Accurate Ventricular Tachyarrhytmia Beats Detection on Low Sample Rate ECG Patch Signals Using 1D U-Net.
ECG signals quality from mobile cardiac telemetry (MCT) wearable is much noisier than Holter or standard twelve leads ECG. Although, there are beats detection algorithms that has been shown to be accurate for MIT-BIH data, their performances degrade when applying to patches data and non sinus rhythms, especially when detecting ventricular beats on ventricular tachyarrhythmia. This paper presents a deep learning approach using convolutional neural network 1D U-net architecture as a core model, accompanied with miniature pre-processing and post-processing. The model consists of contracting path and expanding path. The contracting path is a sequence of multiple convolution layers and max pooling layers while the expanding path is a sequence of multiple convolution layers and up-convolution layers. There are internal connections between the contracting path and the expanding path to avoid gradient vanishing. The output of the model predicts beat probability map which can be converted to beat locations. Performance of the model on patch data gives 98.86% recall and 97.46% F1-score which is better than Pan-Tompkins by 2.48% and 0.33% respectively. For only ventricular beats, the recall is 95.21% which outperform Pan-Tompkins by 3.68%.


Characterization of the angiotensin-converting enzyme inhibitory activity of fermented milks produced with Lactobacillus casei.
Our study assayed angiotensin-converting enzyme (ACE) inhibitory activity and fermentation characteristics of 41 food-originated Lactobacillus casei strains in fermented milk production. Twenty-two of the tested strains produced fermented milks with a high ACE inhibitory activity of over 60%. Two strains (IMAU10408 and IMAU20411) expressing the highest ACE inhibitory activity were selected for further characterization. The heat stability (pasteurization at 63°C for 30 min, 75°C for 25 s, and 85°C for 20 s) and resistance to gastrointestinal proteases (pepsin, trypsinase, and sequential pepsin/trypsinase treatments) of the ACE inhibitory activity in the fermented milks produced with IMAU10408 and IMAU20411 were determined. Interestingly, such activity increased significantly after the heat or protease treatment. Because of the shorter milk coagulation time of L. casei IMAU20411 (vs. IMAU10408), it was selected for optimization experiments for ACE inhibitory activity production. Our results show that fermentation temperature of 37°C, inoculum density of 1 × 106 cfu/g, and fermentation time of 12 h were optimal for maximizing ACE inhibitory activity. Finally, the metabolite profiles of L. casei IMAU20411 after 2 and 42 h of milk fermentation were analyzed by ultra-HPLC electron spray ionization coupled with time-of-flight mass spectrometry. Nine differential abundant metabolites were identified, and 2 of them showed a strong and positive correlation with fermented milk ACE inhibitory activity. To conclude, we have identified a novel ACE inhibitory L. casei strain, which has potential for use as a probiotic in fermented milk production.


The association between disability and unintentional injuries among adolescents in a general education setting: Evidence from a Swedish population-based school survey.
Unintentional injuries are the leading cause of death among adolescents. Adolescents with disabilities may be particularly vulnerable with an increased risk of unintentional injuries. To study the association between a set of disabilities and unintentional injury risks among adolescents, accounting for comorbidity, subjective disability severity and sex. Cross-sectional data from a Swedish national school survey including 4,741 students (15 and 17-year olds) conducted in 2016 was analyzed using log-binomial generalized linear models. We found a 33% increased risk of injury the last 12 months and a 53% increased risk of injury leading to hospitalization for adolescents with any disability compared to their peers with no disability. The differences in injury risk were greater for girls than boys. There was a dose-response relationship between disability severity and injury risk. In analyses adjusted for sociodemographic factors and comorbidity, attention-deficit/hyperactivity disorder (ADHD) and epilepsy were associated with an increased risk of injury the last 12 months, risk ratios [RR] were 1.41 (95% Confidence Interval [CI] 1.08-2.97) and 1.79 (95% CI 1.10-1.81) respectively. Autism spectrum disorder was associated with a decreased injury risk the last 12 months (RR = 0.43, CI 0.2-0.92). ADHD, mobility impairment and visual impairment were associated with hospitalization due to injury during lifetime. There was an increased risk of unintentional injuries for adolescents with disabilities compared to their non-disabled peers, specifically for individuals with ADHD, epilepsy, visual impairment and mobility impairment. Injury prevention strategies may include adapting the physical environment and medical treatment.


A plea for extension of the anatomical nomenclature: Vessels.
This article is the fourth and last part of a series aimed at extending and correcting the anatomical nomenclature. Because of the rapid development of internet and the use of electronic formats in communication in anatomy, embryology, histology, medical education, and clinical medicine, an appropriate, precise, and concise anatomical nomenclature is required. Such tool enables to avoid any potential confusion and possible scientific/medical mistakes. The up-to-date official anatomical terminology, Terminologia Anatomica, is available longer than 20 years and needs to be refined and extended. The authors have collected and listed 210 terms and completed them with definitions and/or explanations. We aimed to start a discussion about their potential incorporation into the new revised version of the Terminologia Anatomica. This article is primarily focused on the vessels of the human body (arteries, veins, and lymphatic system).


Does insomnia mediate the link between childhood trauma and impaired control over drinking, alcohol use, and related problems?
Hyperarousal theory states that stressful negative events can result in a physiological response in the body leading to poor sleep quality. Childhood trauma is associated with many negative health consequences persisting into adulthood such as insomnia. Insomnia itself is a driver of poor physical and psychological health including excessive alcohol use. We examined the direct and indirect relationships between trauma (i.e., physical-neglect, physical, emotional, and sexual abuse) as well as emotionally supportive families on insomnia, impaired control over drinking, alcohol use, and alcohol-related problems. We studied a sample of 941 college students (467 women, 474 men). For our data analysis, we used a structural equation model with model indirect commands and 20,000 iteration bootstrapping with asymmetric confidence intervals in Mplus to obtain our mediated effects. Higher levels of emotional abuse were directly associated with more insomnia. Further, higher levels of physical neglect were directly associated with more impaired control over drinking. We found several mediational pathways from this investigation as well. Higher levels of emotional abuse were indirectly linked to both more alcohol use and alcohol-related problems through increased insomnia and impaired control over drinking. Our results were consistent with Hyperarousability Theory. We suggest that insomnia may contribute to dysregulated drinking and that combating emotional abuse could be a promising therapeutic target of intervention among college student social drinkers.


Causes of death for patients with community-acquired pneumonia: results from the Pneumonia Patient Outcomes Research Team cohort study.
To our knowledge, no previous study has systematically examined pneumonia-related and pneumonia-unrelated mortality. This study was performed to identify the cause(s) of death and to compare the timing and risk factors associated with pneumonia-related and pneumonia-unrelated mortality. For all deaths within 90 days of presentation, a synopsis of all events preceding death was independently reviewed by 2 members of a 5-member review panel (C.M.C., D.E.S., T.J.M., W.N.K., and M.J.F.). The underlying and immediate causes of death and whether pneumonia had a major, a minor, or no apparent role in the death were determined using consensus. Death was defined as pneumonia related if pneumonia was the underlying or immediate cause of death or played a major role in the cause of death. Competing-risk Cox proportional hazards regression models were used to identify baseline characteristics associated with mortality. Patients (944 outpatients and 1343 inpatients) with clinical and radiographic evidence of pneumonia were enrolled, and 208 (9%) died by 90 days. The most frequent immediate causes of death were respiratory failure (38%), cardiac conditions (13%), and infectious conditions (11%); the most frequent underlying causes of death were neurological conditions (29%), malignancies (24%), and cardiac conditions (14%). Mortality was pneumonia related in 110 (53%) of the 208 deaths. Pneumonia-related deaths were 7.7 times more likely to occur within 30 days of presentation compared with pneumonia-unrelated deaths. Factors independently associated with pneumonia-related mortality were hypothermia, altered mental status, elevated serum urea nitrogen level, chronic liver disease, leukopenia, and hypoxemia. Factors independently associated with pneumonia-unrelated mortality were dementia, immunosuppression, active cancer, systolic hypotension, male sex, and multilobar pulmonary infiltrates. Increasing age and evidence of aspiration were independent predictors of both types of mortality. For patients with community-acquired pneumonia, only half of all deaths are attributable to their acute illness. Differences in the timing of death and risk factors for mortality suggest that future studies of community-acquired pneumonia should differentiate all-cause and pneumonia-related mortality.


HIV protease inhibitors block streptolysin S production.
Streptolysin S (SLS) is a post-translationally modified peptide cytolysin that is produced by the human pathogen Streptococcus pyogenes. SLS belongs to a large family of azole-containing natural products that are biosynthesized via an evolutionarily conserved pathway. SLS is an important virulence factor during S. pyogenes infections, but despite an extensive history of study, further investigations are needed to clarify several steps of its biosynthesis. To this end, chemical inhibitors of SLS biosynthesis would be valuable tools to interrogate the various maturation steps of both SLS and biosynthetically related natural products. Such chemical inhibitors could also potentially serve as antivirulence therapeutics, which in theory may alleviate the spread of antibiotic resistance. In this work, we demonstrate that FDA-approved HIV protease inhibitors, especially nelfinavir, block a key proteolytic processing step during SLS production. This inhibition was demonstrated in live S. pyogenes cells and through in vitro protease inhibition assays. A panel of 57 nelfinavir analogs was synthesized, leading to a series of compounds with improved anti-SLS activity while illuminating structure-activity relationships. Nelfinavir was also found to inhibit the maturation of other azole-containing natural products, namely those involved in listeriolysin S, clostridiolysin S, and plantazolicin production. The use of nelfinavir analogs as inhibitors of SLS production has allowed us to begin examining the proteolysis event in SLS maturation and will aid in further investigations of the biosynthesis of SLS and related natural products.


