Potential carcinogenic hazards of non-regulated disinfection by-products: haloquinones, halo-cyclopentene and cyclohexene derivatives, N-halamines, halonitriles, and heterocyclic amines.
Drinking water disinfectants react with natural organic material (NOM) present in source waters used for drinking water to produce a wide variety of by-products. Several hundred disinfections by-products (DBPs) have been identified, but none have been identified with sufficient carcinogenic potency to account for the cancer risks projected from epidemiological studies. In a search for DBPs that might fill this risk gap, the present study projected reactions of chlorine and chloramine that could occur with substructures present in NOM to produce novel by-products. A review of toxicological data on related compounds, supplemented by use of a quantitative structure toxicity relationship (QSTR) program TOPKAT®) identified chemicals with a high probability of being chronically toxic and/or carcinogenic among 489 established and novel DBPs. Classes of DBPs that were specifically examined were haloquinones (HQs), related halo-cyclopentene and cyclohexene (HCP&H) derivatives, halonitriles (HNs), organic N-chloramines (NCls), haloacetamides (HAMs), and nitrosamines (NAs). A review of toxicological data available for quinones suggested that HQs and HCP&H derivatives appeared likely to be of health concern and were predicted to have chronic lowest observed adverse effect levels (LOAELs) in the low μg/kg day range. Several HQs were predicted to be carcinogenic. Some have now been identified in drinking water. The broader class of HNs was explored by considering current toxicological data on haloacetonitriles and extending this to halopropionitriles. 2,2-dichloropropionitrile has been identified in drinking water at low concentrations, as well as the more widely recognized haloacetonitriles. The occurrence of HAMs has been previously documented. The very limited toxicological data on HAMs suggests that this class would have toxicological potencies similar to the dihaloacetic acids. Organic N-halamines are also known to be produced in drinking water treatment and have biological properties of concern, but no member has ever been characterized toxicologically beyond bacterial or in vitro studies of genotoxicity. The documented formation of several nitrosamines from secondary amines from both natural and industrial sources prompted exploration of the formation of additional nitrosamines. N-diphenylnitrosamine was identified in drinking waters. Of more interest, however, was the formation of phenazine (and subsequently N-chorophenazine) in a competing reaction. These are the first heterocyclic amines that have been identified as chlorination by-products. Consideration of the amounts detected of members of these by-product classes and their probable toxicological potency suggest a prioritization for obtaining more detailed toxicological data of HQs>HCP&H derivatives>NCls>HNs. Based upon a ubiquitous occurrence and virtual lack of in vivo toxicological data, NCls are the most difficult group to assign a priority as potential carcinogenic risks. This analysis indicates that research on the general problem of DBPs requires a more systematic approach than has been pursued in the past. Utilization of predictive chemical tools to guide further research can help bring resolution to the DBP issue by identifying likely DBPs with high toxicological potency.


Structural and molecular biology of Sabiá virus.
Brazilian mammarenavirus, or Sabiá virus (SABV), is a New World (NW) arenavirus associated with fulminant hemorrhagic disease in humans and the sole biosafety level 4 microorganism ever isolated in Brazil. Since the isolation of SABV in the 1990s, studies on viral biology have been scarce, with no available countermeasures against SABV infection or disease. Here we provide a comprehensive review of SABV biology, including key aspects of SABV replication, and comparisons with related Old World and NW arenaviruses. SABV is most likely a rodent-borne virus, transmitted to humans, through exposure to urine and feces in peri-urban areas. Using protein structure prediction methods and alignments, we analyzed shared and unique features of SABV proteins (GPC, NP, Z, and L) that could be explored in search of therapeutic strategies, including repurposing intended application against arenaviruses. Highly conserved catalytic activities present in L protein could be targeted for broad-acting antiviral activity among arenaviruses, while protein-protein interactions, such as those between L and the matrix protein Z, have evolved in NW arenaviruses and should be specific to SABV. The nucleoprotein (NP) also shares targetable interaction interfaces with L and Z and exhibits exonuclease activity in the C-terminal domain, which may be involved in multiple aspects of SABV replication. Envelope glycoproteins GP1 and GP2 have been explored in the development of promising cross-reactive neutralizing antibodies and vaccines, some of which could be repurposed for SABV. GP1 remains a challenging target in SABV as evolutive pressures render it the most variable viral protein in terms of both sequence and structure, while antiviral strategies targeting the Z protein remain to be validated. In conclusion, the prediction and analysis of protein structures should revolutionize research on viruses such as SABV by facilitating the rational design of countermeasures while reducing dependence on sophisticated laboratory infrastructure for experimental validation.


Olive oil reduces oxidative damage in a 3-nitropropionic acid-induced Huntington's disease-like rat model.
Free radicals contribute to altered neuronal functions in neurodegenerative diseases and brain aging, by producing lipid- and other molecule-dependent modifications. The Mediterranean diet has been associated with a reduced risk of neurodegenerative disease. This study sought to verify whether extra-virgin olive oil (EVOO) exerted a brain antioxidant effect, protecting the brain against the oxidative stress caused by 3-nitropropionic acid (3NP). 3NP was administered intraperitoneally (i.p.) at a dose of 20 mg/kg body weight over four consecutive days. EVOO (representing 10% of calorie intake in the total standard daily diet of rats) and hydroxytyrosol (HT; 2.5 mg/kg body weight) were administered for 14 days. In all studied samples, 3NP caused a rise in lipid peroxides (LPO) and a reduction in glutathione (GSH) content. While the results showed that EVOO and HT reduces lipid peroxidation product levels and blocks the GSH depletion prompted by 3NP in both striatum and rest of the brain in Wistar rats. In addition, EVOO blocks and reverses the effect of 3NP on succinate dehydrogenase activity. In brief, the data obtained indicate that EVOO and HT act as a powerful brain antioxidant.


Catalytically active telomerase holoenzyme is assembled in the dense fibrillar component of the nucleolus during S phase.
The maintenance of human telomeres requires the ribonucleoprotein enzyme telomerase, which is composed of telomerase reverse transcriptase (TERT), telomerase RNA component, and several additional proteins for assembly and activity. Telomere elongation by telomerase in human cancer cells involves multiple steps including telomerase RNA biogenesis, holoenzyme assembly, intranuclear trafficking, and telomerase recruitment to telomeres. Although telomerase has been shown to accumulate in Cajal bodies for association with telomeric chromatin, it is unclear where and how the assembly and trafficking of catalytically active telomerase is regulated in the context of nuclear architecture. Here, we show that the catalytically active holoenzyme is initially assembled in the dense fibrillar component of the nucleolus during S phase. The telomerase RNP is retained in nucleoli through the interaction of hTERT with nucleolin, a major nucleolar phosphoprotein. Upon association with TCAB1 in S phase, the telomerase RNP is transported from nucleoli to Cajal bodies, suggesting that TCAB1 acts as an S-phase-specific holoenzyme component. Furthermore, depletion of TCAB1 caused an increase in the amount of telomerase RNP associated with nucleolin. These results suggest that the TCAB1-dependent trafficking of telomerase to Cajal bodies occurs in a step separate from the holoenzyme assembly in nucleoli. Thus, we propose that the dense fibrillar component is the provider of active telomerase RNP for supporting the continued proliferation of cancer and stem cells.


Surgical treatment of laryngeal carcinoma with subglottis involvement.
Primary cancers arising in the subglottic region are rare and are characterized by a long asymptomatic phase. More frequently the subglottis is reached by tumors arising in the glottis or even the supraglottis through invasion of the paraglottic space. Involvement of the subglottis is associated with a relatively high frequency of stomal recurrences due to a peculiar lymphatic spread to the paratracheal nodes. We analyzed a retrospective series of 68 patients with squamous cell carcinoma of the larynx extending to the subglottis region submitted to total simple laryngectomy or total laryngectomy enlarged with hemithyroidectomy and dissection of level VI nodes (HT/SPD). Overall median follow-up is 46 months. Subglottic extension was correctly diagnosed before operation in only 13/68 patients, however the resection margins, systematically determined by the pathologist, were in every case negative. Stomal relapses in laryngectomized patients without HT/SPD have been more frequent (0.55% rate per month) than in those treated with laryngectomy and HT/SPD (0.07% rate per month). It is concluded that CT should be routinely applied in preoperative staging in order to estimate the extension of the neoplasia and surgery should always include hemithyroidectomy and dissection of the homolateral paratracheal nodes when there is even minimal involvement of the subglottis. Moreover, the high incidence of second tumors in our series is noteworthy; such patients might benefit from chemopreventive therapy.


The specificity of human anti-cytochrome c autoantibodies that arise in autoimmune disease.
Cytochromes c (cyt c) are among the best characterized model Ag because their amino acid sequences and tertiary structures are well defined. One unique aspect of cyt c as an immunogen is its ability to induce autoantibody responses in animal models, although no pathology resulting from these responses has been reported. In this study, the presence and specificity of autoantibodies to cyt c were investigated in patients with SLE and related connective tissue diseases. Anti-cyt c antibodies were found in approximately 7% of patient sera and were statistically associated with the expression of antimitochondrial antibodies but were not statistically associated with any disease subset among those represented. Anti-cyt c was not associated with the presence of autoantibodies to DNA, histones, Ro, La, or Sm autoantigens. Most of the autoantibodies were specific for native or native-like forms of cyt c but antibodies to denatured forms were also apparent. Autoantibody binding was shown to be directed predominantly at selected sites of evolutionary variability within cyt c. The specificity of the human anti-cyt c autoantibodies appear to be similar to that of mouse anti-human cyt c antibodies and to autoantibodies elicited in mice against rat (mouse) cyt c.


Suppression of plant defenses by a Myzus persicae (green peach aphid) salivary effector protein.
The complex interactions between aphids and their host plant are species-specific and involve multiple layers of recognition and defense. Aphid salivary proteins, which are released into the plant during phloem feeding, are a likely mediator of these interactions. In an approach to identify aphid effectors that facilitate feeding from host plants, eleven Myzus persicae (green peach aphid) salivary proteins and the GroEL protein of Buchnera aphidicola, a bacterial endosymbiont of this aphid species, were expressed transiently in Nicotiana tabacum (tobacco). Whereas two salivary proteins increased aphid reproduction, expression of three other aphid proteins and GroEL significantly decreased aphid reproduction on N. tabacum. These effects were recapitulated in stable transgenic Arabidopsis thaliana plants. Further experiments with A. thaliana expressing Mp55, a salivary protein that increased aphid reproduction, showed lower accumulation of 4-methoxyindol-3-ylmethylglucosinolate, callose and hydrogen peroxide in response to aphid feeding. Mp55-expressing plants also were more attractive for aphids in choice assays. Silencing Mp55 gene expression in M. persicae using RNA interference approaches reduced aphid reproduction on N. tabacum, A. thaliana, and N. benthamiana. Together, these results demonstrate a role for Mp55, a protein with as-yet-unknown molecular function, in the interaction of M. persicae with its host plants.


