Development and Clinical Validation of Discriminatory Multitarget Digital Droplet PCR Assays for the Detection of Hot Spot KRAS and NRAS Mutations in Cell-Free DNA.
Detection and quantification of tumor-derived KRAS and NRAS mutations in plasma cell-free DNA (cfDNA) holds great potential for cancer diagnostics and treatment response monitoring. Because of high sensitivity, specificity, robustness, and affordability, digital droplet PCR (ddPCR) is ideally suited for this application but requires discriminatory multiplexing when used as screening assay. We therefore designed, optimized, and clinically validated mutation-specific locked nucleic acid-based ddPCR assays for 14 commonly occurring KRAS and NRAS mutations and assembled these assays into seven discriminatory multitarget screening assays covering two to six single-nucleotide variants each. Limit of detection, limit of blank, and interassay accuracy were determined. Assay performance and suitability for screening in cfDNA were validated with plasma samples from a clinically fully characterized cohort of pancreatic cancer patients and healthy controls. Limits of detection for single-target assays were between 0.0015% and 0.069% variant allele fraction, and between 0.022% and 0.16% for multitarget assays. Dilution linearity and interassay accuracy were excellent throughout (r2 > 0.99). Multitarget assay screening of cfDNA extracted from pancreatic cancer patients with unknown KRAS mutational status correctly identified single-nucleotide variants in 45 of 45 (100%) of tumor-derived cell-free DNA-positive samples. In summary, we herein present and clinically validate generic single-target and discriminatory multitarget ddPCR assays for KRAS and NRAS hot spot mutations with broad applicability for clinical and translational research.


A Protocol for Decellularizing Mouse Cochleae for Inner Ear Tissue Engineering.
In mammals, mechanosensory hair cells that facilitate hearing lack the ability to regenerate, which has limited treatments for hearing loss. Current regenerative medicine strategies have focused on transplanting stem cells or genetic manipulation of surrounding support cells in the inner ear to encourage replacement of damaged stem cells to correct hearing loss. Yet, the extracellular matrix (ECM) may play a vital role in inducing and maintaining function of hair cells, and has not been well investigated. Using the cochlear ECM as a scaffold to grow adult stem cells may provide unique insights into how the composition and architecture of the extracellular environment aids cells in sustaining hearing function. Here we present a method for isolating and decellularizing cochleae from mice to use as scaffolds accepting perfused adult stem cells. In the current protocol, cochleae are isolated from euthanized mice, decellularized, and decalcified. Afterward, human Wharton's jelly cells (hWJCs) that were isolated from the umbilical cord were carefully perfused into each cochlea. The cochleae were used as bioreactors, and cells were cultured for 30 days before undergoing processing for analysis. Decellularized cochleae retained identifiable extracellular structures, but did not reveal the presence of cells or noticeable fragments of DNA. Cells perfused into the cochlea invaded most of the interior and exterior of the cochlea and grew without incident over a duration of 30 days. Thus, the current method can be used to study how cochlear ECM affects cell development and behavior.


Protective efficacy of a second pneumococcal conjugate vaccine against pneumococcal acute otitis media in infants and children: randomized, controlled trial of a 7-valent pneumococcal polysaccharide-meningococcal outer membrane protein complex conjugate vaccine in 1666 children.
To assess the efficacy of a 7-valent pneumococcal polysaccharide-meningococcal outer membrane protein complex conjugate vaccine (PncOMPC) against acute otitis media (AOM), 1666 infants were randomly assigned to receive either PncOMPC or control vaccine (hepatitis B vaccine) at 2, 4, 6, and 12 months of age. Of the 835 children assigned to receive PncOMPC, 187 received a 23-valent pneumococcal polysaccharide vaccine (PncPS) at 12 months of age instead. Whenever AOM was diagnosed, middle ear fluid was aspirated for bacterial culture. In the PncOMPC and control groups, there were 110 and 250 AOM episodes, respectively, in children between 6.5 and 24 months of age that could be attributed to vaccine serotypes, which indicates a vaccine efficacy of 56% (95% confidence interval, 44%-66%). The serotype-specific efficacy ranged from 37% for 19F to 82% for 9V. The 2 boosters seemed to provide equal protection against AOM, but PncPS induced markedly higher antibody concentrations. The efficacy of PncOMPC was comparable to that of the recently licensed pneumococcal conjugate vaccine.


Cost analysis of an integrated aged care program for residential aged care facilities.
Objective To compare annual costs of an intervention for acutely unwell older residents in residential age care facilities (RACFs) with usual care. The intervention, the Aged Care Emergency (ACE) program, includes telephone clinical support aimed to reduce avoidable emergency department (ED) presentations by RACF residents. Methods This costing of the ACE intervention examines the perspective of service providers: RACFs, Hunter Medicare Local, the Ambulance Service of New South Wales, and EDs in the Hunter New England Local Health District. ACE was implemented in 69 RACFs in the Hunter region of NSW, Australia. Analysis used 14 weeks of ACE and ED service data (June-September 2014). The main outcome measure was the net cost and saving from ACE compared with usual care. It is based on the opportunity cost of implementing ACE and the opportunity savings of ED presentations avoided. Results Our analysis estimated that 981 avoided ED presentations could be attributed to ACE annually. Compared with usual care, ACE saved an estimated A$921214. Conclusions The ACE service supported a reduction in avoidable ED presentations and ambulance transfers among RACF residents. It generated a cost saving to health service providers, allowing reallocation of healthcare resources. What is known about the topic? Residents from RACFs are at risk of further deterioration when admitted to hospital, with high rates of delirium, falls, and medication errors. For this cohort, some conditions can be managed in the RACF without hospital transfer. By addressing avoidable presentations to EDs there is an opportunity to improve ED efficiency as well as providing care that is consistent with the resident's goals of care. RACFs generate some avoidable ED presentations for residents who may be more appropriately treated in situ. What does this paper add? Telephone triaging with nursing support and training is a means by which ED presentations from RACFs can be reduced. One of the consequences of this intervention is 'cost avoided', largely through savings on ambulance costs. What are the implications for practitioners? Unnecessary transfer from RACFs to ED can be avoided through a multicomponent program that includes telephone support with cost-saving implications for EDs and ambulance services.


Real-time MEG neurofeedback training of posterior alpha activity modulates subsequent visual detection performance.
It has been demonstrated that alpha activity is lateralized when attention is directed to the left or right visual hemifield. We investigated whether real-time neurofeedback training of the alpha lateralization enhances participants' ability to modulate posterior alpha lateralization and causes subsequent short-term changes in visual detection performance. The experiment consisted of three phases: (i) pre-training assessment, (ii) neurofeedback phase and (iii) post-training assessment. In the pre- and post-training phases we measured the threshold to covertly detect a cued faint Gabor stimulus presented in the left or right hemifield. During magnetoencephalography (MEG) neurofeedback, two face stimuli superimposed with noise were presented bilaterally. Participants were cued to attend to one of the hemifields. The transparency of the superimposed noise and thus the visibility of the stimuli were varied according to the momentary degree of hemispheric alpha lateralization. In a double-blind procedure half of the participants were provided with sham feedback. We found that hemispheric alpha lateralization increased with the neurofeedback training; this was mainly driven by an ipsilateral alpha increase. Surprisingly, comparing pre- to post-training, detection performance decreased for a Gabor stimulus presented in the hemifield that was un-attended during neurofeedback. This effect was not observed in the sham group. Thus, neurofeedback training alters alpha lateralization, which in turn decreases performances in the untrained hemifield. Our findings suggest that alpha oscillations play a causal role for the allocation of attention. Furthermore, our neurofeedback protocol serves to reduce the detection of unattended visual information and could therefore be of potential use for training to reduce distractibility in attention deficit patients, but also highlights that neurofeedback paradigms can have negative impact on behavioral performance and should be applied with caution.


Bone SPECT/CT has excellent sensitivity and specificity for diagnosis of loosening and patellofemoral problems after total knee arthroplasty.
The primary purpose of this retrospective study with prospectively collected data was to evaluate the clinical value of SPECT/CT, in particular its impact on clinical diagnosis and patient management, in patients with pain, stiffness, or swelling after primary total knee arthroplasty (TKA). The secondary purpose was to determine the sensitivity and the specificity of SPECT/CT for different pathologies such as loosening and patellofemoral overloading. A retrospective analysis was carried out on 214 knees in 202 consecutive patients (mean age 69 ± 11 years) with persistent or recurrent knee pain after TKA. All patients underwent clinical examination, standardised radiographs, and 99mTc-hydroxymethane diphosphonate (HDP) SPECT/CT as part of a routine diagnostic algorithm. The diagnoses before and after SPECT/CT and final treatment were recorded. TKA component position was routinely measured on 3D-reconstructed images. Intensity and anatomical distribution of bone tracer uptake were determined. SPECT/CT changed the clinical diagnosis and final treatment in 139/214 (65%) knees. In 117 (54.7%) out of 214 patients, revision surgery was proposed following SPECT/CT. Early onset of pain was significantly (p = 0.011) correlated with tibiofemoral stress pattern in SPECT/CT. A total of 86 knees (40.2%) were surgically revised using semi-constrained or fully constrained TKA. In 35 patients, a secondary patellar resurfacing was performed. SPECT/CT showed the clinical diagnosis to be incorrect for suspicion of tibial component loosening in 56 patients (26.2%), femoral component loosening in 53 patients (24.8%), and patellofemoral overloading/progressive patellofemoral osteoarthritis in 26 patients (12.1%) and provided different underlying causes of persistent knee pain after TKA. Likewise, SPECT/CT identified tibial component loosening in 8 patients (3.7%), femoral component loosening in 4 patients (1.9%), and patellofemoral OA in 71 patients (33.2%) without prior clinical suspected diagnosis. The sensitivity and specificity of SPECT/CT for detection of patellofemoral OA was 96.5% and 96.2%, respectively. The sensitivity and specificity for detection of tibial component loosening was 96.0% and 100%, respectively. The sensitivity and specificity for detection of femoral component loosening was 95.0% and 100%, respectively. The diagnostic benefits of SPECT/CT in the challenging and complex cohort of patients with pain after primary TKA have been proven. The excellent sensitivity and specificity for detection of tibial or femoral component loosening and diagnosis of patellofemoral OA have been surgically confirmed. Due to the benefits in establishing the correct diagnosis, SPECT/CT should be implemented as part of the routine diagnostic algorithm for patients with pain after primary TKA. II.