Evidence that anaerobic oxidation of toluene in the denitrifying bacterium Thauera aromatica is initiated by formation of benzylsuccinate from toluene and fumarate.
Toluene is degraded anoxically to CO2 by the denitrifying bacterium Thauera aromatica. Toluene first becomes oxidized to benzoyl-CoA by O2-independent reactions. Benzoyl-CoA is then reduced to non-aromatic products by benzoyl-CoA reductase. We set out to study the reactions employed for the initial activation of toluene and its oxidation to the level of benzoate. Evidence is provided for a novel way of toluene degradation based on experiments with cell-free extracts and with whole toluene-grown cells: Cell-free extracts oxidized [14C]toluene to [14C]benzoyl-CoA via several radioactive intermediates. This reaction was strictly dependent on the presence of fumarate, coenzyme A and nitrate as electron acceptor; acetyl-CoA and ATP were not necessary for the reaction. The first product formed in vitro was benzylsuccinate; (2H8)toluene was converted to (2H7)benzylsuccinate. Formation of benzylsuccinate from toluene was independent of coenzyme A and nitrate, but it required the presence of fumarate. Other tricarboxylic acid cycle intermediates were converted to fumarate in cell extracts and therefore could partially substitute for fumarate. [14C]Benzylsuccinate was oxidized further to [14C]benzoyl-CoA and [14C]benzoate in cell extracts if coenzyme A and nitrate were present. No benzyl alcohol and benzaldehyde and no phenylpropionate could be detected as intermediates. In isotope trapping experiments with cell suspensions, two intermediates from [14C]toluene were detected, benzoate and benzylsuccinate. This corroborates the sequence of reactions deduced from in vitro experiments. A hypothetical degradation pathway for the anaerobic oxidation of toluene to benzoyl-CoA via an initial addition of fumarate to the methyl group of toluene and following beta-oxidation of the benzylsuccinate formed is suggested.


Police officers' and paramedics' experiences with overdose and their knowledge and opinions of Washington State's drug overdose-naloxone-Good Samaritan law.
Opioid overdoses are an important public health concern. Concerns about police involvement at overdose events may decrease calls to 911 for emergency medical care thereby increasing the chances than an overdose becomes fatal. To address this concern, Washington State passed a law that provides immunity from drug possession charges and facilitates the availability of take-home-naloxone (the opioid overdose antidote) to bystanders in 2010. To examine the knowledge and opinions regarding opioid overdoses and this new law, police (n = 251) and paramedics (n = 28) in Seattle, WA were surveyed. The majority of police (64 %) and paramedics (89 %) had been at an opioid overdose in the prior year. Few officers (16 %) or paramedics (7 %) were aware of the new law. While arrests at overdose scenes were rare, drugs or paraphernalia were confiscated at 25 % of the most recent overdoses police responded to. Three quarters of officers felt it was important they were at the scene of an overdose to protect medical personnel, and a minority, 34 %, indicated it was important they were present for the purpose of enforcing laws. Police opinions about the immunity and naloxone provisions of the law were split, and we present a summary of the reasons for their opinions. The results of this survey were utilized in public health efforts by the police department which developed a roll call training video shown to all patrol officers. Knowledge of the law was low, and opinions of it were mixed; however, police were concerned about the issue of opioid overdose and willing to implement agency-wide training.


Efficient and self-adaptive in-situ learning in multilayer memristor neural networks.
Memristors with tunable resistance states are emerging building blocks of artificial neural networks. However, in situ learning on a large-scale multiple-layer memristor network has yet to be demonstrated because of challenges in device property engineering and circuit integration. Here we monolithically integrate hafnium oxide-based memristors with a foundry-made transistor array into a multiple-layer neural network. We experimentally demonstrate in situ learning capability and achieve competitive classification accuracy on a standard machine learning dataset, which further confirms that the training algorithm allows the network to adapt to hardware imperfections. Our simulation using the experimental parameters suggests that a larger network would further increase the classification accuracy. The memristor neural network is a promising hardware platform for artificial intelligence with high speed-energy efficiency.


Plaque accumulation on exposed titanium surfaces and peri-implant tissue behavior. A preliminary 1-year clinical study.
The aim of this study was to evaluate plaque accumulation and peri-implant tissue response adjacent to machined and dual acid-etched (DAE) titanium implant surfaces. Two types of implants were used-control implants with a DAE surface in their apical portion and a machined coronal part, and test implants with a DAE surface throughout their entire length. A total of 10 sets of implants were placed in the posterior quadrants of eight patients, with at least 2 implants (1 control and 1 test implant) placed in each site. Machined healing abutments were placed on the control implants and DAE-surfaced healing abutments on the test implants. Plaque Index and bleeding on probing (BOP) were recorded together with histologic and microbiologic analyses of the peri-implant tissues. The healing abutments underwent a scanning electron microscope scan at 5 months postsurgery. Standardized radiographs were also taken at the time of implant placement and 3, 6, and 12 months postsurgery. DAE surfaces accumulated more plaque than machined surfaces (P < .0006) and the plaque was assessed as more difficult to remove (P < .0143). No histologic abnormalities were seen and the test implants showed significantly lower crestal bone resorption than the control (P < .0174). DAE healing abutments showed an increased plaque accumulation, but no significant BOP differences or histologic analyses were found between test and control sites. The test implants showed less interproximal bone resorption than the control ones at the end of a 1-year follow-up evaluation.


Various phosphodiesterase subtypes mediate the in vivo antilipolytic effect of insulin on adipose tissue and skeletal muscle in man.
The antilipolytic effect of insulin on human abdominal subcutaneous adipose tissue and skeletal muscle during local inhibition of cAMP-phosphodiesterases (PDEs) was investigated in vivo, by combining microdialysis with a euglycaemic, hyperinsulinaemic clamp. During hyperinsulinaemia, the glycerol concentration decreased by 40% in fat and by 33% in muscle. Addition of the selective PDE3-inhibitor amrinone abolished the insulin-induced decrease in adipose glycerol concentration, but did not influence the glycerol concentration in skeletal muscle. Nor did the PDE4-selective inhibitor rolipram or the PDE5-selective inhibitor dipyridamole influence the insulin-induced decrease in muscle tissue glycerol. However, the non-selective PDE-inhibitor theophylline counteracted the antilipolytic action of insulin at both sites. The specific activity of PDEs was also determined in both tissues. PDE3-activity was 36.8+/-6.4 pmol x min(-1) x mg(-1) in adipose tissue and 3.9+/-0.5 pmol x min(-1) x mg(-1) in muscle. PDE4-activity in skeletal muscle was high, i.e., 60.7+/-10.2 pmol x min(-1) x mg(-1) but 8.5 pmol x min(-1) x mg(-1) or less in adipose tissue. In conclusion, insulin inhibits lipolysis in adipose tissue and skeletal muscle by activation of different PDEs, suggesting a unique metabolic role of muscle lipolysis.


Isoquinoline thiosemicarbazone displays potent anticancer activity with in vivo efficacy against aggressive leukemias.
A potent class of isoquinoline-based α-N-heterocyclic carboxaldehyde thiosemicarbazone (HCT) compounds has been rediscovered; based upon this scaffold, three series of antiproliferative agents were synthesized through iterative rounds of methylation and fluorination modifications, with anticancer activities being potentiated by physiologically relevant levels of copper. The lead compound, HCT-13, was highly potent against a panel of pancreatic, small cell lung carcinoma, prostate cancer, and leukemia models, with IC50 values in the low-to-mid nanomolar range. Density functional theory (DFT) calculations showed that fluorination at the 6-position of HCT-13 was beneficial for ligand-copper complex formation, stability, and ease of metal-center reduction. Through a chemical genomics screen, we identify DNA damage response/replication stress response (DDR/RSR) pathways, specifically those mediated by ataxia-telangiectasia and Rad3-related protein kinase (ATR), as potential compensatory mechanism(s) of action following HCT-13 treatment. We further show that the cytotoxicity of HCT-13 is copper-dependent, that it promotes mitochondrial electron transport chain (mtETC) dysfunction, induces production of reactive oxygen species (ROS), and selectively depletes guanosine nucleotide pools. Lastly, we identify metabolic hallmarks for therapeutic target stratification and demonstrate the in vivo efficacy of HCT-13 against aggressive models of acute leukemias in mice.


Adalimumab persistence for inflammatory bowel disease in veteran and insured cohorts.
Identify predictors of persistence with adalimumab (ADA) among veterans and privately insured patients with inflammatory bowel disease (IBD) in the United States. Retrospective cohort study. Patients with IBD taking ADA as their first biologic were identified from the Veterans Health Administration (VHA) database from 2009 to 2013 and the Truven Health MarketScan database from 2009 to 2012 with a 12-month follow-up. Persistence was defined as continued use 1 year after initiation. Adherence was assessed by calculating a medication possession ratio, which was dichotomized as greater than 0.86 or less than or equal to 0.86. Multivariable logistic regression was used to evaluate predictors of persistence. There were 1030 patients in the VHA population compared with 3264 patients in the privately insured (MarketScan) cohort. In MarketScan, 1800 patients (55%) remained on ADA compared with 755 (73%) in the VHA cohort. In multivariable analysis, male sex (odds ratio [OR], 1.38; 95% CI, 1.16-1.63; P <.01), Crohn disease (OR, 1.27; 95% CI, 1.02-1.57; P = .03), greater adherence (OR, 1.83; 95% CI, 1.45-2.30; P <.01), and dose escalation (OR, 1.82; 95% CI, 1.42-2.33; P <.01) were associated with higher ADA persistence in the MarketScan cohort; narcotic use (OR, 0.71; 95% CI, 0.58-0.88; P <.01) and hospitalization or new steroid use after initiation (OR, 0.04; 95% CI, 0.03-0.05; P <.01) were associated with lower persistence. In the VHA cohort, only a hospitalization or new steroid use (OR, 0.50; 95% CI, 0.36-0.70; P <.01) was associated with lower persistence. Despite being older and having more comorbidities, patients in the VHA, which is an integrated healthcare system, appear to be more likely to remain on ADA at 1 year than patients in the MarketScan database. Further studies of system differences are needed to understand the reasons behind this discrepancy.


Reproductive endocrine disorders in women with primary generalized epilepsy.
It is known that women suffering from temporal lobe epilepsy may frequently present reproductive endocrine disorders (REDs). We hypothesized that a high occurrence of REDs could be found also in primary generalized epilepsy (PGE), and therefore investigated the hormonal and ovarian echographic profiles in 20 PGE female patients of reproductive age. Fourteen reported normal menstrual cycles, while 6 complained of longstanding menstrual irregularities. All but three patients were receiving antiepileptic drug (AED) therapy. In all subjects, the basal levels of gonadotropins, prolactin, and gonadal steroids were assayed. The response of luteinizing hormone (LH) to gonadotropin-releasing hormone was also investigated and ovarian ultrasonographic findings were evaluated. In five of six patients with menstrual problems (25% of the group), a well-defined RED was diagnosed (polycystic ovarian disease in three cases and hypothalamic ovarian failure in two). The 14 patients with normal menstrual cycles showed an elevation of mean basal follicle-stimulating hormone and prolactin, and a blunting of mean LH response. Our results suggest that a high occurrence of REDs may be found also in PGE. We hypothesize that a neurotransmitter dysfunction might be the common pathogenetic mechanism resulting in both REDs and PGE. The hormonal alterations observed in the patients with normal menstrual cycles seem to support our hypothesis. Previous data seem to rule out a possible AED effect accounting for the hormonal findings observed in our series. However, further studies are needed to confirm our preliminary results.


