressive phenotype was found also in 25-week-old Apoe(-)(/)(-) mice and was associated with reduced atherosclerosis. Alum precipitates recovered from the injection site of Apoe(-)(/)(-) mice contained antigens derived from oxidized LDL. These findings demonstrate that treatment of Apoe(-)(/)(-) mice with Alum results in an increase of regulatory T cells and suggest that these are activated by tolerogenic antigen-presenting cells presenting oxidized LDL antigens. Our findings provide improved mechanistic understanding of the atheroprotective properties of aluminum hydroxide adjuvants but also point to the importance of determining if hypercholesterolemia may compromise the efficacy of Alum-containing vaccines used clinically today.


Clusters in nonsmooth oscillator networks.
For coupled oscillator networks with Laplacian coupling, the master stability function (MSF) has proven a particularly powerful tool for assessing the stability of the synchronous state. Using tools from group theory, this approach has recently been extended to treat more general cluster states. However, the MSF and its generalizations require the determination of a set of Floquet multipliers from variational equations obtained by linearization around a periodic orbit. Since closed form solutions for periodic orbits are invariably hard to come by, the framework is often explored using numerical techniques. Here, we show that further insight into network dynamics can be obtained by focusing on piecewise linear (PWL) oscillator models. Not only do these allow for the explicit construction of periodic orbits, their variational analysis can also be explicitly performed. The price for adopting such nonsmooth systems is that many of the notions from smooth dynamical systems, and in particular linear stability, need to be modified to take into account possible jumps in the components of Jacobians. This is naturally accommodated with the use of saltation matrices. By augmenting the variational approach for studying smooth dynamical systems with such matrices we show that, for a wide variety of networks that have been used as models of biological systems, cluster states can be explicitly investigated. By way of illustration, we analyze an integrate-and-fire network model with event-driven synaptic coupling as well as a diffusively coupled network built from planar PWL nodes, including a reduction of the popular Morris-Lecar neuron model. We use these examples to emphasize that the stability of network cluster states can depend as much on the choice of single node dynamics as it does on the form of network structural connectivity. Importantly, the procedure that we present here, for understanding cluster synchronization in networks, is valid for a wide variety of systems in biology, physics, and engineering that can be described by PWL oscillators.


ATP-driven exchange of histone H2AZ variant catalyzed by SWR1 chromatin remodeling complex.
The conserved histone variant H2AZ has an important role in the regulation of gene expression and the establishment of a buffer to the spread of silent heterochromatin. How histone variants such as H2AZ are incorporated into nucleosomes has been obscure. We have found that Swr1, a Swi2/Snf2-related adenosine triphosphatase, is the catalytic core of a multisubunit, histone-variant exchanger that efficiently replaces conventional histone H2A with histone H2AZ in nucleosome arrays. Swr1 is required for the deposition of histone H2AZ at specific chromosome locations in vivo, and Swr1 and H2AZ commonly regulate a subset of yeast genes. These findings define a previously unknown role for the adenosine triphosphate-dependent chromatin remodeling machinery.


Simple and sensitive technique for alignment of the pinhole of a spatial filter of a high-energy, high-power laser system.
In this paper, a new method for alignment of the pinhole of a spatial filter (SF) has been proposed and demonstrated experimentally. The effect of the misalignment of the pinhole on the laser beam profiles has been calculated for circular and elliptical Gaussian laser beams. Theoretical computation has been carried out to illustrate the effect of an intensity mask, placed before the focusing lens of the SF, on the spatial beam profile after the pinhole of the SF. It is shown, both theoretically and experimentally, that a simple intensity mask, consisting of a black dot, can be used to visually align the pinhole with a high accuracy of 5% of the pinhole diameter. The accuracy may be further improved using a computer-based image processing algorithm. Finally, the proposed technique has been demonstrated to align a vacuum SF of a compact 40 J Nd:phosphate glass laser system.


Aliphatic and chlorinated alkenes and epoxides as inducers of alkene monooxygenase and epoxidase activities in Xanthobacter strain Py2.
The inducible nature of the alkene oxidation system of Xanthobacter strain Py2 has been investigated. Cultures grown with glucose as the carbon source did not contain detectable levels of alkene monooxygenase or epoxidase, two key enzymes of alkene and epoxide metabolism. Upon addition of propylene to glucose-grown cultures, alkene monooxygenase and epoxidase activities increased and after an 11-h induction period reached levels of specific activity comparable to those in propylene-grown cells. Addition of chloramphenicol or rifampin prevented the increase in the enzyme activities. Comparison of the banding patterns of proteins present in cell extracts revealed that polypeptides with molecular masses of 43, 53, and 57 kDa accumulate in propylene-grown but not glucose-grown cells. Pulse-labeling of glucose-grown cells with [35S]methionine and [35S]cysteine revealed that the 43-, 53-, and 57-kDa proteins, as well as two additional polypeptides with molecular masses of 12 and 21 kDa, were newly synthesized upon exposure of cells to propylene or propylene oxide. The addition to glucose-grown cells of a variety of other aliphatic and chlorinated alkenes and epoxides, including ethylene, vinyl chloride (1-chloroethylene), cis- and trans-1,2-dichloroethylene, 1-chloropropylene, 1,3-dichloropropylene, 1-butylene, trans-2-butylene, isobutylene, ethylene oxide, epichlorohydrin (3-chloro-1,2-epoxypropane), 1,2-epoxybutane, cis- and trans-2,3-epoxybutane, and isobutylene oxide stimulated the synthesis of the five propylene-inducible polypeptides as well as increases in alkene monooxygenase and epoxidase activities. In contrast, acetylene, and a range of aliphatic and chlorinated alkanes, did not stimulate the synthesis of the propylene-inducible polypeptides or the increase in alkene monooxygenase and epoxidase activities.