Comparison of Clostripain and Neutral Protease as Supplementary Enzymes for Human Islet Isolation.
Although human islet transplantation has been established as valid and safe treatment for patients with type 1 diabetes, the utilization rates of human pancreases for clinical islet transplantation are still limited and substantially determined by the quality and composition of collagenase blends. While function and integrity of collagenase has been extensively investigated, information is still lacking about the most suitable supplementary neutral proteases. The present study compared islet isolation outcome after pancreas digestion by means of collagenase used alone or supplemented with either neutral protease (NP), clostripain (CP), or both proteases. Decent amounts of islet equivalents (IEQ) were isolated using collagenase alone (3090 ± 550 IEQ/g), or in combination with NP (2340 ± 450 IEQ/g) or CP (2740 ± 280 IEQ/g). Nevertheless, the proportion of undigested tissue was higher after using collagenase alone (21.1 ± 1.1%, P < 0.05) compared with addition of NP (13.3 ± 2.2%) or CP plus NP (13.7 ± 2.6%). Likewise, the percentage of embedded islets was highest using collagenase only (13 ± 2%) and lowest adding NP plus CP (4 ± 1%, P < 0.01). The latter combination resulted in lowest post-culture overall survival (42.7 ± 3.9%), while highest survival was observed after supplementation with CP (74.5 ± 4.8%, P < 0.01). An insulin response toward glucose challenge was present in all experimental groups, but the stimulation index was significantly decreased using collagenase plus NP (2.0 ± 0.12) compared with supplementation with CP (3.16 ± 0.4, P < 0.001). This study demonstrates for the first time that it is possible to isolate significant numbers of human islets combining collagenase only with CP. The supplementation with CP is an effective means to substantially reduce NP activity, which significantly decreases survival and viability after culture. This will facilitate the manufacturing of enzyme blends with less harmful characteristics.


Low birth weight is associated with lower respiratory tract infections in children with hand, foot, and mouth disease.
Low birth weight (LBW) might be a risk factor for acquiring lower respiratory tract infections (LRTIs) associated with disease related complications in early childhood. HFMD, a frequent viral infection in southern China, is a leading cause of lower respiratory tract infections in children. We analyzed whether LBW is a risk factor for children with HFMD to develop lower respiratory tract infections. A total of 298 children with HFMD, admitted to a hospital in Qingyuan city, Guangdong province, were recruited. Demographic data and clinical parameters such as serum glucose level and inflammatory markers including peripheral white blood cell count, serum C-reactive protein, and erythrocyte sedimentation rate were routinely collected on admission. Birth weight data were derived from birth records. Mean birth weight (BW) was 167 g lower in patients with HFMD and LRTIs as compared to patients with solely HFMD (p = 0.022) and the frequency of birth weight below the tenth percentile was significantly higher in patients with HFMD and LRTIs (p = 0.002). The results of the study show that low birth weight is associated with a higher incidence of lower respiratory tract infections in young children with HFMD.


Measurement of macular pigment: Raman spectroscopy versus heterochromatic flicker photometry.
There are several techniques for measuring macular pigment (MP) in vivo, of which Raman spectroscopy (RS) is a recently developed objective This study reports the reproducibility, test-retest variability, and validity of RS MP readings, by comparing them with heterochromatic flicker photometry (HFP). MP was measured with HFP and RS in 120 healthy subjects, and the latter technique was also used on two separate occasions in a sample of 20 subjects to investigate the intersessional variability of readings. Intrasessional reproducibility of RS MP measurements was also calculated. In addition, serum concentrations of lutein (L) and zeaxanthin (Z) were measured and correlated with both RS and HFP MP readings. Mean (+/-SD) MP in the right eye was 0.279 +/- 0.145 and 0.319 +/- 0.155 with RS and HFP, respectively. The differences between corresponding MP readings taken on RS and HFP lay within the Bland-Altman 95% limits of agreement for the two instruments in 93.6% and 94.4% of cases in the right and left eyes, respectively. Intrasessional reproducibility of RS readings, expressed as the coefficient of variation, was 8.42% +/- 7.12%. Ninety-five percent of MP readings taken with RS on two separate occasions lay within the 95% limits of agreement for the two sessions. A positive, but insignificant, relationship was observed between RS and HFP MP readings and serum concentrations of L and Z (RS, P = 0.356; HFP, P = 0.540). RS, an objective method of measuring MP levels in vivo, exhibits acceptable reproducibility and test-retest variability. The results demonstrated good correlation between RS and HFP measurements of MP, thus authenticating RS against a validated psychophysical technique of measuring MP. However, investigators should use only one of these instruments for the duration of any given study because of differences in the scientific rationale, and the factors that influence RS and HFP measurements of MP.


A Cellular Model of Shear-Induced Hemolysis.
A novel model is presented to study red blood cell (RBC) hemolysis at cellular level. Under high shear rates, pores form on RBC membranes through which hemoglobin (Hb) leaks out and increases free Hb content of plasma leading to hemolysis. By coupling lattice Boltzmann and spring connected network models through immersed boundary method, we estimate hemolysis of a single RBC under various shear rates. First, we use adaptive meshing to find local strain distribution and critical sites on RBC membranes, and then we apply underlying molecular dynamics simulations to evaluate damage. Our approach comprises three sub-models: defining criteria of pore formation, calculating pore size, and measuring Hb diffusive flux out of pores. Our damage model uses information of different scales to predict cellular level hemolysis. Results are compared with experimental studies and other models in literature. The developed cellular damage model can be used as a predictive tool for hydrodynamic and hematologic design optimization of blood-wetting medical devices.


Vergence Fusion Sustaining Oscillations.
Previous studies have shown that the slow, or fusion sustaining, component of disparity vergence contains oscillatory behavior as would be expected if fusion is sustained by visual feedback. This study extends the examination of this behavior to a wider range of frequencies and a larger number of subjects. Disparity vergence responses to symmetrical 4.0 deg step changes in target position were recorded in 20 subjects. Approximately three seconds of the late component of each response were isolated using interactive graphics and the frequency spectrum calculated. Peaks in these spectra associated with oscillatory behavior were identified and examined. All subjects exhibited oscillatory behavior with fundamental frequencies ranging between 0.37 and 0.55 Hz; much lower than those identified in the earlier study. All responses showed significant higher frequency components. The relationship between higher frequency components and the fundamental frequency suggest may be harmonics. A correlation was found across subjects between the amplitude of the fundamental frequency and the maximum velocity of the fusion initiating component probably due to the gain of shared neural pathways. Low frequency oscillatory behavior was found in all subjects adding support that the slow, or fusion sustaining, component is mediated by a feedback control.


Viable fibroblast matrix patch induces angiogenesis and increases myocardial blood flow in heart failure after myocardial infarction.
This study examines a viable biodegradable three-dimensional fibroblast construct (3DFC) in a model of chronic heart failure. The viable fibroblasts, cultured on a vicryl mesh, secrete growth factors that stimulate angiogenesis. We ligated the left coronary artery of male Sprague-Dawley rats, implanted the 3DFC 3 weeks after myocardial infarction and obtained end point data 3 weeks later, that is, 6 weeks after myocardial infarction. Implanting the 3DFC increases (p<0.05) myocardial blood flow twofold, microvessel formation (0.02±0.01 vs. 0.07±0.03 vessels/μm2), and ventricular wall thickness (0.53±0.02 to 1.02±0.17mm). The 3DFC shifts the passive pressure volume loop toward the pressure axis but does not alter left ventricular (LV) ejection fraction, systolic displacement, LV end-diastolic pressure/dimension, or LV cavity area. The 3DFC stimulates selected cytokine activation with a decrease in the proinflammatory cascade and increased total protein content stimulated by strained 3DFC in vitro. The 3DFC functions as a cell delivery device providing matrix support for resident cell survival and integration into the heart. The imbedded fibroblasts of the 3DFC release a complex blend of cardioactive cytokines promoting increases in microvessel density and anterior wall blood flow but does not improve ejection fraction or alter LV remodeling.


Capillary supply of the soft palate muscles is reduced in long-term habitual snorers.
Snoring and obstructive sleep apnea (OSA) cause vibration and stretch of the upper airway tissues that may result in neuromuscular damage and changes in the microcirculation. The aim of this investigation was to test whether long-term snoring affects capillary supply in soft palate muscles. Samples from the palatopharyngeus (PP) and uvula (UV) muscles were collected from 8 patients undergoing uvulo-palatopharyngoplasty because of habitual snoring and OSA. Control samples were obtained at autopsy. The muscles were analyzed using immunohistochemistry and morphometry. The patients' palate muscles had a lower capillary density (PP 443 vs. 711 capillaries/mm(2), p < 0.001, and UV 452 vs. 624 capillaries/mm(2), p = 0.009), a lower number of capillaries related to an individual muscle fiber (PP 1.3 vs. 2.7, p = 0.003, and UV 1.0 vs. 1.9, p = 0.03) and a lower number of capillaries related to the fiber size (PP 0.9 vs. 2.1, p = 0.001, and UV 0.6 vs. 1.9, p = 0.002). Our results indicate that reduced capillary supply of palate muscles plays a pathophysiological role in long-term snorers and OSA. The cause of the low capillary supply is unclear, but neuromuscular injury due to repeated vibratory and stretch trauma of the soft palate during snoring is a plausible mechanism.


Cerebral blood flow distribution during exposure to 5 bar oxygen in awake rats.
The regional cerebral blood flow (rCBF) and cardiac output (CO) were measured in conscious rats by the microsphere method during control, after 5 and 60 min at 5 bar O2, and 5 min after decompression to air. The arterial acid-base balance was essentially unchanged during hyperbaric O2 and after decompression, except for a slightly reduced CO2 and HCO3 during the O2 exposure. The heart rate (HR) fell at 1 bar O2, continued to fall during compression, and remained low. A marked HR rise occurred in air after decompression. The systolic arterial pressure (AP) increased, while mean AP was constant during the O2 exposure. The CO and total cerebral blood flow fell in proportion to the arterial O2 content increase. The rCBF was unevenly distributed in control, and fell to a disparate degree and remained low in some regions during O2 exposure. Due to the rCBF fall, the O2 supply was limited, the glucose supply was reduced, and CO2 and heat transport probably were limited, suggesting a labile metabolic state locally in the brain. After decompression, blood flow remained low in several regions, making hypoxia likely for a considerable time in several brain areas, whereas the rest of the brain had normalized or increased blood flow. The HR and systolic AP remained high for at least 30 min after decompression.