UK resilience to a chemical incident.
Following the deliberate chemical attacks in Salisbury last year, a review of UK resilience to chemical incidence was undertaken. The scope was not limited to deliberate attacks, related to terrorism, but was to include non-deliberate and accidental events. Chemical incidents have wide-reaching consequences irrespective of whether they are deliberate or not. The effects of these incidents manifest themselves in terms of immediate health consequences and will also include economic, political and public health effects that may have a much longer impact than the initial disruption. The economic consequences of the Novichok attack were devastating to the local population and businesses in both the short term and the long term, being felt over a year later. This review discusses the effects of these incidents on infrastructure, healthcare provision, law and order, economics and government at a local, regional and national level. The NHS Emergency Preparedness, Resilience and Response Framework was reviewed, and this provides a basic outline of advice for local government to follow in the case of chemical incidents. However, the wider issues of interdepartmental co-operation and how to maintain a response in the long term require further thought. Moreover, the methods by which interagency and regional resilience is maintained in preparing for such an event require a clearer guideline.


Positive psychotherapy for psychosis in Hong Kong: A randomized controlled trial.
Recovery-oriented practice has been advocated in mental health services in Hong Kong since 2009. Well-being has become an important area of focus for mental health services. Positive Psychotherapy for Psychosis (PPP) is a well-being-focused intervention for use in psychosis, with preliminary evidence from a randomized controlled trial in the United Kingdom of impact on well-being and symptomatology. The aim of this study was to test the effectiveness of PPP on the well-being of people with psychosis in Hong Kong. The study was a randomized controlled trial with two-arm parallel groups. Both groups received treatments as usual, and in addition the intervention group received a 13-session intervention based on a Cantonese Chinese translation of the PPP manual. Intention-to-treat analysis was used. The trial was registered (ANZCTR: ACTRN12620000464965). A total of 154 participants (78 intervention, 76 control) were recruited. As compared to control group, intervention group participants showed significant changes over time on the primary outcome of well-being assessed using the Chinese Short Warwick-Edinburgh Mental Well-being Scale (p = 0.001) and on secondary outcomes of hope (Agency subscale: p = 0.029) and self-efficacy (p = 0.001). Positive Psychotherapy for Psychosis was found to be an effective treatment in improving the well-being and other mental health outcomes for people with psychosis. It can be recommended for use in mental health services to promote recovery.


Neuronal gene expression in non-demented individuals with intermediate Alzheimer's Disease neuropathology.
While the clinical and neuropathological characterization of Alzheimer's Disease (AD) is well defined, our understanding of the progression of pathologic mechanisms in AD remains unclear. Post-mortem brains from individuals who did not fulfill clinical criteria for AD may still demonstrate measurable levels of AD pathologies to suggest that they may have presented with clinical symptoms had they lived longer or are able to stave off disease progression. Comparison between such individuals and those clinically diagnosed and pathologically confirmed to have AD will be key in delineating AD pathogenesis and neuroprotection. In this study, we expression profiled laser capture microdissected non-tangle bearing neurons in 6 post-mortem brain regions that are differentially affected in the AD brain from 10 non-demented individuals demonstrating intermediate AD neuropathologies (NDAD; Braak stage of II through IV and CERAD rating of moderate to frequent) and evaluated this data against that from individuals who have been diagnosed with late onset AD as well as healthy elderly controls. We identified common statistically significant expression changes in both NDAD and AD brains that may establish a degenerative link between the two cohorts, in addition to NDAD specific transcriptomic changes. These findings pinpoint novel targets for developing earlier diagnostics and preventative therapies for AD prior to diagnosis of probable AD. We also provide this high-quality, low post-mortem interval (PMI), cell-specific, and region-specific NDAD/AD reference data set to the community as a public resource.


Anti-biofilm Potential of Nanonized Eugenol against Pseudomonas aeruginosa.
The purpose of this study was to synthesize a nanoform of eugenol (an important phytochemical with various pharmacological potentials) and to investigate its anti-biofilm efficacy on Pseudomonas aeruginosa biofilm. Colloidal suspension of eugenol-nanoparticles (ENPs) was synthesized by the simple ultrasonic cavitation method through emulsification of hydrophobic eugenol into hydrophilic gelatin. Thus, the nanonization process made water-insoluble eugenol into water-soluble nano-eugenol, making the nano-form bioavailable. The size of the ENPs was 20-30 nm, entrapment efficiency of eugenol within gelatin was 80% and release of eugenol from gelatin cap was slow and sustained over five-days. Concerning the clinically relevant pathogen Pseudomonas aeruginosa, ENP had higher anti-biofilm (for both formation and eradication) activities than free eugenol. Minimal biofilm inhibitory concentration and minimal biofilm eradication concentration of ENP on P. aeruginosa biofilm were 2.0 and 4.0 mM respectively. In addition, the measurement of P. aeruginosa biofilm biomass, biofilm thickness, amount of biofilm extra-polymeric substance, cell surface hydrophobicity, cell swarming and twitching efficiencies, cellular morphology and biofilm formation in catheter demonstrated that the anti-biofilm efficacy of nano-eugenol was 30-40% higher than that of bulk eugenol. These results signify that future pharmacological and clinical studies are very much required to investigate whether ENP can act as an effective drug against P. aeruginosa biofilm-mediated diseases. Thus, the problem of intrinsic antibiotics-tolerance of biofilm-forming cells may be minimized by ENP. Moreover, ENP may be used as a potential catheter-coating agent to inhibit pseudomonal colonization on catheter surfaces and therefore, to reduce catheter-associated infections and complications.


Evolution of Insect Color Vision: From Spectral Sensitivity to Visual Ecology.
Color vision is widespread among insects but varies among species, depending on the spectral sensitivities and interplay of the participating photoreceptors. The spectral sensitivity of a photoreceptor is principally determined by the absorption spectrum of the expressed visual pigment, but it can be modified by various optical and electrophysiological factors. For example, screening and filtering pigments, rhabdom waveguide properties, retinal structure, and neural processing all influence the perceived color signal. We review the diversity in compound eye structure, visual pigments, photoreceptor physiology, and visual ecology of insects. Based on an overview of the current information about the spectral sensitivities of insect photoreceptors, covering 221 species in 13 insect orders, we discuss the evolution of color vision and highlight present knowledge gaps and promising future research directions in the field.


Monoanionic, Bis(pyrazolyl)methylborate [(Ph3B)CH(3,5-(CH3)2Pz)2)]- as a Supporting Ligand for Copper(I)-ethylene, cis-2-Butene, and Carbonyl Complexes.
The monoanionic bidentate ligand [(Ph3B)CH(3,5-(CH3)2Pz)2)]- has been prepared from lithium bis(pyrazolyl)methanide and triphenylborane. This useful new ligand is closely related to the well-established bis(pyrazolyl)borate and bis(pyrazolyl)methane ligands but has key differences to both analogues as well. The ethylene, cis-2-butene, and carbon monoxide adducts [(Ph3B)CH(3,5-(CH3)2Pz)2]Cu(L) (where L = C2H4, cis-CH3HC═CHCH3, and CO) have been prepared from [(Ph3B)CH(3,5-(CH3)2Pz)2)]Li(THF), copper(I) triflate, and the corresponding coligand. These complexes have been characterized by NMR spectroscopy and X-ray crystallography. In all cases the bis(pyrazolyl) moiety is bound in κ2N fashion with the BPh3 group rotated to sit over the metal center, sometimes coordinating to the metal via phenyl carbons as in [(Ph3B)CH(3,5-(CH3)2Pz)2)]Li(THF) and [(Ph3B)CH(3,5-(CH3)2Pz)2]Cu(CO) or simply hovering above the metal site as in [(Ph3B)CH(3,5-(CH3)2Pz)2)]Cu(C2H4) and [(Ph3B)CH(3,5-(CH3)2Pz)2)]Cu(cis-CH3HC═CHCH3). The 13C and 1H resonances of the ethylene carbon and protons of [(Ph3B)CH(3,5-(CH3)2Pz)2)]Cu(C2H4) appear at δ 81.0 and 3.71 ppm in CD2Cl2, respectively. The characteristic CO frequency for [(Ph3B)CH(3,5-(CH3)2Pz)2]Cu(CO) has been observed at υ̅ 2092 cm-1 by infrared spectroscopy and is lower than that of free CO suggesting moderate M → CO π-back-donation. A detailed analysis of these complexes has been presented herein.


Evidence for "twisted plane" undulations in golden hamster sperm tails.
Motile spermatozoa from the golden hamster have been arrested by rapid freezing and then fixed with glutaraldehyde at low temperature after substitution with ethylene glycol. As far as can be judged, the flagellar waveforms thus stabilized are similar to those seen in living sperm; in contrast, fixation in glutaraldehyde, without prior freezing, induces agonal changes in flagellar conformation. The characteristics waveform after freeze substitution contains three bends. Approx. half of these flagella are entirely planar. The rest are three dimensional, with the third bend displaced in a regular way from the plane containing the proximal two bends. From the geometry of these flagella, it is concluded that the plane of action of a given bending cycle undergoes a clockwise twist (from a forward viewpoint) as the cycle is succeeded by new bending cycles. This "twisted plane" undulation is quite different from helical movement. The twisting seems to occur abruptly, between cycles, as if each bending cycle has a preferred plane of action. The mechanism underlying the twisting is uncertain. However, on the basis of the angular displacements between the preferred planes, and the findings from electron microscopy, the following idea is presented as a working hypothesis: that, if the most proximal plane of bending is topographically determined by peripheral doublet 1, then successive distal planes of action are influenced predominantly by doublets 2, 3, etc., in clockwise sequence. The merits and weaknesses of this hypothesis are discussed.