A syndemic of psychosocial problems places the MSM (men who have sex with men) population at greater risk of HIV infection.
The MSM (Men who have sex with men) population suffers from very high rates of concurrent psychosocial problems. Together, these problems comprise a syndemic that increases the risk of HIV infection for this community. The precise mechanisms through which this syndemic can raise the likelihood of HIV infection warrant further exploration. A total of 522 MSM were enrolled via a multiframe sampling approach and were asked to report psychosocial problems, risky sexual behaviors and HIV test results. A count of psychosocial health problems was calculated to test the additive relationship of these factors on HIV risk. Adjusting analysis and restriction analysis were used to determine a proposed intermediate pathway. Psychosocial health problems are highly concurrent and intercorrelated among urban MSM. Greater numbers of health problems are significantly and positively associated with HIV infection, which is mediated, at least partially, by risky sexual behaviors. MSM experience concurrent psychosocial health problems that correlate with HIV infection in this community. We recommend the development of coping strategies for this population to deal with these psychosocial problems, both in prevention research and health policy.


Neonatal central venous catheter thrombosis: diagnosis, management and outcome.
Thrombotic occlusion of central venous catheters (CVCs) is a common problem in newborns. There is no guideline that systematically addresses the diagnosis, management, and prevention of this complication. The objective of this review is to establish evidence-based guidance for the management of CVC-related thrombosis. A comprehensive search of the scientific literature was conducted from 1948 to 2012. Twenty-six articles fulfilling four criteria - humans, neonates aged below 28 days, CVC insertion, and English language - were included for analysis. The incidence of thrombosis was 9.2% (308/3332). Singly inserted umbilical venous catheters (UVCs) and peripherally inserted central catheters accounted for over 80% of all CVCs. Frequently reported thrombotic sites were the hepatic vein, right atrium, and inferior vena cava. Symptoms included distal swelling of affected areas and thrombocytopenia. Increased length of catheter stay, infusion of blood products and malpositioned UVCs were identified as risk factors. The commonest diagnostic investigations to confirm thrombosis were echocardiography and ultrasonography. Spontaneous resolution may occur in UVC-related thrombosis, but this warrants close monitoring. Thrombolysis with urokinase alone or combined with low-molecular-weight heparin might be effective and well tolerated as treatment strategies. Prophylactic heparin increases the duration of catheter usability (P < 0.005, 95% confidence interval 0.35-0.81), decreases catheter occlusion, but may not uniformly prevent thrombosis. CVL-related thrombosis is an underreported complication because events in the majority occur silently. Currently, solid evidence-based recommendations for diagnosis and treatment are not possible. Well designed prospective studies are urgently required to establish a concrete investigational approach to CVC-related thrombosis and to institute safe therapeutic modalities.


A cohort study and meta-analysis of isolated diastolic hypertension: searching for a threshold to guide treatment.
Whether isolated diastolic hypertension (IDH), as defined by the 2017 American College of Cardiology (ACC)/American Heart Association (AHA) guideline, is associated with cardiovascular disease (CVD) has been disputed. We aimed to further study the associations of IDH with (i) subclinical CVD in the form of coronary artery calcium (CAC), (ii) incident systolic hypertension, and (iii) CVD events. We used multivariable-adjusted logistic and Cox regression to test whether IDH by 2017 ACC/AHA criteria (i.e. systolic blood pressure <130 mmHg and diastolic blood pressure ≥80 mmHg) was associated with the above outcomes in the Multi-Ethnic Study of Atherosclerosis (MESA). In a random-effects meta-analysis of the association between IDH and CVD events, we combined the MESA results with those from seven other previously published cohort studies. Among the 5104 MESA participants studied, 7.5% had IDH by the 2017 ACC/AHA criteria. There was no association between IDH and CAC [e.g. adjusted prevalence odds ratio for CAC >0 of 0.88 (95% CI 0.66, 1.17)]. Similarly, while IDH was associated with incident systolic hypertension, there was no statistically significant associations with incident CVD [hazard ratio 1.19 (95% CI 0.77, 1.84)] or death [hazard ratio 0.94 (95% CI 0.65, 1.36)] over 13 years in MESA. In a stratified meta-analysis of eight cohort studies (10 037 843 participants), the 2017 IDH definition was also not consistently associated with CVD and the relative magnitude of any potential association was noted to be numerically small [e.g. depending on inclusion criteria applied in the stratification, the adjusted hazard ratios ranged from 1.04 (95% CI 0.98, 1.10) to 1.09 (95% 1.03, 1.15)]. The lack of consistent excess in CAC or CVD suggests that emphasis on healthy lifestyle rather than drug therapy is sufficient among the millions of middle-aged or older adults who now meet the 2017 ACC/AHA criteria for IDH, though they require follow-up for incident systolic hypertension. These findings may not extrapolate to adults younger than 40 years, motivating further study in this age group.


