The spontaneous formation of single-molecule junctions via terminal alkynes.
Herein, we report the spontaneous formation of single-molecule junctions via terminal alkyne contact groups. Self-assembled monolayers that form spontaneously from diluted solutions of 1, 4-diethynylbenzene (DEB) were used to build single-molecule contacts and assessed using the scanning tunneling microscopy-break junction technique (STM-BJ). The STM-BJ technique in both its dynamic and static approaches was used to characterize the lifetime (stability) and the conductivity of a single-DEB wire. It is demonstrated that single-molecule junctions form spontaneously with terminal alkynes and require no electrochemical control or chemical deprotonation. The alkyne anchoring group was compared against typical contact groups exploited in single-molecule studies, i.e. amine (benzenediamine) and thiol (benzendithiol) contact groups. The alkyne contact showed a conductance magnitude comparable to that observed with amine and thiol groups. The lifetime of the junctions formed from alkynes were only slightly less than that of thiols and greater than that observed for amines. These findings are important as (a) they extend the repertoire of chemical contacts used in single-molecule measurements to 1-alkynes, which are synthetically accessible and stable and (b) alkynes have a remarkable affinity toward silicon surfaces, hence opening the door for the study of single-molecule transport on a semiconducting electronic platform.


Temperature-controlled assembly of high ordered/disordered dodecylamine layers on HOPG: consequences for DNA patterning.
It is shown that temperature-controlled ordered layers of dodecylamine, self-assembled on highly oriented pyrolytic graphite (HOPG), are an appropriate substrate for aligning individual DNA molecules. High resolution atomic force microscopy (AFM) permits visualization of the lamellar structure of dodecylamine on HOPG. The DNA adsorbed on ordered dodecylamine layers is stretched and oriented along the lamellae, while DNA on disordered dodecylamine layers is found to be in a random conformation. Small DNA bubbles appear in the case of partially denaturated linear double stranded DNA (dsDNA) adsorbed on ordered layers of dodecylamine.


Radiation dosimetry results and safety correlations from 90Y-ibritumomab tiuxetan radioimmunotherapy for relapsed or refractory non-Hodgkin's lymphoma: combined data from 4 clinical trials.
Ibritumomab tiuxetan is an anti-CD20 murine IgG1 kappa monoclonal antibody (ibritumomab) conjugated to the linker-chelator tiuxetan, which securely chelates (111)In for imaging or dosimetry and (90)Y for radioimmunotherapy (RIT). Dosimetry and pharmacokinetic data from 4 clinical trials of (90)Y-ibritumomab tiuxetan RIT for relapsed or refractory B-cell non-Hodgkin's lymphoma (NHL) were combined and assessed for correlations with toxicity data. Data from 179 patients were available for analysis. Common eligibility criteria included <25% bone marrow involvement by NHL, no prior myeloablative therapy, and no prior RIT. The baseline platelet count was required to be > or = 100,000 cells/mm(3) for the reduced (90)Y-ibritumomab tiuxetan administered dose (7.4-11 MBq/kg [0.2-0.3 mCi/kg]) or > or = 150,000 cells/mm(3) for the standard (90)Y-ibritumomab tiuxetan administered dose (15 MBq/kg [0.4 mCi/kg]). Patients were given a tracer administered dose of 185 MBq (5 mCi) (111)In-ibritumomab tiuxetan on day 0, evaluated with dosimetry, and then a therapeutic administered dose of 7.4-15 MBq/kg (0.2-0.4 mCi/kg) (90)Y-ibritumomab tiuxetan on day 7. Both ibritumomab tiuxetan administered doses were preceded by an infusion of 250 mg/m(2) rituximab to clear peripheral B-cells and improve ibritumomab tiuxetan biodistribution. Residence times for (90)Y in blood and major organs were estimated from (111)In biodistribution, and the MIRDOSE3 computer software program was used, with modifications to account for patient-specific organ masses, to calculate radiation absorbed doses to organs and red marrow. Median radiation absorbed doses for (90)Y were 7.42 Gy to spleen, 4.50 Gy to liver, 2.11 Gy to lung, 0.23 Gy to kidney, 0.62 Gy (blood-derived method) and 0.97 Gy (sacral image-derived method) to red marrow, and 0.57 Gy to total body. The median effective blood half-life was 27 h, and the area under the curve (AUC) was 25 h. No patient failed to meet protocol-defined dosimetry safety criteria and all patients were eligible for treatment. Observed toxicity was primarily hematologic, transient, and reversible. Hematologic toxicity did not correlate with estimates of red marrow radiation absorbed dose, total-body radiation absorbed dose, blood effective half-life, or blood AUC. Relapsed or refractory NHL in patients with adequate bone marrow reserve and <25% bone marrow involvement by NHL can be treated safely with (90)Y-ibritumomab tiuxetan RIT on the basis of a fixed, weight-adjusted dosing schedule. Dosimetry and pharmacokinetic results do not correlate with toxicity.


Effects of beta-hydroxybutyrate administration on MK-801-induced schizophrenia-like behaviour in mice.
Impaired cerebral glucose metabolism is a core pathological feature of schizophrenia. We recently demonstrated that a ketogenic diet, causing a shift from glycolysis to ketosis, normalized schizophrenia-like behaviours in an acute N-methyl-D-aspartate (NMDA) receptor antagonist model of the illness. Ketogenic diet produces the ketone body, β-hydroxybutyrate (BHB), which may serve as an alternative fuel source in its own right without a strict dietary regime. We hypothesized that chronic administration of BHB replicates the therapeutic effects of ketogenic diet in an acute NMDA receptor hypofunction model of schizophrenia in mice. C57Bl/6 mice were either treated with acute doses of 2 mmol/kg, 10 mmol/kg, or 20 mmol/kg BHB or received daily intraperitoneal injections of 2 mmol/kg BHB or saline for 3 weeks. Behavioural testing assessed the effect of acute challenge with 0.2 mg/kg MK-801 or saline on open field behaviour, social interaction, and prepulse inhibition of startle (PPI). Acute BHB administration dose-dependently increased BHB plasma levels, whereas the 2 mmol/kg dose increased plasma glucose levels. The highest acute dose of BHB supressed spontaneous locomotor activity, MK-801-induced locomotor hyperactivity and MK-801-induced disruption of PPI. Chronic BHB treatment normalized MK-801-induced hyperlocomotion, reduction of sociability, and disruption of PPI. In conclusion, BHB may present a novel treatment option for patients with schizophrenia by providing an alternative fuel source to normalize impaired glucose metabolism in the brain.


Validation of high throughput sequencing and microbial forensics applications.
High throughput sequencing (HTS) generates large amounts of high quality sequence data for microbial genomics. The value of HTS for microbial forensics is the speed at which evidence can be collected and the power to characterize microbial-related evidence to solve biocrimes and bioterrorist events. As HTS technologies continue to improve, they provide increasingly powerful sets of tools to support the entire field of microbial forensics. Accurate, credible results allow analysis and interpretation, significantly influencing the course and/or focus of an investigation, and can impact the response of the government to an attack having individual, political, economic or military consequences. Interpretation of the results of microbial forensic analyses relies on understanding the performance and limitations of HTS methods, including analytical processes, assays and data interpretation. The utility of HTS must be defined carefully within established operating conditions and tolerances. Validation is essential in the development and implementation of microbial forensics methods used for formulating investigative leads attribution. HTS strategies vary, requiring guiding principles for HTS system validation. Three initial aspects of HTS, irrespective of chemistry, instrumentation or software are: 1) sample preparation, 2) sequencing, and 3) data analysis. Criteria that should be considered for HTS validation for microbial forensics are presented here. Validation should be defined in terms of specific application and the criteria described here comprise a foundation for investigators to establish, validate and implement HTS as a tool in microbial forensics, enhancing public safety and national security.


Molecular Imprinting of Tri-O-Acetyladenosine for the Synthetic Imitation of an ATP-Binding Cleft in Protein Kinases.
A number of diseases, including cancer, diabetes, and inflammation, are linked to deregulation of cell signaling pathways controlled by protein kinases. Inhibition of the kinases involved can interrupt aberrant signaling and have a specific therapeutic effect. Protein kinases are recognized as validated therapeutic targets for the treatment of a number of diseases and there are considerable efforts to discover new kinase inhibitors suitable for drug development. The main goal of this study was to fabricate the synthetic imitations of the adenosine triphosphate (ATP) binding cleft in protein kinases and thus produce polymers suitable for screening and isolation of new protein kinase ATP-mimetic inhibitors from different sources. Such polymers were created by the imprinting of tri-O-acetyladenosine in acrylic polymer matrix with the use of methacrylic acid (MAA) or 3-vinylbenzoic acid (VBA) as a functional monomer and ethylene glycol dimethacrylate as a cross-linking agent. The imprints prepared with the use of VBA demonstrate substantially better binding efficiency than that with MAA and particularly high affinity to the initial template (Kd as low as 1.2 μM), sufficient concentration of binding sites N (up to 32 μmol g-1 ), and pronounced specificity (imprinting factor up to 11). Under flow conditions, the fabricated polymers also demonstrate high capacity and template affinity. The produced imprints reproduce spatially noncovalent interactions present in the ATP binding site of protein kinases and can be anticipated as approximate synthetic imitations of the binding cleft.


Gold-silver@TiO2 nanocomposite-modified plasmonic photoanodes for higher efficiency dye-sensitized solar cells.
In the present investigation, gold-silver@titania (Au-Ag@TiO2) plasmonic nanocomposite materials with different Au and Ag compositions were prepared using a simple one-step chemical reduction method and used as photoanodes in high-efficiency dye-sensitized solar cells (DSSCs). The Au-Ag incorporated TiO2 photoanode demonstrated an enhanced solar-to-electrical energy conversion efficiency of 7.33%, which is ∼230% higher than the unmodified TiO2 photoanode (2.22%) under full sunlight illumination (100 mW cm-2, AM 1.5G). This superior solar energy conversion efficiency was mainly due to the synergistic effect between the Au and Ag, and their surface plasmon resonance effect, which improved the optical absorption and interfacial charge transfer by minimizing the charge recombination process. The influence of the Au-Ag composition on the overall energy conversion efficiency was also explored, and the optimized composition with TiO2 was found to be Au75-Ag25. This was reflected in the femtosecond transient absorption dynamics in which the electron-phonon interaction in the Au nanoparticles was measured to be 6.14 ps in TiO2/Au75:Ag25, compared to 2.38 ps for free Au and 4.02 ps for TiO2/Au100:Ag0. The slower dynamics indicates a more efficient electron-hole separation in TiO2/Au75:Ag25 that is attributed to the formation of a Schottky barrier at the interface between TiO2 and the noble metal(s) that acts as an electron sink. The significant boost in the solar energy conversion efficiency with the Au-Ag@TiO2 plasmonic nanocomposite showed its potential as a photoanode for high-efficiency DSSCs.