[Lymphocytes with antigen receptors in drug allergy].
The data on the possibility of using the rosette-formation tests for the diagnosis of drug allergy are presented. Tests based on changes in the levels of activated T- and B-lymphocytes after their incubation with allergenic drugs have proved to be low informative. The test found to be highly informative is the antigen-specific rosette-formation test based on the detection of lymphocytes, capable of binding allogeneic erythrocytes loaded with antibiotics causing allergy in patients, in the peripheral blood. This test may be of importance not only in diagnosis, but also for prognosis, as it permits the detection of sensitization to a drug before the clinical manifestations of allergy.


Driving blind: instituting SEP-1 without high quality outcomes data.
In 2015, the Centers for Medicare and Medicaid Services (CMS) instituted an all-or-none sepsis performance measure bundle (SEP-1) to promote high-quality, cost-effective care. Systematic reviews demonstrated only low-quality evidence supporting most of SEP-1's interventions. CMS has removed some but not all of these unproven components. The current SEP-1 version requires patients with suspected sepsis have a lactate level, blood cultures, broad-spectrum antibiotics and, if hypotensive, a fixed 30 mL/kg fluid infusion within 3 hours, and a repeat lactate if initially elevated within 6 hours. Experts have continued to raise concerns that SEP-1 remains overly prescriptive, lacks a sound scientific basis and presents risks (overuse of antibiotics and inappropriate fluids not titrated to need). To incentivize compliance with SEP-1, CMS now publicly publishes how often hospitals complete all interventions in individual patients. However, compliance measured across hospitals (5 studies, 48-2,851 hospitals) or patients (three studies, 110-851 patients) has been low (approximately 50%) which is not surprising given SEP-1's lack of scientific basis. The largest observational study (1,738 patients) reporting survival rates employing SEP-1 found they were not significantly improved with the measure (P=0.53) as did the next largest study (851 patients, adjusted survival odds ratio 1.36, 95% CI, 0.85 to 2.18). Two smaller observational studies (158 and 450 patients) reported SEP-1 improved unadjusted survival (P≤0.05) but were confounded either by baseline imbalances or by simultaneous introduction of a code sepsis protocol to improve compliance. Regardless, retrospective studies have well known biases related to non-randomized designs, uncontrolled data collection and failure to adjust for unrecognized influential variables. Such low-quality science should not be the basis for a national mandate compelling care for a rapidly lethal disease with a high mortality rate. Instead, SEP-1 should be based on high quality reproducible evidence from randomized controlled trials (RCT) demonstrating its benefit and thereby safety. Otherwise we risk not only doing harm but standardizing it.


[Precise application of Traditional Chinese Medicine in minimally-invasive techniques].
The miminally-invasive techniques of traditional Chinese medicine(TCM) uses different types of acupuncture needles to treat diseased locations with special techniques. These techniques include different methods of insertion and closed incision (press cutting, sliding cutting, scrape cutting, etc.). This needling technique is based on the traditional Chinese medicine theories of Pi Bu(cutaneous), Jing Jin(sinew), Jing Luo(meridian), Wu Ti(five body structure components) and Zang Fu(organ system). Commonly used needles include: needle Dao, needle with edge, Pi needle, Shui needle, Ren needle, Gou needle, Chang Yuan needle, Bo needle and so on. The principle of this minimally-invasive technique of TCM is to achieve the greatest healing benefit with the least amount of anatomical and physiological intervention. This will result in the highest standard of health care with the lowest rehabilitative need and burden of care. In the past 20 years, through the collaborative research of several hundred hospitals across China, we systemically reviewed the best minimally invasive technique of TCM and the first line treatments for selective conditions. In 2013, the Department of Medical Affairs of the State Administration of Traditional Chinese Medicine created "Traditional Chinese Medicine Technical Manual"(General Version) and released it nationwide, its contents include: (1)Minimally invasive scar tissue release. ¹Suitable for Bi and pain syndromes of neck, shoulder, waist, buttocks and extremities. ²Degeneration causes local hypertrophy and inflammation, creating local tissue adhesion. ³There are two kind incision methods-press cutting and slide cutting. (2)Minimally invasive fascial tension release. ¹Suitable for localized fascial tension caused by trauma, overuse, or wind-cold-dampness, leading to compensatory hyperplasia. ²Long term high-stress stimulation to local fascia creates compensatory hyperplasia, Ashi points, and tissue texture changes (cords, nodules, masses). ³According to the different structural features of the needles, there are two incision methods: penetrating from the outside to the inside and pulling from inside to outside. (3)Minimally invasive decompression technique. ¹Suitable for internal pressure changes within organ cavities caused by trauma, degeneration, inflammation, such as compartment syndrome, bone marrow edema, increased intraluminal pressure in the bone marrow. ²According to the different tissues, it is categorized into soft tissue decompression, and bone decompression. (4)Minimally invasive orthopedic surgery. Applicable to some postural, developmental deformity correction, mainly through the dynamic balance method and/or static balance method. (5)Minimally invasive dissection. Suitable for fractures, tendons injury caused by deep soft tissue adhesion. (6)Minimally invasive separation. ¹Suitable for cutaneous, sinew regions, superficial adhesions due to lesions, and local post-operative incision adhesions. ²According to the structure of the needle tip, the methods are divided into sharp separation and blunt dissection. (7)Minimally invasive sustained pressure technique. ¹Suitable for neuromuscular dysfunction which causes JING(spasm) syndrome and WEI(atrophy) syndrome. ²The needle is applied with sustained pressure, without penetrating select tissue surface. This includes: nerve root sustained pressure technique; peripheral nerve sustained pressure technique; muscle sustained pressure technique; fascial contact procedure; cutaneous sustained pressure technique.(8)Minimally invasive insertion technique. ¹Suitable for systemic regulation to treat disease. ²Different organs are connected to different layers of tissue. Therefore, to treat specific conditions, specific tissues must be targeted. ³For example, back Shu points are used to treat vertigo from cervical spine issues, and spinal degeneration associated digestion issue. ⁴The internal organs can be regulated by the pathways that runs along the different layers of tissue. The types of stimulation include: meridian acupoint stimulation; cutaneous stimulation; fascia stimulation; mucle stimulation; periosteum stimulation. The clinical application of these techniques has enriched the drug-free therapies of traditional Chinese medicine and achieved excellent outcomes, but at the same time it also raises an important question. How can we apply these minimally invasive techniques to clinical practice so it can be safe and effective? In addition, how can practitioners, individually and further develop their understanding of this minimally invasive technique progressive manner? We make the following recommendations. (1)Clear diagnosis and precise application. Any approach has specific indications and choosing the correct technique comes from a comprehensive understanding of its advantages and disadvantages. Moreover, the accurate application of the technique depends the expertise of the practitioner. Through systematic review and clinical observation, we formulated the First Line Treatment, the Second Line Treatment, and the Third Line Treatment for specific conditions. Using the information gathered from research, practitioners can decide on which point is appropriate to use based on the stage of disease progression. For example, common conditions like the nerve ending tension pain(i.e. cutaneous nerve entrapment syndrome) is caused by stress concentration. There are two types of treatment for this condition: ¹Change in the response to stress state (i.e.non-invasive approach such as manual therapy and physiotherapy). ²Change in state of surrounding environment (i.e. invasive approach such as Pi Needle). Before tissue texture changes to pain point, cord, nodules, the former approach is effective. Once tissue texture changes, the latter approach is First Line Treatment. (2)Systematic training and disease progression training. The minimally invasive techniques of traditional Chinese medicine can treat many kinds of disease. To ensure its safety, organization, progressive development, practitioners are trained systematically and manage their treatment approach through disease hierarchy. Moreover, this technique should be conducted according to its technical difficulty, operating conditions, and expertise of the practitioner. The application of minimally invasive techniques of traditional Chinese medicine does not depend on the hospitals' administration system or the regulatory college of medical professionals. The minimally invasive techniques of TCM should be taught from easy to difficult, simple to complicated, and requires gradual progression by the practitioners. Eventually, the minimally invasive techniques of TCM's diagnostic and treatment protocol can be created. These protocols are currently available for reference: ¹Forming diagnosis and differential diagnosis for the conditions below requires expert diagnostic and application skills: cerebral palsy; cervical vertigo; cervical headache; cervical precordial pain; other spine-related diseases. ²The requirements for the diagnosis and differential diagnosis of such techniques are relatively high, and special training is required for the practitioner who performs this technique. The conditions below uses minimally invasive orthopedic surgery and dissection: scar contracture deformity; congenital developmental malformations; cervical Bi-syndrome; shoulder pain syndrome; knee Bi-syndrome; low back pain; cervical spondylosis; lumbar disc herniation; avascular necrosis of the femoral head; ankylosing spondylitis. ³There are no special requirements for the diagnosis and differential diagnosis of such techniques, and special training is required for the practitioner who performs this technique. The technical content is mainly decompression and scar tissue release. a)Muscle strain diseases: levator scapulae, splenius capitis, splenius cervicis, supraspinatus, infraspinatus, teres minor, teres major, serratus posterior superior, serratus posterior inferior, piriformis, gluteus maximus, gluteus medius, and gluteus minimus, erector spinae. b)Joint degenerative disorders: frozen shoulder, tennis elbow, tenosynovitis, knee osteoarthritis, and plantar fascitis. c)JING-JIN PI-BU pain syndrome (cutaneous nerve entrapment syndrome): occipital great nerve entrapment syndrome, occipital small nerve entrapment syndrome, great auricular nerve entrapment syndrome, suprascapular nerve entrapment syndrome, transverse cutaneous nerve of neck entrapment syndrome. (3)People-centred practice. The most attractive feature of the minimally invasive techniques of TCM is that they do not rely on expensive medical equipment and operating conditions. The key to applying this technique is the practitioners' technique, skill, and expertise. The necessary conditions required to successfully apply this technique is ¹practitioner understands disease progression and diagnosis; ²practitioners' skill in applying technique. We require patient-centered approach, which uses evidence based approach as the focus. We aim to seek the truth from facts, to understand the comprehensive picture, to include pertinent details, to be observant, to be goal oriented, from one to another, from outside to inside, from top to the bottom, compare right from left, through active movement and passive movements and weight-bearing movements, and assisted passive movements to determine instantaneous centre to diagnose stress concentration points. The operating technique is based on the response of patient's tissues to this technique. We must pay attention to diagnosis through palpation: layers, structure, texture, deformity, dislocation, movement characteristic, rhythmic changes. To achieve SHOU MO XIN HUI WU WEI: position, quantify, quantity, timing, and pattern. Accurate grasp of timeliness and dose efficiency. Can distinguish between local or systemic effects of treatment. Through comprehensive judgment of hands feeling, acupuncture needle feeling, and inspiration, to achieve the precious treatment requirements as indicated by the "Huangdi Neijing·Suwen": "Puncture the bone without damaging tendons, and puncture tendons without damaging muscles, puncture the muscle without damaging pulse, puncture pulse without damaging skin, puncture skin without damaging muscle. Puncture muscle without damage tendons, puncture tendons without damaging bone... Puncture bone without damaging tendons and it means the needle passes through the tendons and arrives at the bone and work on the bone. Puncture tendons without damaging muscles, and it means the needle passes through the muscles and arrives close to the tendon. Puncture the muscle without damaging pulse and it means the needle passes the pulse and does not touch the muscle. Puncture pulse without damaging skin and it means, the needle passes through the skin without penetrating pulse. Puncture skin without damaging muscle and it means, the disease is in the skin and the needles insert into skin but does not damage muscle. Puncture muscle without damage tendons, and it means, the needle passes through the muscle and arrive on the tendon. Puncture tendons without damaging bone."