A Phylogenetic and Phenotypic Analysis of Salmonella enterica Serovar Weltevreden, an Emerging Agent of Diarrheal Disease in Tropical Regions.
Salmonella enterica serovar Weltevreden (S. Weltevreden) is an emerging cause of diarrheal and invasive disease in humans residing in tropical regions. Despite the regional and international emergence of this Salmonella serovar, relatively little is known about its genetic diversity, genomics or virulence potential in model systems. Here we used whole genome sequencing and bioinformatics analyses to define the phylogenetic structure of a diverse global selection of S. Weltevreden. Phylogenetic analysis of more than 100 isolates demonstrated that the population of S. Weltevreden can be segregated into two main phylogenetic clusters, one associated predominantly with continental Southeast Asia and the other more internationally dispersed. Subcluster analysis suggested the local evolution of S. Weltevreden within specific geographical regions. Four of the isolates were sequenced using long read sequencing to produce high quality reference genomes. Phenotypic analysis in Hep-2 cells and in a murine infection model indicated that S. Weltevreden were significantly attenuated in these models compared to the classical S. Typhimurium reference strain SL1344. Our work outlines novel insights into this important emerging pathogen and provides a baseline understanding for future research studies.


Differences in Cognitive Function of Rats with Traumatic Brain Injuries Following Hyperbaric Oxygen Therapy.
BACKGROUND Hyperbaric oxygen (HBO) is a historical therapeutic option in the treatment of various types of brain damage. At present, clinical treatment of hypoxic-ischemic injury is giving priority to cognitive training. The effects of HBO on cognitive dysfunction were observed in a controlled cortical impact (CCI) rat model. MATERIAL AND METHODS Seventy male SD rats were randomly divided into control (n=10) and intervention (n=60) groups. All rats underwent baseline water maze testing 1 day before modeling, and were retested 8 weeks after modeling. The percentage of residence time during escape latency in the target quadrant and the total time were recorded. Data were analyzed by SPSS 16.0 software. P<0.05 was considered statistically significant. RESULTS After 8 weeks, no statistical difference (P>0.05) existed in spatial learning ability in the 3-day and 5-day groups when compared with baseline. The other groups were statistically different by auto-comparison (P<0.05). No statistical difference (P>0.05) in spatial memory existed in the 5-day and 1-week groups when compared with baseline, while a significant difference was noted in the other groups by self-comparison (P<0.05). No statistical difference (P>0.05) was noted in the level of expression of growth-associated protein-43 (GAP-43) and synaptophysin (Syn) in the 1-day group compared with the control group. The remaining groups and the control group were statistically different (P<0.05), while the level of expression of GAP-43 and Syn in the 5-day, 1-week, and 2-week groups was significantly different compared with that in the control group (P<0.01). CONCLUSIONS If HBO therapy was provided 5-7 days after craniocerebral trauma, there was apparent improvement in cognitive function and neuroplasticity.


Generation of functional dendritic cells for potential use in the treatment of acute lymphoblastic leukemia.
Immunotherapy of malignant diseases mediated by dendritic cells (DC) pulsed with tumor antigens ex vivo is a promising new tool in the individual treatment of malignant diseases. The present study focuses on the problem of how to optimize in vitro culture conditions and induce the maturation of DC with the capacity to induce antitumor immunity toward leukemic cells. DC were generated from peripheral mononuclear cells by co-cultivation with granulocyte/macrophage-colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). Tumor antigens were added for 2 h after 7 days in culture. Irradiated leukemic blasts, blast lysate, apoptotic cells from the Jurkat cell line (T ALL) and their lysate were used in various concentrations for antigen pulsing. Harvested DC were phenotyped by flow cytometry, and viability was assessed using trypan blue exclusion (Annexin test). After the cells had been pulsed with tumor antigens and co-cultured with autologous lymphocytes, the production of interferon-gamma (IFN-gamma) and IL-12 was analyzed, and lymphocyte proliferative response and cytotoxicity against the target tumor cell line were assessed. The cultivation of monocytes under the described conditions led to the expression of surface markers typical of DC (i.e. CD83, CD86, HLA-DR, CD11c and CD40). Pulsation by antigens from leukemic cells further increased the cell populations expressing these markers. Antigen pulsation decreased the viability of generated DC depending on the increase in concentration of tumor antigens. Pulsed DC-lymphocyte interaction increased the proliferative response of lymphocytes and IFN-gamma production depending on the type of tumor antigens used for pulsation. The highest proliferative response was detected with DC pulsed with Jurkat cell-line lysate. Similarly to the proliferation assay, cytotoxic testing showed the highest efficiency of DC pulsed with Jurkat cell-line lysate in killing the target malignant cells. Our results show that an appropriate antigen concentration used for DC pulsing is one of the crucial factors in an effective treatment strategy, as high concentrations of tumor antigens induce apoptosis of DC, thereby rendering them non-functional. Under optimal conditions, pulsation by lysate from leukemic blasts induced the maturation of DC and led to an increase in the proliferation of autologous lymphocytes, to the production of Th1-cytokines and to the induction of cytotoxicity toward the leukemic cell line. These results are encouraging for the possible application of pulsed DC in the therapy of acute lymphoblastic leukemia.