Maternal consumption of coffee and tea during pregnancy and risk of childhood ALL: a pooled analysis from the childhood Leukemia International Consortium.
The early onset of childhood acute lymphoblastic leukemia (ALL) suggests that critical exposures occurring during pregnancy may increase risk. We investigated the effects of maternal coffee and tea consumption during pregnancy on ALL risk by pooling data from eight case-control studies participating in the Childhood Leukemia International Consortium. Data on maternal coffee intake were available for 2,552 cases and 4,876 controls, and data on tea intake were available for 2,982 cases and 5,367 controls. Coffee and tea intake was categorized into 0, > 0-1, > 1-2, and > 2 cups/day, and covariates were combined and harmonized. Data on genetic variants in NAT2, CYP1A1, and NQO1 were also available in a subset. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using unconditional logistic regression, and linear trends across categories were assessed. No association was seen with 'any' maternal coffee consumption during pregnancy, but there was evidence of a positive exposure-response; the pooled OR for > 2 cups/day versus none was 1.27 (95% CI 1.09-1.43), p trend = 0.005. No associations were observed with tea consumption. No interactions were seen between coffee or tea intake and age, maternal smoking or genotype, and there was little or no evidence that associations with coffee or tea differed among cases with and without chromosomal translocations. Despite some limitations, our findings suggest that high coffee intake during pregnancy may increase risk of childhood ALL. Thus, current advice to limit caffeine intake during pregnancy to reduce risk of preterm birth may have additional benefits.


Evaluation of the PTW microDiamond in edge-on orientation for dosimetry in small fields.
The PTW microDiamond has an enhanced spatial resolution when operated in an edge-on orientation but is not typically utilized in this orientation due to the specifications of the IAEA TRS-483 code of practice for small field dosimetry. In this work the suitability of an edge-on orientation and advantages over the recommended face-on orientation will be presented. The PTW microDiamond in both orientations was compared on a Varian TrueBeam linac for: machine output factor (OF), percentage depth dose (PDD), and beam profile measurements from 10 × 10 cm2 to a 0.5 × 0.5 cm2 field size for 6X and 6FFF beam energies in a water tank. A quantification of the stem effect was performed in edge-on orientation along with tissue to phantom ratio (TPR) measurements. An extensive angular dependence study for the two orientations was also undertaken within two custom PMMA plastic cylindrical phantoms. The OF of the PTW microDiamond in both orientations agrees within 1% down to the 2 × 2 cm2 field size. The edge-on orientation overresponds in the build-up region but provides improved penumbra and has a maximum observed stem effect of 1%. In the edge-on orientation there is an angular independent response with a maximum of 2% variation down to a 2 × 2 cm2 field. The PTW microDiamond in edge-on orientation for TPR measurements agreed to the CC01 ionization chamber within 1% for all field sizes. The microDiamond was shown to be suitable for small field dosimetry when operated in edge-on orientation. When edge-on, a significantly reduced angular dependence is observed with no significant stem effect, making it a more versatile QA instrument for rotational delivery techniques.


Comparison of intravenous anesthetic induction doses and physiologic effects of ketamine or alfaxalone in goats undergoing surgery with isoflurane anesthesia.
To compare IV doses of alfaxalone and ketamine needed to facilitate orotracheal intubation and assess effects of each treatment on selected physiologic variables in goats undergoing orthopedic surgery with isoflurane anesthesia. 18 healthy adult goats. Behavior was assessed before and after sedation with midazolam (0.1 mg/kg, IV) for IV catheter placement. Anesthesia was induced with additional midazolam (0.1 mg/kg, IV) and alfaxalone (n = 9) or ketamine (9) at 2 mg/kg, IV, over 30 seconds. An additional dose of alfaxalone or ketamine (1 mg/kg) was given IV if needed for intubation; anesthesia was maintained with isoflurane in oxygen and IV fluids with ketamine (0.5 to 1 mg/kg/h). Direct systolic (SAP), diastolic (DAP), and mean (MAP) arterial blood pressures; heart rate; and respiratory rate were recorded before induction, immediately after intubation, and during surgery. Qualitative anesthetic induction and recovery characteristics were assessed. Variables were compared within and between groups by statistical methods. No preinduction variables differed significantly between groups. Postintubation and 30-minute intraoperative SAP, DAP, and MAP were higher for the ketamine group than for the alfaxalone group; within the alfaxalone group, postintubation SAP, MAP, and respiratory rate prior to mechanical ventilation were lower than respective preinduction values. All alfaxalone-group goats were intubated after 1 dose of the induction agent; 5 of 9 ketamine-group goats required an additional (1-mg/kg) dose. Postoperative recovery was good to excellent for all animals. Both drugs were suitable for induction of anesthesia after sedation with midazolam, but most goats required higher doses of ketamine to allow intubation. For situations in which alfaxalone administration is appropriate, the potential for decreased arterial blood pressures and respiratory rate should be considered.


Activation of brain macrophages/microglia cells in hepatitis C infection.
Hepatitis C virus (HCV) infection is commonly associated with cognitive dysfunction. Viral sequences and proteins were previously found in brain macrophage/microglia cells. The aim of the current study was to determine whether HCV infection affects the expression of key cytokines and chemokines in these cells. Autopsy brain tissue from 15 patients was studied; 7 patients were HCV positive and 8 were HCV negative. Cryostat sections of frontal cortex and subcortical white matter were stained with monoclonal antibodies specific for microglia/macrophages (anti-CD68) and separated by laser capture microscopy. Transcripts representing 25 various cytokines and chemokines were measured by real-time quantitative PCR. Compared with HCV-negative controls, HCV-positive patients demonstrated significantly higher levels of proinflammatory cytokines interleukin 1α (IL-1α), IL-1β, tumour necrosis factor α (TNFα), IL-12 and IL-18. HCV infection was also associated with increased transcription of chemokines IL-8, IL-16 and interferon-inducible protein 10 (IP-10). Type 1 interferon (IFN) activation was suggested by increased concentrations of IFNβ and myxovirus resistance protein A (MxA) transcripts. Similar results were obtained when CD68-positive/HCV-positive cells were compared with CD68-positive/HCV-negative cells in each of the 7 HCV-infected patients. Evidence was found for activation of brain macrophages/microglia cells in autopsy brain tissue from HCV-positive patients. These findings could relate to the common presence of neurocognitive dysfunction among patients with chronic hepatitis C.


Synthesis of mitochondrial DNA in permeabilised human cultured cells.
The mechanisms that underlie the maintenance of and increase in mutant mitochondrial DNA (mtDNA) are central to our understanding of mitochondrial disease. We have therefore developed a technique based on saponin permeabilisation that allows the study of mtDNA synthesis in intact cells. Permeabilisation of cells has been extensively used in an established method both for studying transcription and DNA replication in the nucleus and for measuring respiratory chain activities in mitochondria. We have quantitatively studied incorporation of radiolabelled DNA precursors into mtDNA in human cell lines derived from controls and from patients with mitochondrial DNA disease. Total cell DNA is extracted, restriction digested and Southern blotted, newly synthesised mtDNA being proportional to the label incorporated in each restriction band. A rate of synthesis can then be derived by estimating the relative steady-state mtDNA after probing with full-length mtDNA. Where co-existing mutant and wild-type mtDNA (heteroplasmy) can be distinguished using restriction digestion, their rates of synthesis can be compared within a single cell line. This will be particularly useful in elucidating the pathophysiology of mtDNA diseases in which the distribution of mutant and wild-type mtDNA in cell lines in patient tissues may evolve with time.


Autism and epilepsy: A population-based nationwide cohort study.
To investigate the risk of autism spectrum disorder (ASD) in individuals with epilepsy and in their first-degree relatives to determine shared etiology. Through the Swedish Patient Register, we identified 85,201 individuals with epilepsy, as well as all their siblings (n = 80,511) and offspring (n = 98,534). Each individual with epilepsy was compared with 5 controls, matched for age, sex, calendar period, and county, while siblings and offspring were compared with siblings and offspring of controls. We excluded siblings and offspring with epilepsy. Using Cox regression, we calculated hazard ratios (HRs) for future diagnosis of ASD. Logistic regression was applied to calculate odds ratios (ORs) for prior diagnosis of ASD. During follow-up, 1,381 (1.6%) individuals with epilepsy and 700 (0.2%) controls were diagnosed with ASD. Individuals with epilepsy were therefore at increased risk of future ASD (HR 10.49, 95% confidence interval [CI] 9.55-11.53), with the highest risk seen in individuals diagnosed with epilepsy in childhood. Both siblings (HR 1.62, 95% CI 1.43-1.83) and offspring (HR 1.64, 95% CI 1.46-1.84) of epilepsy patients were at increased risk of ASD. The risk in the offspring was particularly high in mothers with epilepsy (HR 1.91; 95% CI 1.63-2.23). Epilepsy was also associated with a prior diagnosis of ASD (OR 4.56, 95% CI 4.02-5.18). Individuals with epilepsy are at increased risk of ASD, especially if epilepsy appears in childhood. Further, ASD is more common in the siblings and offspring of individuals with epilepsy, suggesting shared etiology.


Honey - a potential agent against Porphyromonas gingivalis: an in vitro study.
Honey has been discussed as a therapeutic option in wound healing since ancient time. It might be also an alternative to the commonly used antimicrobials in periodontitis treatment. The in-vitro study was aimed to determine the antimicrobial efficacy against Porphyromonas gingivalis as a major periodontopathogen. One Manuka and one domestic beekeeper honey have been selected for the study. As a screening, MICs of the honeys against 20 P. gingivalis strains were determined. Contents of methylglyoxal and hydrogen peroxide as the potential antimicrobial compounds were determined. These components (up to 100 mg/l), propolis (up to 200 mg/l) as well as the two honeys (up to 10% w/v) were tested against four P. gingivalis strains in planktonic growth and in a single-species biofilm. 2% of Manuka honey inhibited the growth of 50% of the planktonic P. gingivalis, the respective MIC50 of the German beekeeper honey was 5%. Manuka honey contained 1.87 mg/kg hydrogen peroxide and the domestic honey 3.74 mg/kg. The amount of methylglyoxal was found to be 2 mg/kg in the domestic honey and 982 mg/kg in the Manuka honey. MICs for hydrogen peroxide were 10 mg/l - 100 mg/l, for methylglyoxal 5 - 20 mg/l, and for propolis 20 mg/l - 200 mg/l. 10% of both types of honey inhibited the formation of P. gingivalis biofilms and reduced the numbers of viable bacteria within 42 h-old biofilms. Neither a total prevention of biofilm formation nor a complete eradication of a 42 h-old biofilm by any of the tested compounds and the honeys were found. Honey acts antibacterial against P. gingivalis. The observed pronounced effects of Manuka honey against planktonic bacteria but not within biofilm can be attributed to methylglyoxal as the characteristic antimicrobial component.