Cutaneous Apocrine Carcinoma With an In Situ Component and Histiocytoid and Signet-Ring Cells.
We present a case of cutaneous apocrine carcinoma arising in the axilla of a 71-year-old man. The tumor had a significant component of histiocytoid and signet-ring cells as well as in situ carcinoma within the apocrine glands. The cells expressed GATA3, gross cystic disease fluid protein 15, androgen receptor, and E-cadherin. Estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 were negative. Clinical correlation was required to rule out a metastasis from the breast or the gastrointestinal tract. Although most cutaneous apocrine carcinomas do not behave aggressively, our patient developed bone metastases and eventually died of his disease. It is debated whether histiocytoid and signet-ring cell cutaneous carcinomas should be classified as apocrine neoplasm. The presence of in situ carcinoma associated with this kind of tumor has been reported only once in the literature. This characteristic and the immunohistochemical profile are in favor of apocrine differentiation.


Insulin resistance and metabolic syndrome in the pediatric population.
The metabolic syndrome is a constellation of specific anthropometric, physiological, and biochemical abnormalities predisposing affected individuals to the development of diabetes and cardiovascular disease. The syndrome is well described in the adult literature. However, its description in the pediatric literature is more limited. Due in large part to the normal physiological changes that occur in children and adolescents with respect to growth and puberty, investigators have also struggled to establish a standard definition of the syndrome in the pediatric age group, hindering coordinated research efforts. However, whatever definition of the syndrome is used, the prevalence of the metabolic syndrome in the pediatric age group has increased worldwide. Insulin resistance is the principal metabolic abnormality that is common to the development of the metabolic syndrome in both children and adults. This review summarizes current research regarding the pathophysiology of insulin resistance and how this may contribute to specific abnormalities seen in children and adolescents with the metabolic syndrome. Specifically, insulin resistance in pediatric patients is correlated with cardiovascular risk factors such as elevated blood pressure, dyslipidemia, and type 2 diabetes mellitus, all of which are significant risk factors for adult disease. In addition, current treatment and prevention strategies, including lifestyle modifications, pharmacologic agents, and certain surgical therapies, are highlighted. The need for collaborative changes at the family, school, city, state, and national levels to address the growing prevalence of the metabolic syndrome in the pediatric age group is also reviewed.


[Investigation of Pneumocystis jirovecii pneumonia and colonization in iatrogenically immunosuppressed and immunocompetent patients].
Pneumocystis pneumonia (PCP) is a potentially life-threatening infection for the immunocompromized patients. However, Pneumocystis jirovecii colonization can also be detected in healthy individuals and in patients with various underlying lung diseases. The aim of this study was to evaluate the immunocompetent and iatrogenically immunosuppressed patients in terms of PCP and P.jirovecii colonization. A total of 92 patients (66 male, 26 female; age range: 18-93 years, median: 58.5) who underwent bronchoscopy due to various pulmonary symptoms between January 2011-April 2014, were included in the study. Of these patients, 65 were under immunosuppressive therapy (38 were treated with anti-cancer drugs, 15 with anti-rejection/immunomodulatory drugs and 12 with corticosteroids), while 27 were immunocompetent. Bronchoalveolar lavage (BAL) fluids were evaluated for the presence of P.jirovecii mitochondrial gene coding ribosomal large subunit (mtLSUrRNA) with nested PCR (nPCR) method. All of the samples were also examined by Giemsa and Gomori's methenamine silver (GMG) staining methods. P.jirovecii DNA was detected in 31 (33.7%) out of 92 BAL samples by nPCR. Although six immunosuppressed patients were positive in the first round of amplification, 26 of 65 (40%) immunosuppressed and five of 27 (18.5%) immunocompetent patients were positive with nPCR. P.jirovecii cysts and trophozoites were detected in only five (16.1%) of the 31 nPCR positive samples. The probability of being immunosuppressive among nPCR positive cases was statistically higher than nPCR negative cases (χ²= 3.940; p= 0.047). This difference was more significant in organ transplant recipients and patients under anti-rejection/immunomodulatory treatment (χ²= 6.715, p= 0.01; χ²= 5.550, p= 0.018, respectively). When clinical, laboratory and radiological findings of nPCR positive patients were considered, five patients (2 kidney transplant, 1 bone marrow transplant, 1 interstitial lung disease and 1 lung cancer case) in immunosuppressed group were interpreted as "definite PCP" and eight patients (2 kidney transplant, 1 leukemia, 1 connective tissue disease, 1 Wegener's granulomatosus, 2 rheumatoid arthritis and 1 lung cancer case) were interpreted as "probable PCP". Other 18 (19.6%) nPCR positive patients, of them 13 were immunosuppressive and five were immunocompetent, were considered as "P.jirovecii colonization". The colonization rate was determined as 50% (13/26) in immunosuppressive patients, and was mostly detected in patients with hematological malignancies (4/13), followed by patients with solid tumors (3/13) and organ transplantations (3/13). On the other hand, all of the nPCR positive immunocompetent patients (5/5) were evaluated as colonization. In this study significant data was obtained about P.jirovecii epidemiology in our country. Our results also showed that iatrogenically immunosuppressed patients are under risk of PCP and nPCR method is more sensitive than conventional PCR and classical staining methods in the diagnosis of these patients.


Relation of serum cholesterol, lipid, serotonin and tryptophan levels to severity of depression and to suicide attempts.
To determine if there is a relation to low serum cholesterol, lipoprotein, serotonin or tryptophan levels in patients with depression who have recently attempted suicide. Biochemical and behavioural study. Inpatient and outpatient treatment at the Instituto Mexicano de Psiquiatría. Thirty-three patients with a diagnosis of major depressive episode. Eighteen of these patients had attempted suicide in the month before the start of the study; 15 patients had never attempted suicide. Serum levels of total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides, serotonin (5-HT) and tryptophan. Scores on Hamilton Depression Rating Scale, Carroll Depression Rating Scale, Beck Hopelessness Scale and Beck Suicide Attempt Severity Scale. There were no significant differences between patients who had attempted suicide and those who had not in terms of serum cholesterol, HDL, LDL and triglyceride levels. Serum levels of 5-HT and tryptophan were significantly lower in patients with depression who had a recent suicide attempt than in those patients who had never attempted suicide. A comparison of patients not taking antidepressant medication found serum 5-HT levels to be more than 3 times lower in those patients with a recent suicide attempt than in patients with no history of suicide attempt. The study found no difference in lipid profiles between patients who had attempted suicide and those who had not. Low serum levels of 5-HT may increase the risk of suicide attempt in patients who are depressed.


Combining physiological threshold knowledge to species distribution models is key to improving forecasts of the future niche for macroalgae.
Species distribution models (SDM) are a useful tool for predicting species range shifts in response to global warming. However, they do not explore the mechanisms underlying biological processes, making it difficult to predict shifts outside the environmental gradient where the model was trained. In this study, we combine correlative SDMs and knowledge on physiological limits to provide more robust predictions. The thermal thresholds obtained in growth and survival experiments were used as proxies of the fundamental niches of two foundational marine macrophytes. The geographic projections of these species' distributions obtained using these thresholds and existing SDMs were similar in areas where the species are either absent-rare or frequent and where their potential and realized niches match, reaching consensus predictions. The cold-temperate foundational seaweed Himanthalia elongata was predicted to become extinct at its southern limit in northern Spain in response to global warming, whereas the occupancy of southern-lusitanic Bifurcaria bifurcata was expected to increase. Combined approaches such as this one may also highlight geographic areas where models disagree potentially due to biotic factors. Physiological thresholds alone tended to over-predict species prevalence, as they cannot identify absences in climatic conditions within the species' range of physiological tolerance or at the optima. Although SDMs tended to have higher sensitivity than threshold models, they may include regressions that do not reflect causal mechanisms, constraining their predictive power. We present a simple example of how combining correlative and mechanistic knowledge provides a rapid way to gain insight into a species' niche resulting in consistent predictions and highlighting potential sources of uncertainty in forecasted responses to climate change.


The N-terminal domain of human DNA helicase Rtel1 contains a redox active iron-sulfur cluster.
Human telomere length regulator Rtel1 is a superfamily II DNA helicase and is essential for maintaining proper length of telomeres in chromosomes. Here we report that the N-terminal domain of human Rtel1 (RtelN) expressed in Escherichia coli cells produces a protein that contains a redox active iron-sulfur cluster with the redox midpoint potential of -248 ± 10 mV (pH 8.0). The iron-sulfur cluster in RtelN is sensitive to hydrogen peroxide and nitric oxide, indicating that reactive oxygen/nitrogen species may modulate the DNA helicase activity of Rtel1 via modification of its iron-sulfur cluster. Purified RtelN retains a weak binding affinity for the single-stranded (ss) and double-stranded (ds) DNA in vitro. However, modification of the iron-sulfur cluster by hydrogen peroxide or nitric oxide does not significantly affect the DNA binding activity of RtelN, suggesting that the iron-sulfur cluster is not directly involved in the DNA interaction in the N-terminal domain of Rtel1.