Hierarchical natural move Monte Carlo refines flexible RNA structures into cryo-EM densities.
Ribonucleic acids (RNAs) play essential roles in living cells. Many of them fold into defined three-dimensional (3D) structures to perform functions. Recent advances in single-particle cryo-electron microscopy (cryo-EM) have enabled structure determinations of RNA to atomic resolutions. However, most RNA molecules are structurally flexible, limiting the resolution of their structures solved by cryo-EM. In modeling these molecules, several computational methods are limited by the requirement of massive computational resources and/or the low efficiency in exploring large-scale structural variations. Here we use hierarchical natural move Monte Carlo (HNMMC), which takes advantage of collective motions for groups of nucleic acid residues, to refine RNA structures into their cryo-EM maps, preserving atomic details in the models. After validating the method on a simulated density map of tRNA, we applied it to objectively obtain the model of the folding intermediate for the specificity domain of ribonuclease P from Bacillus subtilis and refine a flexible ribosomal RNA (rRNA) expansion segment from the Mycobacterium tuberculosis (Mtb) ribosome in different conformational states. Finally, we used HNMMC to model atomic details and flexibility for two distinct conformations of the complete genomic RNA (gRNA) inside MS2, a single-stranded RNA virus, revealing multiple pathways for its capsid assembly.


Speciation over the edge: gene flow among non-human primate species across a formidable biogeographic barrier.
Many genera of terrestrial vertebrates diversified exclusively on one or the other side of Wallace's Line, which lies between Borneo and Sulawesi islands in Southeast Asia, and demarcates one of the sharpest biogeographic transition zones in the world. Macaque monkeys are unusual among vertebrate genera in that they are distributed on both sides of Wallace's Line, raising the question of whether dispersal across this barrier was an evolutionary one-off or a more protracted exchange-and if the latter, what were the genomic consequences. To explore the nature of speciation over the edge of this biogeographic divide, we used genomic data to test for evidence of gene flow between macaque species across Wallace's Line after macaques colonized Sulawesi. We recovered evidence of post-colonization gene flow, most prominently on the X chromosome. These results are consistent with the proposal that gene flow is a pervasive component of speciation-even when barriers to gene flow seem almost insurmountable.


Measuring patient satisfaction for quality improvement.
Surveys used for health plan quality reporting are generally administered annually to health plan enrollees to assess satisfaction with both the health plan and health care services. Therefore, surveys may lack sensitivity to measure the effects of patient-focused, quality improvement initiatives that could demonstrate results in a shorter time period. We describe the development and testing of a multidimensional, visit-specific measure of satisfaction with primary care that may be used in quality improvement. Conducted in five adult and pediatric primary care sites serving a commercial, largely managed-care population, the survey includes the Medical Outcomes Study Visit-Specific Questionnaire, the American Board of Internal Medicine Patient Satisfaction Questionnaire, and locally developed items. We assessed the instrument's reliability, validity, and utility for quality improvement. For both adult and pediatric samples, three factors emerged: satisfaction with the provider, satisfaction with access, and satisfaction with the office. Satisfaction with the provider and with the office were independently correlated with overall satisfaction in both samples; satisfaction with access was significantly correlated with overall satisfaction only for adults. For adults, patients who disenrolled from the health plan were less satisfied with the office compared with patients who remained with the health plan. Finally, for adults, we detected significant differences across practice sites in terms of satisfaction with office and access; for children, there were intersite differences in terms of satisfaction with provider, office, and access. We have support for the reliability and validity of this instrument that has identified differences in satisfaction between practice sites that may be used for quality improvement.


Removal efficiency of propolis paste dressing from the root canal.
The aim of this study was to investigate through scanning electron microscopy (SEM) the cleaning of root canal walls after the use of experimental propolis or calcium hydroxide root canal dressings. Twenty single-rooted teeth were used. After conventional cleaning and shaping procedures and removal of the smear layer with 17% EDTA, the teeth were divided into four groups according to the medication used (N=5): Group I (control) - No drug, Group II - Calcium hydroxide dressing, Group III - Propolis paste A70D and Group IV - Propolis paste D70D. The medications were introduced into the root canals and maintained for 7 days, then removed with a K-file and 5 mL of 1% sodium hypochlorite irrigation. Finally, the canals were flushed with 2 mL of 17% EDTA for 3 min. For SEM analysis, the roots were cleaved and microphotographs from the middle third of the root canal were taken at 750x. The cleaning of the root canal walls was determined by the number of open dentinal tubules as verified with the software Image Tool 3.1. The statistical analysis was performed by ANOVA and Tukey's test (p<0.05). The results showed no statistically significant difference between the calcium hydroxide and propolis groups. The experimental propolis pastes presented acceptable physical characteristics to be used as intracanal medicaments.