The influence of dietary fat on liver fat accumulation.
Obesity is a known risk factor for the development of non-alcoholic fatty liver disease (NAFLD); however, it has been suggested that dietary fat, both amount and composition, may play a pivotal role in its development, independent of body fatness. Studies that have investigated the role of dietary fat on liver fat accumulation are reasonably sparse. We review here the available work that has investigated the impact of dietary fat: amount, composition and frequency, on liver fat accumulation in human observational and intervention studies. Overall, it would seem that total calorie consumption, rather than dietary fat composition, is an important factor in the development of fatty liver disease in humans.


Hyperbaric Oxygen Therapy in Liver Diseases.
Hyperbaric oxygen therapy (HBOT) is an efficient therapeutic option to improve progress of lots of diseases especially hypoxia-related injuries, and has been clinically established as a wide-used therapy for patients with carbon monoxide poisoning, decompression sickness, arterial gas embolism, problematic wound, and so on. In the liver, most studies positively evaluated HBOT as a potential therapeutic option for liver transplantation, acute liver injury, nonalcoholic steatohepatitis, fibrosis and cancer, especially for hepatic artery thrombosis. This might mainly attribute to the anti-oxidation and anti-inflammation of HBOT. However, some controversies are existed, possibly due to hyperbaric oxygen toxicity. This review summarizes the current understandings of the role of HBOT in liver diseases and hepatic regeneration. Future understanding of HBOT in clinical trials and its in-depth mechanisms may contribute to the development of this novel adjuvant strategy for clinical therapy of liver diseases.


HIV-1 integrase variants retarget viral integration and are associated with disease progression in a chronic infection cohort.
Distinct integration patterns of different retroviruses, including HIV-1, have puzzled virologists for over 20 years. A tetramer of the viral integrase (IN) assembles on the two viral cDNA ends, docks onto the target DNA (tDNA), and catalyzes viral genome insertion into the host chromatin. We identified the amino acids in HIV-1 IN that directly contact tDNA bases and affect local integration site sequence selection. These residues also determine the propensity of the virus to integrate into flexible tDNA sequences. Remarkably, natural polymorphisms INS119G and INR231G retarget viral integration away from gene-dense regions. Precisely these variants were associated with rapid disease progression in a chronic HIV-1 subtype C infection cohort. These findings link integration site selection to virulence and viral evolution, but also to the host immune response and antiretroviral therapy, since HIV-1 IN119 is under selection by HLA alleles and integrase inhibitors.


Associations of breakfast skipping with obesity and health-related quality of life: evidence from a national survey in Taiwan.
This study investigated the associations of breakfast skipping with obesity and health-related quality of life (QOL). We also tested the hypothesis that there is a dose-dependent relationship between frequency of breakfast consumption and prevalence of obesity. This cross-section study used a national representative sample (n=15 340) from the 2005 Taiwan National Health Interview Survey. Breakfast skippers were defined as those who ate breakfast about once a week or less often and those who never ate breakfast. Individuals were classified as 'obese' if their body mass index was >or=27. Health-related QOL was assessed using the Medical Outcome Studies 36-Item Short-Form (SF-36) Health Survey. Logistic regression was used to examine the odds ratio of obesity and associated 95% confidence intervals (CIs) in breakfast skippers compared with breakfast eaters. Multivariable logistic regression modeling was used to adjust all risk estimates for covariates. The unadjusted odds ratio of obesity in breakfast skippers was 1.23 (95% CI: 1.06, 1.43). The odds of developing obesity for breakfast skippers was 1.34 (95% CI: 1.15, 1.56) controlling for age, sex, marital status, educational level, monthly income, smoking, alcohol, betel nut chewing and exercise habit. The Cochran-Armitage trend test revealed that the prevalence rate of obesity decreased as the frequency of breakfast consumption increased (P=0.005). Breakfast skippers had significantly worse health-related QOL than breakfast eaters (P<0.001). Moreover, breakfast skippers had significantly lower scores in 5 out of 8 domain scores of the SF-36, namely general health perceptions (P<0.001), vitality (P<0.001), social functioning (P=0.036), emotional role (P<0.001) and mental health (P<0.001). The findings from this study add support to the potential role of breakfast eating in obesity prevention.


Prospective analysis of long term control of mild hypertension on cerebral blood flow.
A group of 12 otherwise normal elderly volunteers (mean age = 69.8 years), were detected to have mild hypertension. Cerebral blood flow (CBF) values were measured using 133Xe inhalation method prior to initiating medical treatment and repeated at 6, 12, 24 and 36 months after BP was adequately controlled and restored to normal (below 150/90). Results indicate that CBF values increased markedly during follow-up intervals at 6, 12 and 24 months but not at 36 months. Hypertension is known to be a risk factor for stroke and 4 of the 12 subjects subsequently developed symptoms of cerebrovascular disease (stroke, multi-infarct dementia or transient ischemic attacks) despite control of hypertension. Analyses separating asymptomatic and symptomatic groups indicated that the eight asymptomatic patients continued to maintain increased CBF levels throughout the entire three year interval, whereas the 4 symptomatic patients developed declines in CBF which began, and progressively decreased below the initial pretreatment values, during the second and third years.


Impact of Warm-Up on Muscle Temperature and Athletic Performance.
Purpose: We performed two studies to investigate: the minute-by-minute changes in muscle temperature following a 20-min warm-up routine (Study-1) and the impact of the typical post-warm-up period of inactivity on the performance of basketball athletes (Study-2). Method: In Study-1, 26 males (age: 23.6 ± 6.2 yr; BMI: 24.1 ± 3.1 kg/m2) performed a 20-min cycling warm-up and then rested for 20 min. Tibialis anterior muscle temperature was assessed throughout. In Study-2, six male professional basketball players (age: 24.9 ± 4.6 yr; BMI: 25.5 ± 1.8 kg/m2) performed a series of basketball performance tests after a 20-min warm-up, as well as 9-min and 23-min into a post-warm-up period of inactivity. Results: On average, muscle temperature increased by 0.1°C every minute during warm-up and dropped by the same amount every minute during inactivity. The increase during warm-up and the decrease during inactivity were higher at the start of each period. A 9-min inactivity period is accompanied by 3.8 ± 0.6% reduction in countermovement jump (p = .046). A 23-min inactivity period is accompanied by 7.3 ± 0.7% reduction in lay-up points (p = .027). Conclusion: These two studies show that a 20-min warm-up routine increases muscle temperature but this benefit is lost after a typical post-warm-up inactivity period in high-level basketball, leading to reductions in certain aspects of athletic performance.


Saccharomyces boulardii improves intestinal cell restitution through activation of the α2β1 integrin collagen receptor.
Intestinal epithelial cell damage is frequently seen in the mucosal lesions of inflammatory bowel diseases such as ulcerative colitis or Crohn's disease. Complete remission of these diseases requires both the cessation of inflammation and the migration of enterocytes to repair the damaged epithelium. Lyophilized Saccharomyces boulardii (Sb, Biocodex) is a nonpathogenic yeast widely used as a therapeutic agent for the treatment and prevention of diarrhea and other gastrointestinal disorders. In this study, we determined whether Sb could accelerate enterocyte migration. Cell migration was determined in Sb force-fed C57BL6J mice and in an in vitro wound model. The impact on α2β1 integrin activity was assessed using adhesion assays and the analysis of α2β1 mediated signaling pathways both in vitro and in vivo. We demonstrated that Sb secretes compounds that enhance the migration of enterocytes independently of cell proliferation. This enhanced migration was associated with the ability of Sb to favor cell-extracellular matrix interaction. Indeed, the yeast activates α2β1 integrin collagen receptors. This leads to an increase in tyrosine phosphorylation of cytoplasmic molecules, including focal adhesion kinase and paxillin, involved in the integrin signaling pathway. These changes are associated with the reorganization of focal adhesion structures. In conclusion Sb secretes motogenic factors that enhance cell restitution through the dynamic regulation of α2β1 integrin activity. This could be of major importance in the development of novel therapies targeting diseases characterized by severe mucosal injury, such as inflammatory and infectious bowel diseases.


Biochemical and structural characterization of a protein complex containing a hyaluronidase and a CRISP-like protein isolated from the venom of the spider Acanthoscurria natalensis.
Spider venoms are composed of a complex mixture of bioactive molecules. The structural and functional characterization of these molecules in the venom of the Brazilian spider Acanthoscurria natalensis, has been little explored. The venom was fractionated using reversed-phase liquid chromatography. The fraction with hyaluronidase activity was named AnHyal. The partial sequencing of AnHyal revealed the presence of a CRISP-like protein, in addition to hyaluronidase, comprising 67% coverage for hyaluronidase from Brachypelma vagans and 82% for CRISP-like protein from Grammostola rosea. 1D BN-PAGE zymogram assays of AnHyal confirmed the presence of enzymatically active 53 kDa monomer and 124 and 178 kDa oligomers. The decomposition of the complexes by 2D BN/SDS-PAGE zymogram assays showed two subunits, 53 (AnHyalH) and 44 kDa (AnHyalC), with sequence similarity to hyaluronidase and CRISP proteins, respectively. The secondary structure of AnHyal is composed by 36% of α-helix. AnHyal presented maximum activity at pH between 4.0 and 6.0 and 30 and 60 °C, showed specificity to hyaluronic acid substrate and presented a KM of 617.9 μg/mL. Our results showed that hyaluronidase and CRISP proteins can form a complex and the CRISP protein may contribute to the enzymatic activity of AnHyalH.


Low HDL Cholesterol is Associated with Lower Gray Matter Volume in Cognitively Healthy Adults.
Dyslipidemia is common in adults and contributes to high rates of cardiovascular disease and may be linked to subsequent neurodegenerative and neurovascular diseases. This study examined whether lower brain volumes and cognition associated with dyslipidemia could be observed in cognitively healthy adults, and whether apolipoprotein E (APOE) genotype or family history of Alzheimer's disease (FHAD) alters this effect. T1-weighted magnetic resonance imaging was used to examine regional brain gray matter (GM) and white matter (WM) in 183 individuals (58.4 +/- 8.0 years) using voxel-based morphometry. A non-parametric multiple linear regression model was used to assess the effect of high-density lipoprotein (HDL) and non-HDL cholesterol, APOE, and FHAD on regional GM and WM volume. A post hoc analysis was used to assess whether any significant correlations found within the volumetric analysis had an effect on cognition. HDL was positively correlated with GM volume in the bilateral temporal poles, middle temporal gyri, temporo-occipital gyri, and left superior temporal gyrus and parahippocampal region. This effect was independent of APOE and FHAD. A significant association between HDL and the Brief Visuospatial Memory Test was found. Additionally, GM volume within the right middle temporal gyrus, the region most affected by HDL, was significantly associated with the Controlled Oral Word Association Test and the Center for Epidemiological Studies Depression Scale. These findings suggest that adults with decreased levels of HDL cholesterol may be experiencing cognitive changes and GM reductions in regions associated with neurodegenerative disease and therefore, may be at greater risk for future cognitive decline.