Dissolution difference between acidic and neutral media of acetaminophen tablets containing a super disintegrant and a soluble excipient. II.
The disintegration and dissolution of acetaminophen tablets containing sucrose and Ac-Di-Sol/Primojel was significantly different between acidic and neutral media. The purpose of this study was to investigate the mechanism of this phenomenon and to propose a way of reducing the dissolution difference between the two media. Tablets of different combinations of active ingredient, sucrose, and Ac-Di-Sol/Primojel were prepared and their dissolution in various media was evaluated. The dissolution differences were found to be largely related to the hydrophobicity of the active ingredient and pH difference of the two media. This difference was even more evident under the condition where acetaminophen, sucrose, and Primojel were combined. The dissolution difference was therefore attributed to the depressed function of Primojel in the acidic medium, the stronger binding of sucrose, the hydrophobicity of the active ingredient and pH difference of the two media. Increasing the concentration of Primojel or incorporating the surfactant in the tablet can thus greatly decrease the dissolution difference between acidic and neutral media.


Disruption of maternal behaviour by acute conspecific interaction induces selective activation of the lateral periaqueductal grey.
Maternal behaviour is sensitive to stress and opioidergic activation. The periaqueductal grey (PAG) is involved in coping strategies to stress, whereas morphine inhibition of maternal behaviour depends on the activation of the PAG. The aim of this study was to investigate whether the PAG is activated by disrupting maternal behaviour. Lactating Wistar rats were assigned to four groups: C (control); E1 (acute exposure to a male rat); E2 (daily 2-h exposure to another lactating female and a male rat from Day 3 to 6 of lactation); and E1 + 2 (treated first as E2 and, on Day 9, as E1). Maternal behaviour was recorded on Day 9 of lactation and analysed for 1 h. The E1 group spent more time retrieving their pups, took longer to initiate nursing, had shorter nursing bouts and spent more time in non-maternal activities compared with control. Rats submitted to E2 or E1 + 2 did not differ from the control. In another experiment, lactating rats were treated as above, except that 90 min after the end of the observation period the rats were killed and their brains were processed for immunohistochemical detection of Fos protein in the PAG. Fos increased in the lateral PAG only in the E1 group. We also observed that neurons activated by acute conspecific interaction in the PAG could be responsible for an opioid-dependent decrease in maternal behaviour as this effect was reversed by a microinjection of naltrexone, nor-binaltorphimine or naloxonazine into the lateral PAG. Chronic conspecific interaction alters the way this circuitry responds to acute conspecific interaction.


Validation of a model of feline ureteral obstruction as a tool for teaching microsurgery to veterinary surgeons.
To assess the content and face validity of a model of an obstructed feline ureter as a tool for teaching ureteral microsurgery. Prospective, experimental study. Seven expert and 11 novice microsurgeons. The model was created from latex rubber with an inner diameter of 0.8 mm and an outer diameter of 5 mm. The "ureter" was created with an inner compartment, a thin wall, and a soft, outer layer mimicking periureteral fat. A "ureterolith" 0.8-1.2 mm in diameter was placed inside the inner compartment by using a blunt-tipped cannula. A standardized "ureterotomy" was performed by 7 expert and 11 novice microsurgeons. Both groups completed questionnaires evaluating the content and face validity of the model using subjective measures and a 5-point Likert scale. Reliability was analysed by calculation of Cronbach's α for all questions to ensure α ≥ .7. The median responses to each question were compared between groups with a nonparametric independent samples median test. P < .05 was considered significant. The Cronbach's α for the experts and the novices for content validity questions was .7 and .9, respectively, and for the face validity questions it was .7 and .8, respectively. The model was rated to have excellent content validity and very good face validity. The model elicited positive responses from expert and novice microsurgeons and can be recommended as a tool for teaching ureteral microsurgery. A model validated by face and content measures should next be scrutinized by determination of construct, concurrent, and predictive validity by using objective measures.


Carotid artery occlusion and collateral circulation in C57Black/6J mice detected by synchrotron radiation microangiography.
Using monochromatic synchrotron radiation, we performed microangiography in C57BL/6J mice and investigated their vasculature after unilateral and bilateral carotid artery occlusion. Bilateral occlusion of the carotid artery was made by a ligation of the left common carotid artery followed by a ligation of the right internal carotid artery (ICA) two days later (n=12). Five days after the second surgery, angiography was performed. Unilateral occlusion was made by clipping the right ICA and then angiography was performed immediately (n=5). The control mice did not undergo any occlusion (n=5). We removed the brain of the bilateral occlusion mice after angiography and examined the infarction area. The cerebral microvessels in all animals were clearly visualized. In the control mice, the posterior communicating artery (Pcom) was not visualized. In the unilateral occlusion mice, the anastomosis of the pterygopalatine artery (PPA) and the external carotid artery (ECA) were recognized. The PPA is thus considered to play a role in the collateral vessel between the ICA and the ECA. The Pcom was not visualized. In the bilateral occlusion mice, the Pcom was observed either unilaterally (n=5) or bilaterally (n=5). The Pcom supplied blood flow to the anterior circulation from the vertebrobasilar arteries. The bilateral occlusion mice that had at least one visualized Pcom did not have any infarction. We could successfully visualize the cerebral vasculature of normal mice and carotid artery occluded mice in an in vivo study. Microangiography can demonstrate the development of vasculature and the blood flow dynamics in mice.


Cycloaddition reactions of acrylonitrile on the Si(100)-2 x 1 surface.
Multi-reference as well as single-reference quantum mechanical methods were adopted to study the potential energy surface along three possible surface reaction mechanisms of acrylonitrile on the Si(100)-2 x 1 surface. All three reactions occur via stepwise radical mechanisms. According to the computed potential energy surfaces, both [4+2] and [2+2](CN) cycloaddition products resulting from the reactions of surface dimers with the C[triple bond]N of acrylonitrile are expected, due to the negligible activation barriers at the surface. Another possible surface product, [2+2](CC), requires a 16.7 kcal/mol activation energy barrier. The large barrier makes this route much less favorable kinetically, even though this route produces the thermodynamically most stable products. Isomerization reactions among the surface products are very unlikely due to the predicted large activation barriers preventing thermal redistributions of the surface products. As a result, the distribution of the final surface products is kinetically controlled leading to a reinterpretation of recent experiments. An intermediate Lewis acid-base type complex appears in both the [4+2] and [2+2](CN) cycloadditions entrance channels, indicating that the surface may act as an electrophile/Lewis acid toward a strong Lewis base substrate.


Engineering a circularly permuted GFP scaffold for peptide presentation.
The use of peptides as in vivo and in vitro ligand binding agents is hampered by the high flexibility, low stability and lack of intrinsic detection signal of peptide aptamers. Recent attempts to overcome these limitations included the integration of the binding peptide into a stable protein scaffold. In this paper, we present the optimization and testing of a circularly permuted variant of the green fluorescent protein (GFP). We examined the ability of the optimized scaffold to accept peptide insertions at three different regions. The three regions chosen are localized in close spatial proximity to each other and support different conformations of the inserted peptides. In all the three regions peptides with a biased, but still comprehensive, amino acid repertoire could be presented without disturbing the function of the optimized GFP-scaffold.


Shifts in Alcohol-Related Diagnoses After the Introduction of International Classification of Diseases, Tenth Revision, Clinical Modification Coding in U.S. Hospitals: Implications for Epidemiologic Research.
In October 2015, the United States transitioned healthcare diagnosis codes from the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM), to the Tenth Revision (ICD-10-CM). Trend analyses of alcohol-related stays could show discontinuities solely from the change in classification systems. This study examined the impact of the ICD-10-CM coding system on estimates of hospital stays involving alcohol-related diagnoses. This analysis used 2014 to 2017 administrative data from the Agency for Healthcare Research and Quality Healthcare Cost and Utilization Project State Inpatient Databases for 17 states. Quarterly ICD-9-CM data from second quarter 2014 through third quarter 2015 were concatenated with ICD-10-CM data from fourth quarter 2015 through first quarter 2017. Quarterly counts of alcohol-related stays were examined overall and then by 6 diagnostic subgroups: withdrawal, abuse, dependence, alcohol-induced mental disorders (AIMD), nonpsychiatric alcohol-induced disease, and intoxication or toxic effects. Within each group, we calculated the difference in the average number of stays between ICD-9-CM and ICD-10-CM coding periods. On average, the number of stays involving any alcohol-related diagnosis in the 6 quarters before and after the ICD-10-CM transition was stable. However, substantial shifts in stays occurred for alcohol abuse, AIMD, and intoxication or toxic effects. For example, the average quarterly number of stays involving AIMD was 170.7% higher in the ICD-10-CM period than in the ICD-9-CM period. This increase was driven in large part by 1 ICD-10-CM code, Alcohol use, unspecified with unspecified alcohol-induced disorder. Researchers conducting trend analyses of inpatient stays involving alcohol-related diagnoses should consider how ongoing modifications in the ICD-10-CM code system and coding guidelines might affect their work. An advisable approach for trend analyses across the ICD-10-CM transition is to aggregate diagnosis codes into broader, clinically meaningful groups-including a single global group that encompasses all alcohol-related stays-and then to select diagnostic groupings that minimize discontinuities between the 2 coding systems while providing useful information on this important indicator of population health.


Longitudinal bone growth is impaired by direct involvement of caffeine with chondrocyte differentiation in the growth plate.
We showed previously that caffeine adversely affects longitudinal bone growth and disrupts the histomorphometry of the growth plate during the pubertal growth spurt. However, little attention has been paid to the direct effects of caffeine on chondrocytes. Here, we investigated the direct effects of caffeine on chondrocytes of the growth plate in vivo and in vitro using a rapidly growing young rat model, and determined whether they were related to the adenosine receptor signaling pathway. A total of 15 male rats (21 days old) were divided randomly into three groups: a control group and two groups fed caffeine via gavage with 120 and 180 mg kg-1 day-1 for 4 weeks. After sacrifice, the tibia processed for the analysis of the long bone growth and proliferation of chondrocytes using tetracycline and BrdU incorporation, respectively. Caffeine-fed animals showed decreases in matrix mineralization and proliferation rate of growth plate chondrocytes compared with the control. To evaluate whether caffeine directly affects chondrocyte proliferation and chondrogenic differentiation, primary rat chondrocytes were isolated from the growth plates and cultured in either the presence or absence of caffeine at concentrations of 0.1-1 mm, followed by determination of the cellular proliferation or expression profiles of cellular differentiation markers. Caffeine caused significant decreases in extracellular matrix production, mineralization, and alkaline phosphatase activity, accompanied with decreases in gene expression of the cartilage-specific matrix proteins such as aggrecan, type II collagen and type X. Our results clearly demonstrate that caffeine is capable of interfering with cartilage induction by directly inhibiting the synthetic activity and orderly expression of marker genes relevant to chondrocyte maturation. In addition, we found that the adenosine type 1 receptor signaling pathway may be partly involved in the detrimental effects of caffeine on chondrogenic differentiation, specifically matrix production and mineralization, as evidenced by attenuation of the inhibitory effects of caffeine by blockade of this receptor. Thus, our study provides novel information on the integration of caffeine and adenosine receptor signaling during chondrocyte maturation, extending our understanding of the effect of caffeine at a cellular level on chondrocytes of the growth plate.