Prospective Nonrandomized Analytical Comparative Study of Clinicoradiological Relationship and Quality of Life between Lumbar Interbody Fusion and Posterolateral Lumbar Fusion.
This prospective, nonrandomized, analytic comparative study analyzed the outcome of 100 patients who underwent spinal fusion surgeries (Posterolateral fusion (PLF) and posterior lumbar interbody fusion [PLIF]) with 6 months of follow-up. The aim of our study was to compare clinicoradiological outcome of the lumbar spinal fusion surgeries (PLF vs. PLIF/transforaminal lumbar interbody fusion [TLIF]), to assess the quality of life pre- and post-operatively in all patients, and to compare the results within the groups. We retrieved the articles related to posterolateral fusion (PLF) and PLIF/TLIF through computer-assisted PubMed and Cochrane database search. Most of the studies in previous literature did not show any significant difference in the success of fusion between the two groups. However, the global outcome in terms of clinical and radiological parameters was good in all the studies. Senior neurosurgeons who are part of the study treated 100 patients presented with degenerative disc disease (DDD). Patients underwent two types of spinal fusion surgeries: Posterolateral fusion (PLF) and PLIF/TLIF, and those two groups were compared for clinicoradiological outcome, successful fusion, and quality of life at 6 months of follow-up. Results were analyzed statistically using SPSS version 21.0 and presented in terms of frequency, percentage, mean, and standard deviation. Measurements were compared between groups using the Student's t-test (independent and paired) for normally distributed variables. Percentage was compared with Chi-square test in various parameters. Statistical significance was determined with P values; P < 0.05 was considered statistically significant. Patients aged between 30and 65 years with lumbar DDD who met the inclusion criteria were included in the study. Patients were operated for 1-3 vertebral levels and showed clinical satisfactory outcome on visual analog scale (P = 0.004) and modified-Oswestry low back pain disability questionnaire (Oswestry disability index) (P = 0.000) at 6 months as compared to the preoperative data, which was statistically significant. Radiological outcome in terms of lumbar lordotic angle (LLA, P = 0.000) and ratio of disc space height (DSH) and height of immediate superior vertebral body (P = 0.000) at 3 months of follow-up was also statistically significant. All of our patients showed a well-placed implant (screws and cage) in the follow-up period. Our patients showed successful fusion more in the PLIF group (81.25%) as compared to the PLF group (67.30%), but those results are not statistically significant (P = 0.112), and all of our patients showed statistically significant difference (P = 0.000) in quality of life score at 6 months of follow-up when compared with the preoperative score. We concluded from our study and after the review of literature that the patients with lumbar DDD should undergo spinal instrumentation surgery either PLF or PLIF as per the requirement as these surgeries provide good clinical and radiological outcomes in terms of pain, disability index, LLA, and maintenance of DSH. Although both the groups have few degrees of risks and complications, these are not major one and can be managed easily. The fusion rates are similar in both the groups (PLF and PLIF). Quality of life index showed significant difference within 1 week, after 3 months, and after 6 months of surgery in all of our patients.


Adversarial Deep Learning in EEG Biometrics.
Deep learning methods for person identification based on electroencephalographic (EEG) brain activity encounters the problem of exploiting the temporally correlated structures or recording session specific variability within EEG. Furthermore, recent methods have mostly trained and evaluated based on single session EEG data. We address this problem from an invariant representation learning perspective. We propose an adversarial inference approach to extend such deep learning models to learn session-invariant person-discriminative representations that can provide robustness in terms of longitudinal usability. Using adversarial learning within a deep convolutional network, we empirically assess and show improvements with our approach based on longitudinally collected EEG data for person identification from half-second EEG epochs.


Report from a WHO Working Group: standard method for detecting upper respiratory carriage of Streptococcus pneumoniae.
Numerous studies evaluating the efficacy of conjugate pneumococcal vaccines are being conducted or planned throughout the world. Some of these studies are evaluating the effect of vaccine on nasopharyngeal (NP) carriage. The World Health Organization established a Working Group comprised of representatives from these trials and other NP colonization experts to establish core, standardized methods for the study of pneumococcal NP colonization that could be used in these trials. The intent was to reduce or eliminate variability in key methods which themselves could contribute to variability of observed pneumococcal NP colonization. In this way variability of vaccine effects between trials on NP colonization could more easily be analyzed for population or vaccine differences without the confounding effect caused by differences in study methodology. This paper presents the evidence base supporting the need for standardized NP colonization study methods, the methods themselves (Core Consensus Methods), including collection techniques, culture media, equipment, serotyping, storage of specimens and transport of isolates agreed on by the Working Group as well as a discussion of research priorities. The Core Consensus Methods provide a common methodology to conduct pneumococcal NP colonization studies with minimum interstudy method variability. The intention is to allow more meaningful comparisons of study results from conjugate pneumococcal vaccine trials.