The variation in biological effectiveness of X-rays and gamma rays with energy.
The ICRP has attributed the same relative risk for all low-LET (linear energy transfer) radiations, including X and gamma radiations of all energies. However, very low energy X-rays are expected to be more biologically effective, per unit absorbed dose, than high energy X-rays or gamma rays due to the production of lower energy secondary electrons, with a correspondingly higher LET. This increase in relative biological effectiveness (RBE) is also seen experimentally for a range of biological end-points, however, a wide range of RBE values have been reported. The assessment of risks is particularly important due to the use of low energy X-rays for mammography screening. A review of the published data on the variation in biological effectiveness with energy is presented here.


Hypothalamic inhibition of neurones in the nucleus tractus solitarius of the cat is GABA mediated.
1. In pentobarbitone-anaesthetized cats extracellular activity of neurones in the vicinity of the nucleus tractus solitarius receiving inputs from the carotid sinus nerve (SN) and/or vagus nerve (VN) during stimulation of the hypothalamic defence area (HDA) and application of gamma-aminobutyric acid (GABA) and glycine and their antagonists have been studied. 2. A total of forty neurones have been tested, of which twenty-four only had an input from the SN, one only from the VN, twelve from both nerves and three had neither SN or VN inputs. 3. Short trains of stimuli to the HDA inhibited both the ongoing activity (if present) and evoked discharge in thirty-nine of the forty neurones tested. 4. In the forty cells tested ionophoretic application of GABA reduced (4) or totally inhibited (35) neuronal discharge whilst in the thirty-eight tested with glycine discharge was totally (25) or partially (12) suppressed. 5. Ionophoresis of bicuculline totally (14) or partially (6) antagonized the inhibitory actions of GABA in the twenty-five cells tested, and in eighteen of these the ongoing and/or evoked activity was simultaneously increased. In eighteen of the nineteen cells tested this level of bicuculline also antagonized the inhibitory actions of HDA stimuli whereas in none of the sixteen cells tested did it affect glycine-evoked inhibitions. 6. Ionophoretic application of strychnine antagonized the inhibitory effects of glycine in eight of nine cells tested but in these eight cells strychnine had no effect on ongoing or evoked discharges, GABA- or HDA-evoked inhibitions. 7. In a chloralose-anaesthetized cat five neurones receiving SN inputs (three also receiving VN inputs) were recorded. All could be inhibited by HDA stimuli and by application of GABA. In the three of four cells in which bicuculline antagonized GABA inhibitions, the effects of HDA stimuli were simultaneously antagonized whereas glycine-evoked inhibitions were unaffected. 8. In two neurones, in addition to inhibiting neuronal discharge HDA stimulation also evoked activity in the cells. In a further four neurones similar excitatory responses were uncovered when the HDA inhibitory effects were antagonized by bicuculline. 9. The importance of these observations in cardiovascular control and in the functioning of the baroreceptor reflex is discussed.


Cause specific mortality among employees engaged in the manufacture, formulation, or packaging of 2,4-dichlorophenoxyacetic acid and related salts.
Mortality is reported to the end of 1982 for 878 chemical workers potentially exposed to 2,4-dichlorophenoxyacetic acid (2,4-D) at any time between 1945 and 1983. Observed mortality was compared with expected levels based on adjusted rates for United States white men and for other male employees from this manufacturing location who were not exposed to 2,4-D. Because of a recently reported increased incidence of astrocytomas in male rats fed the highest dose level of 2,4-D, special attention was given to deaths from brain neoplasms in the cohort. None was observed. The absence of an increased risk of brain cancer in people exposed to 2,4-D is supported by studies of other exposed populations and those studies are briefly reviewed. Moreover, in the present study, analyses by production area, duration of exposure, and cumulative dose showed no patterns suggestive of a causal association between 2,4-D exposure and any other particular cause of death.


Long-Term Risk of Infections After Treatment of Childhood Leukemia: A Population-Based Cohort Study Using Administrative Health Data.
Infections are a frequent complication during childhood leukemia treatment. Little is known about the infectious risk in survivors. We compared the relative rate (RR) of infections after treatment completion between pediatric leukemia survivors and the general population. We performed a retrospective, population-based cohort study of children diagnosed with leukemia between 1992 and 2015 in Ontario, Canada, who were alive and relapse free 30 days after treatment completion (index date). Leukemia survivors were matched 5:1 with the general population by year of birth, sex, and rural status and stratified by initial treatment, including and excluding hematopoietic stem-cell transplantation (HSCT). The primary outcome was time to infections, as identified using validated diagnostic codes from administrative databases. Individuals were censored at the earliest of death, first relapse, loss to follow-up, or end of study. A total of 2,204 leukemia survivors were included and matched with 11,020 controls. The rate of infections was elevated after treatment completion compared with controls (RR, 1.51; 95% CI, 1.45 to 1.57) and at less than 1 year (RR, 1.77; 95% CI, 1.69 to 1.86); 1 to 4.99 years (RR, 1.66; 95% CI, 1.62 to 1.71), and 5 or more years (RR, 1.29; 95% CI, 1.22 to 1.36) from the index date. Among those whose initial treatment excluded HSCT, the rate remained elevated more than 5 years from the index date (RR, 1.29; 95% CI, 1.23 to 1.35). Infection-related death was significantly increased in leukemia survivors both among the entire cohort (hazard ratio, 149.3; 95% CI, 20.4 to 1,091.9) and among those without HSCT (hazard ratio, 92.7; 95% CI, 12.4 to 690.7). A significant association was found between a history of leukemia therapy and an increased risk of infections. Additional study is needed to establish which exposures in patients with leukemia lead to late infections.


Reputation management: evidence for ability but reduced propensity in autism.
Previous research has reported that autistic adults do not manage their reputation, purportedly due to problems with theory of mind [Izuma, Matsumoto, Camerer, & Adolphs]. The current study aimed to test alternative explanations for this apparent lack of reputation management. Twenty typical and 19 autistic adults donated to charity and to a person, both when alone and when observed. In an additional manipulation, for half of the participants, the observer was also the recipient of their donations, and participants were told that this observer would subsequently have the opportunity to donate to them (motivation condition). This manipulation was designed to encourage an expectation of a reciprocal "tit-for-tat" strategy in the participant, which may motivate participants to change their behavior to receive more donations. The remaining participants were told that the person watching was just observing the procedure (no motivation condition). Our results replicated Izuma et al.'s finding that autistic adults did not donate more to charity when observed. Yet, in the motivation condition, both typical and autistic adults donated significantly more to the observer when watched, although this effect was significantly attenuated in autistic individuals. Results indicate that, while individuals with autism may have the ability to think about reputation, a reduced expectation of reciprocal behavior from others may reduce the degree to which they engage in reputation management.


Reconstructing each cell's genome within complex microbial communities-dream or reality?
As the vast majority of microorganisms have yet to be cultivated in a laboratory setting, access to their genetic makeup has largely been limited to cultivation-independent methods. These methods, namely metagenomics and more recently single-cell genomics, have become cornerstones for microbial ecology and environmental microbiology. One ultimate goal is the recovery of genome sequences from each cell within an environment to move toward a better understanding of community metabolic potential and to provide substrate for experimental work. As single-cell sequencing has the ability to decipher all sequence information contained in an individual cell, this method holds great promise in tackling such challenge. Methodological limitations and inherent biases however do exist, which will be discussed here based on environmental and benchmark data, to assess how far we are from reaching this goal.


Characterization of the deoxycytidine kinase promoter in human lymphoblast cell lines.
Deoxycytidine kinase (dCK) phosphorylates 2'-deoxycytidine, as well as the purine deoxyribonucleosides and a number of nucleoside analogues that are important in the chemotherapy of leukemias. The enzyme is highly expressed in the thymus relative to other tissues and may play an important role in the T cell depletion associated with adenosine deaminase and purine nucleoside phosphorylase deficiencies. To characterize the dCK promoter region and to determine whether it mediates higher levels of gene expression in T lymphoblasts, we have analyzed a 700-bp genomic fragment encompassing 548 bp of 5' flanking region for functional activity and for transcription factor binding using T and B lymphoblast cell lines and nuclear extracts. The regions of the promoter that were defined as important to its function include a 5' GC box, and E box, a 3' GC box, and an E2F site. The transcription factor Sp1 binds to both GC boxes, activating at the 5' site but repressing at the 3' site. MLTF/USF activates transcription through the E box, whereas E2F activates through the E2F site, but binds weakly to this site in vitro and does not appear to mediate cell cycle-specific expression of dCK in vivo. No significant differences in promoter activity or transcription factor binding were observed between Jurkat T and Raji B lymphoblasts. The promoter of the dCK gene is thus regulated by a number of ubiquitously expressed transcription factors. DCK expression in cultured lymphoblast cell lines is not solely a function of the T or B lineage derivation.


Polymorphic pemphigoid.
We describe 20 patients with a chronic polymorphic eruption; they shared clinical, histopathological, and therapeutic features of both dermatitis herpetiformis and bullous pemphigoid (BP). In 14 of these 20 cases, direct and indirect immunofluorescence studies corresponded to BP. The remaining six patients showed IgA deposits in a linear pattern at the basement membrane zone, and two of these six showed IgA pemphigoid antibodies in their sera as well. No significant clinical and histological differences were detected in the patients, in connection with the immunological findings. Furthermore, one patient's condition, which was studied by repeated immunofluorescence examinations, changed from a linear IgA pattern and a negative indirect test to a linear IgG pattern and a positive reaction for IgG pemphigoid antibodies. We concluded that these cases constitute a polymorphic variant of BP.


NCCN Guidelines Insights: Thyroid Carcinoma, Version 2.2018.
The NCCN Guidelines for Thyroid Carcinoma provide recommendations for the management of different types of thyroid carcinoma, including papillary, follicular, Hürthle cell, medullary, and anaplastic carcinomas. These NCCN Guidelines Insights summarize the panel discussion behind recent updates to the guidelines, including the expanding role of molecular testing for differentiated thyroid carcinoma, implications of the new pathologic diagnosis of noninvasive follicular thyroid neoplasm with papillary-like nuclear features, and the addition of a new targeted therapy option for BRAF V600E-mutated anaplastic thyroid carcinoma.