Fate of Salmonella enterica in a mixed ingredient salad containing lettuce, cheddar cheese, and cooked chicken meat.
Food service and retail sectors offer consumers a variety of mixed ingredient salads that contain fresh-cut vegetables and other ingredients such as fruits, nuts, cereals, dairy products, cooked seafood, cooked meat, cured meats, or dairy products obtained from external suppliers. Little is known about the behavior of enteric bacterial pathogens in mixed ingredient salads. A model system was developed to examine the fate of Salmonella enterica (inoculum consisting of S. enterica serovars Agona, Typhimurium, Enteritidis, Brandenberg, and Kentucky) on the surface of romaine lettuce tissues incubated alone and in direct contact with Cheddar cheese or cooked chicken. S. enterica survived but did not grow on lettuce tissues incubated alone or in contact with Cheddar cheese for 6 days at either 6 or 14°C. In contrast, populations increased from 2.01 ± 0.22 to 9.26 ± 0.22 CFU/cm(2) when lettuce washed in water was incubated in contact with cooked chicken at 14°C. Populations on lettuce leaves were reduced to 1.28 ± 0.14 CFU/cm(2) by washing with a chlorine solution (70 ppm of free chlorine) but increased to 8.45 ± 0.22 CFU/cm(2) after 6 days at 14°C. Experimentation with a commercial product in which one third of the fresh-cut romaine lettuce was replaced with inoculated lettuce revealed that S. enterica populations increased by 4 log CFU/g during storage for 3 days at 14°C. These findings indicate that rapid growth of bacterial enteric pathogens may occur in mixed ingredient salads; therefore, strict temperature control during the manufacture, distribution, handling, and storage of these products is critical.


Effect of alpha-tocopherol and beta-carotene supplementation on macrovascular complications and total mortality from diabetes: results of the ATBC Study.
To determine whether alpha-tocopherol or beta-carotene supplementation affects diabetic macrovascular complications and total mortality. This study was carried out as part of the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study, a double-blind, randomized trial with a 2x2 factorial design. A total of 29,133 middle-aged male smokers received either vitamin E 50 mg/day or beta-carotene 20 mg/day, or both, or placebo for a median of 6.1 years. At base-line, 1700 men had type 2 diabetes. Of these men, 662 were diagnosed with first-ever macrovascular complication, and 1142 died during the 19-year follow-up. Neither supplementation affected the risk of macrovascular complication or total mortality during the intervention period. For the alpha-tocopherol-supplemented versus no alpha-tocopherol-supplemented, and beta-carotene-supplemented versus no beta-carotene-supplemented we found relative risk (RR) 0.84 (95% confidence interval (CI) 0.65-1.10) and RR 1.15 (95% CI 0.89-1.50) for macrovascular complication, respectively, and RR 1.00 (95% CI 0.80-1.25) and RR 1.06 (95% CI 0.85-1.33) for total mortality, respectively. No essential changes were found in these effects when the follow-up was extended up to 19 years. Alpha-tocopherol or beta-carotene supplementation has no protective effect on macrovascular outcomes or total mortality of diabetic male smokers.


Stillbirth: fetal disorders.
Fetal disorders (including congenital malformations) are among the most frequent of causes of intrauterine death. Assessment to detect fetal processes is straightforward: history (prenatal, perinatal, and family); external clinical examination; photographs; whole body radiographs; cytogenetic investigation; and internal necropsy. Unfortunately such investigations are profoundly underused. When appropriately and nonselectively assessed, about one-fifth of all stillborn infants will be found to have a fetal cause of their death. Fetal diagnoses are markedly heterogeneous, with no single process accounting for more than 1.5% of all fetal deaths. Identifying a fetal cause has marked implications for the family and changes some elements of counseling and care in more than half of those whose stillborn infants were adequately evaluated. Currently unmet needs include research needs including efforts to understand the variability of intrauterine effects of fetal disorders and their causes, and clinical needs, particularly related to funding for evaluation of stillborns and development of geographically dispersed multidisciplinary commitment and expertise devoted to stillbirth evaluation.


On the optimal reconstruction and control of adaptive optical systems with mirror dynamics.
In adaptive optics (AO) the deformable mirror (DM) dynamics are usually neglected because, in general, the DM can be considered infinitely fast. Such assumption may no longer apply for the upcoming Extremely Large Telescopes (ELTs) with DM that are several meters in diameter with slow and/or resonant responses. For such systems an important challenge is to design an optimal regulator minimizing the variance of the residual phase. In this contribution, the general optimal minimum-variance (MV) solution to the full dynamical reconstruction and control problem of AO systems (AOSs) is established. It can be looked upon as the parent solution from which simpler (used hitherto) suboptimal solutions can be derived as special cases. These include either partial DM-dynamics-free solutions or solutions derived from the static minimum-variance reconstruction (where both atmospheric disturbance and DM dynamics are neglected altogether). Based on a continuous stochastic model of the disturbance, a state-space approach is developed that yields a fully optimal MV solution in the form of a discrete-time linear-quadratic-Gaussian (LQG) regulator design. From this LQG standpoint, the control-oriented state-space model allows one to (1) derive the optimal state-feedback linear regulator and (2) evaluate the performance of both the optimal and the sub-optimal solutions. Performance results are given for weakly damped second-order oscillatory DMs with large-amplitude resonant responses, in conditions representative of an ELT AO system. The highly energetic optical disturbance caused on the tip/tilt (TT) modes by the wind buffeting is considered. Results show that resonant responses are correctly handled with the MV regulator developed here. The use of sub-optimal regulators results in prohibitive performance losses in terms of residual variance; in addition, the closed-loop system may become unstable for resonant frequencies in the range of interest.


Advax augments B and T cell responses upon influenza vaccination via the respiratory tract and enables complete protection of mice against lethal influenza virus challenge.
Administration of influenza vaccines via the respiratory tract has potential benefits over conventional parenteral administration, inducing immunity directly at the site of influenza exposure as well as being needle free. In this study, we investigated the suitability of Advax™, a stable particulate polymorph of inulin, also referred to as delta inulin, as a mucosal adjuvant for whole inactivated influenza vaccine (WIV) administered either as a liquid or dry powder formulation. Spray freeze-drying produced Advax-adjuvanted WIV powder particles in a size range (1-5 μm) suitable for inhalation. The physical and biological characteristics of both WIV and Advax remained unaltered both by admixing WIV with Advax and by spray freeze drying. Upon intranasal or pulmonary immunization, both liquid and dry powder formulations containing Advax induced significantly higher systemic, mucosal and cellular immune responses than non-adjuvanted WIV formulations. Furthermore, pulmonary immunization with Advax-adjuvanted WIV led to robust memory B cell responses along with an increase of lung localization factors i.e. CXCR3, CD69, and CD103. A less pronounced but still positive effect of Advax was seen on memory T cell responses. In contrast to animals immunized with WIV alone, all animals pulmonary immunized with a single dose of Advax-adjuvanted WIV were fully protected with no visible clinical symptoms against a lethal dose of influenza virus. These data confirm that Advax is a potent mucosal adjuvant that boosts vaccine-induced humoral and cellular immune responses both in the lung and systemically with major positive effects on B-cell memory and complete protection against live virus. Hence, respiratory tract immunization, particularly via the lungs, with Advax-adjuvanted WIV formulation as a liquid or dry powder is a promising alternative to parenteral influenza vaccination.


Personalized exposure assessment: promising approaches for human environmental health research.
New technologies and methods for assessing human exposure to chemicals, dietary and lifestyle factors, infectious agents, and other stressors provide an opportunity to extend the range of human health investigations and advance our understanding of the relationship between environmental exposure and disease. An ad hoc Committee on Environmental Exposure Technology Development was convened to identify new technologies and methods for deriving personalized exposure measurements for application to environmental health studies. The committee identified a "toolbox" of methods for measuring external (environmental) and internal (biologic) exposure and assessing human behaviors that influence the likelihood of exposure to environmental agents. The methods use environmental sensors, geographic information systems, biologic sensors, toxicogenomics, and body burden (biologic) measurements. We discuss each of the methods in relation to current use in human health research; specific gaps in the development, validation, and application of the methods are highlighted. We also present a conceptual framework for moving these technologies into use and acceptance by the scientific community. The framework focuses on understanding complex human diseases using an integrated approach to exposure assessment to define particular exposure-disease relationships and the interaction of genetic and environmental factors in disease occurrence. Improved methods for exposure assessment will result in better means of monitoring and targeting intervention and prevention programs.


Global molecular effects of tocilizumab therapy in rheumatoid arthritis synovium.
To investigate the global molecular effects of tocilizumab (TCZ) in comparison with methotrexate (MTX) treatment in synovial biopsy tissue obtained from patients with previously untreated rheumatoid arthritis (RA) before therapy (T0) and 12 weeks after the initiation of therapy (T12), and to compare the results with previous gene expression data obtained in synovial biopsy tissue from adalimumab (ADA)- and rituximab (RTX)-treated patients with RA. Paired synovial biopsy samples were obtained at T0 and T12 from the affected knee of TCZ-treated RA patients and MTX-treated RA patients. Gene expression studies were performed using GeneChip Human Genome U133 Plus 2.0 microarrays, and confirmatory quantitative real-time reverse transcription-polymerase chain reaction experiments were performed on selected transcripts. The effects of TCZ and MTX on synovial cell populations and histologic characteristics were assessed by immunohistochemistry. Gene expression studies showed that blockade of the interleukin-6 receptor (IL-6R) gene (IL6R) using TCZ induced a significant decrease in the expression of numerous chemokine and T cell activation genes in the RA synovium. These effects strongly correlated with the molecular effects of MTX and RTX therapy on RA synovial tissue, but differed from the molecular changes induced by ADA (decreased expression of genes involved in cell proliferation). The molecular similarities between the effects of TCZ, RTX, and MTX therapies in the RA synovium indicate that B cell- and IL-6-dependent pathways play synergistic roles in the pathogenesis of the disease, in particular through activation of T cell responses. Moreover, these results open perspectives for the individualization of therapeutic decisions, based on a better knowledge of the synovial molecular effects of each type of RA therapy.


