In vitro messenger properties of a satellite RNA of cucumber mosaic virus.
The nucleotide sequence of a naturally occurring satellite RNA (satRNA) of cucumber mosaic virus strain F (CMV-F) isolated from Petasites japonicum Miq. has been determined. F-satRNA is 338 nucleotides long and possesses a single open reading frame (ORF; residues 11 to 91) in its 5-terminal region. Analysis of in vitro translation products of the RNA revealed that the major peptide synthesized had an Mr of 2800, corresponding to the value (Mr 2874) calculated from the amino acid sequence predicted from the ORF. Furthermore, a ribosome-binding fragment of the RNA in the initiation step of an in vitro translation system was found to contain the first AUG codon (residues 11 to 13). These results suggest that of the F-satRNA sequence, the coding region for the peptide can be assigned to the region from AUG to GGU (residues 11 to 91), accompanied by a termination codon UGA.


Autoimmune diseases associated with Neuromyelitis Optica Spectrum Disorders: A literature review.
Neuromyelitis Optica (NMO) is an autoimmune inflammatory demyelinating disease of the central nervous system (CNS) which predominantly involves optic nerves and spinal cord. Since the introduction of Neuromyelitis Optica Spectrum Disorders (NMOSD) as a separate entity, there have been many reports on its association with other disorders including systemic and organ-specific autoimmune diseases. Here, we reviewed other immune-mediated diseases associated with NMOSD and tried to categorize them. The present review was conducted using the PUBMED database based on papers from 1976 (i.e., since the first NMO comorbidity with SLE was reported) to 2017. We included all articles published in English. The keywords utilized included Neuromyelitis optica, Neuromyelitis Optica Spectrum Disorders, Devic's disease, in combination with comorbidity or comorbidities. Diseases with immune-based pathogenesis are the most frequently reported co-morbidities associated with NMOSD, most of which are antibody-mediated diseases. According to literature, Sjogren's Syndrome (SS) and Systemic Lupus Erythematosus (SLE) are the most frequently reported diseases associated with NMOSD among systemic autoimmune diseases. Further, myasthenia gravis in neurological and autoimmune thyroid diseases in non-neurological organ-specific autoimmune diseases are the most reported comorbidities associated with NMOSD in the literature. NMOSD may be associated with a variety of different types of autoimmune diseases. Therefore, systemic or laboratory signs which are not typical for NMOSD should be properly investigated to exclude other associated comorbidities. These comorbidities may affect the treatment strategy and may improve the patients' care and management.


Coexpression of pyruvate decarboxylase and alcohol dehydrogenase genes in Lactobacillus brevis.
Lactobacillus brevis ATCC367 was engineered to express pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) genes in order to increase ethanol fermentation from biomass-derived residues. First, a Gram-positive Sarcina ventriculi PDC gene (Svpdc) was introduced into L. brevis ATCC 367 to obtain L. brevis bbc03. The SvPDC was detected by immunoblot using an SvPDC oligo peptide antiserum, but no increased ethanol was detected in L. brevis bbc03. Then, an ADH gene from L. brevis (Bradh) was cloned behind the Svpdc gene that generated a pdc/adh-coupled ethanol cassette pBBC04. The pBBC04 restored anaerobic growth and conferred ethanol production of Escheirichia coli NZN111 (a fermentative defective strain incapable of growing anaerobically). Approximately 58 kDa (SvPDC) and 28 kDa (BrADH) recombinant proteins were observed in L. brevis bbc04. These results indicated that the Gram-positive ethanol production genes can be expressed in L. brevis using a Gram-positive promoter and pTRKH2 shuttle vector. This work provides evidence that expressing Gram-positive ethanol genes in pentose utilizing L. brevis will further aid manipulation of this microbe toward biomass to ethanol production.