Intentional modulation of emotional responding to unpleasant pictures: an ERP study.
Intentionally altering responses to unpleasant stimuli affects physiological and hemodynamic activity associated with emotional and cognitive processing. In the present experiment, we measured the late-positive potential (LPP) of the visually evoked event-related brain potential to examine the effects of intentional emotion modulation on electrophysiological correlates of emotional and cognitive processing. Seventeen participants received instructions to view, suppress, and enhance emotional responses to unpleasant stimuli. Results revealed significantly decreased electrophysiological activity during suppression of emotional responses beginning around 250 ms poststimulus and lasting several hundred milliseconds. These data suggest that ERPs are sensitive to emotion modulation/regulation processes.


Full endoscopic versus open discectomy for sciatica: randomised controlled non-inferiority trial.
To assess whether percutaneous transforaminal endoscopic discectomy (PTED) is non-inferior to conventional open microdiscectomy in reduction of leg pain caused by lumbar disc herniation. Multicentre randomised controlled trial with non-inferiority design. Four hospitals in the Netherlands. 613 patients aged 18-70 years with at least six weeks of radiating leg pain caused by lumbar disc herniation. The trial included a predetermined set of 125 patients receiving PTED who were the learning curve cases performed by surgeons who did not do PTED before the trial. PTED (n=179) compared with open microdiscectomy (n=309). The primary outcome was self-reported leg pain measured by a 0-100 visual analogue scale at 12 months, assuming a non-inferiority margin of 5.0. Secondary outcomes included complications, reoperations, self-reported functional status as measured with the Oswestry Disability Index, visual analogue scale for back pain, health related quality of life, and self-perceived recovery. Outcomes were measured until one year after surgery and were longitudinally analysed according to the intention-to-treat principle. Patients belonging to the PTED learning curve were omitted from the primary analyses. At 12 months, patients who were randomised to PTED had a statistically significantly lower visual analogue scale score for leg pain (median 7.0, interquartile range 1.0-30.0) compared with patients randomised to open microdiscectomy (16.0, 2.0-53.5) (between group difference of 7.1, 95% confidence interval 2.8 to 11.3). Blood loss was less, length of hospital admission was shorter, and timing of postoperative mobilisation was earlier in the PTED group than in the open microdiscectomy group. Secondary patient reported outcomes such as the Oswestry Disability Index, visual analogue scale for back pain, health related quality of life, and self-perceived recovery, were similarly in favour of PTED. Within one year, nine (5%) in the PTED group compared with 14 (6%) in the open microdiscectomy group had repeated surgery. Per protocol analysis and sensitivity analyses including the patients of the learning curve resulted in similar outcomes to the primary analysis. PTED was non-inferior to open microdiscectomy in reduction of leg pain. PTED resulted in more favourable results for self-reported leg pain, back pain, functional status, quality of life, and recovery. These differences, however, were small and may not reach clinical relevance. PTED can be considered as an effective alternative to open microdiscectomy in treating sciatica. NCT02602093ClinicalTrials.gov NCT02602093.


The Characterization of Varicella Zoster Virus-Specific T Cells in Skin and Blood during Aging.
Reactivation of the varicella zoster virus (VZV) increases during aging. Although the effects of VZV reactivation are observed in the skin (shingles), the number and functional capacity of cutaneous VZV-specific T cells have not been investigated. The numbers of circulating IFN-γ-secreting VZV-specific CD4(+) T cells are significantly decreased in old subjects. However, other measures of VZV-specific CD4(+) T cells, including proliferative capacity to VZV antigen stimulation and identification of VZV-specific CD4(+) T cells with an major histocompatibility complex class II tetramer (epitope of IE-63 protein), were similar in both age groups. The majority of T cells in the skin of both age groups expressed CD69, a characteristic of skin-resident T cells. VZV-specific CD4(+) T cells were significantly increased in the skin compared with the blood in young and old subjects, and their function was similar in both age groups. In contrast, the number of Foxp3(+) regulatory T cells and expression of the inhibitory receptor programmed cell death -1 PD-1 on CD4(+) T cells were significantly increased in the skin of older humans. Therefore, VZV-specific CD4(+) T cells in the skin of older individuals are functionally competent. However, their activity may be restricted by multiple inhibitory influences in situ.


Subintimal angioplasty in the management of critical lower-extremity ischemia: value in limb salvage.
The interest in and overall usage of endovascular procedures for the treatment of lower extremity ischemia continues to grow at a rapid pace. An increasing number of centers throughout the world are gaining experience with subintimal angioplasty. Promising results have been reported and the application of the technique has been expanded to include the iliac and crural arteries. The technique of subintimal angioplasty and several variations are discussed. Although primary patency rates compared with bypass are relatively low for patients undergoing subintimal angioplasty, limb salvage rates remain high. When a subintimal angioplasty fails, it frequently does so without the recurrence of symptoms, especially when a gangrenous lesion or ulcer has healed. Given the many advantages of this technique, which include reduced anesthesia requirements, a minimally invasive approach, and potential reductions in length of stay and cost, subintimal angioplasty will continue to have a role in the treatment of lower extremity ischemia. When applied judiciously, bypass options may be preserved. This does not, however, mean that the availability of this technique should be used as justification to lower the threshold for the treatment.


Association between neighborhood deprivation and fruits and vegetables consumption and leisure-time physical activity: a cross-sectional multilevel analysis.
Most studies of the association between neighborhood socioeconomic deprivation and individual lifestyles leading to cardiovascular disease focused on a single cardiovascular risk factor. The concomitant assessment of more than one risk factor may provide clues to specific mechanisms linking neighborhood disadvantage to individual lifestyles. We investigated the association of neighborhood deprivation with fruits and vegetables consumption and leisure-time physical activity in adults living in an urban center in Portugal. In 1999-2003, we assembled a random sample of 2081 adult residents in the city of Porto. Data on sociodemographic characteristics were collected by trained interviewers using structured questionnaires. Fruits and vegetables consumption was estimated using a validated 82-item semiquantitative food frequency questionnaire covering the previous year and expressed in portions per day. Physical activity was evaluated using a questionnaire exploring leisure-time activities over the previous year and expressed in metabolic equivalents (MET).minute/day. Self-reported address was used to place individuals in neighborhoods. Neighborhoods' socioeconomic characterization was based on aggregated data at the census block level provided by the 2001 National Census. Latent class analysis models were used to identify three discrete socioeconomic classes of neighborhoods. Random effects models with random intercepts at the neighborhood level were used to explore clustering and contextual effects of neighborhood deprivation on each of the outcomes. We found evidence of neighborhood clustering of fruits and vegetables consumption and leisure-time physical activity that persisted after adjustment for neighborhood deprivation only among women. Women living in the most deprived neighborhoods presented a consumption increase of 0.43 (95% CI: -0.033 to 0.89) portions of fruits and vegetables per day and a decrease in leisure-time physical activity of 47.8 (95% CI: -91.8 to 1.41) MET.minute/day, when compared to those living in the most affluent neighborhoods. Among men, no contextual neighborhood deprivation effects were observed. Overall, neighborhood deprivation had a small effect on the consumption of fruits and vegetables and leisure-time physical activity. Neighborhood factors other than socioeconomic deprivation may still impact on the studied outcomes among women. This study provides relevant information for the design of interventions directed to neighborhood characteristics in the prevention of cardiovascular diseases.


29 gauge needles improve patient satisfaction over 27 gauge needles for daily glatiramer acetate injections.
Using three surveys, a comparative assessment of needle performance and patient preference for 27-gauge (G) and 29G needles for glatiramer acetate administration for multiple sclerosis therapy was performed. Eligible patients participated in a specialty pharmacy program and administered glatiramer acetate for ≥1 month. In Survey 1 on the 27G needle, 545 (82.70%) patients reported no needle problems, 106 (16.09%) cited one type (dull, bent, or broken), five (0.76%) cited two types, and three (0.46%) cited all three types. In Survey 2 on the 29G needle, 553 (98.05%) indicated no problems, two (0.35%) cited dull needles, and nine (1.60%) cited bent needles. On the 29G needles versus 27G needles pain comparison, 219 (38.83%) reported the 29G needle was a little better, and 155 (27.48%) reported it was a lot better than the 27G. For injection-site experiences, 515 patients (91.31%) reported no, very slight, or mild reactions with the 29G needle. In Survey 3, over 76% of patients preferred the 29G to the 27G needle and significantly fewer patients reported one or more problems with the 29G needle compared to patients reporting problems with the 27G needle (P < 0.00001). In conclusion, significantly fewer patients reported problems after 30 days of use of the 29G than the 27G needle. Fewer injection-site experiences occurred with the 29G needle and the 29G needle was preferred overall.


Safety of zoledronic acid and incidence of osteonecrosis of the jaw (ONJ) during adjuvant therapy in a randomised phase III trial (AZURE: BIG 01-04) for women with stage II/III breast cancer.
The AZURE trial is an ongoing phase III, academic, multi-centre, randomised trial designed to evaluate the role of zoledronic acid (ZOL) in the adjuvant therapy of women with stage II/III breast cancer. Here, we report the safety and tolerability profile of ZOL in this setting. Eligible patients received (neo)adjuvant chemotherapy and/or endocrine therapy and were randomised to receive neither additional treatment nor intravenous ZOL 4 mg. ZOL was administered after each chemotherapy cycle to exploit potential sequence-dependent synergy. ZOL was continued for 60 months post-randomisation (six doses in the first 6 months, eight doses in the following 24 months and five doses in the final 30 months). Serious (SAE) and non-serious adverse event (AE) data generated during the first 36 months on study were analysed for the safety population. 3,360 patients were recruited to the AZURE trial. The safety population comprised 3,340 patients (ZOL 1,665; control 1,675). The addition of ZOL to standard treatment did not significantly impact on chemotherapy delivery. SAE were similar in both treatment arms. No significant safety differences were seen apart from the occurrence of osteonecrosis of the jaw (ONJ) in the ZOL group (11 confirmed cases; 0.7%; 95% confidence interval 0.3-1.1%). ZOL in the adjuvant setting is well tolerated, and can be safely administered in addition to adjuvant therapy including chemotherapy. The adverse events were consistent with the known safety profile of ZOL, with a low incidence of ONJ.