Diisopropylammonium bromide is a high-temperature molecular ferroelectric crystal.
Molecular ferroelectrics are highly desirable for their easy and environmentally friendly processing, light weight, and mechanical flexibility. We found that diisopropylammonium bromide (DIPAB), a molecular crystal processed from aqueous solution, is a ferroelectric with a spontaneous polarization of 23 microcoulombs per square centimeter [close to that of barium titanate (BTO)], high Curie temperature of 426 kelvin (above that of BTO), large dielectric constant, and low dielectric loss. DIPAB exhibits good piezoelectric response and well-defined ferroelectric domains. These attributes make it a molecular alternative to perovskite ferroelectrics and ferroelectric polymers in sensing, actuation, data storage, electro-optics, and molecular or flexible electronics.


A new nerve block procedure for the suprascapular nerve based on a cadaveric study.
Shoulder pain after rotator cuff repair surgery is sometimes very severe. Suprascapular nerve block (SSNB) is a method used to relieve this shoulder pain, but is not always completely effective. The purpose of this study is to develop the new effective suprascapular nerve block procedure. To obtain an effective SSNB, we investigated an anatomic lesion of the sensory branches of the SSN in 8 cadavers and developed a new procedure. We evaluated its effectiveness by recording visual analog scale (VAS) scores in 8 patients experiencing severe pain after rotator cuff repair surgery before SSNB and for up to 24 hours postoperatively. All cadaver shoulders had 2 or 3 sensory branches that just passed the scapular notch. The branches originated from the SSN, pursued the base of the coracoid process, perforated the supraspinatus muscle, and extended toward the subacromial bursa. The average VAS score of the 8 patients postoperatively was 5.4 +/- 2.7 before SSNB and improved to 2.8 +/- 1.6 immediately after SSNB, to 1.2 +/- 0.6 at 1 hour, and to 0.8 +/- 0.8 at 4 hours. Effective pain control was achieved, and SSNB presented no complications. Our SSNB method is safe and effective for pain control in patients after open cuff repair. We expect it will be a useful management tool for pain relief after rotator cuff repair in the future. Basic science study.


Completing the Pain Circuit: Recent Advances in Imaging Pain and Inflammation beyond the Central Nervous System.
This review describes some of the recent developments in imaging aspects of pain in the periphery. It is now possible to image nerves in the cornea non-invasively, to image receptor level expression and inflammatory processes in injured tissue, to image nerves and alterations in nerve properties, to image astrocyte and glial roles in neuroinflammatory processes, and to image pain conduction functionally in the trigeminal ganglion. These advances will ultimately allow us to describe the pain pathway, from injury site to behavioral consequence, in a quantitative manner. Such a development could lead to diagnostics determining the source of pain (peripheral or central), objective monitoring of treatment progression, and, hopefully, objective biomarkers of pain.


Cognitive therapy versus exposure and applied relaxation in social phobia: A randomized controlled trial.
A new cognitive therapy (CT) program was compared with an established behavioral treatment. Sixty-two patients meeting Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994) criteria for social phobia were randomly assigned to CT, exposure plus applied relaxation (EXP = AR), or wait-list (WAIT). CT and EXP = AR were superior to WAIT on all measures. On measures of social phobia, CT led to greater improvement than did EXP = AR. Percentages of patients who no longer met diagnostic criteria for social phobia at posttreatment-wait were as follows: 84% in CT, 42% in EXP = AR, and 0% in WAIT. At the 1-year follow-up, differences in outcome persisted. In addition, patients in EXP = AR were more likely to have sought additional treatment. Therapist effects were small and nonsignificant. CT appears to be superior to EXP = AR in the treatment of social phobia.


FAT/CD36 is localized in sarcolemma and in vesicle-like structures in subsarcolemma regions but not in mitochondria.
The primary aim of the present study was to investigate in which cellular compartments fatty acid trans-locase CD36 (FAT/CD36) is localized. Intact and fully functional skeletal muscle mitochondria were isolated from lean and obese female Zucker rats and from 10 healthy male individuals. FAT/CD36 could not be detected in the isolated mitochondria, whereas the mitochondrial marker F(1)ATPase-beta was clearly detected using immunoblotting. Lack of markers for other membrane structures indicated that the mitochondria were not contaminated with membranes known to contain FAT/CD36. In addition, fluorescence immunocytochemistry was performed on single muscle fibers dissected from soleus muscle of lean and obese Zucker rats and from the vastus lateralis muscle from humans. Costaining against FAT/CD36 and MitoNEET clearly show that FAT/CD36 is highly present in sarcolemma and it also associates with some vesicle-like intracellular compartments. However, FAT/CD36 protein was not detected in mitochondrial membranes, supporting the biochemical findings. Based on the presented data, FAT/CD36 seems to be abundantly expressed in sarcolemma and in vesicle-like structures throughout the muscle cell. However, FAT/CD36 is not present in mitochondria in rat or human skeletal muscle. Thus, the functional role of FAT/CD36 in lipid transport seems primarily to be allocated to the plasma membrane in skeletal muscle.


The effect of combining plant sterols, soy protein, viscous fibers, and almonds in treating hypercholesterolemia.
Reductions in low-density lipoprotein-cholesterol (LDL-C) result from diets containing almonds, or diets that are either low in saturated fat or high in viscous fibers, soy proteins, or plant sterols. We have therefore combined all of these interventions in a single diet (portfolio diet) to determine whether cholesterol reductions could be achieved of similar magnitude to those reported in recent statin trials which reduced cardiovascular events. Twenty-five hyperlipidemic subjects consumed either a portfolio diet (n=13), very low in saturated fat and high in plant sterols (1.2 g/1,000 kcal), soy protein (16.2 g/1,000 kcal), viscous fibers (8.3 g/1,000 kcal), and almonds (16.6 g/1,000 kcal), or a low-saturated fat diet (n=12) based on whole-wheat cereals and low-fat dairy foods. Fasting blood, blood pressure, and body weight were obtained at weeks 0, 2, and 4 of each phase. LDL-C was reduced by 12.1% +/- 2.4% (P<.001) on the low-fat diet and by 35.0% +/- 3.1% (P<.001) on the portfolio diet, which also reduced the ratio of LDL-C to high-density lipoprotein-cholesterol (HDL-C) significantly (30.0% +/- 3.5%; P<.001). The reductions in LDL-C and the LDL:HDL-C ratio were both significantly lower on the portfolio diet than on the control diet (P<.001 and P<.001, respectively). Mean weight loss was similar on test and control diets (1.0 kg and 0.9 kg, respectively). No difference was seen in blood pressure, HDL-C, serum triglycerides, lipoprotein(a) [Lp(a)], or homocysteine concentrations between diets. Combining a number of foods and food components in a single dietary portfolio may lower LDL-C similarly to statins and so increase the potential effectiveness of dietary therapy.


Disruption of hippocampus-regulated behavioural and cognitive processes by heterozygous constitutive deletion of SynGAP.
The brain-specific Ras/Rap-GTPase activating protein (SynGAP) is a prime candidate linking N-methyl-d-aspartate receptors to the regulation of the ERK/MAP kinase signalling cascade, suggested to be essential for experience-dependent synaptic plasticity. Here, we evaluated the behavioural phenotype of SynGAP heterozygous knockout mice (SG(+/-)), expressing roughly half the normal levels of SynGAP. In the cognitive domain, SG(+/-) mice demonstrated severe working and reference memory deficits in the radial arm maze task, a mild impairment early in the transfer test of the water maze task, and a deficiency in spontaneous alternation in an elevated T-maze. In the non-cognitive domain, SG(+/-) mice were hyperactive in the open field and appeared less anxious in the elevated plus maze test. In contrast, object recognition memory performance was not impaired in SG(+/-) mice. The reduction in SynGAP thus resulted in multiple behavioural traits suggestive of aberrant cognitive and non-cognitive processes normally mediated by the hippocampus. Immunohistochemical evaluation further revealed a significant reduction in calbindin-positive interneurons in the hippocampus and doublecortin-positive neurons in the dentate gyrus of adult SG(+/-) mice. Heterozygous constitutive deletion of SynGAP is therefore associated with notable behavioural as well as morphological phenotypes indicative of hippocampal dysfunction. Any suggestion of a possible causal link between them however remains a matter for further investigation.


Accurate rates of the complex mechanisms for growth and dissolution of minerals using a combination of rare-event theories.
Mineral growth and dissolution are often treated as occurring via a single reversible process that governs the rate of reaction. We show that multiple distinct intermediate states can occur during both growth and dissolution. Specifically, we used metadynamics, a method for efficiently exploring the free-energy landscape of a system, coupled to umbrella sampling and reactive flux calculations to examine the mechanism and rates of attachment and detachment of a barium ion onto a stepped barite (BaSO(4)) surface. The activation energies calculated for the rate-limiting reactions, which are different for attachment and detachment, precisely match those measured experimentally during both growth and dissolution. These results can potentially explain anomalous non-steady-state mineral reaction rates observed experimentally and will enable the design of more efficient growth inhibitors and facilitate an understanding of the effect of impurities.


On utilizing search methods to select subspace dimensions for kernel-based nonlinear subspace classifiers.
In Kernel-based Nonlinear Subspace (KNS) methods, the subspace dimensions have a strong influence on the performance of the subspace classifier. In order to get a high classification accuracy, a large dimension is generally required. However, if the chosen subspace dimension is too large, it leads to a low performance due to the overlapping of the resultant subspaces and, if it is too small, it increases the classification error due to the poor resulting approximation. The most common approach is of an ad hoc nature, which selects the dimensions based on the so-called cumulative proportion computed from the kernel matrix for each class. In this paper, we propose a new method of systematically and efficiently selecting optimal or near-optimal subspace dimensions for KNS classifiers using a search strategy and a heuristic function termed the Overlapping criterion. The rationale for this function has been motivated in the body of the paper. The task of selecting optimal subspace dimensions is reduced to finding the best ones from a given problem-domain solution space using this criterion as a heuristic function. Thus, the search space can be pruned to very efficiently find the best solution. Our experimental results demonstrate that the proposed mechanism selects the dimensions efficiently without sacrificing the classification accuracy.