A multicenter evaluation of diagnostic tools to define endpoints for programs to eliminate bancroftian filariasis.
Successful mass drug administration (MDA) campaigns have brought several countries near the point of Lymphatic Filariasis (LF) elimination. A diagnostic tool is needed to determine when the prevalence levels have decreased to a point that MDA campaigns can be discontinued without the threat of recrudescence. A six-country study was conducted assessing the performance of seven diagnostic tests, including tests for microfilariae (blood smear, PCR), parasite antigen (ICT, Og4C3) and antifilarial antibody (Bm14, PanLF, Urine SXP). One community survey and one school survey were performed in each country. A total of 8,513 people from the six countries participated in the study, 6,443 through community surveys and 2,070 through school surveys. Specimens from these participants were used to conduct 49,585 diagnostic tests. Each test was seen to have both positive and negative attributes, but overall, the ICT test was found to be 76% sensitive at detecting microfilaremia and 93% specific at identifying individuals negative for both microfilariae and antifilarial antibody; the Og4C3 test was 87% sensitive and 95% specific. We conclude, however, that the ICT should be the primary tool recommended for decision-making about stopping MDAs. As a point-of-care diagnostic, the ICT is relatively inexpensive, requires no laboratory equipment, has satisfactory sensitivity and specificity and can be processed in 10 minutes-qualities consistent with programmatic use. Og4C3 provides a satisfactory laboratory-based diagnostic alternative.


Density functional theory as a tool for the structure determination of radiation-induced bioradicals.
The use of density functional methods for the elucidation of the structure of radiation-induced bio-radicals by comparison of computed and experimental EPR properties is discussed. Three case studies, radiation induced radicals of the amino acid alanine, steroid hormones and beta-D-fructose, with increasing degree of uncertainty about the proposed radical structures, are investigated. Next to the analysis of the isotropic and anisotropic components of the hyperfine tensor, also the direction cosines of the principal axes of this tensor were investigated in greater detail in the case of the beta-D-fructose radicals. Since all radicals considered in this contribution are formed in a solid matrix, also the question as to how to incorporate the effect of the molecular environment is addressed. It is concluded that the methodology outlined represents a powerful tool to aid experimentalists in the assignment of the contributions of various radicals contributing to the observed EPR spectra.


Enzyme inactivation via disulphide-thiol exchange as catalysed by a rat liver membrane protein.
1. The inactivation of cytosol enzymes by a rat liver membrane protein was studied with crude microsomal fraction, plasma membranes or a partially purified preparation of inactivation factor. 2. Complete inactivation of (125)I-labelled glucose 6-phosphate dehydrogenase (EC 22.214.171.124) by membranes did not result in any detectable change in molecular weight when the products were analysed by gradient polyacrylamide-gel electrophoresis. 3. Inactivation of radioactive enzyme was not accompanied by extensive binding to the membrane surface. The maximum extent of binding was 15% of the total enzyme labelled, and bound radioactivity was released only slowly, mainly as trichloroacetic acid-insoluble material. 4. Treatment of membranes with dithiothreitol destroyed the inactivation capacity, whereas the thiol-alkylating agent iodoacetamide had no effect. Partial restoration of the inactivation capacity of reduced membranes after exposure to air was prevented by membrane alkylation with iodoacetamide. 5. Modification of enzyme thiol groups during inactivation was determined by measuring a decrease in iodoacetamide-reactive groups in purified glucose 6-phosphate dehydrogenase. 6. Incubation of membrane-inactivated glucose 6-phosphate dehydrogenase with dithiothreitol resulted in a partial restoration of enzyme activity. 7. As a result of these experiments it is concluded that inactivation proceeds by a disulphide-thiol exchange mechanism. The proposal that this reaction could be involved in the initial step of enzyme degradation is discussed.


Growth arrest specific protein (GAS) 6: a role in the regulation of proliferation and functional capacity of the perinatal rat beta cell.
Maternal low-protein (LP) diet during gestation results in a reduced beta cell mass in the offspring at birth and this may hamper the ability to adapt to high-energy food and sedentary lifestyle later in life. To investigate the biology behind the LP-offspring phenotype, this study aimed to identify differentially expressed genes in the pancreas and their potential role in the fetal programming. Wistar rats were given either an LP diet or normal-chow (NC) diet during gestation and differentially expressed genes in the offspring around the time of birth were identified using RNA microarray and quantitative PCR. The role of a differentially expressed gene, growth arrest specific protein 6 (GAS6), was evaluated in vitro using neonatal rat islets. The mRNA level of Gas6, known to be mitogenic in other tissues, was reduced in LP offspring. The mRNA content of Mafa was increased in LP offspring suggesting an early maturation of beta cells. When applied in vitro, GAS6 increased proliferation of neonatal pancreatic beta cells, while reducing glucose-stimulated insulin secretion without changing the total insulin content of the islets. In addition, GAS6 decreased the mRNA content of Mafa. We propose a role for GAS6 in the regulation of pancreatic beta cells in the critical period around the time of birth. Our results support the hypothesis that the reduced beta cell mass seen in LP offspring is caused by a change in the intra-uterine environment that favours premature maturation of the beta cells.