Quick phases of infantile nystagmus show the saccadic inhibition effect.
Infantile nystagmus (IN) is a pathological, involuntary oscillation of the eyes consisting of slow, drifting eye movements interspersed with rapid reorienting quick phases. The extent to which quick phases of IN are programmed similarly to saccadic eye movements remains unknown. We investigated whether IN quick phases exhibit 'saccadic inhibition,' a phenomenon typically related to normal targeting saccades, in which the initiation of the eye movement is systematically delayed by task-irrelevant visual distractors. We recorded eye position from 10 observers with early-onset idiopathic nystagmus while task-irrelevant distractor stimuli were flashed along the top and bottom of a large screen at ±10° eccentricity. The latency distributions of quick phases were measured with respect to these distractor flashes. Two additional participants, one with possible albinism and one with fusion maldevelopment nystagmus syndrome, were also tested. All observers showed that a distractor flash delayed the execution of quick phases that would otherwise have occurred approximately 100 ms later, exactly as in the standard saccadic inhibition effect. The delay did not appear to differ between the two main nystagmus types under investigation (idiopathic IN with unidirectional and bidirectional jerk). The presence of the saccadic inhibition effect in IN quick phases is consistent with the idea that quick phases and saccades share a common programming pathway. This could allow quick phases to take on flexible, goal-directed behavior, at odds with the view that IN quick phases are stereotyped, involuntary eye movements.


Evaluation of flavonoids and enterolactone in overnight urine as intake biomarkers of fruits, vegetables and beverages in the Inter99 cohort study using the method of triads.
Since collection of 24 h urine samples is very time consuming and difficult to obtain, epidemiological studies typically only obtain spot urine samples. The aim of the present study was to evaluate whether flavonoids and enterolactone in overnight urine could substitute flavonoids and enterolactone in 24 h urine as an alternative and more feasible biomarker of fruit, vegetable and beverage intake. A total of 191 individuals in the Inter99 cohort in Denmark completed the validation study. Concentrations of nine urinary flavonoid aglycones (quercetin, isorhamnetin, tamarixetin, kaempferol, hesperetin, naringenin, eriodictyol, phloretin and apigenin) and enterolactone were determined in overnight and 24 h urine samples, and their validity as biomarkers of fruit, vegetable and beverage intake was evaluated in relation to two independent reference methods (Inter99 FFQ data and plasma carotenoids) by using the method of triads. The intakes of fruit, juice, vegetables and tea reported in the FFQ were reflected by the flavonoid biomarker both in overnight and 24 h urine samples. Validity coefficients for the flavonoid biomarker in overnight urine ranged from 0·39 to 0·49, while the corresponding validity coefficients for the biomarker in 24 h urine ranged from 0·43 to 0·66. Although the validity coefficients were lower for overnight urine than for the 24 h urine flavonoid biomarker, they were still of acceptable magnitude. In conclusion, the results indicate that flavonoids and enterolactone in overnight urine samples may be used as a more feasible biomarker than 24 h urine for the assessment and validation of fruit, juice, vegetable and tea intakes in epidemiological studies.


Predicting protein decomposition: the case of aspartic-acid racemization kinetics.
The increase in proportion of the non-biological (D-) isomer of aspartic acid (Asp) relative to the L-isomer has been widely used in archaeology and geochemistry as a tool for dating. the method has proved controversial, particularly when used for bones. The non-linear kinetics of Asp racemization have prompted a number of suggestions as to the underlying mechanism(s) and have led to the use of mathematical transformations which linearize the increase in D-Asp with respect to time. Using one example, a suggestion that the initial rapid phase of Asp racemization is due to a contribution from asparagine (Asn), we demonstrate how a simple model of the degradation and racemization of Asn can be used to predict the observed kinetics. A more complex model of peptide bound Asx (Asn + Asp) racemization, which occurs via the formation of a cyclic succinimide (Asu), can be used to correctly predict Asx racemization kinetics in proteins at high temperatures (95-140 degrees C). The model fails to predict racemization kinetics in dentine collagen at 37 degrees C. The reason for this is that Asu formation is highly conformation dependent and is predicted to occur extremely slowly in triple helical collagen. As conformation strongly influences the rate of Asu formation and hence Asx racemization, the use of extrapolation from high temperatures to estimate racemization kinetics of Asx in proteins below their denaturation temperature is called into question. In the case of archaeological bone, we argue that the D:L ratio of Asx reflects the proportion of non-helical to helical collagen, overlain by the effects of leaching of more soluble (and conformationally unconstrained) peptides. Thus, racemization kinetics in bone are potentially unpredictable, and the proposed use of Asx racemization to estimate the extent of DNA depurination in archaeological bones is challenged.


Curvature coding is tuned for motion direction.
We have investigated the global and local motion tuning properties of curvature coding mechanisms using two shape after-effects believed to be mediated by curvature-sensitive mechanisms: the shape-frequency after-effect, or SFAE, and the shape-amplitude after-effect, or SAAE. The SFAE and SAAE are the phenomena in which adaptation to a sine-wave-shaped contour causes a shift in respectively the apparent shape-frequency and apparent shape-amplitude of a test contour in a direction away from that of the adapting stimulus. In the global motion condition the sinusoidal-shaped contours were made to drift within a fixed stimulus window in the direction of their axis of modulation. In the local motion condition the contour was constructed from a string of Gabors, and their carriers but not envelopes moved. We investigated selectivity to motion direction by using adaptor and test contours that moved either in the same or opposite directions. We found that in the global motion condition both the SFAE and SAAE showed selectivity to motion direction, and that for the same-motion-direction condition, both after-effects increased with shape temporal frequency. We then examined the effect of luminance spatial frequency and luminance temporal frequency on global motion direction selectivity. Luminance temporal frequency accounted for some of the increase in after-effect magnitude with shape temporal frequency, but shape temporal frequency also contributed. The local motion after-effects on the other hand were neither selective to motion direction nor increased with luminance temporal frequency. Taken together, the results are best understood by supposing that curvature is encoded by mechanisms that are selective to motion direction and that the directional selectivity best manifests itself psychophysically when there is sufficient spatio-temporal coverage of the stimulus to stimulate the full array of potentially responsive curvature-coding mechanisms.


Cardiovascular effects of air pollution.
Air pollution is a heterogeneous mixture of gases, liquids and PM (particulate matter). In the modern urban world, PM is principally derived from fossil fuel combustion with individual constituents varying in size from a few nanometres to 10 microm in diameter. In addition to the ambient concentration, the pollution source and chemical composition may play roles in determining the biological toxicity and subsequent health effects. Nevertheless, studies from across the world have consistently shown that both short- and long-term exposures to PM are associated with a host of cardiovascular diseases, including myocardial ischaemia and infarctions, heart failure, arrhythmias, strokes and increased cardiovascular mortality. Evidence from cellular/toxicological experiments, controlled animal and human exposures and human panel studies have demonstrated several mechanisms by which particle exposure may both trigger acute events as well as prompt the chronic development of cardiovascular diseases. PM inhaled into the pulmonary tree may instigate remote cardiovascular health effects via three general pathways: instigation of systemic inflammation and/or oxidative stress, alterations in autonomic balance, and potentially by direct actions upon the vasculature of particle constituents capable of reaching the systemic circulation. In turn, these responses have been shown to trigger acute arterial vasoconstriction, endothelial dysfunction, arrhythmias and pro-coagulant/thrombotic actions. Finally, long-term exposure has been shown to enhance the chronic genesis of atherosclerosis. Although the risk to one individual at any single time point is small, given the prodigious number of people continuously exposed, PM air pollution imparts a tremendous burden to the global public health, ranking it as the 13th leading cause of morality (approx. 800,000 annual deaths).


Dynamics of cross-bridge cycling, ATP hydrolysis, force generation, and deformation in cardiac muscle.
Despite extensive study over the past six decades the coupling of chemical reaction and mechanical processes in muscle dynamics is not well understood. We lack a theoretical description of how chemical processes (metabolite binding, ATP hydrolysis) influence and are influenced by mechanical processes (deformation and force generation). To address this need, a mathematical model of the muscle cross-bridge (XB) cycle based on Huxley's sliding filament theory is developed that explicitly accounts for the chemical transformation events and the influence of strain on state transitions. The model is identified based on elastic and viscous moduli data from mouse and rat myocardial strips over a range of perturbation frequencies, and MgATP and inorganic phosphate (Pi) concentrations. Simulations of the identified model reproduce the observed effects of MgATP and MgADP on the rate of force development. Furthermore, simulations reveal that the rate of force re-development measured in slack-restretch experiments is not directly proportional to the rate of XB cycling. For these experiments, the model predicts that the observed increase in the rate of force generation with increased Pi concentration is due to inhibition of cycle turnover by Pi. Finally, the model captures the observed phenomena of force yielding suggesting that it is a result of rapid detachment of stretched attached myosin heads.


Characterization of the active site and calcium binding in cytochrome c nitrite reductases.
The decahaem homodimeric cytochrome c nitrite reductase (NrfA) is expressed within the periplasm of a wide range of Gamma-, Delta- and Epsilon-proteobacteria and is responsible for the six-electron reduction of nitrite to ammonia. This allows nitrite to be used as a terminal electron acceptor, facilitating anaerobic respiration while allowing nitrogen to remain in a biologically available form. NrfA has also been reported to reduce nitric oxide (a reaction intermediate) and sulfite to ammonia and sulfide respectively, suggesting a potential secondary role as a detoxification enzyme. The protein sequences and crystal structures of NrfA from different bacteria and the closely related octahaem nitrite reductase from Thioalkalivibrio nitratireducens (TvNir) reveal that these enzymes are homologous. The NrfA proteins contain five covalently attached haem groups, four of which are bis-histidine-co-ordinated, with the proximal histidine being provided by the highly conserved CXXCH motif. These haems are responsible for intraprotein electron transfer. The remaining haem is the site for nitrite reduction, which is ligated by a novel lysine residue provided by a CXXCK haem-binding motif. The TvNir nitrite reductase has five haems that are structurally similar to those of NrfA and three extra bis-histidine-coordinated haems that precede the NrfA conserved region. The present review compares the protein sequences and structures of NrfA and TvNir and discusses the subtle differences related to active-site architecture and Ca2+ binding that may have an impact on substrate reduction.


SmI2-mediated reductive cyclization of β-arylthio ketones: a facile and diastereoselective synthesis of thiochroman derivatives.
SmI2-mediated reductive cyclization of β-arylthio ketones to generate thiochroman derivatives is not a generally observed process and the reported examples are limited to geminal disubstitution in the substrates. The results of the current study show that the cyclization also occurs when other substitution patterns are present, affording a general approach to dihydrothiochroman-ols in good yields and high degrees of diastereoselectivity. Besides, the halogen substitution on β-aryl is tolerated in most cases here although reductive dehalogenation has been reported to predominate in the reductive cyclization process. Dihydrothiochroman-4-ols were readily oxidized to thiochroman-4-ols in almost quantitative yields.