Morphogenesis of bacteriophage phi 80: identification of the cistron 13 product.
An in vitro complementation reaction leading to the assembly of bacteriophage phi80 tails from component proteins is described. Tail assembly occurs when a lysate of any mutant in cistron 13 is mixed with a second lysate of a mutant in any of the other cistrons involved in tail formation. Lysates of mutants that are blocked in tail formation contain phage heads that can unite with free tails to form infective particles. The rate of the complementation reaction shows little dependence upon temperature, suggesting that the assembly depends largely upon the kinetic encounter of the interacting components. The tail component missing in cistron 13 mutant lysates was purified approximately 55-fold and shown to be, at least in part, a protein having a molecular weight of approximately 22,000. This protein was also released from highly purified infective phi80 particles after osmotic shock followed by heattreatment, suggesting that it most probably is an integral structural protein of the phage tail. Lysates of mutants of bacteriophage lambda that are defective in tail formation were shown to contain a tail component identical with or similar to the phi80 cistron 13 product.


TelCoVis: Visual Exploration of Co-occurrence in Urban Human Mobility Based on Telco Data.
Understanding co-occurrence in urban human mobility (i.e. people from two regions visit an urban place during the same time span) is of great value in a variety of applications, such as urban planning, business intelligence, social behavior analysis, as well as containing contagious diseases. In recent years, the widespread use of mobile phones brings an unprecedented opportunity to capture large-scale and fine-grained data to study co-occurrence in human mobility. However, due to the lack of systematic and efficient methods, it is challenging for analysts to carry out in-depth analyses and extract valuable information. In this paper, we present TelCoVis, an interactive visual analytics system, which helps analysts leverage their domain knowledge to gain insight into the co-occurrence in urban human mobility based on telco data. Our system integrates visualization techniques with new designs and combines them in a novel way to enhance analysts' perception for a comprehensive exploration. In addition, we propose to study the correlations in co-occurrence (i.e. people from multiple regions visit different places during the same time span) by means of biclustering techniques that allow analysts to better explore coordinated relationships among different regions and identify interesting patterns. The case studies based on a real-world dataset and interviews with domain experts have demonstrated the effectiveness of our system in gaining insights into co-occurrence and facilitating various analytical tasks.


Religiosity and participation in mutual-aid support groups for addiction.
Mutual-aid support groups play a vital role in substance abuse treatment in the United States. A national survey of mutual-aid support groups for addiction was conducted to identify key differences between participants in recovery groups. Survey data indicate that active involvement in support groups significantly improves one's chances of remaining clean and sober, regardless of the group in which one participates. Respondents whose individual beliefs better matched those of their primary support groups showed greater levels of group participation, resulting in better outcomes as measured by increased number of days clean and sober. Religious respondents were more likely to participate in 12-step groups and Women for Sobriety. Nonreligious respondents were significantly less likely to participate in 12-step groups. Religiosity had little impact on SMART Recovery participation but actually decreased participation in Secular Organizations for Sobriety. These results have important implications for treatment planning and matching individuals to appropriate support groups.


Benzidine dihydrochloride: risk assessment.
Benzidine, recognized as a bladder carcinogen in man and as a liver carcinogen in experimental animals, is the chemical basis of as many as 200 commercial dyes. Physiological processes can metabolize these dyes to release benzidine, thereby creating a potential exposure hazard. To assess this hazard, both sexes of F1 hybrid (genetically homogeneous) and monohybrid (genetically heterogeneous) mice from a BALB/c male and C57BL/6 female cross were exposed for their respective lifespans to benzidine dihydrochloride in their drinking water at concentrations of 0, 30, 40, 60, 80, 120, and 160 ppm for males, and 0, 20, 30, 40, 60, 80, and 120 ppm for females. Animals were removed from the study when they were dead or moribund. This study was terminated after 33 months of exposure. Using the endpoint of hepatocellular adenomas and carcinomas, the Armitage Doll multistage model was used to describe the tumor rates in the experimental dose range and to obtain the upper confidence level on tumor rates. Linear interpolation was used between zero dose and the upper confidence level of the lowest experimental dosage for predicting potential low dose tumor rates. Dose-response effects on body weight, survival, and liver neoplasms were noted in both stocks. For each of the endpoints, the females were more susceptible than males and the F1 (homogeneous) stock was more susceptible than the monohybrid cross (heterogeneous). The calculated virtually "safe" dose predicted to produce less than one per million F1 female mice with a liver tumor is 0.045 ppb. One part per billion of benzidine dihydrochloride in the drinking water of these mice is estimated to produce liver tumors in less than 2.23 mice per 100,000 population.


Music interventions in disorders of consciousness (DOC) - a systematic review.
The effects of music interventions including active and receptive music therapy for people living with neurogenic disorders of consciousness (DOC) have been subject to empirical studies in the past. The aim of this systematic review was to find and analyse the current research about the effects of musical interventions on people with DOC. For this purpose, studies with music interventions and patients with DOC from the year 1900 to 2017 were searched in English, German, and French in different databases. Risk-of-bias-analyses were conducted for each study that fulfilled the inclusion criteria. Twenty-two quantitative studies (three randomised controlled trials with more than 10 participants) were found eligible for review. They include a total of 329 participants living with either coma, unresponsive wakefulness syndrome, or minimally conscious state. Music interventions were associated with favourable behavioural and physiological responses in several studies, but methodological quality and outcomes were heterogeneous. More studies with a larger number of participants are needed as well as a consensus on key characteristics of effective short-term and long-term music interventions for DOC.


Intermittent Preventive Treatment for Malaria in Pregnancy: Optimization of Target Concentrations of Dihydroartemisinin-Piperaquine.
Dihydroartemisinin-piperaquine (DHA-PQ) is highly efficacious as intermittent preventive therapy for malaria during pregnancy (IPTp). Determining associations between piperaquine (PQ) exposure, malaria risk, and adverse birth outcomes informs optimal dosing strategies. Human immunodeficiency virus-uninfected pregnant women (n = 300) were enrolled in a placebo-controlled trial of IPTp at 12-20 weeks' gestation and randomized to sulfadoxine-pyrimethamine every 8 weeks, DHA-PQ every 8 weeks, or DHA-PQ every 4 weeks during pregnancy. Pharmacokinetic sampling for PQ was performed every 4 weeks, and an intensive pharmacokinetic substudy was performed in 30 women at 28 weeks' gestation. Concentration-effect relationships were assessed between exposure to PQ; the prevalence of Plasmodium falciparum infection during pregnancy; outcomes at delivery including placental malaria, low birth weight, and preterm birth; and risks for toxicity. Simulations of new dosing scenarios were performed. Model-defined PQ target venous plasma concentrations of 13.9 ng/mL provided 99% protection from P. falciparum infection during pregnancy. Each 10-day increase in time above target PQ concentrations was associated with reduced odds of placental parasitemia, preterm birth, and low birth weight, though increases in PQ concentrations were associated with QT interval prolongation. Modeling suggests that daily or weekly administration of lower dosages of PQ, compared to standard dosing, will maintain PQ trough levels above target concentrations with reduced PQ peak levels, potentially limiting toxicity. The protective efficacy of IPTp with DHA-PQ was strongly associated with higher drug exposure. Studies of the efficacy and safety of alternative DHA-PQ IPTp dosing strategies are warranted. NCT02163447.


Alkali Metal Ion Partitioning with Calix[4]arene-benzo-crown-6 Ionophore in Acidic Medium: Insights from Experiments, Statistical Mechanical Framework, and Molecular Dynamics Simulations.
The current work reports the experimental and predicted interfacial behavior of metal ion extraction from aqueous phase-diluent system using a newly synthesized calix-benzo-crown-6 (CBCBGA) ionophore. Conductor-like screening model for real solvents was used to predict the selectivity at infinite dilution for the metal ion complexes in both aqueous and diluent phases. The selectivity for Cs+-CBCBGA extraction was found to be higher than that of other metal ions, namely, K+, Na+, and Rb+. This was confirmed by the experimental distribution coefficients obtained in the diluents system at 3 M HNO3 along with 0.01 M CBCBGA/organic solvents. The high selectivity of Cs+-CBCBGA complex over other complexes (K+, Rb+, and Na+) in nitrobenzene was also confirmed and validated by the highest occupied molecular orbital-lowest unoccupied molecular orbital energy gap (i.e., 0.13114 > 0.12411 > 0.11719 > 0.11561 eV) and interaction energy (i.e., -68.25 > -57.11 > -55.52 > -52.37 kcal/mol). The interaction and free energies of the extraction were found to increase with the dielectric constant of the organic solvents, namely, nitrobenzene > o-nitrophenyl hexyl ether > 1-octanol > chloroform. Overall, a higher selectivity of Cs+ ion over that of other metal ions (K+, Na+, and Rb+) was obtained for the newly synthesized CBCBGA ionophore in a radioactive waste solution.


Long-term vitamin E supplementation fails to reduce lipid peroxidation in people at cardiovascular risk: analysis of underlying factors.
BACKGROUND: Antioxidant supplementation with vitamin E had no effect in the prevention of cardiovascular diseases (CVD) in three recent large, randomized clinical trials. In order to reassess critically the role of vitamin E in CVD prevention, it is important to establish whether these results are related to a lack of antioxidant action. METHODS: We examined the in vivo antioxidant effect of vitamin E (300 mg/day for about three years) in 144 participants in the Primary Prevention Project (females and males, aged >/= 50 y, with at least one major CV risk factor, but no history of CVD). Urinary 8-epi-PGF2alpha (isoprostane F2alpha-III or 15-F2t-isoP), a validated biomarker of lipid peroxidation, was measured by mass spectrometry. RESULTS: Urinary excretion of 8-epi-PGF2alpha [pg/mg creatinine, median (range)] was 141 (67-498) in treated and 148 (76-561) in untreated subjects (p = 0.10). Taking into account possible confounding variables, multiple regression analysis confirmed that vitamin E had no significant effect on this biomarker. Levels of 8-epi-PGF2alpha were in the normal range for most subjects, except smokers and those with uncontrolled blood pressure or hyperglycemia. CONCLUSIONS: Prolonged vitamin E supplementation did not reduce lipid peroxidation in subjects with major cardiovascular risk factors. The observation that the rate of lipid peroxidation was near normal in a large proportion of subjects may help explain why vitamin E was not effective as an antioxidant in the PPP study and was ineffective for CVD prevention in large scale trials.