Frontoparietal and Cingulo-opercular Networks Play Dissociable Roles in Control of Working Memory.
We used magnetoencephalography to characterize the spatiotemporal dynamics of cortical activity during top-down control of working memory (WM). fMRI studies have previously implicated both the frontoparietal and cingulo-opercular networks in control over WM, but their respective contributions are unclear. In our task, spatial cues indicating the relevant item in a WM array occurred either before the memory array or during the maintenance period, providing a direct comparison between prospective and retrospective control of WM. We found that in both cases a frontoparietal network activated following the cue, but following retrocues this activation was transient and was succeeded by a cingulo-opercular network activation. We also characterized the time course of top-down modulation of alpha activity in visual/parietal cortex. This modulation was transient following retrocues, occurring in parallel with the frontoparietal network activation. We suggest that the frontoparietal network is responsible for top-down modulation of activity in sensory cortex during both preparatory attention and orienting within memory. In contrast, the cingulo-opercular network plays a more downstream role in cognitive control, perhaps associated with output gating of memory.


Quantifying the impact of different approaches for handling continuous predictors on the performance of a prognostic model.
Continuous predictors are routinely encountered when developing a prognostic model. Investigators, who are often non-statisticians, must decide how to handle continuous predictors in their models. Categorising continuous measurements into two or more categories has been widely discredited, yet is still frequently done because of its simplicity, investigator ignorance of the potential impact and of suitable alternatives, or to facilitate model uptake. We examine three broad approaches for handling continuous predictors on the performance of a prognostic model, including various methods of categorising predictors, modelling a linear relationship between the predictor and outcome and modelling a nonlinear relationship using fractional polynomials or restricted cubic splines. We compare the performance (measured by the c-index, calibration and net benefit) of prognostic models built using each approach, evaluating them using separate data from that used to build them. We show that categorising continuous predictors produces models with poor predictive performance and poor clinical usefulness. Categorising continuous predictors is unnecessary, biologically implausible and inefficient and should not be used in prognostic model development. © 2016 The Authors. Statistics in Medicine published by John Wiley & Sons Ltd.


Smad2 and Smad3 are redundantly essential for the suppression of iNOS synthesis in macrophages by regulating IRF3 and STAT1 pathways.
Although transforming growth factor (TGF)-β1 is a well-known immunosuppressive cytokine, little is known about the role of its downstream transcription factors, Smad2 and Smad3, in the suppression of macrophage activation. Previous studies have demonstrated that Smad3 is critical for the suppression of LPS-mediated inducible nitric oxide (NO) synthase (iNOS) induction, although the role of Smad2 remains to be investigated. In this study, we found that iNOS induction was enhanced in Smad2-deficient bone marrow-derived macrophages (BMDMs) and peritoneal macrophages in vitro and tumor-associated macrophages in vivo, compared with wild-type (WT) macrophages. However, TGF-β1 still suppressed iNOS induction in Smad2-deficient macrophages. In Smad2/3 double knockout (KO) (Smad2/3 DKO) BMDMs, LPS-mediated NO/iNOS induction was more strongly elevated than in Smad2 or Smad3 single KO BMDMs, and its suppression by exogenous TGF-β1 was severely impaired. These data suggest that Smad2 and Smad3 redundantly regulate iNOS induction. Similarly, the production of IL-6 and TNFα, but not IL-10 was augmented in Smad2/3 DKO BMDMs, suggesting that Smad2 and Smad3 also redundantly suppressed some cytokines production. In Smad2/3 DKO macrophages, TLR3- as well as TLR4-mediated IRF3 activation and IFN-β production were strongly augmented, which resulted in hyper STAT1 phosphorylation. Furthermore, IFN-β- and IFN-γ-induced iNOS induction in the absence of TLR signaling and STAT1 transcriptional activity were augmented in Smad2/3 DKO BMDMs. These results suggest that Smad2 and Smad3 negatively regulate iNOS induction in macrophages by suppressing multiple steps in the IRF3-IFN-β-STAT1 pathway.