Qiang-Xin 1 Formula Prevents Sepsis-Induced Apoptosis in Murine Cardiomyocytes by Suppressing Endoplasmic Reticulum- and Mitochondria-Associated Pathways.
Sepsis is reported to be an unusual systemic reaction to infection, accompanied by multiple-organ failure. Sepsis-induced cardiomyopathy (SIC), defined as damages and dysfunction of the heart, is essential in the pathogenesis of sepsis. Traditional Chinese formula, which has long been used to improve the situation of patients through multitarget regulation, is now gradually being used as complementary therapy. The present study aimed to investigate the effect of Qiang-Xin 1 (QX1) formula, a traditional Chinese herbal medicine designed for cardiac dysfunction, on cecal ligation puncture (CLP)-induced heart damage and its underlying mechanisms in mice. Survival test first showed that an oral administration of QX1 formula significantly increased the 7-days survival of septic mice from 22 to 40%. By estimating the secretion of serum cytokines, QX1 treatment dramatically inhibited the excessive production of interleukin-1β and tumor necrosis factor-α. Immunohistochemical staining illustrated that the expression of c-Jun N-terminal kinase, caspase-12, and high-mobility group box 1 was downregulated in cardiomyocytes of the QX1-treated group compared with that of the CLP surgery group. Western blotting confirmed that the activation of essential caspase family members, such as caspase-3, caspase-9, and caspase-12, was prohibited by treatment with QX1. Moreover, the abnormal expression of key regulators of endoplasmic reticulum (ER) and mitochondria-associated apoptosis in cardiomyocytes of septic mice, including CHOP, GRP78, Cyt-c, Bcl-2, Bcl-XL, and Bax, was effectively reversed by treatment with QX1 formula. This study provided a new insight into the role of QX1 formula in heart damage and potential complementary therapeutic effect of traditional Chinese medicine on sepsis.


Intramembrane proteolysis by rhomboids: catalytic mechanisms and regulatory principles.
Rhomboids are intramembrane serine proteases that cleave membrane proteins within the bilayer, and which control a wide variety of biological processes. Recent structures of Escherichia coli rhomboids in complex with mechanism-based inhibitors provide insight into their catalytic mechanism. The inhibitor structures also reveal potential substrate-binding sites within the enzyme and provide a template for modeling substrate binding at the active site. The regulation of rhomboid activity exploits the different membrane compartments in cells to segregate enzyme and substrate. Catalytically inactive rhomboid-like proteins called iRhoms provide another form of regulation, by interacting with rhomboid substrates and preventing their cleavage. Extramembranous domains of rhomboids may play an as yet unexplored role in substrate recognition and regulation.


[Risk factors for cardiovascular disease in children with type 1 diabetes in 2000-2010 in Podlasie Province].
There has been an increase over the last decade in the number of young patients with type 1 diabetes treated with a continuous subcutaneous insulin infusion (CSII). The accelerated development of atherosclerosis is closely linked to metabolic control and traditional risk factors. Analysis of changes in the treatment and clinical picture of type 1 diabetes in children over the years 2000-2010, with emphasis on risk factors for cardiovascular disease. The study included 567 children diagnosed with type 1 diabetes. 251 children who were treated in 2000 were compared with 316 children treated in 2010. The study analyzed anthropometric parameters, laboratory tests and data obtained using questionnaires. In 2010, there was an increase in the percentage of children treated with CSU (up to 60.1%) and a decrease in the percentage of children using traditional insulin In favour of insulin analogues. An increase in HbA1c was observed from 7.4% to 8% (p < 0.001) and an increase in the percentage of patients with HbA1c>7.5%. There was an increase in the percentage of children with obesity from 5.2% to 13.7% (p =0.004) and an increase in BMI SDS. The number of children with hypertension was comparable in both groups (17.9% vs 15.8%), as was the percentage of children with dyslipidemią (48.6% ys 513%). Antihypertensive drugs were used in 97.8% vs. 70% of children with hypertension, metformin in 15.4% vs. 14% of children with obesity, and lipid-lowering drugs only in 3.3% vs; 2.5% of patients with dyslipidemia. It has been shown that nowadays children live in families burdened with risk factors for atherosclerosis. Contemporary patients frequently have excessive body weight and live in families burdened with risk factors for atherosclerosis. Despite the use of modern technology, metabolic control is not satisfactory.


Ubiquitination of K-Ras enhances activation and facilitates binding to select downstream effectors.
The guanosine triphosphate (GTP)--loaded form of the guanosine triphosphatase (GTPase) Ras initiates multiple signaling pathways by binding to various effectors, such as the kinase Raf and phosphatidylinositol 3-kinase (PI3K). Ras activity is increased by guanine nucleotide exchange factors that stimulate guanosine diphosphate release and GTP loading and is inhibited by GTPase-activating proteins that stimulate GTP hydrolysis. KRAS is the most frequently mutated RAS gene in cancer. Here, we report that monoubiquitination of lysine-147 in the guanine nucleotide-binding motif of wild-type K-Ras could lead to enhanced GTP loading. Furthermore, ubiquitination increased the binding of the oncogenic Gly12Val mutant of K-Ras to the downstream effectors PI3K and Raf. Thus, monoubiquitination could enhance GTP loading on K-Ras and increase its affinity for specific downstream effectors, providing a previously unidentified mechanism for Ras activation.


T helper lymphocytes rescue CTL from activation-induced cell death.
T cell activation is characterized by a vast expansion of Ag-specific T cells followed by an equally extensive reduction in T cell numbers. This decline is due, in part, to activation-induced apoptosis of the responding T cells during repeated encounter with Ag. In the current study, we used solid-phase MHC class I/peptide monomers to cause activation-induced cell death (AICD) of previously activated CD8 T cells in an Ag-specific manner. AICD occurred rapidly and was mediated primarily by Fas-FasL interactions. Most interestingly, we observed that Th cells could provide survival signals to CTL significantly reducing the level of AICD. Both Th1 and Th2 subsets were capable of protecting CTL from AICD, and a major role for soluble factors in this protection was ruled out, as cell-to-cell contact was an essential component of this Th-mediated protection. Upon encounter with Ag-expressing tumor cells, CTL underwent significant apoptosis. However, in the presence of Th cells, the CTL not only were protected against death, but also had significantly greater lytic ability. In vivo tumor protection studies using peptide immunization showed that the activation of Ag-specific Th cells was crucial for optimal protection, but did not affect the magnitude of the CTL response in the lymphoid tissues. In this study, we examine the type of help that CD4 T cells may provide and propose a model of Th cell-CTL interaction that reduces CTL death. Our results show a novel role for Th cells in the maintenance of CTL responses.


Variability in physicians' decisions on caring for chronically ill elderly patients: an international study.
To determine what treatment decisions physicians will make when faced with an incompetent elderly patient with life-threatening gastrointestinal bleeding and to identify the factors that affect their decisions. Survey. Family practice, medical and geriatrics rounds in academic medical centres and community hospitals in seven countries. Physicians who regularly cared for incompetent elderly patients. A self-administered questionnaire containing three case vignettes. Each provided the same details on an incompetent elderly patient; however, one gave no information about the wishes of the patient and his family (no directive), the second provided a do-not-resuscitate (DNR) request, and the third included a detailed therapeutic and resuscitative effort chart (DTREC) requesting maximum therapeutic care without admission to the intensive care unit (ICU). The four treatment options were supportive care only, limited therapeutic care, maximum therapeutic care without admission to the ICU and maximum care with admission to the ICU. Treatment decisions varied and were systematically related to age, level of training and country (p less than 0.001). The older physicians and those in family medicine were less likely than the others to choose aggressive treatment options. Brazilian and US physicians were the most aggressive; Australian physicians were the most conservative. The DNR request resulted in a significant decrease in the number of physicians choosing aggressive options (p less than 0.001). The DTREC resulted in a move toward more aggressive treatment, as outlined in the directive (p less than 0.001). Overall, however, about 40% of the physicians chose a level of care different from what had been requested. Furthermore, over 10% would have tried cardiopulmonary resuscitation despite the DNR request. Treatment of incompetent elderly patients with life-threatening illness varies widely within and between countries. Uniform standards should be developed on the basis of societal values and be communicated to physicians.


Neurotoxicology: principles and considerations of in vitro assessment.
Neurotoxicology is an exciting area of science, not only because of the importance of toxic injury to the nervous system in human disease, but also because specific toxicants have served as invaluable tools for the advancement of our knowledge of "normal" neurobiological processes. In fact, much of our understanding of the organisation and function of the nervous system is based on observations derived from the actions of neurotoxicants. This paper addresses various physiological aspects behind the exquisite sensitivity of the nervous system to toxic agents, including the privileged status of the nervous system vis-a-vis blood-brain barrier function, the extensions of the nervous system over space and the requirements of cells with such a complex geometry, and the transmission of information across extracellular space. In addition, in vitro models and their utility in the assessment of neurotoxicological outcome are discussed, with reference to both their advantages and disadvantages.


Spatiotemporal structure of femtosecond Bessel beams from spatial light modulators.
We numerically investigate the spatiotemporal structure of Bessel beams generated with spatial light modulators (SLMs). Grating-like phase masks enable the spatial filtering of undesired diffraction orders produced by SLMs. Pulse front tilt and temporal broadening effects are investigated. In addition, we explore the influence of phase wrapping and show that the spatiotemporal structure of SLM-generated femtosecond Bessel beams is similar to Bessel X-pulses at short propagation distance and to subluminal pulsed Bessel beams at long propagation distance.


Aloesin as a medical food ingredient for systemic oxidative stress of diabetes.
Diabetes is a chronic disease that requires a long term management where oxidative stress plays a pivotal role in disease progression and intensifying secondary complications. In spite of all the research on diabetes and recent advances in diabetes treatments, the reality is that there is no cure for diabetes and its devastating complications. While currently available anti-diabetic therapies are effective in reducing blood glucose level, they are not without associated side effects when they are used for a long term applications. As a result, physicians and patients are inclining more towards to a safer therapy with less serious side effects in the form of medicinal foods and botanical alternatives that are suitable for chronic usage. Aloesin, an Aloe chromone, has previously been formulated with an aloe polysaccharide to give a composition called Loesyn, where it showed significant impact in reducing glycosylated hemoglobin, fasting blood glucose, fructosamine and plasma insulin level in humans. Radical scavenging activities of chromones and polysaccharides from Aloe have also been reported. Here we rationalize the relevance of use of Aloesin alone or in a standardized blend with Aloe polysaccharides, as a potential medical food to manage systemic oxidative stress and/or high blood glucose of diabetes.