New Insights Into the Lineage-Specific Expansion and Functional Diversification of Lamprey AID/APOBEC Family.
The AID/APOBEC family which converts cytidine to uridine on RNA or DNA experienced dynamic expansion in primates in order to resist exogenous viruses and endogenous retrotransposons. Recently, expansion of AID/APOBEC-like homologs has also been observed in the extant jawless vertebrate lamprey. To reveal what causes such expansion and leads to the functional diversification of lamprey cytosine deaminases (CDAs), we reassessed the CDA genes in Lethenteron japonicum (Lj). We first confirmed the expansion of LjCDA1L1 (CDA1-like 1) genes and found the expression correlation of LjCDA2 and LjCDA1L2 with LjVLRs (variable lymphocyte receptors). Among up to 14 LjCDA1L1 proteins, LjCDA1L1_4a has an extremely high deamination activity on ssDNA and buDNA and, unexpectedly, on dsDNA. LjCDA1L1s can also restrict the infection of HSV-1 particles. Thus, the arms race between the host and pathogens along with the recruitment by VLR assembly may participate together to form a driving force in the expansion and diversification of the lamprey AID/APOBEC family.


Ultrasonically treated liquid interfaces for progress in cleaning and separation processes.
Ultrasound and acoustic cavitation enable ergonomic and eco-friendly treatment of complex liquids with outstanding performance in cleaning, separation and recycling of resources. A key element of ultrasonic-based technology is the high speed of mixing by streams, flows and jets (or shock waves), which is accompanied by sonochemical reactions. Mass transfer across the phase boundary with a great variety of catalytic processes is substantially enhanced through acoustic emulsification. Encapsulation, separation and recovery of liquids are fast with high production yield if applied by ultrasound. Here we discuss the state of knowledge of these processes by ultrasound and acoustic cavitation from a perspective of a physico-chemical model in order to predict and control the outcome. We focus on the physical interpretation and quantification of ultrasonic parameters and properties of liquids to understand the chemistry of liquid/liquid interfaces in acoustic fields. The roles of thermodynamic enthalpy and entropy (incl. Laplace and osmotic pressure) in the context of sonochemical reactions (separation, catalysis, degradation, cross-linking, ion exchange and phase transfer) are outlined. The synergy of ultrasound and electric fields or continuous flow chemistry for cleaning and separation via emulsification is highlighted by specific strategies involving polymers and ultrasonic membranes.


Isolation and metabolic characteristics of previously uncultured members of the order aquificales in a subsurface gold mine.
Culture-dependent and -independent techniques were combined to characterize the physiological properties and the ecological impacts of culture-resistant phylotypes of thermophiles within the order Aquificales from a subsurface hot aquifer of a Japanese gold mine. Thermophilic bacteria phylogenetically associated with previously uncultured phylotypes of Aquificales were successfully isolated. 16S ribosomal DNA clone analysis of the entire microbial DNA assemblage and fluorescence in situ whole-cell hybridization analysis indicated that the isolates dominated the microbial population in the subsurface aquifer. The isolates were facultatively anaerobic, hydrogen- or sulfur/thiosulfate-oxidizing, thermophilic chemolithoautotrophs utilizing molecular oxygen, nitrate, ferric iron, arsenate, selenate, and selenite as electron acceptors. Their versatile energy-generating systems may reflect the geochemical conditions of their habitat in the geothermally active subsurface gold mine.


Monitoring Legionella species in hospital water systems. Link with disease and evaluation of different detection methods.
The aim of this work was to evaluate three currently available isolation methods for Legionella using water samples and swabs of a single pediatric hospital water system. Additionally, high risk patients were screened for the presence of Legionella pneumophila antigen in urine. Fifteen water samples and 11 swab samples were collected from distal sites at 18 sampling locations. The International Standard Method (PN-ISO11731-2) based on membrane filtration and direct culture of bacteria on selective media were compared with amoebic co-culture. The numbers of legionellae detected exceeded 10(2) cfu/100 ml in 50% of the samples. All the positive samples contained L. pneumophila SGs 2-14. Urine samples were obtained from 57 immunosuppressed children and screened for the presence of L. pneumophila serogroup (SG) 1 antigen by Legionella urinary antigen EIA. Of the 57 urine samples tested for the presence of Legionella pneumophila SG 1 antigen, none were positive. Our results highlight the value of combined membrane filtration and amoebic co-culture methods in detecting viable L. pneumophila strains. Direct plating of 0.2 ml water is a useful screening method for samples containing large bacterial amount.


Low-dose photon irradiation alters cell differentiation via activation of hIK channels.
To understand the impact of ionizing irradiation from diagnostics and radiotherapy on cells, we examined K(+) channel activity before and immediately after exposing cells to X-rays. Already, low dose in the cGy range caused in adenocarcinoma A549 cells within minutes a hyperpolarization following activation of the human intermediate-conductance Ca(2+)-activated K(+) channel (hIK). The response was specific for cells, which functionally expressed hIK channels and in which hIK activity was low before irradiation. HEK293 cells, which do not respond to X-ray irradiation, accordingly develop a sensitivity to this stress after heterologous expression of hIK channels. The data suggest that hIK activation involves a Ca(2+)-mediated signaling cascade because channel activation is suppressed by a strong cytosolic Ca(2+) buffer. The finding that an elevation of H2O2 causes an increase in the concentration of cytosolic Ca(2+) suggests that radicals, which emerge early in response to irradiation, trigger this Ca(2+) signaling cascade. Inhibition of hIK channels by specific blockers clotrimazole and TRAM-34 slowed cell proliferation and migration in "wound" scratch assays; ionizing irradiation, in turn, stimulated the latter process presumably via its activation of the hIK channels. These data stress an indirect radiosensitivity of hIK channels with an impact on cell differentiation.


Triazole linked picolylimine conjugate of calix[6]arene as a sequential sensor for La3+ followed by F-.
A new 1,3,5-tris-triazole linked picolylimine conjugate of calix[6]arene (L) has been shown to be selective toward La(3+) by turn on fluorescence with ∼70-fold enhancement and emits blue, with a minimal detection limit of 65 ± 5 ppb (490 nM). The species of recognition has been modeled computationally to have a monocapped twisted square antiprism with a N(6)O(3) binding core about La(3+). The in situ complex of L with La(3+) recognizes F(-) via fluorescence quenching. The reversible response of sensing La(3+) and F(-) sequentially by L has been demonstrated.


Low pacemaker incidence with continuous-sutured valves: a retrospective analysis.
Background Permanent pacemaker implantation after surgical aortic valve replacement depends on patient selection and risk factors for conduction disorders. We aimed to identify risk criteria and obtain a selected group comparable to patients assigned to transcatheter aortic valve implantation. Methods Isolated sutured aortic valve replacements in 994 patients treated from 2007 to 2015 were reviewed. Demographics, hospital stay, preexisting conduction disorders, surgical technique, and etiology in patients with and without permanent pacemaker implantation were compared. Reported outcomes after transcatheter aortic valve implantation were compared with those of a subgroup including only degenerative valve disease and first redo. Results The incidence of permanent pacemaker implantation was 2.9%. Longer hospital stay ( p = 0.01), preexisting rhythm disorders ( p < 0.001), complex prosthetic endocarditis ( p = 0.01), and complex redo ( p < 0.001) were associated with permanent pacemaker implantation. Although prostheses were sutured with continuous monofilament in the majority of cases (86%), interrupted pledgetted sutures were used more often in the pacemaker group ( p = 0.002). In the subgroup analysis, the incidence of permanent pacemaker implantation was 2%; preexisting rhythm disorders and the suture technique were still major risk factors. Conclusion Permanent pacemaker implantation depends on etiology, preexisting rhythm disorders, and suture technique, and the 2% incidence compares favorably with the reported 5- to 10-fold higher incidence after transcatheter aortic valve implantation. Cost analysis should take this into account. Often dismissed as minor complication, permanent pacemaker implantation increases the risks of endocarditis, impaired myocardial recovery, and higher mortality if associated with prosthesis regurgitation.


Analysis of Intrasession Repeatability of Ocular Aberrometric Measurements and Validation of Keratometry Provided by a New Integrated System in Mild to Moderate Keratoconus.
To evaluate the intrasession repeatability and validity of corneal curvature measurements provided by a new multifunctional device and to assess the intrasession repeatability of its ocular aberrometric measures. This prospective study comprises 37 mild to moderate keratoconus eyes of 37 patients. In all cases, 3 repeated measures of corneal topography and ocular aberrometry were performed with the iDesign 2 system (iD2; Johnson & Johnson Vision Care Inc, Jacksonville, FL) and one keratometric measurement was performed with the intraocular lens-(IOL) Master 500 (Carl Zeiss Meditec Inc, Dublin, CA) and Sirius (CSO) (SIR) systems. The within-subject SD (Sw) was <0.50 D for all iD2 keratometric measurements, with intraclass correlation coefficient (ICC) >0.980. Sw and ICC for the keratometric axis were 2.60 degrees and 0.992, respectively. Concerning aberrations, all Sw values for high-order Zernike terms were ≤0.11 μm (ICC > 0.900), and all Sw values for refraction data were <0.75 D (ICC > 0.95), except for J45. No statistically significant differences were found between the devices in any keratometric parameter evaluated (P ≥ 0.222), but the limits of agreement between the devices were clinically relevant. The magnitude of K readings and astigmatism correlated significantly with the difference in these parameters between iD2 and SIR (0.432 ≤ r ≤ 0.489, P ≤ 0.041). The iD2 system provides consistent measures of keratometry and ocular aberrometry in eyes with mild to moderate keratoconus. Keratometric data obtained with this system in these eyes cannot be considered as interchangeable with data provided by intraocular lens-Master 500 and SIR.