Reactive High-Valent Iron Intermediates in Enhancing Treatment of Water by Ferrate.
Efforts are being made to tune the reactivity of the tetraoxy anion of iron in the +6 oxidation state (FeVIO42-), commonly called ferrate, to further enhance its applications in various environmental fields. This review critically examines the strategies to generate highly reactive high-valent iron intermediates, FeVO43- (FeV) and FeIVO44- or FeIVO32- (FeIV) species, from FeVIO42-, for the treatment of polluted water with greater efficiency. Approaches to produce FeV and FeIV species from FeVIO42- include additions of acid (e.g., HCl), metal ions (e.g., Fe(III)), and reductants (R). Details on applying various inorganic reductants (R) to generate FeV and FeIV from FeVIO42- via initial single electron-transfer (SET) and oxygen-atom transfer (OAT) to oxidize recalcitrant pollutants are presented. The common constituents of urine (e.g., carbonate, ammonia, and creatinine) and different solids (e.g., silica and hydrochar) were found to accelerate the oxidation of pharmaceuticals by FeVIO42-, with potential mechanisms provided. The challenges of providing direct evidence of the formation of FeV/FeIV species are discussed. Kinetic modeling and density functional theory (DFT) calculations provide opportunities to distinguish between the two intermediates (i.e., FeIV and FeV) in order to enhance oxidation reactions utilizing FeVIO42-. Further mechanistic elucidation of activated ferrate systems is vital to achieve high efficiency in oxidizing emerging pollutants in various aqueous streams.


Malnutrition in patients with COVID-19: assessment and consequences.
COVID-19 disease often presents with malnutrition and nutrition impact symptoms, such as reduced appetite, nausea and loss of taste. This review summarizes the most up-to-date research on nutritional assessment in relation to mortality and morbidity risk in patients with COVID-19. Numerous studies have been published on malnutrition, muscle wasting, obesity, and nutrition impact symptoms associated with COVID-19, mostly observational and in hospitalized patients. These studies have shown a high prevalence of symptoms (loss of appetite, nausea, vomiting, diarrhea, dysphagia, fatigue, and loss of smell and taste), malnutrition, micronutrient deficiencies and obesity in patients with COVID-19, all of which were associated with increased mortality and morbidity risks. Early screening and assessment of malnutrition, muscle wasting, obesity, nutrition impact symptoms and micronutrient status in patients with COVID-19, followed by pro-active nutrition support is warranted, and expected to contribute to improved recovery. There is limited research on nutritional status or nutrition impact symptoms in patients living at home or in residential care. RCTs studying the effects of nutrition intervention on clinical outcomes are lacking. Future research should focus on these evidence gaps.


Effect of alkalinisation of lignocaine for propofol injection pain: a prospective, randomised, double-blind study.
The aim of this study was to determine whether pretreatment with alkalinised lignocaine reduced the incidence and severity of pain during propofol injection. This prospective, randomised, double-blind study included 300 adult, American Society of Anesthesiologists physcial status I to II patients undergoing elective surgery. Patients were randomly allocated to one of three groups: Group L received 0.05 ml/kg of 1% lignocaine (5 ml normal saline + 5 ml 2% lignocaine), Group A received 0.05 ml/kg alkalinised lignocaine (5 ml 2% lignocaine + 1 ml 8.4% NaHCO3 + 4 ml normal saline), and Group S, the control group, was given the same amount of normal saline (NaCl 0.9%). All drugs were given as a bolus over 20 seconds before propofol administration. A blinded researcher assessed the patient's pain level using a four-point scale. The pain score [median (range)] and the incidence of pain in Group A (6%) was significantly lower than in groups L (41%) and S (88%, P <0.001). In addition, the pain score and the incidence of pain were found to be significantly different between Group L and Group S (P <0.001). The incidence of moderate and severe pain were greater in Group S when compared with groups A and L (P <0.001). Intravenous pretreatment with alkalinised lignocaine appears to be effective in reducing the pain during propofol injection.


Postoperative autologous blood transfusion drain or no drain in primary total hip arthroplasty? A randomised controlled trial.
Postoperative maintenance of high haemoglobin (Hb) levels and avoidance of homologous blood transfusions is important in total hip arthroplasty (THA). The introduction of a postoperative drainage autologous blood transfusion (ABT) system or no drainage following THA has resulted in reduction of homologous blood transfusion requirements compared with closed-suction drains. The purpose of this study was to examine which regimen is superior following THA. A randomised controlled blinded prospective single-centre study was conducted in which 100 THA patients were randomly allocated to ABT or no drainage. The primary endpoint was the Hb level on the first postoperative day. The postoperative collected drained blood loss was 274 (±154) ml in the ABT group, of which 129 (±119) ml was retransfused (0-400). There was no statistical difference in Hb levels on the first postoperative day (ABT vs no drainage: Hb 11.0 vs 10.9 g/dl), on consecutive days (day 3: Hb 10.7 vs 10.2, p = 0.08) or in total blood loss (1,506 vs 1,633 ml), homologous transfusions, pain scores, Harris Hip Score, SF-36 scores, length of hospital stay or adverse events. The use of a postoperative autologous blood retransfusion drain did not result in significantly higher postoperative Hb levels or in less total blood loss or fewer homologous blood transfusions compared with no drain.


Mismatch between BDNF mRNA and protein expression in the developing visual cortex: the role of visual experience.
Brain-derived neurotrophic factor (BDNF) messenger RNA (mRNA) expression in the rat visual cortex of young and postnatal day 90 (P90) animals is developmentally regulated and influenced by visual experience. In the present paper we compared the expression of BDNF mRNA to the actual changes of BDNF protein occurring during postnatal development and verified whether BDNF protein distribution is controlled by visual activity. To achieve this aim we analysed BDNF mRNA and/or BDNF protein cellular distribution in the rat visual cortex at different postnatal ages by using immunohistochemistry and highly sensitive in situ hybridization. We found that before eye opening (P13), in all cortical layers a large number of visual cortical neurons contain BDNF mRNA with no detectable amount of BDNF protein. At later ages (P23 and P90), the number of BDNF-immunostained cells increases; most neurons are double labelled for BDNF mRNA and protein, and a small group of neurons is labelled only for BDNF protein. The cellular increase of BDNF immunolabelling is blocked in animals deprived of visual experience from birth (dark rearing), with a large population of neurons containing BDNF mRNA but not BDNF protein. This is similar to what is observed before eye opening. Exposure of dark-reared rats to a brief period (2 h) of light restores a good match between BDNF mRNA and BDNF protein cellular expression. We propose that visual experience controls the neuronal content of BDNF mRNA and BDNF protein in developing visual cortex.


Screening Tumor-Related Genes of Gallbladder Cancer Based on AR-Based Tumor Expression Profile Gene Chip.
The rapid development of molecular biology and gene chip technology has produced a large amount of gene expression profile data. The main research in this article is to screen the tumor-related genes of gallbladder cancer based on AR-based tumor expression profile gene chip. First, convert the chip data into an expression matrix pattern that can be analyzed, and then standardize and normalize all the data. Run ReliefF, GA, and IReliefF-GA on the data set, record the size of the feature subset, and use the tenfold cross-validation method to obtain the classification accuracy, specificity, and sensitivity of each method on the classifier. The target genes used in the chip were amplified by PCR with the universal primers used in cDNA library construction, and the quality of PCR was monitored by agarose gel electrophoresis. The gene chip data of gallbladder cancer was processed with missing values, singular values, and so forth, and 22294 transcripts were obtained. After statistical testing, there were 9483 transcripts with statistically significant differences. The results show that as the number of clusters increases, the network can be better reconstructed through decomposition modeling.


Root caries prevalence in a group of Brazilian adult dental patients.
The aim of this study was to measure the caries prevalence in the root surface in a group of Brazilian adults. The prevalence and their intraoral distribution of caries lesions of root surface were assessed in dental patients ranging in age from 35 to 44 and from 50 to 59 years of age of both sexes (n=360). A total of 98.9% had root surfaces with gingival recession and 78.1% had at least one root caries lesion. The prevalence of the disease was analyzed using the Root Caries Index (RCI). The average value was greater for women (18.6%) than for men (13.4%) (p<0.01) and similar in both age groups (p>0.01). The maxillary canines and first premolars and the mandibular molars presented the greatest RCI values. The RCI was greater in the proximal surface of the maxilla and buccal surface of the mandible. We conclude that caries of the root surface are present in the Brazilian population and deserve attention concerning their actual role in the epidemiology of principal oral diseases of the adult population.


High-resolution SNP scan of chromosome 6p21 in pooled samples from patients with complex diseases.
We apply a high-throughput protocol of chip-based mass spectrometry (matrix-assisted laser desorption/ionization time-of-flight; MALDI-TOF) as a method of screening for differences in single-nucleotide polymorphism (SNP) allele frequencies. Using pooled DNA from individuals with asthma, Crohn's disease (CD), schizophrenia, type 1 diabetes (T1D), and controls, we selected 534 SNPs from an initial set of 1435 SNPs spanning a 25-Mb region on chromosome 6p21. The standard deviations of measurements of time of flight at different dots, from different PCRs, and from different pools indicate reliable results on each analysis step. In 90% of the disease-control comparisons we found allelic differences of <10%. Of the T1D samples, which served as a positive control, 10 SNPs with significant differences were observed after taking into account multiple testing. Of these 10 SNPs, 5 are located between DQB1 and DRB1, confirming the known association with the DR3 and DR4 haplotypes whereas two additional SNPs also reproduced known associations of T1D with DOB and LTA. In the CD pool also, two earlier described associations were found with SNPs close to DRB1 and MICA. Additional associations were found in the schizophrenia and asthma pools. They should be confirmed in individual samples or can be used to develop further quality criteria for accepting true differences between pools. The determination of SNP allele frequencies in pooled DNA appears to be of value in assigning further genotyping priorities also in large linkage regions.