Tolerance optimization of a mobile phone camera lens system.
In the manufacturing process for the lens system of a mobile phone camera, various types of assembly and manufacturing tolerances, such as tilt and decenter, should be appropriately allocated. Because these tolerances affect manufacturing cost and the expected optical performance, it is necessary to choose a systematic design methodology for determining optimal tolerances. In order to determine the tolerances that minimize production cost while satisfying the reliability constraints on important optical performance indices, we propose a tolerance design procedure for a lens system. A tolerance analysis is carried out using Latin hypercube sampling for evaluating the expected optical performance. The tolerance optimization is carried out using a function-based sequential approximate optimization technique that can reduce the computational burden and smooth numerical noise occurring in the optimization process. Using the proposed design approach, the optimal production cost was decreased by 28.3% compared to the initial cost while satisfying all the constraints on the expected optical performance. We believe that the tolerance analysis and design procedure presented in this study can be applied to the tolerance optimization of other systems.


Three-dimensional reconstruction of the 70S Escherichia coli ribosome in ice: the distribution of ribosomal RNA.
A reconstruction, at 40 A, of the Escherichia coli ribosome imaged by cryo-electron microscopy, obtained from 303 projections by a single-particle method of reconstruction, shows the two subunits with unprecedented clarity. In the interior of the subunits, a complex distribution of higher mass density is recognized, which is attributed to ribosomal RNA. The masses corresponding to the 16S and 23S components are linked in the region of the platform of the small subunit. Thus the topography of the rRNA regions responsible for protein synthesis can be described.


Genetic susceptibility to lung cancer as determined by lymphocytic chromosome analysis.
Chromosomal anomalies were analyzed in the lymphocyte cultures among 96 untreated lung cancer patients and 74 clinically normal comparison subjects. The analysis revealed that >15% of the lung cancer patients showed structural or numerical rearrangements in chromosomes 1,3,5,7,9,12,14, and 21. A case control comparison showed that these aberrations were significantly higher in chromosome 7 [odds ratio (OR) = 2.32; 95% confidence interval (CI), 1.14 and 4.82], chromosome 9 (OR = 2.61; 95% CI, 1.27 and 5.48), chromosome 12 (OR = 4.10; 95% CI, 1.40 and 14.54), and chromosome 21 (OR = 7.75; 95% CI, 1.73 and 70.80) of the patients than in the controls. However, only chromosome 9 (OR = 3.57; 95% CI, 1.33 and 9.46) and chromosome 21 (OR = 6.94; 95% CI, 3.15 and 9.98) retained significance after stratifying on smoking status. Among the lung cancer patients, the breakpoints cluster in specific regions of some of these chromosomes. These regions are 1p13-q21, 3q21-q13, 7p12-q12, 7q12-q12,7q22, 7q32, 9p13-q13, 12p13, 14q11, and 14q32. The distribution of lung cancer patients, according to histological types, showed that aberrations in chromosomes 1,7, and 9 dominated the scenario of chromosomal changes in non-small cell lung carcinomas. Thus, the data on lymphocytic chromosomal rearrangements in lung cancer patients not only indicate the importance of specific genetic changes in the etiology of lung cancer but also emphasizes the putative role of such analysis in determining primary genetic abnormalities in the large heterogeneous group of lung cancers.


Induction of apoptosis by the retinoid inducible growth regulator RIG1 depends on the NC motif in HtTA cervical cancer cells.
Retinoid-inducible gene 1 (RIG1), also known as tazarotene-induced gene 3 or retinoic-acid receptor responder 3, is a growth regulator, which induces apoptosis and differentiation. RIG1 is classified into the NC protein family. This study investigated functional domains and critical amino acids associated with RIG1-mediated cell death and apoptosis. Using enhanced green fluorescence protein (EGFP)-tagged RIG1 variants, RIG1 proteins with deletion at the NC domain significantly decreased cell death induced by RIG1, and fusion variants containing only the NC domain significantly induced apoptosis of HtTA cervical cancer cells. The EGFP-RIG1-induced apoptosis was significantly decreased in cells expressing N(112)C(113) motif double- (NC-->FG) or triple- (NCR-->FGE) mutated RIG1 variants. Using dodecapeptides, nuclear localization and profound cell death was observed in HtTA cells expressing wild type RIG1(111-123) or Leu121-mutated RIG1(111-123):L--> C peptide, but peptides double- or triple-mutated at the NC motif alone, RIG1(111-123):NC-->FG or RIG1(111-123):NCR-->FGE, were cytoplasmically localized and did not induce apoptosis. The RIG1(111-123) also induced apoptosis of A2058 melanoma cells but not normal human fibroblasts. The NC domain, especially the NC motif, plays the major role in RIG1-mediated pro-apoptotic activity. The RIG1(111-123) dodecapeptide exhibited strong pro-apoptotic activity and has potential as an anticancer drug.


Recall and recognition memory for spider information.
Two experiments were devised to examine the effect of anxiety on memory for threatening information in those with spider phobia. Those with spider phobia, control participants, and those with other phobias were shown video clips of spiders. In the recognition task, participants were asked to state whether they remembered having seen specific video clips of spiders that had been presented to them previously. In the recall task, they were asked to remember and note as much detail as possible about each of a second series of video clips of spiders. Those with spider phobia were no different from the other two groups in their ability to recognize spider stimuli or in the amount of detail they were able to recall.


Efficacy of faecal microbiota transplantation for patients with irritable bowel syndrome in a randomised, double-blind, placebo-controlled study.
Faecal microbiota transplantation (FMT) from healthy donors to patients with irritable bowel syndrome (IBS) has been attempted in two previous double-blind, placebo-controlled studies. While one of those studies found improvement of the IBS symptoms, the other found no effect. The present study was conducted to clarify these contradictory findings. This randomised, double-blind, placebo-controlled study randomised 165 patients with IBS to placebo (own faeces), 30 g FMT or 60 g FMT at a ratio of 1:1:1. The material for FMT was obtained from one healthy, well-characterised donor, frozen and administered via gastroscope. The primary outcome was a reduction in the IBS symptoms at 3 months after FMT (response). A response was defined as a decrease of 50 or more points in the total IBS symptom score. The secondary outcome was a reduction in the dysbiosis index (DI) and a change in the intestinal bacterial profile, analysed by 16S rRNA gene sequencing, at 1 month following FMT. Responses occurred in 23.6%, 76.9% (p<0.0001) and 89.1% (p<00.0001) of the patients who received placebo, 30 g FMT and 60 g FMT, respectively. These were accompanied by significant improvements in fatigue and the quality of life in patients who received FMT. The intestinal bacterial profiles changed also significantly in the groups received FMT. The FMT adverse events were mild self-limiting gastrointestinal symptoms. FMT is an effective treatment for patients with IBS. Utilising a well-defined donor with a normal DI and favourable specific microbial signature is essential for successful FMT. The response to FMT increases with the dose. Trial registration www.clinicaltrials.gov (NCT03822299) and www.cristin.no (ID657402).


Sulfate assimilation mediates tellurite reduction and toxicity in Saccharomyces cerevisiae.
Despite a century of research and increasing environmental and human health concerns, the mechanistic basis of the toxicity of derivatives of the metalloid tellurium, Te, in particular the oxyanion tellurite, Te(IV), remains unsolved. Here, we provide an unbiased view of the mechanisms of tellurium metabolism in the yeast Saccharomyces cerevisiae by measuring deviations in Te-related traits of a complete collection of gene knockout mutants. Reduction of Te(IV) and intracellular accumulation as metallic tellurium strongly correlated with loss of cellular fitness, suggesting that Te(IV) reduction and toxicity are causally linked. The sulfate assimilation pathway upstream of Met17, in particular, the sulfite reductase and its cofactor siroheme, was shown to be central to tellurite toxicity and its reduction to elemental tellurium. Gene knockout mutants with altered Te(IV) tolerance also showed a similar deviation in tolerance to both selenite and, interestingly, selenomethionine, suggesting that the toxicity of these agents stems from a common mechanism. We also show that Te(IV) reduction and toxicity in yeast is partially mediated via a mitochondrial respiratory mechanism that does not encompass the generation of substantial oxidative stress. The results reported here represent a robust base from which to attack the mechanistic details of Te(IV) toxicity and reduction in a eukaryotic organism.


Glucagon-stimulated copeptin measurements in the differential diagnosis of diabetes insipidus: a double-blind, randomized, placebo-controlled study.
The differential diagnosis of diabetes insipidus is challenging. The most reliable approaches are copeptin measurements after hypertonic saline infusion or arginine, which is a known growth hormone secretagogue but has recently also been shown to stimulate the neurohypophysis. Similar to arginine, glucagon stimulates growth hormone release, but its effect on the neurohypophysis is poorly studied. Double-blind, randomized, placebo-controlled trial including 22 healthy participants, 10 patients with central diabetes insipidus, and 10 patients with primary polydipsia at the University Hospital Basel, Switzerland. Each participant underwent the glucagon test (s.c. injection of 1 mg glucagon) and placebo test. The primary objective was to determine whether glucagon stimulates copeptin and to explore whether the copeptin response differentiates between diabetes insipidus and primary polydipsia. Copeptin levels were measured at baseline, 30, 60, 90, 120, 150, and 180 min after injection. In healthy participants, glucagon stimulated copeptin with a median increase of 7.56 (2.38; 28.03) pmol/L, while placebo had no effect (0.10 pmol/L (-0.70; 0.68); P < 0.001). In patients with diabetes insipidus, copeptin showed no relevant increase upon glucagon, with an increase of 0.55 (0.21; 1.65) pmol/L, whereas copeptin was stimulated in patients with primary polydipsia with an increase of 15.70 (5.99; 24.39) pmol/L. Using a copeptin cut-off level of 4.6pmol/L had a sensitivity of 100% (95% CI: 100-100) and a specificity of 90% (95% CI: 70-100) to discriminate between diabetes insipidus and primary polydipsia. Glucagon stimulates the neurohypophysis, and glucagon-stimulated copeptin has the potential for a safe, novel, and precise test in the differential diagnosis of diabetes insipidus.


Metal-organic frameworks for the removal of toxic industrial chemicals and chemical warfare agents.
Owing to the vast diversity of linkers, nodes, and topologies, metal-organic frameworks can be tailored for specific tasks, such as chemical separations or catalysis. Accordingly, these materials have attracted significant interest for capture and/or detoxification of toxic industrial chemicals and chemical warfare agents. In this paper, we review recent experimental and computational work pertaining to the capture of several industrially-relevant toxic chemicals, including NH3, SO2, NO2, H2S, and some volatile organic compounds, with particular emphasis on the challenging issue of designing materials that selectively adsorb these chemicals in the presence of water. We also examine recent research on the capture and catalytic degradation of chemical warfare agents such as sarin and sulfur mustard using metal-organic frameworks.