[Skull metastasis of hepatocellular carcinoma: case report].
We report a case of skull metastasis from a hepatocellular carcinoma (HCC). A 68-year-old male with hepatitis C virus-associated cirrhosis and HCC presented with painless subcutaneous mass in the left frontotemporal region. Radiologically, skull X-ray showed an osteolytic lesion, and non-contrast CT displayed a hyperdense tumor that homogeneously enhanced on contrast CT. Cerebral angiography demonstrated a hypervascular mass with the feeding arteries from the external carotid artery. The tumor was shown as an iso-intensity mass with flow voids on T1WI, as iso- to hypo- intensity on T2WI and FLAIR images, and as hypo-intensity on DWI. The lesion was markedly enhanced by Gd-DTPA. Previous reports have described a low incidence of skull metastasis in HCC patients. However, the incidence is gradually increasing due to the prolonged survival rates of HCC patients, indicating the significance of the disease as a differencial diagnosis for skull tumors. Recent reports describe characteristic MRI findings of skull metastasis from HCC (hypointensity on T2WI with flow void); however, owing to the rarity of this disease, more studies are needed to establish the typical MRI pathognomonic findings.


Casuistry: an alternative or complement to principles?
Casuistry is a traditional method of interpreting and resolving moral problems. It focuses on the circumstances of particular cases rather than on the application of ethical theories and principles. After a brief history of casuistry, the method is explained and its relation to theory and principles is discussed.


Clinical evaluation of the enhanced Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test for rapid diagnosis of tuberculosis in prison inmates.
The reliability of the enhanced Amplified Mycobacterium Tuberculosis Direct Test (E-MTD; Gen-Probe, Inc., San Diego, Calif.) for rapid diagnosis of pulmonary tuberculosis (TB) was evaluated by testing 1, 004 respiratory specimens from 489 Texas prison inmates. Results were compared to those of mycobacterial culture (BACTEC TB 460 and Middlebrook 7H11 biplates), smear for acid-fast bacilli (AFB; auramine O), and clinical course. After chart review, three patients (nine specimens) who were on antituberculosis therapy before the study began were excluded from final analysis. Of the remaining 995 specimens, 21 were AFB smear positive: 13 grew Mycobacterium tuberculosis complex (MTBC), 6 grew nontuberculous mycobacteria, and 2 (from two patients diagnosed with TB and started on therapy after the study began) were culture negative. Twenty-eight specimens (20 patients) were positive for MTBC by culture and E-MTD. Seven specimens (seven patients) were positive by culture alone; three were from patients who had other E-MTD-positive specimens, two were false-positive cultures, and two were false-negative E-MTD results. Eight specimens were positive by E-MTD only; four specimens (four patients) were false-positive E-MTD results, and four specimens were from two patients with earlier E-MTD-positive specimens that grew MTBC. Thus, there were 22 patients with TB (10 smear positive and 12 smear negative). The sensitivity and specificity of the AFB smear for diagnosis of TB, by patient, were 45.5 and 98.9%, respectively. After resolving discrepancies, these same values for E-MTD were 90.9 and 99.1% overall, 100 and 100% for the smear-positive patients, and 83.3 and 99.1% for the smear-negative patients. Excluding the one smear-negative patient whose E-MTD-negative, MTBC culture-positive specimen contained inhibitory substances, the sensitivity of E-MTD was 95.2% overall and 90.9% in smear-negative patients. The specificity and positive predictive value of E-MTD can be improved, without altering other performance characteristics, by modifying the equivocal zone recommended by the manufacturer. These data suggest that E-MTD is a reliable method for rapid diagnosis of pulmonary TB, irrespective of the AFB smear result. Guidelines for the most appropriate use of E-MTD with smear-negative patients are needed.


Differential Impact of Body Mass Index in Hip Arthroscopy: Obesity Does Not Impact Outcomes.
Background: Hip arthroscopy is commonly used for the treatment of hip pathologies. As population obesity rates continue to increase, elucidating the impact of body mass index (BMI) on hip arthroscopy outcomes is essential. This investigation was conducted to quantify the effects of BMI on hip arthroscopy outcomes. Methods: We conducted a retrospective medical records review of 459 patients undergoing hip arthroscopy at a single center from 2008 to 2016. The Harris Hip Score (HHS) and 2 component scores of the 12-Item Short Form Survey-the physical component score (PCS-12) and the mental component score (MCS-12)-were used to measure outcomes. Patients were stratified into 4 cohorts based on their BMI: underweight (BMI <18.5 kg/m2), normal weight (BMI 18.5 to 24.9 kg/m2), overweight (BMI 25.0 to 29.9 kg/m2), and obese (BMI ≥30.0 kg/m2). Results: At 1 and 2 years postoperatively, all cohorts experienced statistically significant improvements in the HHS and PCS-12. At 3 years postoperatively, statistically significant improvements were seen in the HHS for all cohorts; in the PCS-12 for the normal weight, overweight, and obese cohorts; and in the MCS-12 for the normal weight cohort. Intercohort differences were not statistically significant at 1, 2, or 3 years postoperatively. Conclusion: In our population, BMI did not have statistically significant effects on patient outcome scores following hip arthroscopy. All patient cohorts showed postoperative improvements, and differences between BMI cohorts were not statistically significant at any postoperative time point.


The effect of high and low doses of cycloheximide on nucleolar ribonucleic acid synthesis.
In resting rat liver a dose of cycloheximide (1 mg. per kg.) inhibits leucine incorporation by 80 per cent whereas doses above 15 mg. per kg. inhibit it more than 90 per cent, when tested at 90 minutes following intraperitoneal injection. Initial stimulation of incorporation of orotate into total cellular, nuclear, and nucleolar fractions was seen at all doses tested. After 3 hours, however, marked suppression of incorporation particularly into nucleolar RNA was seen at 5 mg. per kg. and above while 1 mg. per kg. continued to stimulate. At 5 hours there was continued marked inhibition of RNA synthesis by 30 mg. per kg. and slight depression of RNA synthesis even by 1 mg. per kg. Preliminary ultrastructural studies failed to show objective nucleolar alterations even after 5 hours of cycloheximide treatment at a dose of 30 mg. per kg. No direct effect of several concentrations of cycloheximide on RNA synthesis was seen in an in vitro nucleolar system.


Preliminary findings on the possible role of B-lymphocyte stimulator (BLyS) on diabetes-related periodontitis.
The possible role of B-cell growth and differentiation-related cytokines on the pathogenesis of diabetes-related periodontitis has not been addressed so far. The aim of this study was to evaluate the effects of diabetes mellitus (DM) on the gene expression of proliferation-inducing ligand (APRIL) and B-lymphocyte stimulator (BLyS), two major cytokines associated to survival, differentiation and maturation of B cells in biopsies from gingival tissue with periodontitis. Gingival biopsies were obtained from subjects with periodontitis (n = 17), with periodontitis and DM (n = 19) as well as from periodontally and systemically healthy controls (n = 10). Gene expressions for APRIL, BLyS, RANKL, OPG, TRAP and DC-STAMP were evaluated using qPCR. The expressions APRIL, BLyS, RANKL, OPG, TRAP and DC-STAMP were all higher in both periodontitis groups when compared to the control group (p < 0.05). Furthermore, the expressions of BLyS, TRAP and RANKL were significantly higher in the subjects with periodontitis and DM when compared to those with periodontitis alone (p < 0.05). The mRNA levels of BLyS correlated positively with RANKL in the subjects with periodontitis and DM (p < 0.05). BLyS is overexpressed in periodontitis tissues of subjects with type 2 DM, suggesting a possible role of this cytokine on the pathogenesis DM-related periodontitis.


[Mumps myocarditis in a small child].
The clinical history and post-mortem findings in a five-year-old girl are reported. She died in consequence of a chronic mumps myocarditis 13 weeks after having fallen ill with mumps. The antigen specific for mumps virus could be demonstrated in the myocardium by means of immunofluorescence.


An evaluation of analgesic efficacy and clinical acceptability of intravenous tramadol as an adjunct to propofol sedation for third molar surgery.
This article details a double-blind, randomized, placebo-controlled pilot study evaluating the analgesic efficacy and clinical acceptability of intravenous tramadol in patients undergoing surgical removal of an impacted third molar tooth under local anesthesia and intravenous sedation with propofol. Forty-five ASA status 1 dental outpatients were randomly allocated to 2 groups of 22 (group A) and 23 (group B) patients each (n = 45). Group A (T/P) received intravenous tramadol 1.5 mg/kg injected over 2 minutes, followed by a bolus dose of intravenous propofol 0.4 mg/ kg. Maintenance consisted of a continuous infusion of propofol 3 mg/kg/h, with an additional bolus dose of 0.4 mg/kg intravenously 2-3 minutes prior to the infiltration of the local anesthetic solution. Group B (P/P) patients received no tramadol but instead a saline placebo solution and an identical amount of propofol. Overall, in this study, postoperative pain was much better controlled in the group receiving tramadol 1.5 mg/kg intravenously despite there being no significant difference in the dose of propofol administered in both groups. Intravenous tramadol, when given with propofol, did not affect the cardiovascular, respiratory, and sedative effects of propofol. Following tramadol, despite being an opioid, no nausea and vomiting were reported in the early postoperative period, indicating the value of using tramadol with propofol. Thus, this pilot study demonstrated the potential use of intravenous tramadol with propofol in day-case dento-alveolar surgery.


How will the introduction of primary certificate training programs change vascular surgery training programs?
New requirements for vascular surgery training allow several routes to Board eligibility in the specialty. Individuals can enter vascular residency directly from medical school, after 3 years of surgical residency, or after completion of the traditional 5 years of surgery training. Vascular surgery program directors will be faced with the challenges of obtaining institutional support, designing an acceptable educational program, and working closely with the general surgery program director to ensure both programs are successful. Faculty in the vascular program may find working with residents right out of medical school or after only 3 years of surgery training to be a challenge, especially in terms of developing the requisite technical skills. Residents must be able to demonstrate mastery of the six competencies in addition to the skills of vascular surgery. Because, in some ways, this new vascular training scheme is an experiment in redesigning all surgical education, the vascular community will need to carefully evaluate the results by monitoring the practices of those who graduate from these programs.