Evaluation of Two Strategies for Community-Based Safety Monitoring during Seasonal Malaria Chemoprevention Campaigns in Senegal, Compared with the National Spontaneous Reporting System.
Seasonal malaria chemoprevention (SMC) using sulfadoxine-pyrimethamine plus amodiaquine has been introduced in 12 African countries. Additional strategies for safety monitoring are needed to supplement national systems of spontaneous reporting that are known to under represent the incidence of adverse reactions. This study aimed to determine if adverse event (AE) reporting could be improved using a smartphone application provided to village health workers, or by active follow-up using a symptom card provided to caregivers. Two strategies to improve reporting of AEs during SMC campaigns were evaluated, in comparison with the national system of spontaneous reporting, in 11 health post areas in Senegal. In each health post, an average of approximately 4000 children under 10 years of age received SMC treatment each month for 3 months during the 2015 malaria transmission season-a total of 134,000 treatments. In three health posts (serving approximately 14,000 children), caregivers were encouraged to report any adverse reactions to the nurse at the health post or to a community health worker (CHW) in their village, who had been trained to use a smartphone application to report the event (enhanced spontaneous reporting). In two health posts (approximately 10,000 children), active follow-up of children at home was organized after each SMC campaign to ask about AEs that caregivers had been asked to record on a symptom card (active surveillance). Six health posts (approximately 23,000 children) followed the national system of spontaneous reporting using the national reporting (yellow) form. Each AE report was assessed by a panel to determine likely association with SMC drugs. The incidence of reported AEs was 2.4, 30.6, and 21.6 per 1000 children treated per month, using the national system, enhanced spontaneous reporting, and active surveillance, respectively. The most commonly reported symptoms were vomiting, fever, and abdominal pain. The incidence of vomiting, known to be caused by amodiaquine, was similar using both innovative methods (10/1000 in the first month, decreasing to 2.5/1000 in the third month). Despite increased surveillance, no serious adverse drug reactions were detected. Training CHWs in each village and health facility staff to report AEs using a mobile phone application led to much higher reporting rates than through the national system. This approach is feasible and acceptable, and could be further improved by strengthening laboratory investigation and the collection of control data immediately prior to SMC campaigns.


Photodynamic action of proflavine on coliphage T3. 3. Damages to the deoxyribonucleic acid associated with Rxl and Rx2.
Ultracentrifugational studies with deoxyribonucleic acid (DNA) extracted from phage exposed to light in the presence of either 0.25 or 8.5 mug of proflavine per ml reveal that the lethal damage of Rx1 renders DNA alkali-labile, with lethality resulting from damage that occurs singly on either strand or simultaneously on both strands. Apparently nonlethal damages temporally associated with Rx1 consist of (i) heat and alkali-labile cross-links (which produce undenatured DNA that migrates at 45S) and (ii) heat-labile bonds. The formation of 45S material is a linear function of light dose, and the production of this material ceases when Rx2 appears at the higher dye concentration. No tendency to plateau is seen at 0.25 mug of dye per ml. The nature of the lethal damage of Rx2 could not be determined. Damages that were temporally associated with Rx2 at 8.5 mug of dye per ml were heat-labile, alkali-stable cross-links (undenatured DNA, 38 to 41S; alkali-denatured-reneutralized DNA, 85 to 89S) and some double-stranded breaks. No such changes were seen at 0.25 mug of dye per ml.


Impact of the Southern Oscillation Index, Temperature, and Precipitation on Eastern Equine Encephalitis Virus Activity in Florida.
Eastern equine encephalitis virus (EEEV), an Alphavirus from family Togaviridae, is a highly pathogenic arbovirus affecting the eastern United States, especially Florida. Effects of the Southern Oscillation Index (SOI), precipitation, and cooling degree days on EEEV horse case data in Florida from 2004 to 2018 were modeled using distributed lag nonlinear models (DLNMs). The analysis was conducted at statewide and regional scales. DLNMs were used to model potential delayed effects of the covariates on monthly counts of horse cases. Both models confirmed a seasonal trend in EEEV transmission and found that precipitation, cooling degree days, and the SOI were all predictors of monthly numbers of horse cases. EEEV activity in horses was associated with higher amounts of rainfall during the month of transmission at the statewide scale, as well as the prior 3 mo at the regional scale, fewer cooling degree days during the month of transmission and the preceding 3 mo and high SOI values during the month and the previous 2 mo, and SOI values in the prior 2 to 8 mo. Horse cases were lower during El Niño winters but higher during the following summer, while La Niña winters were associated with higher numbers of cases and fewer during the following summer. At the regional scale, extremely low levels of precipitation were associated with a suppression of EEEV cases for 3 mo. Given the periodicity and potential predictability of El Niño Southern Oscillation (ENSO) cycles, precipitation, and temperature, these results may provide a method for predicting EEEV risk potential in Florida.


Introduction to the article collection 'Translation in healthcare: ethical, legal, and social implications'.
New technologies are transforming and reconfiguring the boundaries between patients, research participants and consumers, between research and clinical practice, and between public and private domains. From personalised medicine to big data and social media, these platforms facilitate new kinds of interactions, challenge longstanding understandings of privacy and consent, and raise fundamental questions about how the translational patient pathway should be organised.This editorial introduces the cross-journal article collection "Translation in healthcare: ethical, legal, and social implications", briefly outlining the genesis of the collection in the 2015 Translation in healthcare conference in Oxford, UK and providing an introduction to the contemporary ethical challenges of translational research in biology and medicine accompanied by a summary of the papers included in this collection.


Intramyocardial bone marrow cell injection for chronic myocardial ischemia: a randomized controlled trial.
Previous studies have suggested that bone marrow cell injection may improve myocardial perfusion and left ventricular (LV) function in patients with chronic myocardial ischemia. To investigate the effect of intramyocardial bone marrow cell injection on myocardial perfusion and LV function in patients with chronic myocardial ischemia. Randomized, double-blind, placebo-controlled trial at a Netherlands university hospital, May 1, 2005-March 3, 2008 (6-month follow-up ended September 2008) of 50 patients with chronic myocardial ischemia (mean age [SD], 64 [8] years; 43 men). severe angina pectoris despite optimal medical therapy and myocardial ischemia. All patients were ineligible for conventional revascularization. Intramyocardial injection of 100 x 10(6) autologous bone marrow-derived mononuclear cells or placebo solution. Primarily, the summed stress score, a 17-segment score for stress myocardial perfusion assessed by Tc-99m tetrofosmin single-photon emission computed tomography (SPECT). Secondary included LV ejection fraction (LVEF), Canadian Cardiovascular Society (CCS) class, and Seattle Angina Questionnaire quality-of-life score (mean difference >5% considered clinically significant). After 3-month follow-up, the summed stress score (mean [SD]) improved from 23.5 (4.7) to 20.1 (4.6) (P < .001) in the bone marrow cell group, compared with a decrease from 24.8 (5.5) to 23.7 (5.4) (P = .004) in the placebo group. In the bone marrow cell-treated patients who underwent magnetic resonance imaging (MRI), a 3% absolute increase in LVEF was observed at 3 months (95% CI, 0.5% to 4.7%; n = 18), but the placebo group showed no improvement. CCS angina score improved significantly in the bone marrow cell group (6-month absolute difference, -0.79; 95% CI, -1.10 to -0.48; P < .001) compared with no significant improvement in the placebo group. Quality-of-life score increased from 56% (9%) to 64% (12%) at 3 months and 69% (12%) at 6 months in bone marrow cell-treated patients, compared with a smaller increase in the placebo group from 57% (11%) to 61% (14%) to 64% (17%). The improvements in CCS class and quality of life score were significantly greater in bone marrow cell-treated patients than in placebo-treated patients (P = .03 and P = .04, respectively). In this short-term study of patients with chronic myocardial ischemia refractory to medical treatment, intramyocardial bone marrow cell injection resulted in a statistically significant but modest improvement in myocardial perfusion compared with placebo. Further studies are required to assess long-term results and efficacy for mortality and morbidity. trialregister.nl Identifier: NTR400 and isrctn.org Identifier: ISRCTN58194927.


Developing and evaluating interventions to reduce inappropriate prescribing by general practitioners of antibiotics for upper respiratory tract infections: a randomised controlled trial to compare paper-based and web-based modelling experiments.
Much implementation research is focused on full-scale trials with little evidence of preceding modelling work. The Medical Research Council Framework for developing and evaluating complex interventions has argued for more and better theoretical and exploratory work prior to a trial as a means of improving intervention development. Intervention modelling experiments (IMEs) are a way of exploring and refining an intervention before moving to a full-scale trial. They do this by delivering key elements of the intervention in a simulation that approximates clinical practice by, for example, presenting general practitioners (GPs) with a clinical scenario about making a treatment decision. The current proposal will run a full, web-based IME involving 250 GPs that will advance the methodology of IMEs by directly comparing results with an earlier paper-based IME. Moreover, the web-based IME will evaluate an intervention that can be put into a full-scale trial that aims to reduce antibiotic prescribing for upper respiratory tract infections in primary care. The study will also include a trial of email versus postal invitations to participate. More effective behaviour change interventions are needed and this study will develop one such intervention and a system to model and test future interventions. This system will be applicable to any situation in the National Health Service where behaviour needs to be modified, including interventions aimed directly at the public. ClinicalTrials (NCT): NCT01206738.


[Investigation of aflatoxins,mycobiota,and toxigenic fungi during post-harvest handling of Ziziphi Spinosae Semen].
Ziziphi Spinosae Semen is one of the Chinese herbal medicine being susceptible to aflatoxins contamination. To investigate the sources of aflatoxins contamination and toxigenic fungi species on Ziziphi Spinosae Semen,32 samples were collected from multiple steps during the post-harvest processing in this study. Aflatoxins in these samples were determined by immunoaffinity column and HPLC coupled with post-column photochemical derivatization. The dilution-plate method was applied to the fungi isolation. The isolated fungi strains were identified by morphological characterization and molecular approaches. The results showed that aflatoxins were detected in 28 samples from every step during the processing of Ziziphi Spinosae Semen. Three samples were detected with aflatoxin B_1 and 2 samples with both aflatoxin B_1 and total aflatoxin exceeding the limit of Chinese Pharmacopoeia. Especially the samples from the washing step,with the highest detected amounts of AFB_1 and AFs were reached 94. 79,121. 43 μg·kg~(-1),respectively. All 32 samples were contaminated by fungi. The fungal counts on the newly harvested samples were 2. 20 × 10~2 CFU·g~(-1). Moreover,it increased as tphreocessing progresses,and achieved 1. 16×10~6 CFU·g~(-1) after washing. A total of 321 isolates were identified to 17 genera. Aspergillus flavus was the main source of aflatoxins during the processing and storage of Ziziphi Spinosae Semen. One isolate of A. flavus was confirmed producing AFB_1 and AFB_2. The fungal count was significantly increased by composting,and Aspergillus was the predominant genus after shell breaking. The contamination level of aflatoxins was increased by composting and washing.