Dyadic advance care planning: systematic review of patient-caregiver interventions and effects.
Family caregiver's involvement in advance care planning (ACP) is essential to provide high-quality end-of-life (EOL) care and to ease the surrogate decision-making burden. However, no systematic review has focused on existing ACP interventions involving patients and their families. To systematically summarise current ACP interventions involving patients and their families. Five English and two Chinese databases were searched from inception to September 2022. The eligible studies were experimental studies describing original data. The Joanna Briggs Institute critical appraisal tools assessed the methodological quality. Narrative synthesis was conducted for data analysis. In total, twenty-eight articles were included. Fifteen studies were randomised controlled trials, and the rest 13 studies were quasi-experimental studies. The data synthesis identified: (1) Key intervention components: strategies to promote ACP, ACP discussion and follow-up, as well as the role of family caregivers; (2) Effects on intended outcomes: interventions have shown benefit on completion of ACP actions, while inconsistent findings were found on the process outcomes and quality of EOL care. In addition, a logic model for patient-caregiver dyadic ACP was created, and the underlying mechanisms of action included well-preparation, open discussion and adequate support for plan/action. This review provides comprehensive evidence about patient-caregiver dyadic ACP, a promising intervention to better prepare for EOL communication and decision-making. A logic model has been mapped to give a preliminary indication for future implementation. More empirical studies are needed to improve this model and culturally adapt it in a real-world setting.


Trends and influential factors of high ecological status mobility in Irish Rivers.
The decline in high ecological water status in rivers is a significant concern in European countries. It is thus important to investigate the factors that cause sites to lose high status in order to undertake measures to protect and restore high status water quality. Analysis of 20 years of water quality data reveals strong mobility between high status and non-high status (especially good status) rivers. Associations between this mobility and socio-economic and physical environmental variables were estimated by multinomial logistic regression at national scale and regional scale. Based on reported changes in water quality status cross across 1990, 2000 and 2010, four classes of the mobility of high status were defined in this study: those sites that maintain high status (maintain), enter high status (enter), fluctuate between high and non-high status (fluctuate) and exit from high status (exit). The national results indicate that agricultural activity as indicated by variables representing intensity of livestock farming (organic nitrogen) and tillage farming (cereal share) and elevation had significant negative impacts on high status rivers. Meanwhile, significant differences in population density and septic tank density between 'exit', 'maintain', 'fluctuate' and 'enter' classes indicate that these factors played important roles in the stability of high status rivers. The regional outcomes reveal differential significant pressures across regions. For example, rainfall and elevation had positive impacts on high status rivers in the north-west region, while organic nitrogen had a negative effect in the south-west. This paper demonstrates the challenge in achieving the Water Framework Directive goal of maintaining high status rivers, given the sensitive and highly differentiated nature of areas that have lost high status or fluctuated in and out of high status. This paper also suggests the necessity for localised policies and mitigation measures.


Identification of chemical mixtures to which women are exposed through the diet: Results from the French E3N cohort.
Due to the large number of chemical food contaminants, consumers are exposed simultaneously to a wide range of chemicals which can interact and have a negative impact on health. Nevertheless, due to the multitude of possible chemical combinations it is unrealistic to test all combined toxicological effects. It is therefore essential to identify the most relevant mixtures to which the population is exposed through the diet and investigate their impact on heath. The present study aims to identify and describe the main chemical mixtures to which women enrolled in the E3N study, a large French prospective cohort, are chronically exposed through the diet. 74522 women who had answered a validated semi-quantitative food frequency questionnaire in 1993, were included in the present study. Dietary exposure to chemical contaminates was estimated based on the food contamination measured in 186 core food in France collected between 2007 and 2009 by the French agency for food, environment and occupational health, and safety (ANSES) in the framework of the second French total diet study (2TDS). The sparse non-negative matrix under-approximation (SNMU) was used to identify mixtures of chemical substances. A k-means clustering classification of the whole study population was then performed to define clusters with similar co-exposure profiles. Overall, 8 mixtures which explained 83% of the total variance, were retained. The first mixture, entitled "Minerals, inorganic contaminants, and furans", explained the highest proportion of the total variance (38%), and was correlated in particular with the consumption of "Offal" (rho = 0.22), "Vegetables except roots" (rho = 0.20), and "Eggs" (rho = 0.19). The other seven mixtures explained between 17% and 1% of the variance. Finally, 5 clusters were identified based on the adherence to the 8 mixtures. This study, being the largest ever conducted to identify dietary exposure to chemical mixtures, represents a concrete attempt to prioritize mixtures for which it is essential to investigate combined health effects based on exposure.