Pathways and gene networks mediating the regulatory effects of cannabidiol, a nonpsychoactive cannabinoid, in autoimmune T cells.
Our previous studies showed that the non-psychoactive cannabinoid, cannabidiol (CBD), ameliorates the clinical symptoms in mouse myelin oligodendrocyte glycoprotein (MOG)35-55-induced experimental autoimmune encephalomyelitis model of multiple sclerosis (MS) as well as decreases the memory MOG35-55-specific T cell (TMOG) proliferation and cytokine secretion including IL-17, a key autoimmune factor. The mechanisms of these activities are currently poorly understood. Herein, using microarray-based gene expression profiling, we describe gene networks and intracellular pathways involved in CBD-induced suppression of these activated memory TMOG cells. Encephalitogenic TMOG cells were stimulated with MOG35-55 in the presence of spleen-derived antigen presenting cells (APC) with or without CBD. mRNA of purified TMOG was then subjected to Illumina microarray analysis followed by ingenuity pathway analysis (IPA), weighted gene co-expression network analysis (WGCNA) and gene ontology (GO) elucidation of gene interactions. Results were validated using qPCR and ELISA assays. Gene profiling showed that the CBD treatment suppresses the transcription of a large number of proinflammatory genes in activated TMOG. These include cytokines (Xcl1, Il3, Il12a, Il1b), cytokine receptors (Cxcr1, Ifngr1), transcription factors (Ier3, Atf3, Nr4a3, Crem), and TNF superfamily signaling molecules (Tnfsf11, Tnfsf14, Tnfrsf9, Tnfrsf18). "IL-17 differentiation" and "IL-6 and IL-10-signaling" were identified among the top processes affected by CBD. CBD increases a number of IFN-dependent transcripts (Rgs16, Mx2, Rsad2, Irf4, Ifit2, Ephx1, Ets2) known to execute anti-proliferative activities in T cells. Interestingly, certain MOG35-55 up-regulated transcripts were maintained at high levels in the presence of CBD, including transcription factors (Egr2, Egr1, Tbx21), cytokines (Csf2, Tnf, Ifng), and chemokines (Ccl3, Ccl4, Cxcl10) suggesting that CBD may promote exhaustion of memory TMOG cells. In addition, CBD enhanced the transcription of T cell co-inhibitory molecules (Btla, Lag3, Trat1, and CD69) known to interfere with T/APC interactions. Furthermore, CBD enhanced the transcription of oxidative stress modulators with potent anti-inflammatory activity that are controlled by Nfe2l2/Nrf2 (Mt1, Mt2a, Slc30a1, Hmox1). Microarray-based gene expression profiling demonstrated that CBD exerts its immunoregulatory effects in activated memory TMOG cells via (a) suppressing proinflammatory Th17-related transcription, (b) by promoting T cell exhaustion/tolerance, (c) enhancing IFN-dependent anti-proliferative program, (d) hampering antigen presentation, and (d) inducing antioxidant milieu resolving inflammation. These findings put forward mechanism by which CBD exerts its anti-inflammatory effects as well as explain the beneficial role of CBD in pathological memory T cells and in autoimmune diseases.


Elution of loosely bound acid phosphatase from Staphylococcus aureus.
Strains of Staphylococcus aureus from the International-Blair and the Seto-Wilson series of phage propagating strains were examined for acid phosphatase activity. This enzyme was found to occur in varying amounts in three different fractions: free (6 to 60%), loosely bound (25 to 82%), and firmly bound (0 to 46%). Propagating strain 3A, because of its high activity, was chosen for further study. The rate of enzyme production paralleled cell growth in Trypticase Soy Broth, but followed a biphasic pattern in a semisynthetic casein acid-hydrolysate medium with glyceryl phosphate. Maximal elution of acid phosphatase in the loosely bound fraction, presumably from the surface of cells, occurred in the alkaline pH range. From log-phase cells, elution was maximally effected with buffered 1.0 M KCl (pH 7.5), but stationary-phase cells required twice the concentration of KCl.


Early detection and intervention for young children with early developmental disabilities in Western Uganda: a mixed-methods evaluation.
Early support for children with developmental disabilities is crucial but frequently unavailable in low-resource settings. We conducted a mixed-methods evaluation to assess the feasibility, acceptability, and impact of a programme of early detection and intervention for young children with developmental disabilities in Western Uganda. Early child development training for healthcare workers (HCWs) was implemented in three rural districts, and attendance was tracked. HCW knowledge and confidence were assessed pre-/post-intervention, and referral numbers tracked to evaluate impact. Facilitators were trained and mentored to deliver a participatory, group, early intervention programme (EIP) for young children with developmental disabilities and their families. Facilitators were tracked as they were identified, trained, and delivered the intervention, and attendance of families was tracked. Pre-/post-intervention assessments evaluated changes in family quality of life (PedsQL 2.0, Family Impact Module), and child nutritional outcomes. Focus group discussions with stakeholders also assessed feasibility, acceptability and impact. Overall, 93 HCWs from 45 healthcare facilities received training. In the pre-/post-evaluation, median knowledge and confidence scores increased significantly (from 4.0 to 7.0 and from 2.7 to 4.7, respectively (p < 0.001)). HCWs reported feeling empowered to refer and offer care for families with a young child with disability. Referral rates increased significantly from 148 to 251 per annum (70%; p = 0.03). Eleven EIP facilitators were trained, and all delivered the intervention; 84 families were enrolled, of which 78% attended at least 6 out of 10 modules. Amongst those with paired pre-/post-intervention data (n = 48), total family quality of life scores increased significantly (21%, p < 0.001). Improvements were seen across all domains of quality of life, with the largest impacts on emotional functioning and social functioning (p < 0.001). The programme was acceptable to caregivers and facilitators. Caregivers reported improved knowledge, family relationships, hope, emotional wellbeing, and reduced self-stigma. A programme of early detection and intervention for children with early developmental disabilities and their families was feasible and acceptable in a rural community-based Ugandan setting. HCW training positively impacted knowledge, confidence, attitudes, and referral rates. Families enrolled to the EIP reported significant improvements in quality of life. Important programmatic barriers identified included geographical spread, poverty, gender inequality, and stigma.


[Safety and efficacy of olanzapine versus perphenazine in patients with schizophrenia: results of multicenter, 18-week, double-blind clinical trial].
The primary objective of the study was to evaluate the severity of extrapyramidal symptoms during treatment with olanzapine (10-20 mg) versus perphenazine (8-40 mg) using the Simpson Angus Scale (SAS). The secondary objective was to assess the safety profile and clinical efficacy of the investigated drugs. A total of 95 patients with schizophrenia who met the criteria for DSM-IV were randomized to a double-blind, 18 week prospective comparative trail conducted in Poland. The tolerance of treatment was assessed with the use of scales: BAS, SAS and UKU. The efficacy of treatment was evaluated with BPRS, PANSS and CGOI scales. For olanzapine patients, the severity of extrapyramidal symptoms improved after 3 first weeks of treatment, and significantly decreased from the baseline to endpoint. Perphenazine patients showed an increase of extrapyramidal symptoms. The difference of the SAS scores change was statistically significant between olanzapine and perphenazine groups. Akathisia symptoms decreased significantly in the olanzapine group during the treatment period, whereas symptoms of akathisia increased in the perphenazine group. Statistically significant differences of mean change of BAS total score from baseline to endpoint were noted between treatment groups Treatment--emergent adverse events occurred more frequently in patients receiving perphenazine (46%), than in patients receiving olanzapine (17%). The proportion of patients complying with improvement criteria for CGI scale score was statistically greater in the olanzapine group (72.7%) than in the perphenazine group (47.9%). Results of this study showed that the tolerance profile in patients taking olanzapine is superior to perphenazine. Olanzapine was better tolerated than perphenazine. After olanzapine treatment more subjects fulfilled the criterion of improvement and schizophrenic symptoms were less severe than in patients treated with perphenazine.


Serum and pleural adenosine deaminase. Correlation with lymphocytic populations.
This study attempts to correlate levels of ADA in tuberculous and neoplastic pleural exudates with the different immunologic cellular expressions that follow these clinical situations. Seventy-three patients with pleural effusion were studied in order to assess ADA activity (pleural and serum); in 25 of them, a study of delayed cellular immunity (pleural and sanguineous) was performed through B, CD3, CD4, and CD8 lymphocytic populations. The activity of ADA was determined, and the study of lymphocytic populations was made through the use of monoclonal antibodies. The data obtained showed the following: levels of ADA were significantly (p less than 0.0005) higher in the pleural fluid and the serum of tuberculous effusions compared to neoplastic effusions; percentages of CD3 and CD4 T-cells were significantly (p less than 0.05 and p less than 0.0005, respectively) greater in tuberculous effusions. The statistical study of the levels of ADA activity and the percentage of CD4 T-cells in pleural exudates produced a significant regression curve (r = 0.612 and p less than 0.0001) which showed a positive correlation between these two parameters. The pathogenic implications of these results suggest the possibility that ADA could be a new marker of cell-mediated immune activity.


Fabrication of multiple heterojunctions with tunable visible-light-active photocatalytic reactivity in BiOBr-BiOI full-range composites based on microstructure modulation and band structures.
The fabrication of multiple heterojunctions with tunable photocatalytic reactivity in full-range BiOBr-BiOI composites based on microstructure modulation and band structures is demonstrated. The multiple heterojunctions are constructed by precipitation at room temperature and characterized systematically. Photocatalytic experiments indicate that there are two types of heterostructures with distinct photocatalytic mechanisms, both of which can greatly enhance the visible-light photocatalytic performance for the decomposition of organic pollutants and generation of photocurrent. The large separation and inhibited recombination of electron-hole pairs rendered by the heterostructures are confirmed by electrochemical impedance spectra (EIS) and photoluminescence (PL). Reactive species trapping, nitroblue tetrazolium (NBT, detection agent of (•)O2(-)) transformation, and terephthalic acid photoluminescence (TA-PL) experiments verify the charge-transfer mechanism derived from the two types of heterostructures, as well as different enhancements of the photocatalytic activity. This article provides insights into heterostructure photocatalysis and describes a novel way to design and fabricate high-performance semiconductor composites.


[Regression analysis of cerebral blood perfusion and cognitive function in patients with mild cognitive impairment and Alzheimer's disease].
Objective: To analyze the relationships between cerebral blood perfusion and impairments of cognitive function in patients with Alzheimer's disease (AD) and mild cognitive impairment (MCI). Methods: A total of 21 patients with AD, 18 patients with MCI and 21 healthy adults as control group (NC group) matched in age, gender and education were recruited between November 2016 and November 2017 from Northern Jiangsu People's Hospital. All cases were performed routine head MRI, arterial spin labeling (ASL). Cognitive function evaluation including simple mental state scale (MMSE) and improved Addenbrooke's cognitive examination revised (ACE-R) of patients were also assessed. The difference between cerebral blood perfusion signals and cognitive impairment of the three groups were analyzed. Results: The subtest scores of attention and orientation, memory, language fluency, language and visual space of ACE-R scores were significantly reduced (P<0.001,uncorrected) for AD when compared with MCI. Using REST1.8 software to perform single factor ANOVA, and adjusted for P<0.05. The AD group showed a decrease in total cerebral perfusion compared to the NC group, a decrease and in the frontotemporal lobe compared to MCI group; compared with the normal group, MCI group also showed decreased perfusion in frontotemporal cortex. Pearson correlation analysis was performed on the cerebral blood flow and ACE-R scores in the AD group and the MCI group,the decrease degree of CBF in multiple brain regions was positively correlated with the total score of ACE-R's total scores, and the subtest score of ACE-R including attention and orientation, memory, language fluency, language and visual space also showed positive correlation. Conclusion: ASL technology can detect the changes of cerebral blood flow metabolism in AD and MCI patients, and reveal association relationships between brain function damage and cerebral blood flow metabolism in AD and MCI patients.