Mismatch repair-dependent G2 checkpoint induced by low doses of SN1 type methylating agents requires the ATR kinase.
S(N)1-type alkylating agents represent an important class of chemotherapeutics, but the molecular mechanisms underlying their cytotoxicity are unknown. Thus, although these substances modify predominantly purine nitrogen atoms, their toxicity appears to result from the processing of O(6)-methylguanine ((6Me)G)-containing mispairs by the mismatch repair (MMR) system, because cells with defective MMR are highly resistant to killing by these agents. In an attempt to understand the role of the MMR system in the molecular transactions underlying the toxicity of alkylating agents, we studied the response of human MMR-proficient and MMR-deficient cells to low concentrations of the prototypic methylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). We now show that MNNG treatment induced a cell cycle arrest that was absolutely dependent on functional MMR. Unusually, the cells arrested only in the second G(2) phase after treatment. Downstream targets of both ATM (Ataxia telangiectasia mutated) and ATR (ATM and Rad3-related) kinases were modified, but only the ablation of ATR, or the inhibition of CHK1, attenuated the arrest. The checkpoint activation was accompanied by the formation of nuclear foci containing the signaling and repair proteins ATR, the S(*)/T(*)Q substrate, gamma-H2AX, and replication protein A (RPA). The persistence of these foci implied that they may represent sites of irreparable damage.


Search for Low-Mass Quark-Antiquark Resonances Produced in Association with a Photon at sqrt[s]=13 TeV.
A search for narrow low-mass resonances decaying to quark-antiquark pairs is presented. The search is based on proton-proton collision events collected at 13 TeV by the CMS detector at the CERN LHC. The data sample corresponds to an integrated luminosity of 35.9 fb^{-1}, recorded in 2016. The search considers the case where the resonance has high transverse momentum due to initial-state radiation of a hard photon. To study this process, the decay products of the resonance are reconstructed as a single large-radius jet with two-pronged substructure. The signal would be identified as a localized excess in the jet invariant mass spectrum. No evidence for such a resonance is observed in the mass range 10 to 125 GeV. Upper limits at the 95% confidence level are set on the coupling strength of resonances decaying to quark pairs. The results obtained with this photon trigger strategy provide the first direct constraints on quark-antiquark resonance masses below 50 GeV obtained at a hadron collider.


[The surgical treatment of gunshot wounds on the neck].
The author summarizes the original experience with the surgical treatment of gunshot wounds on the neck with reference to the specific approaches for the purpose dictated by the marked variability of the clinical course. An active surgical strategy is proposed depending on the character of tissue lesions, the form and direction of the wound canal.


Risk factors of lower limb cellulitis in a level-two healthcare facility in Cameroon: a case-control study.
Cellulitis is a common infection of the skin and subcutaneous tissues. It is associated with significant morbidity from necrosectomies and amputations especially in sub-Saharan Africa. We aimed at identifying the risk factors and burden of lower limb cellulitis to inform preventive strategies in Cameroon. This was a hospital-based case-control study carried out in the Bamenda Regional Hospital (BRH) between September 2015 and August 2016. Cases were defined as consenting adults admitted to the surgical unit who presented with a localised area of lower limb erythema, warmth, oedema and pain, associated with fever (temperature ≥ 38 °C) and/or chills of sudden onset. Controls were adults hospitalised for diseases other than cellulitis, necrotising fasciitis, myositis, abscess or other variants of dermo-hypodermitis. Cases and controls were matched (1:2) for age and sex. Of the 183 participants (61 cases of cellulitis and 122 controls) included in the study, the median age was 52 years [Interquartile range (IQR): 32.5-74.5]. After controlling for potential confounders, obesity [adjusted odds ratio (AOR) = 4.7, 95% CI (1.5-14.7); p = 0.009], history of skin disruption [AOR = 12.4 (3.9-39.1); p < 0.001], and presence of toe-web intertrigo [AOR = 51.4 (11.7-225.6); p < 0.001] were significantly associated with cellulitis. Median hospital stay was longer (14 days [IQR: 6-28]) in cases compared to the controls (3 days [IQR: 2-7]). Among the cases, Streptococci species were the most frequent (n = 50, 82%) isolated germ followed by staphylococci species (n = 9, 15%). Patients with cellulitis were more likely to undergo necrosectomy (OR: 21.2; 95% CI: 7.6-59.2). Toe-web intertrigo had the highest (48.9%) population attributable risk for cellulitis, followed by history of disruption of skin barrier (37.8%) and obesity (20.6%). This study showed a high disease burden among patients with cellulitis. While risk factors identified are similar to prior literature, this study provides a contextual evidence-base for clinicians in this region to be more aggressive in management of these risk factors to prevent disease progression and development of cellulitis.


Two dimensional Dirac carbon allotropes from graphene.
Using a structural search method in combination with first-principles calculations, we found lots of low energy 2D carbon allotropes and examined all possible Dirac points around their Fermi levels. Three amazing 2D Dirac carbon allotropes have been discovered, named as S-graphene, D-graphene and E-graphene. By analyzing the topology correlations among S-, T, net W graphene and graphene, we found that a general rule is valuable for constructing 2D carbon allotropes that are keen to possess Dirac cones in their electronic structures. Based on this rule, we have successfully designed many new 2D carbon allotropes possessing Dirac cones. Their energy order can be well described by an Ising-like model, and some allotropes are energetically more stable than those recently reported. The related electronic structures of these Dirac allotropes are anisotropy distinguished from those of graphene. Moreover, the fact that D- and E-graphene present Dirac cones suggests that sp hybridization or sp(3) hybridization could not suppress the emerging of Dirac features. Our results demonstrate that the Dirac cone and carrier linear dispersion is a very common feature in 2D carbon allotropes and can exist beyond the limitations of fundamental structure features of graphene.


Deaths and medical visits attributable to environmental pollution in the United Arab Emirates.
This study estimates the potential health gains achievable in the United Arab Emirates (UAE) with improved controls on environmental pollution. The UAE is an emerging economy in which population health risks have shifted rapidly from infectious diseases to chronic conditions observed in developed nations. The UAE government commissioned this work as part of an environmental health strategic planning project intended to address this shift in the nature of the country's disease burden. We assessed the burden of disease attributable to six environmental exposure routes outdoor air, indoor air, drinking water, coastal water, occupational environments, and climate change. For every exposure route, we integrated UAE environmental monitoring and public health data in a spatially resolved Monte Carlo simulation model to estimate the annual disease burden attributable to selected pollutants. The assessment included the entire UAE population (4.5 million for the year of analysis). The study found that outdoor air pollution was the leading contributor to mortality, with 651 attributable deaths (95% confidence interval [CI] 143-1,440), or 7.3% of all deaths. Indoor air pollution and occupational exposures were the second and third leading contributors to mortality, with 153 (95% CI 85-216) and 46 attributable deaths (95% CI 26-72), respectively. The leading contributor to health-care facility visits was drinking water pollution, to which 46,600 (95% CI 15,300-61,400) health-care facility visits were attributed (about 15% of the visits for all the diseases considered in this study). Major study limitations included (1) a lack of information needed to translate health-care facility visits to quality-adjusted-life-year estimates and (2) insufficient spatial coverage of environmental data. Based on international comparisons, the UAE's environmental disease burden is low for all factors except outdoor air pollution. From a public health perspective, reducing pollutant emissions to outdoor air should be a high priority for the UAE's environmental agencies.


Consequences, characteristics, and causes of mathematical learning disabilities and persistent low achievement in mathematics.
The goals of the review are threefold: (a) to highlight the educational and employment consequences of poorly developed mathematical competencies; (b) overview the characteristics of children with mathematical learning disability (MLD) and with persistently low achievement (LA) in mathematics; and (c) provide a primer on cognitive science research that is aimed at identifying the cognitive mechanisms underlying these learning disabilities and associated cognitive interventions. Literatures on the educational and economic consequences of poor mathematics achievement were reviewed and integrated with reviews of epidemiological, behavioral genetic, and cognitive science studies of poor mathematics achievement. Poor mathematical competencies are common among adults and result in employment difficulties and difficulties in many common day-to-day activities. Among students, ∼ 7% of children and adolescents have MLD and another 10% show persistent LA in mathematics, despite average abilities in most other areas. Children with MLD and their LA peers have deficits in understanding and representing numerical magnitude, difficulties retrieving basic arithmetic facts from long-term memory, and delays in learning mathematical procedures. These deficits and delays cannot be attributed to intelligence but are related to working memory deficits for children with MLD, but not LA children. These individuals have identifiable number and memory delays and deficits that seem to be specific to mathematics learning. Interventions that target these cognitive deficits are in development and preliminary results are promising.


Visual cues used by ball-rolling dung beetles for orientation.
Ball rolling by dung beetles is considered to be a derived behaviour that evolved under pressure for space, and from competitors at the dung pat. Straight-line orientation away from the pat using a celestial cue should be the most successful rolling strategy to move dung to an unknown burial site. We tested this hypothesis in the field and the laboratory by presenting five species of ball-rolling beetles with different orientation tasks, involving reaction to obstacles as well as to reflected sunlight and artificial light sources. Beetles were found to consistently orientate along a chosen route, usually in the direction of the sun. Beetles rolling dung balls successfully negotiated barriers and returned to the original path as did beetles falling from ramps, or rotated about a fixed point while rolling a ball. The sun was found to be the main orientation cue, which could be substituted by reflected or artificial light. However, beetles reoriented themselves less accurately in response to lights in the laboratory, than they did to the reflected sun in the field. It is probable that phototactic orientation using the sun, which is widespread amongst arthropods, has been incorporated in the straight-line foraging behaviour that has evolved in ball-rolling dung beetles.