The natural history of sclerosteosis.
Sclerosteosis (SCL) is a severe, progressive, autosomal-recessive craniotubular hyperostosis (MIM 269500). The determinant gene (SOST) has been isolated, and genotype-phenotype correlations, as well as the elucidation of pathogenetic mechanisms, are dependent upon the documentation of the natural history of the condition. For this reason, the course and complications in 63 affected individuals in South Africa, seen over a 38-year period, have been analyzed. Thirty-four of these persons died during the course of the survey, 24 from complications related to elevation of intracranial pressure as a result of calvarial overgrowth. The mean age of death in this group of individuals was 33 years, with an even gender distribution. Facial palsy and deafness, as a result of cranial nerve entrapment, developed in childhood in 52 (82%) affected persons. Mandibular overgrowth was present in 46 (73%) adults and syndactyly in 48 (76%). In South Africa in 2002, 29 affected persons were alive, 10 being < or =20 years of age. It is evident that sclerosteosis is a severe disorder which places a considerable burden upon affected individuals and their families.


Fludarabine-based induction therapy does not overcome the negative effect of ABCG2 (BCRP) over-expression in adult acute myeloid leukemia patients.
Over-expression of multidrug resistance (MDR) proteins PGP and BCRP has a negative prognostic impact in acute myeloid leukemia (AML) patients. Inclusion of fludarabine in induction chemotherapy increases remission rate in PGP over-expressing cases. We investigated the role of BCRP in 138 adult AML patients receiving induction therapy with fludarabine. None of the MDR-related proteins influenced complete remission attainment. Conversely, high levels of BCRP significantly affected disease-free survival, as higher relapse rates (48.5% vs 28.5%) and earlier relapse occurred in BCRP+ patients. Also overall survival was affected by BCRP positivity, and survival significantly worsened in case of concomitant PGP and BCRP over-expression.


Direct radiation damage to crystalline DNA: what is the source of unaltered base release?
The radiation chemical yields of unaltered base release have been measured in three crystalline double-stranded DNA oligomers after X irradiation at 4 K. The yields of released bases are between 10 and 20% of the total free radical yields measured at 4 K. Using these numbers, we estimate that the yield of DNA strand breaks due to the direct effect is about 0.1 micromol J(-1). The damage responsible for base release is independent of the base type (C, G, A or T) and is not scavenged by anthracycline drugs intercalated in the DNA. For these reasons, reactions initiated by the hydroxyl radical have been ruled out as the source of base release. Since the intercalated anthracycline scavenges electrons and holes completely but does not inhibit base release, the possibility for damage transfer from the bases to the sugars can also be ruled out. The results are consistent with a model in which primary radical cations formed directly on the sugar-phosphate backbone react by two competing pathways: deprotonation, which localizes the damage on the sugar, and hole tunneling, which transfers the damage to the base stack. Quantitative estimates indicate that these two processes are approximately equally efficient.


Chemometric Exploration of APPI(+)-FT-ICR MS Data Sets for a Comprehensive Study of Aromatic Sulfur Compounds in Gas Oils.
Sulfur content in gas oils is strictly regulated by legal specifications for environmental reasons. Gas oils are composed of various aromatic sulfur compounds, and some of them are known to be very refractory for sulfur removal reactions. Thus, an accurate analysis of sulfur compounds is important to find the appropriate operating conditions of the gas oil hydrotreating processes. Aromatic sulfur compounds contained in 23 gas oils samples were analyzed using APPI(+)-FT-ICR MS considering six replicates. Significant differences were spotted within several processed gas oils. A comparison of one feed and its corresponding effluents also confirmed the well-known refractory character of sulfur compounds such as polyalkylated dibenzothiophenes. To go deeper in the molecular exploration, chemometric tools were applied on this spectral data set including principal component analysis (PCA) and hierarchical cluster analysis (HCA). A unique data rearrangement was performed directly inspired on DBE vs carbon number plots that are systematically used in petroleomics studies. Then, these chemometric tools provided a successful classification of each type of gas oils. The PCA model has also been validated on mixed blends allowing us to conclude that it could be applied to unknown samples in order to identify the process used to produce them. Moreover, the exploration of the generated loadings revealed key types of molecules driving the classification such as C3-DBT which is a dibenzothiophene core with three additional carbon atoms. Indeed, it is known to remain mainly in deeply hydrotreated samples, validating previous observations regarding its potential refractory character. The ability of chemometric tools to extract specific molecular information from ultra-high resolution MS spectra reveals its huge potential for an exhaustive study of highly complex mixtures such as crude oils.


Context-specific emergence and growth of the SARS-CoV-2 Delta variant.
The SARS-CoV-2 Delta (Pango lineage B.1.617.2) variant of concern spread globally, causing resurgences of COVID-19 worldwide1,2. The emergence of the Delta variant in the UK occurred on the background of a heterogeneous landscape of immunity and relaxation of non-pharmaceutical interventions. Here we analyse 52,992 SARS-CoV-2 genomes from England together with 93,649 genomes from the rest of the world to reconstruct the emergence of Delta and quantify its introduction to and regional dissemination across England in the context of changing travel and social restrictions. Using analysis of human movement, contact tracing and virus genomic data, we find that the geographic focus of the expansion of Delta shifted from India to a more global pattern in early May 2021. In England, Delta lineages were introduced more than 1,000 times and spread nationally as non-pharmaceutical interventions were relaxed. We find that hotel quarantine for travellers reduced onward transmission from importations; however, the transmission chains that later dominated the Delta wave in England were seeded before travel restrictions were introduced. Increasing inter-regional travel within England drove the nationwide dissemination of Delta, with some cities receiving more than 2,000 observable lineage introductions from elsewhere. Subsequently, increased levels of local population mixing-and not the number of importations-were associated with the faster relative spread of Delta. The invasion dynamics of Delta depended on spatial heterogeneity in contact patterns, and our findings will inform optimal spatial interventions to reduce the transmission of current and future variants of concern, such as Omicron (Pango lineage B.1.1.529).


An automated image processing method to quantify collagen fibre organization within cutaneous scar tissue.
Standard approaches to evaluate scar formation within histological sections rely on qualitative evaluations and scoring, which limits our understanding of the remodelling process. We have recently developed an image analysis technique for the rapid quantification of fibre alignment at each pixel location. The goal of this study was to evaluate its application for quantitatively mapping scar formation in histological sections of cutaneous burns. To this end, we utilized directional statistics to define maps of fibre density and directional variance from Masson's trichrome-stained sections for quantifying changes in collagen organization during scar remodelling. Significant increases in collagen fibre density are detectable soon after burn injury in a rat model. Decreased fibre directional variance in the scar was also detectable between 3 weeks and 6 months after injury, indicating increasing fibre alignment. This automated analysis of fibre organization can provide objective surrogate endpoints for evaluating cutaneous wound repair and regeneration.


Actin binding domain of filamin distinguishes posterior from anterior actin filaments in migrating Dictyostelium cells.
Actin filaments in different parts of a cell interact with specific actin binding proteins (ABPs) and perform different functions in a spatially regulated manner. However, the mechanisms of those spatially-defined interactions have not been fully elucidated. If the structures of actin filaments differ in different parts of a cell, as suggested by previous in vitro structural studies, ABPs may distinguish these structural differences and interact with specific actin filaments in the cell. To test this hypothesis, we followed the translocation of the actin binding domain of filamin (ABDFLN) fused with photoswitchable fluorescent protein (mKikGR) in polarized Dictyostelium cells. When ABDFLN-mKikGR was photoswitched in the middle of a polarized cell, photoswitched ABDFLN-mKikGR rapidly translocated to the rear of the cell, even though actin filaments were abundant in the front. The speed of translocation (>3 μm/s) was much faster than that of the retrograde flow of cortical actin filaments. Rapid translocation of ABDFLN-mKikGR to the rear occurred normally in cells lacking GAPA, the only protein, other than actin, known to bind ABDFLN. We suggest that ABDFLN recognizes a certain feature of actin filaments in the rear of the cell and selectively binds to them, contributing to the posterior localization of filamin.


How to handle genetic information: a comparison of attitudes among patients and the general population.
So far there are no studies comparing the attitudes of patients with hereditary conditions to the attitudes of the general public on how to handle genetic risk information which mutation carriers refuse to disclose to relevant family members. The purpose of the present study was to investigate whether such patients and members of the general public want to be informed about the existence of hereditary conditions within their family, and under which conditions they want healthcare providers to breach confidentiality. It was hypothesized that the desire to be informed would be influenced by characteristics of both the disease and the individual. Systematically varying 3 disease characteristics (fatality, penetrance and treatment availability) yielded 8 versions of a questionnaire, which was administered to general population samples in Norway and Sweden (N = 3,207) and to patient samples in both countries (N = 822). Individual differences in uncertainty avoidance, coping style and consideration for future consequences were also assessed. A majority of both patients and the general public want to be informed about the existence of hereditary conditions within their family. However, patients are more positive towards being informed, both with and without the relative's consent, than the general public. The main predictor of the desire to be informed was uncertainty avoidance in both samples.


Biological clocks and regulation of seasonal reproduction and migration in birds.
Timekeeping is important at two levels: to time changes in physiology and behavior within each day and within each year. For the former, birds have a system of at least three independent circadian clocks present in the retina of the eyes, the pineal gland, and the hypothalamus. This differs from the situation in mammals in which the input, pacemaker, and output are localized in different structures. Each bird clock interacts with at least one other clock, and together, they appear to form a centralized clock system that keeps daily time. These clocks have a powerful endogenous component, and the daily light-dark cycle entrains them to 24 h. The timing and duration of life history stages that make up annual cycle of an individual must also be controlled by some form of timekeeping. However, evidence for the existence of an equivalent endogenous circannual clock is less clear. Environmental cues, particularly photoperiod, appear to have a more direct role than simply entraining the clock to calendar time. For example, the timing of migration is probably greatly influenced by photoperiod, but its manifestation each day, as Zugunruhe, appears to be under circadian control. Migration involves marked changes in physiology to cope with the energetic demands. There is still much that we do not know about how organisms' timekeeping systems respond to their natural environment, particularly how salient signals from the environment are perceived and then transduced into appropriately timed biological functions. However, given that changes in environmental input affects the clock, increasing human disturbance of the environment is likely to adversely affect these systems.