The need for a national cancer policy in Malawi.
Cancer is causing a lot of suffering and death in Africa but is not considered a major health problem in Africa. This needs to change. Cancer should be given equal emphasis to HIV/AIDS, tuberculosis (TB) and Malaria. A national cancer policy is required in Malawi to develop and improve evidence-based cancer prevention, early diagnosis, curative and palliative therapy. A national cancer policy is crucial to ensure a priotised, clear, coordinated and sustained fight against cancer. When no policy exists, events are likely to be random, stakeholders and practitioners in the fight against cancer may not agree on how to proceed, may duplicate efforts or may neglect areas that would have greater nationwide impact resulting in poor quality activities and haphazard development.


Integrating Case Detection of Visceral Leishmaniasis and Other Febrile Illness with Vector Control in the Post-Elimination Phase in Nepal.
Nepal has completed the attack phase of visceral leishmaniasis (VL) elimination and now needs active case detection (ACD) and vector control methods that are suitable to the consolidation and maintenance phases. We evaluated different ACD approaches and vector control methods in Saptari district. We assessed 1) mobile teams deployed in villages with VL cases in 2015 to conduct combined camps for fever and skin lesions to detect VL/PKDL (post-kala-azar dermal leishmaniasis) and other infections; 2) an incentive approach by trained female community health volunteers (FCHVs) in villages with no VL cases in 2015. Both were followed by house-to-house visits. For vector control, four villages were randomly allocated to insecticide impregnation of bednets, insecticide wall painting, indoor residual spraying (IRS), and control. Sandfly density (by CDC light traps, The John W. Hock Company, USA) and mortality (World Health Organization cone bioassay) were assessed in randomly selected households. One VL, three tuberculosis, one leprosy, and one malaria cases were identified among 395 febrile cases attending the camps. Post-camp house-to-house screening involving 7,211 households identified 679 chronic fever and 461 skin lesion cases but no additional VL/PKDL. No VL/PKDL case was found by FCHVs. The point prevalence of chronic fever in camp and FCHV villages was 242 and 2 per 10,000 populations, respectively. Indoor residual spraying and bednet impregnation were effective for 1 month versus 12 months with insecticidal wall paint. Twelve-month sandfly mortality was 23%, 26%, and 80%, respectively, on IRS, bednet impregnation, and insecticide wall painting. In Nepal, fever camps and insecticidal wall paint prove to be alternative, sustainable strategies in the VL post-elimination program.


Growth inhibitory effects of kimchi (Korean traditional fermented vegetable product) against Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus.
Kimchi is a unique Korean traditional vegetable product that is fermented by lactic acid bacteria (LAB) and is mainly consumed as a side dish with boiled rice. Its main ingredients are brined Chinese cabbage, red pepper powder, and fermented fish sauce, and these are combined with many spices such as garlic, green onion, ginger, and some seaweed. The relationship between the concentration of LAB or the pH and the growth of three gram-positive foodborne pathogens (Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus) was evaluated. Heat treatment (HT; 85 degrees C for 15 min) or neutralization treatment (NT; pH 7.0) was conducted on day 0 (0-D group) and day 3 (3-D group) of incubation. The pH in the control group and the NT group dropped sharply to 4.12 to 4.30 after 2 days of incubation and slightly decreased thereafter, whereas the pH in the control group and HT group stayed at 7.0 during incubation. LAB were not detected in the HT kimchi during incubation. B. cereus in the NT-0-D, NT-3-D, and HT-3-D groups was reduced by 1.5 to 3.1 log CFU/ml but increased slightly in the HT-0-D group. L. monocytogenes in HT-3-D and NT-3-D groups disappeared after 5 days of incubation, and S. aureus in the NT-0-D group disappeared after 4 days. These findings indicate that growth of all the foodborne pathogens was inhibited by NT-0-D, HT-3-D, and NT-3-D, but B. cereus was not inhibited by HT-0-D. Thus, growth of LAB in kimchi is an important factor in the control of foodborne pathogens.


Entrapment of intracytosolic bacteria by septin cage-like structures.
Actin-based motility is used by various pathogens for dissemination within and between cells. Yet host factors restricting this process have not been identified. Septins are GTP-binding proteins that assemble as filaments and are essential for cell division. However, their role during interphase has remained elusive. Here, we report that septin assemblies are recruited to different bacteria that polymerize actin. We observed that intracytosolic Shigella either become compartmentalized in septin cage-like structures or form actin tails. Inactivation of septin caging increases the number of Shigella with actin tails and enhances cell-to-cell spread. TNF-α, a host cytokine produced upon Shigella infection, stimulates septin caging and restricts actin tail formation and cell-to-cell spread. Finally, we show that septin cages entrap bacteria targeted to autophagy. Together, these results reveal an unsuspected mechanism of host defense that restricts dissemination of invasive pathogens.


Soluble (pro)renin receptor: a novel ligand for angiotensin II type 1 receptor?
This commentary highlights the study entitled 'Soluble (pro)renin receptor induces endothelial dysfunction and hypertension in mice with diet-induced obesity via activation of angiotensin II type 1 receptor' presented by Fu et al. published in Clinical Science (Clin Sci (Lond) (2021) 135(6), https://doi.org/10.1042/CS20201047). The authors evaluated the role of the soluble (pro)renin receptor (sPRR), a cleavage product of the prorenin receptor (PRR) by the site 1 protease, as a ligand for angiotensin II type 1 receptor (AT1R). They presented for the first time that sPRR directly interacts with AT1R, causing nuclear factor-κB activation, inflammation, apoptosis, and endothelial dysfunction in primary human umbilical vein endothelial cells (HUVECs). Furthermore, the interaction between sPRR and AT1R was responsible for endothelial dysfunction and hypertension in diet-induced obesity mice. These results provide a potential mechanism for obesity-induced endothelial dysfunction and hypertension. Thus, the sPRR/AT1R complex may be a novel therapeutic target for cardiovascular diseases associated with endothelial dysfunction.


Characterization of the Duffy-Binding-Like Domain of Plasmodium falciparum Blood-Stage Antigen 332.
Studies on Pf332, a major Plasmodium falciparum blood-stage antigen, have largely been hampered by the cross-reactive nature of antibodies generated against the molecule due to its high content of repeats, which are present in other malaria antigens. We previously reported the identification of a conserved domain in Pf332 with a high degree of similarity to the Duffy-binding-like (DBL) domains of the erythrocyte-binding-like (EBL) family. We here describe that antibodies towards Pf332-DBL are induced after repeated exposure to P. falciparum and that they are acquired early in life in areas of intense malaria transmission. Furthermore, a homology model of Pf332-DBL was found to be similar to the structure of the EBL-DBLs. Despite their similarities, antibodies towards Pf332-DBL did not display any cross-reactivity with EBL-proteins as demonstrated by immunofluorescence microscopy, Western blotting, and peptide microarray. Thus the DBL domain is an attractive region to use in further studies on the giant Pf332 molecule.


Chiral meta-molecules consisting of gold nanoparticles and genetically engineered tobacco mosaic virus.
We demonstrate a chiral meta-molecule in the ultraviolet (UV) and visible (VIS) regions using a complex of Au nanoparticles (NPs) and rod-shaped tobacco mosaic virus (TMV). Au NPs five nm in diameter are uniformly formed on peptide-modified TMV. The peptide-modified TMV with uniform-sized Au NPs has improved dispersion in solution. A negative circular dichroism (CD) peak is produced around 540 nm, at plasmonic resonance wavelength of Au NPs. Additionally, modification of a CD peak in the UV region is observed. Attaching NPs to a virus causes the enhancement and modification of CD peaks in both the UV and VIS regions. Our results open a new avenue for the preparation of three dimensional chiral metamaterials at optical frequencies.


Antifibrotic effect of Pirfenidone on orbital fibroblasts of patients with thyroid-associated ophthalmopathy by decreasing TIMP-1 and collagen levels.
The aim of this study was to determine the antifibrotic effects of pirfenidone in orbital fibroblasts of patients with thyroid-associated ophthalmopathy (TAO). The effects of interleukin (IL)-1beta and of fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), and transforming growth factor (TGF)-beta on the induction of tissue inhibitors of metalloproteinases (TIMP)-1 were assessed in orbital fibroblasts of TAO patients. TIMP-1 protein levels were measured by ELISA and Western blot analyses, and TIMP-1 activity was assessed by reverse zymography. The effect of pirfenidone on TIMP-1 induction in orbital fibroblasts was evaluated with the same methods using dexamethasone as a reference agent. A hydroxyproline assay was used to determine the effect of pirfenidone and dexamethasone on collagen production in orbital fibroblasts, and the tetrazolium-based MTT assay was used to assess pirfenidone cytotoxicity. IL-1beta strongly and dose dependently increased the level of active TIMP-1 protein, whereas FGF, PDGF, and TGF-beta did not significantly induce TIMP-1 protein. Pirfenidone was more effective than dexamethasone in inhibiting IL-1beta-induced increases in TIMP-1, reducing TIMP-1 levels to less than those in untreated controls at a minimal concentration (5 mM). Moreover, pirfenidone effectively decreased hydroxyproline levels in orbital fibroblasts, whereas dexamethasone had no effect on hydroxyproline levels. Pirfenidone exhibited no toxicity in orbital fibroblasts at the concentrations used. These results indicate that nontoxic concentrations of pirfenidone have significant antifibrotic effects on orbital fibroblasts from patients with TAO.


Radiation Therapy for Small-Cell Lung Cancer: ASCO Guideline Endorsement of an ASTRO Guideline.
The American Society for Radiation Oncology (ASTRO) produced an evidence-based guideline on radiation therapy (RT) for small-cell lung cancer (SCLC). Because of the relevance of this topic to ASCO membership, ASCO reviewed the guideline, applying a set of procedures and policies used to critically examine guidelines developed by other organizations. The ASTRO guideline on RT for SCLC was reviewed for developmental rigor by methodologists. Then, an ASCO Expert Panel reviewed the content and the recommendations. The ASCO Expert Panel determined that the recommendations from ASTRO guideline on RT for SCLC, published in June 2020, are clear, thorough, and based upon the most relevant scientific evidence. ASCO endorsed ASTRO guideline on RT for SCLC with a few discussion points. Recommendations addressed thoracic radiotherapy for limited-stage SCLC, role of stereotactic body radiotherapy in stage I or II node-negative SCLC, prophylactic cranial radiotherapy, and thoracic consolidation for extensive-stage SCLC.Additional information is available at www.asco.org/thoracic-cancer-guidelines.