Causal attributions in King-Kopetzky syndrome.
At least 10% of people who present for help with hearing difficulties will be found to have normal hearing thresholds. These cases are clinically categorized as King-Kopetzky syndrome (KKS), obscure auditory dysfunction (OAD), or auditory processing disorder (APD). While recent research has focussed on the possible mechanistic basis for these difficulties, the perceptions of the hearing difficulties that lead people to seek help have not hitherto been identified. This study presents findings from an observational survey of causal attributions of hearing difficulties from 100 people with KKS. The findings suggest that participants regard immunity and risk related causes of hearing difficulties as pre-dominant. Psychological factors were not considered to be causal for hearing difficulties. These factors were not affected by diagnostic classification. These findings inform audiologists about their patient beliefs for the first time. The authors suggest that clinicians take care to ensure that their counselling is responsive to these beliefs.


A Neuroeconomic Framework for Creative Cognition.
Neuroeconomics is the study of the neurobiological bases of subjective preferences and choices. We present a novel framework that synthesizes findings from the literatures on neuroeconomics and creativity to provide a neurobiological description of creative cognition. We propose that value-based decision-making processes and activity in the locus ceruleus-norepinephrine (LC-NE) neuromodulatory system underlie creative cognition, as well as the large-scale brain network dynamics shown to be associated with creativity. This reconceptualization leads to several falsifiable hypotheses that can further understanding of creativity, decision making, and brain network dynamics.


Glomerular injury associated with hepatitis C infection: a correlation with blood and tissue HCV-PCR.
Membranoproliferative glomerulonephritis, with or without cryoglobulinemia, and membranous glomerulonephritis are the best characterized glomerulonephropathies associated with hepatitis C virus (HCV) infection. Other more unusual patterns of glomerular injury, including IgA nephropathy, focal segmental glomerulosclerosis, crescentic glomerulonephritis, fibrillary glomerulonephritis, immunotactoid glomerulopathy, and thrombotic microangiopathy, have also been associated with HCV infection, but primarily on an anecdotal basis. It remains uncertain whether the patterns of glomerular injury seen in HCV infected patients, particularly the unusual patterns, represent a disease process specifically related to HCV infection or whether they represent nonspecific patterns of injury due to other causes that happen to occur in HCV-infected patients. We examine this issue by reviewing the epidemiological and pathological evidence in the literature that either supports or refutes a specific relationship between HCV and the pattern of glomerular injury. We also include our experience with 31 HCV-infected patients. In addition, the pathogenesis of HCV-associated glomerulonephropathies is discussed with an emphasis on the significance of detecting HCV in renal biopsies by reverse transcriptase-polymerase chain reaction.


Two-year outcomes after vaginal prolapse reconstruction with mesh pelvic floor repair system.
The aim of this study was to assess anatomical and functional outcomes 2 years after prolapse repair using vaginal mesh repair system. Women enrolled in a 12-month observational study of outcomes after transvaginal mesh-augmented prolapse repair were invited to participate in an extended follow-up. Subjects completed questionnaires assessing pelvic symptoms, quality of life, global satisfaction, and a pelvic examination for anatomical support and mesh complications. Of 118 eligible women, 85 enrolled, 82 provided subjective data at 24 months, and pelvic examination/Pelvic Organ Prolapse Quantification data are available from 79 women. Total, anterior, and posterior Prolift kits were used in 47 (55%), 25 (29%), and 13 (15%), respectively. At baseline, most of the women had stage III prolapse (75%), with the anterior compartment constituting the leading edge in 71% of subjects. At 24 months, Pelvic Organ Prolapse Quantification measures were significantly improved from baseline in all compartments, with 51 (65%) stage 0/I, 25 (31%) stage II, 3 (4%) and stage III (P < 0.001), as were quality of life scores (P < 0.001), with the exception of sexual function. Symptomatic prolapse was reported by 7 (8.5%) women, of which 4 demonstrated prolapse in the nonoperated compartment. Three subjects (4%) reported persistent pelvic pain. The 2-year mesh exposure incidence was at least 13% (11/85). The proportion reporting dyspareunia was 28.9% (13/45) and was unchanged from baseline. The median global satisfaction was 9.3 (range 2.0-10.0). Anatomical support, symptom relief, and satisfaction are high 24 months after mesh-augmented vaginal prolapse repair, although mesh exposure and new onset prolapse of the nonoperated compartment are not uncommon.


Grapevine Subtilase Family: Update on New Sequences and Nomenclature Proposal.
In grapevine, serine peptidases from the subtilase family were recently associated to Plasmopara viticola resistance. This family in grapevine, first characterized in 2014, was re-analyzed last year and 82 subtilase genes were identified. However, in November of 2016, the National Center for Biotechnology Information database (NCBI) made a new public release of the grapevine genome annotation based on new sequencing data and better prediction algorithms. As a consequence, some gene annotations and lengths changed. Here we present an update to the grapevine subtilase gene family sequences (SBT), namely sequence identifiers, bioinformatic predictions and recommend a nomenclature for the grapevine SBT genes. Our results show that grapevine subtilase gene family is now constituted by 87 subtilase genes encoding for 109 subtilase proteins and, despite the reported alterations, expression data on subtilases associated to grapevine resistance to P. viticola pathosystem did not suffer any alteration.


Rhizosphere selection of Pseudomonas putida KT2440 variants with increased fitness associated to changes in gene expression.
As the interface between plant roots and soil, the rhizosphere is a complex environment where nutrients released by the plant promote microbial growth. Increasing evidences indicate that the plant also exerts a selective pressure on microbial populations in the rhizosphere, favouring colonization by certain groups. In this work, we have designed an experimental setup to begin analysing the evolution of a specific bacterial population in the rhizosphere, using Pseudomonas putida KT2440 as model organism. After several rounds of selection without passage through laboratory growth conditions, derivatives of this strain with increased fitness in the rhizosphere were isolated. Detailed analysis of one of these clones indicated that this effect is specific for rhizosphere conditions and derives from changes in its transcriptional profile in this environment, with 43 genes being differentially expressed with respect to the parental strain. Several of these genes belong to functional categories which could affect stress adaptation and availability of particular nutrients. By inactivating two genes identified as upregulated in the selected clone (coding for a stress-response protein and a rRNA modifying protein), these functions were shown to contribute to rhizosphere fitness. Our data also suggest the existence of different evolutionary pathways leading to increased rhizosphere fitness.


Capsaicin inhibits the in vitro binding of peptides selective for mu- and kappa-opioid, and nociceptin-receptors.
Capsaicin inhibited the equilibrium specific binding of endogenous opioid-like peptide ligands such as endomorphin-1, nociceptin, and dynorphin((1-17)) in rat brain membrane preparations. We studied the in vitro effect of capsaicin (1-10 microM) on homologous and heterologous competitive binding of opioid ligands, using unlabeled synthetic peptides and the following tritiated compounds: [(3)H]endomorphin-1, [(3)H]endomorphin-2, [(3)H]nociceptin((1-17)) and [(3)H]dynorphin((1-17)). Capsaicin-dependent inhibition was also observed in [(35)S]GTPgammaS stimulation assays in the presence of certain opioid peptides. The inhibition of opioid binding was further investigated using other synthetic and natural mu-opioid ligands such as [D-Ala(2),(NMe)Phe(4),Gly(5)-ol]enkephalin (DAMGO), morphine and naloxone. The decrease in opioid ligand affinity upon capsaicin treatments was most apparent with endomorphin-1, followed by nociceptin and dynorphin. The binding of other investigated opioids were not affected in the presence of capsaicin. In [(3)H]endomorphin-1 binding assays, capsazepine antagonized the inhibitory effect of capsaicin in rat brain membranes suggesting the involvement of TRPV1 receptors. In Chinese hamster ovary (CHO) cells stably expressing mu-opioid receptors, but lacking vanilloid receptors, the inhibition by capsaicin on the binding of [(3)H]endomorphin-1 was not present. It is concluded that the inhibitory effect of capsaicin on the receptor binding affinity of endogenous opioid peptides in brain membrane preparations seems not to be a direct effect, it is rather a negative feedback interaction with opioid receptors.


The accuracy of the bedside swallowing evaluation for detecting aspiration in survivors of acute respiratory failure.
Dysphagia with subsequent aspiration occurs in up to 60% of acute respiratory failure (ARF) survivors. Accurate bedside tests for aspiration can reduce aspiration-related complications while minimizing delay of oral nutrition. In a cohort of ARF survivors, we determined the accuracy of the bedside swallowing evaluation (BSE) and its components for detecting aspiration. Patients who were extubated after at least 24hours of mechanical ventilation were eligible for enrollment. Within 3 days of extubation, patients underwent comprehensive BSE including 3-oz water swallowing test (3-WST), followed by a criterion standard test for aspiration, flexible endoscopic evaluation of swallowing (FEES). Forty-five patients were included in the analysis. Median patient age was 55years (interquartile range, 47-65). Median duration of mechanical ventilation was 3.3days (interquartile range 1.8-6.0). Fourteen patients (31%) aspirated on FEES. Physical examination findings on BSE and abnormal swallowing during trials of different consistencies were variably associated with aspiration. Compared with FEES, the 3-WST yielded a sensitivity of 77% (95% confidence interval [CI], 50%-92%), specificity of 65% (95% CI, 47%-79%), and an area under the receiver operating characteristic curve (AUC) of 0.71; a speech language pathologist's recommendation for altered diet yielded a sensitivity of 86% (95% CI, 60%-96%), a specificity of 52% (95% CI, 35%-68%), and an AUCof 0.69; an speech language pathologist's recommendation for nil per os (nothing by mouth) yielded a sensitivity of 50% (95% CI, 27%-73%), a specificity of 94% (95% CI, 79%-98%), and an AUCof 0.72. The BSE and its components, including the 3-WST, demonstrated variable accuracy for aspiration in survivors of ARF. Investigation to determine the optimal noninvasive test for aspiration in ARF survivors is warranted. ClinicalTrials.gov identifier: NCT02363686, Aspiration in Acute Respiratory Failure Survivors.