Investigation of a grating-based stretcher/compressor for carrier-envelope phase stabilized fs pulses.
In this work, we experimentally investigate the effect of a grating based pulse stretcher/compressor on the carrier-envelope phase stability of femtosecond pulses. Grating based stretcher-compressor (SC) setups have been avoided in past demonstrations of chirped pulse amplification (CPA) of carrier envelope phase (CEP) stabilized femtosecond pulses, because they were expected to introduce significantly stronger CEP fluctuations than material-based SC systems. Using a microstructure fiber-based detection setup, we measure CEP fluctuations of PhiCE,SC = 340 milliradians rms for a frequency range from 63 mHz to 102 kHz for pulses propagating through the SC setup. When bypassing the beam path through the SC, we find CEP fluctuations of PhiCE,bypass = 250 milliradians rms. These values contain significant contributions from amplitude-to-phase conversion in our microstructure fiber-based detection setup for PhiCE. Hence, we do not unambiguously measure any added CEP noise intrinsic to the SC setup. To distinguish between intrinsic SC effects and amplitude-to-phase conversion, we introduce controlled beam pointing fluctuations alpha and again compare the phase noise introduced when passing through / bypassing the SC. Our measurements do not reveal any intrinsic effects of the SC system, but allow us to place an upper limit on the sensitivity of our SC system of PhiCEintrinsic,SC / alpha < 13000 rad/rad. Our results demonstrate experimentally that there is not a strong coupling mechanism between CEP and beam pointing through a stretcher/compressor , as well as measuring significantly smaller CEP fluctuations than experimental results reported previously.


The SRS-Schwab adult spinal deformity classification: assessment and clinical correlations based on a prospective operative and nonoperative cohort.
The SRS-Schwab classification of adult spinal deformity (ASD) is a validated system that provides a common language for the complex pathology of ASD. Classification reliability has been reported; however, correlation with treatment has not been assessed. To assess the clinical relevance of the SRS-Schwab classification based on correlations with health-related quality of life (HRQOL) measures and the decision to pursue operative vs nonoperative treatment. Prospective analysis of consecutive ASD patients (18 years of age and older) collected through a multicenter group. The SRS-Schwab classification includes a curve type descriptor and 3 sagittal spinopelvic modifiers (sagittal vertical axis, pelvic tilt, pelvic incidence/lumbar lordosis mismatch). Differences in demographics, HRQOL (Oswestry Disability Index, SRS-22, Short Form-36), and classification between operative and nonoperative patients were evaluated. A total of 527 patients (mean age, 52.9 years; range, 18.4-85.1 years) met inclusion criteria. Significant differences in HRQOL were identified based on SRS-Schwab curve type, with thoracolumbar and primary sagittal deformities associated with greater disability and poorer health status than thoracic or double curve deformities. Operative patients had significantly poorer grades for each of the sagittal spinopelvic modifiers, and progressively higher grades were associated with significantly poorer HRQOL (P < .05). Patients with worse sagittal spinopelvic modifier grades were significantly more likely to require major osteotomies, iliac fixation, and decompression (P ≤ .009). The SRS-Schwab classification provides a validated language to describe and categorize ASD. This study demonstrates that the SRS-Schwab classification reflects severity of disease state based on multiple measures of HRQOL and significantly correlates with the important decision of whether to pursue operative or nonoperative treatment.


Prediction of forearm muscle activity during gripping.
Occupational exposure is typically assessed by measuring forces and body postures to infer muscular loading. Better understanding of workplace muscle activity levels would aid in indicating which muscles may be at risk for overexertion and injury. However, electromyography collection in the workplace is often not practical. Therefore, a set of equations was developed and validated using data from two separate days to predict forearm muscle activity (involving six wrist and finger muscles) from grip force and posture of the wrist (flexed, neutral and extended) and forearm (pronated, neutral, supinated). The error in predicting activation levels of each forearm muscle across the range of grip forces, using the first day data (root mean square error; RMSEmodel), ranged from 8.9% maximal voluntary electrical activation (MVE) (flexor carpi radialis) to 11% MVE (extensor digitorum communis). Grip force was the main contributor to predicting muscle activity levels, explaining over 70% of the variance in flexor activation levels and up to 60% in extensor activation levels, respectively. Inclusion of gender as a variable in the model improved estimates of flexor but not extensor activity. While posture itself explained minimal variance in activation without grip force (< 10% MVE), wrist and forearm posture were required (with grip force) to explain over 70% of the variance of all six muscles. The validation process indicated good day-to-day reliability of each equation, with similar error for flexor muscle models but slightly higher error in the extensor models when predicting activity levels for the second day of data (RMSEvalid ranging from 8.9% to 12.7% MVE). Detailed error analysis during validation revealed that inclusion of posture in the model effectively decreased error at grip forces above 25% maximum, but was detrimental at very low grip forces. This study presents a potential new tool to estimate forearm muscle loading in the workplace using grip force and posture, as a surrogate to use of a complex biomechanical model.


Behavioral neurophysiology: insights into seeing and grasping.
One marvels at a batter's ability to hit a baseball traveling at 150 kilometers per hour or a monkey's skill in snatching a flying insect. Indeed, the ability of many animals to reach out, grasp, and manipulate objects is a feat of biological engineering unmatched by even state-of-the-art robots. But how are the objects of our attention chosen and how are the eyes and hands directed to it? Recent progress in behavioral neurophysiology has clarified some of the brain mechanisms at work.