Clinical training in self-regulation and practical pediatric hypnosis: what pediatricians want pediatricians to know.
A survey was taken of graduates of the Society for Developmental and Behavioral Pediatrics' (SDBP) Pediatric Hypnosis Course to describe the impact of the course on participants' practical knowledge of self-regulation and hypnosis, the acquisition through direct experience of a realistic understanding of hypnosis, their estimation of the value of the course to their practice of pediatrics, their use of techniques learned, what they would communicate to peers in pediatrics about training in this area, and their beliefs concerning barriers to professional interest in learning hypnotic techniques for primary care. Forty-eight percent (126) of course graduates responded to the survey. Sixty-five percent of these were physicians; almost all were pediatricians. The overwhelming positive response of these pediatricians to the value of the course (4.4 on a scale of 5), the utility of hypnosis, its ease of use, and its practicality for primary care were conveyed richly in survey comments. Sixty-three percent reported that their practice satisfaction increased as a result of this training, and 64% recommended a course like this to all general pediatricians. Course graduates were concerned that the widespread mythology and strong feelings surrounding the term "hypnosis" eclipsed its empirical validation and its practical uses and was a significant barrier to interest in learning hypnotic techniques; 75% thought that the term should be de-emphasized in continuing medical education communications.


Two distinct β-sheet structures in Italian-mutant amyloid-beta fibrils: a potential link to different clinical phenotypes.
Most Alzheimer's disease (AD) cases are late-onset and characterized by the aggregation and deposition of the amyloid-beta (Aβ) peptide in extracellular plaques in the brain. However, a few rare and hereditary Aβ mutations, such as the Italian Glu22-to-Lys (E22K) mutation, guarantee the development of early-onset familial AD. This type of AD is associated with a younger age at disease onset, increased β-amyloid accumulation, and Aβ deposition in cerebral blood vessel walls, giving rise to cerebral amyloid angiopathy (CAA). It remains largely unknown how the Italian mutation results in the clinical phenotype that is characteristic of CAA. We therefore investigated how this single point mutation may affect the aggregation of Aβ1-42 in vitro and structurally characterized the resulting fibrils using a biophysical approach. This paper reports that wild-type and Italian-mutant Aβ both form fibrils characterized by the cross-β architecture, but with distinct β-sheet organizations, resulting in differences in thioflavin T fluorescence and solvent accessibility. E22K Aβ1-42 oligomers and fibrils both display an antiparallel β-sheet structure, in comparison with the parallel β-sheet structure of wild-type fibrils, characteristic of most amyloid fibrils described in the literature. Moreover, we demonstrate structural plasticity for Italian-mutant Aβ fibrils in a pH-dependent manner, in terms of their underlying β-sheet arrangement. These findings are of interest in the ongoing debate that (1) antiparallel β-sheet structure might represent a signature for toxicity, which could explain the higher toxicity reported for the Italian mutant, and that (2) fibril polymorphism might underlie differences in disease pathology and clinical manifestation.


High serum D-lactate in patients on continuous ambulatory peritoneal dialysis.
As abnormally high serum D-lactate levels may cause neurological impairment, we determined whether patients undergoing continuous ambulatory peritoneal dialysis (CAPD) with lactate-containing fluids have increased serum D-lactate concentrations. D- and L-lactate concentrations were determined in peritoneal dialysis fluids and in serum from control subjects (n = 10), haemodialysis patients (n = 10), and CAPD patients (n = 30) before and after 1 h of dialysis. We found the median D-lactate concentration in Dianeal CAPD fluid to be 26 mM (range 19-27), whereas it was less than 0.5 mM in DPCA2 fluid. Control, haemodialysis, and CAPD (DPCA2) patient median serum D-lactate concentrations were below 0.07 mM. However, CAPD (Dianeal) patient serum D-lactate concentrations were 4-fold higher than controls (P < 0.0001), at 0.28 mM, an hour after instillation of D-lactate-containing fluid. Three patients, whose serum D-lactate averaged 0.59 mM, were found to have D-lactate concentrations at 0.22 mM after overnight cessation of dialysis. We conclude that CAPD with D-lactate-containing fluids raises serum D-lactate to abnormal levels.


Method for measuring microbial growth in rumen content.
Radioactive sodium sulfide was used to label the sulfide pool of rumen contents in vitro. Microbial protein synthesis was calculated from the size and rate of dilution of label in the sulfide pool, and from the radioactivity incorporated into protein together with a conversion factor specifying the nitrogen-sulfur ratio determined for microbial protein. The microbial cell yield, calculated on the basis of the adenosine triphosphate (ATP) available from fermentation, was 13 to 14 g (dry weight) per mole of ATP, which is in good agreement with the values obtained for pure cultures of bacteria. Calculation of microbial protein yield per kilogram of ration agreed fairly well with previous estimates for similar rations.


Tumor-derived exosomes in oncogenic reprogramming and cancer progression.
In multicellular organisms, effective communication between cells is a crucial part of cellular and tissue homeostasis. This communication mainly involves direct cell-cell contact as well as the secretion of molecules that bind to receptors at the recipient cells. However, a more recently characterized mode of intercellular communication-the release of membrane vesicles known as exosomes-has been the subject of increasing interest and intensive research over the past decade. Following the discovery of the exosome-mediated immune activation, the pathophysiological roles of exosomes have been recognized in different diseases, including cancer. In this review, we describe the biogenesis and main physical characteristics that define exosomes as a specific population of secreted vesicles, with a special focus on their role in oncogenic transformation and cancer progression.


The Alvarez and Lohmann refractive lenses revisited.
Alvarez and Lohmann lenses are variable focus optical devices based on lateral shifts of two lenses with cubic-type surfaces. I analyzed the optical performance of these types of lenses computing the first order optical properties (applying wavefront refraction and propagation) without the restriction of the thin lens approximation, and the spot diagram using a ray tracing algorithm. I proposed an analytic and numerical method to select the most optimum coefficients and the specific configuration of these lenses. The results show that Lohmann composite lens is slightly superior to Alvarez one because the overall thickness and optical aberrations are smaller.


The impact of eyewitness expert evidence and judicial instruction on juror ability to evaluate eyewitness testimony.
It has been argued that psychologists should provide expert evidence to help jurors discriminate between accurate and inaccurate eyewitness identifications. In this article we compare the effects of judicial instruction with expert evidence that is either congruent or incongruent with the ground truth, focusing on juror ability to evaluate "real" eyewitness evidence. In contrast to studies which have employed "fictional" eyewitness designs, we found no appreciable effect of either congruent or incongruent expert evidence on participant-juror sensitivity to eyewitness accuracy. We discuss the role of methodology on the inferences and conclusions that can be made regarding the impact of eyewitness expert evidence.


Validating a knowledge transfer framework in health services.
The study of knowledge transfer (KT) has been proceeding in parallel but independently in health services and in business, presenting an opportunity for synergy and sharing. This paper uses a survey of 32 empirical KT studies with their 96 uniquely named determinants of KT success to identify ten unique determinants for horizontal knowledge transfer success. These determinants, the outcome measure of Knowledge Use, and separate explicit and tacit transfer flows constitute the KT Framework, extending the work of previous KT framework authors. Our Framework was validated through a case study of the transfer of clinical practice guideline knowledge between the cardiac teams of selected Ontario hospitals, using a survey of senders and receivers developed from the KT literature. The study findings were: 8 of 10 determinants were supported by the Successful Transfer Hospitals; and 4 of 10 determinants were found to a higher degree in the Successful than non-Successful transfer hospitals. Taken together, the results show substantive support for the KT Framework determinants, indicating aggregate support of 9 of these determinants, but not the 10th - Knowledge Complexity. The transfer of tacit knowledge was found to be related to the transfer of the explicit knowledge and expressed as the transfer or recreation of resource profile and internal process tacit knowledge, where this tacit transfer did not require interactions between Sender and Receiver. This study provides managers with the building blocks to assess and improve the success rates of their knowledge transfers.


The role of aerobic fitness in cortical thickness and mathematics achievement in preadolescent children.
Growing evidence suggests that aerobic fitness benefits the brain and cognition during childhood. The present study is the first to explore cortical brain structure of higher fit and lower fit 9- and 10-year-old children, and how aerobic fitness and cortical thickness relate to academic achievement. We demonstrate that higher fit children (>70th percentile VO2max) showed decreased gray matter thickness in superior frontal cortex, superior temporal areas, and lateral occipital cortex, coupled with better mathematics achievement, compared to lower fit children (<30th percentile VO2max). Furthermore, cortical gray matter thinning in anterior and superior frontal areas was associated with superior arithmetic performance. Together, these data add to our knowledge of the biological markers of school achievement, particularly mathematics achievement, and raise the possibility that individual differences in aerobic fitness play an important role in cortical gray matter thinning during brain maturation. The establishment of predictors of academic performance is key to helping educators focus on interventions to maximize learning and success across the lifespan.


Complementary functions of the flippase ATP8B1 and the floppase ABCB4 in maintaining canalicular membrane integrity.
Progressive familial intrahepatic cholestasis can be caused by mutations in ABCB4 or ATP8B1; each encodes a protein that translocates phospholipids, but in opposite directions. ABCB4 flops phosphatidylcholine from the inner to the outer leaflet, where it is extracted by bile salts. ATP8B1, in complex with the accessory protein CDC50A, flips phosphatidylserine in the reverse direction. Abcb4(-/-) mice lack biliary secretion of phosphatidylcholine, whereas Atp8b1-deficient mice have increased excretion of phosphatidylserine into bile. Each system is thought to have a role protecting the canalicular membrane from bile salts. To investigate the relationship between the mechanisms of ABCB4 and ATP8B1, we expressed the transporters separately and together in cultured cells and studied viability and phospholipid transport. We also created mice with disruptions in ABCB4 and ATP8B1 (double knockouts) and studied bile formation and hepatic damage in mice fed bile salts. Overexpression of ABCB4 was toxic to HEK293T cells; the toxicity was counteracted by coexpression of the ATP8B1-CDC50A complex. In Atp8b1-deficient mice, bile salts induced extraction of phosphatidylserine and ectoenzymes from the canalicular membrane; this process was not observed in the double-knockout mice. ATP8B1 is required for hepatocyte function, particularly in the presence of ABCB4. This is most likely because the phosphatidylserine flippase complex of ATP8B1-CDC50A counteracts the destabilization of the membrane that occurs when ABCB4 flops phosphatidylcholine. Lipid asymmetry is therefore important for the integrity of the canalicular membrane; ABCB4 and ATP8B1 cooperate to protect hepatocytes from bile salts.