Survey of Cherry necrotic rusty mottle virus and Cherry green ring mottle virus incidence in Korea by Duplex RT-PCR.
The incidence of Cherry necrotic rusty mottle virus (CNRMV) and Cherry green ring mottle virus (CGRMV) have recently been occurred in Korea, posing a problem for sweet cherry cultivation. Since infected trees have symptomless leaves or ring-like spots on the pericarp, it is difficult to identify a viral infection. In this study, the incidence of CNRMV and CGRMV in sweet cherry in Gyeongbuk province was surveyed using a newly developed duplex reverse transcriptase polymerase chain reaction (RT-PCR) method that can detect both viruses in a single reaction. CNRMV and CGRMV co-infection rates were 29.6%, 53.6%, and 17.6%, respectively, in samples collected from three different sites (Daegu, Gyeongju and Gyeongsan) in Gyeongbuk province during 2012 and 2013. This duplex RT-PCR method offers a simple, rapid, and effective way of identifying CNRMV and CGRMV simultaneously in sweet cherry trees, which can aid in the management of viral infections that could undermine yield.


Structural basis for multiple ligand specificity of the periplasmic lysine-, arginine-, ornithine-binding protein.
The substrate-binding site of a protein with multiple specificity should satisfy geometric and energetic complementarity for several different substrates. The structural basis of the multiple ligand specificity of the periplasmic lysine-, arginine-, ornithine-binding protein (LAO) was investigated by determining and analyzing the structures of the protein unliganded and liganded with each of the three high-affinity ligands (L-lysine, L-arginine, and L-ornithine) and with one low-affinity ligand (L-histidine). The geometric complementarity is achieved primarily by virtue of the large size of the ligand-binding site which can accommodate the maximum common volume of the four ligands plus three water molecules. The optimization of energetic complementarity is achieved by the relocation of protein-bound water molecules and by the movement of the Asp-11 side chain. The structure of the LAO-histidine complex indicates that the 30-fold reduced affinity of the protein for histidine is primarily due to unavailability of one ionic interaction of the histidine side chain with the protein which is present in the other three complexes.


Activation of CO2 by Vanadium Nitrogenase.
The reduction of CO2 into useful products, including hydrocarbon fuels, is an ongoing area of particular interest due to efforts to mitigate buildup of this greenhouse gas. While the industrial Fischer-Tropsch process can facilitate the hydrogenation of CO2 with H2 to form short-chain hydrocarbon products under high temperatures and pressures, a desire to perform these reactions under ambient conditions has inspired the use of biological approaches. Particularly, enzymes offer insight into how to activate and reduce CO2 , but only one enzyme, nitrogenase, can perform the multielectron, multiproton reduction of CO2 into hydrocarbons. The vanadium-containing variant, V-nitrogenase, displays especial reactivity towards the hydrogenation of CO and CO2 . This Focus Review discusses recent progress towards the activation and reduction of CO2 with three primary V-nitrogenase systems. These systems span both ATP-dependent and ATP-independent processes and utilize approaches with whole cells, isolated proteins, and extracted cofactors.


The Distance Between Forests and Crops Affects the Abundance of Drosophila suzukii During Fruit Ripening, But Not During Harvest.
Drosophila suzukii (Matsumura; Diptera: Drosophilidae) is an invasive pest with the ability to reproduce not only in various soft fruit crops, but also in numerous wild hosts. Forests and forest edges harbor many wild hosts, provide suitable microclimatic conditions and are therefore thought to enhance the abundance of D. suzukii. Although the comprehension of pest activity based on specific landscape elements is important to implement efficient management strategies, knowledge of how forests affect the abundance of D. suzukii in nearby crops is very limited. We conducted a monitoring study with liquid baited traps across different crops at different distance from the forests. During fruit ripening, more flies were captured in crops closer to forests (22.21 % decrease per 500 m distance), whereas there was no significant relationship during harvest. Since color can affect the efficiency of D. suzukii traps, we have used traps either with a red or black lid. Acquired data suggest that traps with black lids capture significantly more flies than traps with red lids. We provide a quantitative estimation of how and when distance from adjacent forests affects the abundance of D. suzukii in crop fields. Our results can help consultants and farmers to estimate the pest pressure of D. suzukii in crop fields near forested, noncrop areas and to implement appropriate control strategies when D. suzukii populations increase and fruit becomes susceptible to infestation.


Effectiveness of an online mental health strengthening module to build resilience and overcome stress for transitional aged medical students.
Transitional-aged youths (17-to-24-years-old) are prone to mental-health problems. Students in higher education, especially medical students, are more exposed to stressors and thus need training to increase resilience. However, there have been limited mental-health strengthening modules specifically developed for medical students of transitional age, and none in Indonesia. This study intends to test the effectiveness of an online mental-health strengthening module in altering resilience. A pragmatic randomized trial with repeated measurements was employed to evaluate biopsychosocial outcomes of resilience. The intervention module was delivered in 4 weeks to 105 eligible students. Participants were divided into intervention group (n = 52) and control group (n = 53). Outcomes were measured in the 4th, 8th, and 12th weeks. Primary outcome was resilience level as measured by Connor-Davidson Resilience Scale (CD-RISC). Perceived Stress Scale (PSS), Depression Anxiety Stress Scale (DASS) were utilized to measure stress, depression and anxiety. Knowledge and attitude toward mental-health were also measured through validated questionnaires. Stress levels of participants were measured biologically by measuring salivary cortisol and alpha-amylase levels at the baseline and 12th-week. Compared to the control group, there were no significant difference in resilience score of the intervention group compared to control group [F(1, 103) = 2.243, P = .137]; however, there was a significant main effect of time [F(3, 309) = 18.191, P < .001] and interaction effect between intervention and time in resilience score [F(3, 309) = 5.056, P = .002]. Additionally, compared to the control group, there were significant increases in knowledge [F(1, 103) = 66.805, P < .001], attitudes and behavior towards mental-health [F(1, 103) = 5.191, P = .025], and a significant decrease in stress perception score [F(1, 103) = 27.567, P < .001]. The mean salivary delta cortisol during pre-test and post-test at week 12 in the intervention group showed significant difference (P < .001). However, there was no significant difference in the mean delta salivary alpha-amylase between pre-test and post-test at week 12, both in the intervention and control groups. The mental-health strengthening module was accepted and applicable to first-year medical students and was found to be effective in increasing resilience from various biopsychosocial aspects. It is also advisable to have similar modules throughout the medical school to maintain sustainability.


Effectiveness of Manual Ventilation in Intubated Helicopter Emergency Services-Transported Trauma Patients.
Helicopter Emergency Medical Services agencies frequently transport intubated patients to definitive care. No evidence exists to determine the type of ventilation in this population. Practice varies amongst programs from bag-valve-mask to mechanical ventilation. Evaluate the effectiveness of bag-valve ventilation in intubated trauma patients. We hypothesized manual ventilation provides adequate support to maintain physiologic ETCO2. From June to December 2015, twenty patients were enrolled in this prospective, observational study. Included were endotracheally intubated trauma patients transported by this HEMS program. Excluded were interfacility transports, non-scene calls, and patients with supraglottic devices. ETCO2 was recorded every 30 seconds during the flight. As a descriptive pilot study, power was not considered. 20 patients provided over 500 cumulative minutes of manual ventilation data. The percentage of cumulative time spent with adequate oxygen saturations was 83.6%. The percentage of cumulative time spent with adequate ETCO2 was 48.7%, with 34.6% of time spent under and 16.7% above this range. Manual ventilation maintained a physiologic ETCO2 only 16.7% of the time. Significant variability existed, resulting in intermittent hypoxia and hyperventilation. Prior research linked such events to increased morbidity and mortality. Further studies are warranted to compare manual against mechanically ventilated patients.


P052 National Registration of Patients With Inflammatory Bowel Disease in Brazil on Behalf of GEDIIB.
The IBD National Patient Registry is an initiative of the GEDIIB (Brazilian Study Group of Inflammatory Bowel Disease) who aims to survey the epidemiological profile of IBD patients through the creation of a centralized registry with data on patients monitored in public and private health services which will allow the planning of actions by the GEDIIB to facilitate the diagnosis and access to treatment of IBD, enabling the implementation of actions of the GEDIIB and the partnership with government agencies to improve care and, consequently, the quality of life of patients with IBD. This study aims to show the results of the IBD National Patient Registry. A cohort study was performed. Data were collected from July 2020 to August 2021. Data were obtained from medical records and/or from patients during the regular follow-up visit and stored in pre-established records for further analysis. Only patients with an established diagnosis of CD and UC were included. The study was approved by the local ethical committees and all patients signed the consent form. In total, 797 patients were included, 60% with UC and 40% with CD; 52.9% from University Hospitals. The mean age was 44.75 ± 16.11 (12 - 92y), 59.9% female, 59.3% married, 76.4% Caucasian, 85.1% non-smokers, 30.5% completed higher education, 14.9% presented familial history of IBD. The age of onset of symptoms ranged from 3 - 79 years (32.94 ± 14.22) and 33.2% presented diarrhea as an initial manifestation. The age of diagnosis ranged from 4 - 81 years (35.07 ± 14.60) and the time from symptoms to diagnosis ranged from 1 to 2 years. The Montreal classification of CD patients were A1: 6.3%, A2: 59.9%, A3: 33.8%; L1: 38%, L2: 16.7%, L3: 43.9%, B1: 51.5%, B2: 27.8%, B3: 7.8%; perianal 12.8%. In UC, 47.8% presented pancolitis, 30.3% left-sided and 21.8% distal colitis. EIMs were present in 45.7% of patients, the most frequent being rheumatological 21.8%. Comorbidities were present in 72%, the most frequent were high blood pressure (15.3%) and diabetes (6.3%); 50% were with BMI > 25 Kg/m2. Most of the patients were in use of medical therapy (95.5%), of which 81.3% salicylate, 70.3% biological therapy, 49% immunosuppressor, 25.6% corticosteroid and 1.2% tofacitinib. Regarding biological therapy, the following medications were used: infliximab 47.6%, adalimumab 28.4%, vedolizumab 9.5%, ustekinumab 7.5%, certolizumab 2.2% and golimumab 1.3%. Eleven women used the medication during pregnancy. IBD surgery-related was performed in 69.7%, 77.2% abdominal and 22.8% perianal. Almost 30% performed more than one surgery. In 62% of patients, at least one complication was reported; most of them were infective disorders, demanding prolonged hospitalizations. To date, there is no IBD epidemiologic study covering the entire Brazilian territory. The results found with the registry will be fundamental to show the epidemiology of a country with continental dimensions such as Brazil. The greater the number of researchers included and from different regions of the country, the greater the representativeness of the data and may even help direct government actions on behalf of IBD patients.