Functional connectivity analysis and prediction of cognitive change after carotid artery stenting.
The neurocognitive course of patients who have undergone cerebral revascularization has been the subject of many studies, and the reported effects of carotid artery stenting (CAS) on cognitive function have varied from study to study. The authors hypothesized that cognitive amelioration after CAS is associated with alteration of the default mode network (DMN) connectivity, and they investigated the correlation between functional connectivity (FC) of the DMN and post-CAS changes in cognitive function in order to find a clinical marker that can be used to predict the effect of cerebral revascularization on patients' cognitive function in this preliminary exploratory study. The authors examined post-CAS changes in cognitive function in relation to FC in patients treated for unilateral carotid artery stenosis. Resting-state functional MRI (rs-fMRI) was performed with a 3-T scanner before and 6 months after CAS in 8 patients. Neuropsychological tests (Wechsler Adult Intelligence Scale III and Wechsler Memory Scale-Revised) were administered to each patient before and 6 months after CAS. The DMN was mapped for each patient through independent component analysis of the rs-fMR images, and the correlation between FC of the DMN and post-CAS change in cognitive function was analyzed on a voxel level. Multivariable regression analysis was performed to identify preoperative factors associated with a post-CAS change in cognitive function. Post-CAS cognitive function varied between patients and between categories of neuropsychological tests. Although there was no significant overall improvement in Working Memory scores after CAS, post-CAS Working Memory scores changed in negative correlation with changes in FC between the DMN and the precentral/superior frontal gyrus and between the DMN and the middle frontal gyrus. In addition, the preoperative FC between those areas correlated positively with the post-CAS improvement in working memory. FC between the DMN and working memory-related areas is closely associated with improvement in working memory after CAS. Preoperative analysis of FC of the DMN may be useful for predicting postoperative improvement in the working memory of patients being treated for unilateral stenosis of the extracranial internal carotid artery.Clinical trial registration no.: UMIN000020045 (www.umin.ac.jp/ctr/index.htm).


CPR decision-making conversations in the UK: an integrative review.
Do Not Attempt Cardio-Pulmonary Resuscitation (DNACPR) discussions with patients and their caregivers have been subjected to intense ethical and legal debate in recent years. Legal cases and national guidelines have tried to clarify the best approach to DNACPR discussions; however, there is little evidence of how best to approach them from the patient, family or caregiver perspective. This paper describes published accounts of patient, family and caregiver experiences of discussions about advance cardiopulmonary resuscitation (CPR) decision making. An integrative review of the UK literature between 2000 and 2016 including qualitative and quantitative studies was conducted. Worldwide, 773 abstracts were identified, and 20 papers from the UK were included in the final analysis. Patient, family and caregivers prefer discussions to be initiated by someone trusted, and wishes for family involvement vary depending on the context. Timing of discussions should be individualised, though discussions earlier in the illness are often preferable. Discussions held in the acute setting are suboptimal. CPR decisions should be part of a wider discussion about future care and adequate communication skills training is important. The findings of this review are at odds with the current statutory framework and potentially challenging for medical professionals who are working in a stretched health service, with pressure to discuss DNACPR decisions at the earliest opportunity. With increasing focus on person-centred care and realistic medicine, patient narratives must be considered by doctors and policy makers alike, to minimise harm.


Thermostability of native and pegylated Myceliophthora thermophila laccase in aqueous and mixed solvents.
A commercial preparation of laccase (EC 126.96.36.199), cloned from Myceliophthora thermophila and expressed in Aspergillus oryzae (MtL), was purified and modified by conjugation with poly(ethylene glycol) (M(r) = 5000) and is labeled PEG-MtL. Native enzyme was found to have a molecular mass of 80 kDa, as determined by gel filtration, and 110 kDa, by SDS-PAGE. The oxidative dimerization of 2,6-dimethoxyphenol (DMP) to produce the corresponding dibenzoquinone was catalyzed by MtL in a manner comparable to that for a diffusion-controlled reaction (k(cat)/K(M) approximately = 10(8) M(-)(1) s(-)(1) and E(a) approximately = 18 kJ M(-)(1)). PEG-MtL was found, by TNBS titration, to have blocked 54% of lysine groups; its hydrodynamic and charge properties were different from those of MtL. Catalytic efficiency (k(cat)/K(M)) of PEG-MtL was similar to that of MtL with DMP as substrate; however, k(cat)/K(M) was 2-fold reduced for the reaction in which 2',2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) is oxidized to form a radical cation. E(a) values were similar in both enzyme preparations when assayed in buffered solutions. Far-UV CD spectra were similar for MtL and PEG-MtL and consistent with a protein rich in beta-sheet structure with negligible content of alpha-helices. A blue shift of near-UV CD spectrum for PEG-MtL as compared to MtL was consistent with the decreased polarity of the tyrosyl side chains upon PEG conjugation. Also the blue band of the copper active site was shifted from lambda approximately 610 nm (MtL) to lambda approximately 575 nm (PEG-MtL). Scanning microcalorimetry showed small denaturation enthalpies (6.3 and 7.5 J g(-)(1) for MtL and PEG-MtL, respectively), indicating the high stability of the beta-sheet folding pattern of laccases. However, PEG-MtL proved to be more stable, its half-denaturation temperature being 2 degrees C higher than that of MtL. In 30% alcohol, pegylated laccase showed slower enzyme-activity decay rates than the unmodified enzyme; this behavior was caused by a decrease in the activation entropy of the denaturation reaction. Results can be explained by entropic stabilization by PEG conjugation because of the restricted motion of some surface amino acid side chains, which results in a more stable active site.


Monitoring the formation and decay of transient photosensitized intermediates using pump-probe UV resonance Raman spectroscopy. II: Kinetic modeling and multidimensional least-squares analysis.
Analysis of transient excited-state Raman spectra is a challenging spectroscopic measurement since transient spectral features are often overlapped with dominant ground-state and solvent bands. In the previous manuscript, resolution of component Raman spectra from the time-resolved amine quenching of excited-triplet benzophenone was accomplished using self-modeling curve resolution, a model-free factor analysis technique that relies on correlation in the data along a changing composition dimension. The results are consistent with the production of diphenylketyl radicals by H-atom abstraction from the amine and subsequent free-radical decay by recombination reactions. A kinetic model for this chemistry is developed in the present work, based on the observed Raman scattering data and the structures of product species confirmed by mass spectral analysis. The model is applied to the analysis of the time-dependent Raman scattering data using multidimensional least-squares methods, and it yielded well-resolved spectra of benzophenone excited-triplet states, diphenyl ketyl radical, and the solvent and ground-state precursors. The best-fit kinetic parameters agree well with the time-dependent triplet-state and ketyl-radical concentration profiles.


Optimizing Sowing and Flooding Depth for Anaerobic Germination-Tolerant Genotypes to Enhance Crop Establishment, Early Growth, and Weed Management in Dry-Seeded Rice (Oryza sativa L.).
Poor crop establishment, high weed infestation, and consequent yield loss are major concerns for dry-seeded rice (DSR). Flooding after seeding helps in managing weeds but reduces seed germination and crop stand. Anaerobic germination (AG)-tolerant rice genotypes could overcome these problems in DSR. Screenhouse experiments were established to evaluate the effect of seed sowing depth (SD) (0.5 cm, 1 cm, and 2 cm) and flooding depth (FD) (saturated, 2 cm, and 5 cm) on crop establishment, early growth, and weed competitiveness in DSR using AG-tolerant genotypes (Khao Hlan On, Ma-Zhan Red, IR64+AG1, and IR64). Echinochloa crus-galli, Ludwigia hyssopifolia, and Cyperus difformis were used in the weedy treatment. Rice plants reached maximum emergence 9-13 days later under flooding compared with saturated conditions. Crop emergence decreased by 12-22% at 0.5 and 1 cm SD and by 48-60% at 2 cm SD, when combined with 2 or 5 cm FD compared with saturated conditions. The 2 cm SD reduced seedling emergence by 23-42% in Khao Hlan On and Ma-Zhan Red, by 62-70% in IR64+AG1, and by 90-92% in IR64 under flooding. Initial growth in rice plant height was slow under flooding but increased progressively after the seedlings emerged from water and the final height was not affected by FD. Leaf area, total shoot biomass, tiller density, and leaf number per pot of rice were higher at 1 cm SD (P< 0.05), but decreased drastically at 2 cm SD under flooding. The emergence of E. crus-galli and L. hyssopifolia decreased by 53-65% and 89-95%, respectively, but increased by 49-68% in C. difformis under 2 and 5 cm FD, respectively, compared with saturated conditions. The shoot biomass of the weeds followed the same trend. Khao Hlan On showed the highest weed-competitive ability under all FD while the biomass of IR64+AG1 and IR64 decreased by 10-14% due to weed competition under 2 cm FD. The 1 cm SD showed better growth for all genotypes under different FD. The 2 cm FD is sufficient to have a significant control of problematic weed species. The tolerance of AG of rice genotypes should be further enhanced to increase their weed-competitive ability.


Interleukin-1beta enhanced action of kinins on extracellular matrix of spontaneous hypertensive rat cardiac fibroblasts.
Interaction between an enhanced action of kinins and cytokines is accepted as important to the cardioprotective effect of angiotensin-converting-enzyme inhibitors. Kinins mediate their effects through B1 and B2 subtype receptors that may be modulated by cytokines including interleukin (IL)-1beta. We examined expression of kinin receptors and the effects of bradykinin (B2 agonist) and des-Arg10-kallidin (B1 agonist) on extracellular matrix components of adult rat cardiac fibroblasts with or without prior exposure to IL-1beta. We compared responses of cells cultured from spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) hearts. mRNA levels of kinin receptors, procollagens, promatrix metalloproteinases (proMMP-2 and proMMP-9), and tissue inhibitors of metalloproteinase (TIMP-1 and TIMP-2) were all assessed by a semiquantitative RT-PCR. In the absence of IL-1beta, SHR cells expressed more B2 receptor, procollagen alpha1(I), procollagen alpha1(III), and proMMP-9 mRNA than WKY cells. IL-1beta exposure enhanced B1, B2, proMMP-2, and proMMP-9 mRNA in cells of both strains to equivalent levels. Zymographic studies confirmed the results of proMMPs. Following IL-1beta treatment, bradykinin attenuated procollagens alpha1(I) and alpha1(III) mRNA expression in SHR but not WKY cells. In contrast, des-Arg10-kallidin did not show any significant effects in either SHR or WKY cells. Our findings indicate greater extracellular matrix turnover in cultured SHR cardiac fibroblasts than WKY under basal conditions, an IL-1beta stimulation of turnover in cells from both strains, and a strain-differential effect of bradykinin following cytokine treatment. These results imply a genetically determined response of cardiac extracellular matrix and the potential of direct enhancement of the efficacy of kinins by the local release of IL-1beta in hearts genetically programmed to exhibit excessive remodeling to injury.