A Community-Embedded Implementation Model for Mental-Health Interventions: Reaching the Hardest to Reach.
The mental-health-care treatment gap remains very large in low-resource communities, both within high-income countries and globally in low- and middle-income countries. Existing approaches for disseminating psychological interventions within health systems are not working well enough, and hard-to-reach, high-risk populations are often going unreached. Alternative implementation models are needed to expand access and to address the burden of mental-health disorders and risk factors at the family and community levels. In this article, we present empirically supported implementation strategies and propose an implementation model-the community-embedded model (CEM)-that integrates these approaches and situates them within social settings. Key elements of the model include (a) embedding in an existing, community-based social setting; (b) delivering prevention and treatment in tandem; (c) using multiproblem interventions; (d) delivering through lay providers within the social setting; and (e) facilitating relationships between community settings and external systems of care. We propose integrating these elements to maximize the benefits of each to improve clinical outcomes and sustainment of interventions. A case study illustrates the application of the CEM to the delivery of a family-based prevention and treatment intervention within the social setting of religious congregations in Kenya. The discussion highlights challenges and opportunities for applying the CEM across contexts and interventions.


Facilitation of recovery from ischemic brain damage in rabbits by polymeric prostaglandin PGBx, a mitochondrial protective agent.
Administration of polymeric prostaglandin PGBx to rabbits after a 20-minute period of almost total obstruction of blood flow to the brain resulted in marked improvement of recovery from brain damage as measured by recovery of brain function. This beneficial effect of PGBx on recovery from acute ischemic tissue damage, analogous to that observed in cardiac tissue of monkeys, may be due to the positive effect of PGBx on phosphorylation in damaged mitochondria as observed previously in vitro. A convenient surgical technique for producing reversible cerebral ischemia in rabbits was developed for and used in this pharmacological study.


Overexpression of MEKK3 confers resistance to apoptosis through activation of NFkappaB.
Many cancers have constitutively activated NFkappaB, the elevation of which contributes to cancer cell resistance to chemotherapeutic agent-induced apoptosis. Although mitogen-activated protein kinase/extracellular-regulated kinase kinase kinase-3 (MEKK3) has been shown to participate in the activation of NFkappaB, its relations to apoptosis and cancer are unclear. In this study, we established cell model systems to examine whether stable expression of MEKK3 could lead to increased NFkappaB activity and confer resistance to apoptosis. In addition, we investigated in breast and ovarian cancers whether MEKK3 expression may be altered and correlated with aberrant NFkappaB activity. We show that stable cell lines overexpressing MEKK3 not only had elevated levels of NFkappaB binding activity but also were more responsive to cytokine stimulation. These stable cells showed 2-4-fold higher basal expression of Bcl-2 and xIAP than the parental cells. Consistent with this increased expression of cell survival genes, MEKK3 stable cells showed reduced activation of caspases 3 and 8 and poly(ADP-ribose) polymerase cleavage and dramatically increased resistance to apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand, doxorubicin, daunorubicin, camptothecin, and paclitaxel. Intriguingly, analysis of human breast and ovarian cancers showed that a significant fraction of these samples have elevated MEKK3 protein levels with corresponding increases in NFkappaB binding activities. Thus, our results established that elevated expression of MEKK3 appears to be a frequent occurrence in breast and ovarian cancers and that overexpression of MEKK3 in cells leads to increased NFkappaB activity and increased expression of cell survival factors and ultimately contributes to their resistance to apoptosis. As such, MEKK3 may serve as a therapeutic target to control cancer cell resistance to cytokine- or drug-induced apoptosis.


Clinical superinfection and its attendant symptomatic changes in pediatrics.
Studies were conducted on the effects of antibiotics on intestinal bacterial flora and symptomatic changes associated with possible superinfection following antibiotic treatment. Following the administration of oral antibiotics, there were no marked changes in the intestinal flora. After second and third-generation cephems were injected, most bacteria, excluding Streptococcus faecalis which is resistant to them, decreased and fungi increased. The incidences of diarrhea after administering oral antibiotics were high for amoxicillin and amoxicillin + clavulanic acid. In some patients with depressed immunity, such as leukemic patients and neonates, decreases in intestinal bacteria after doses of antibiotics led to increases in pathogenic bacteria. They invaded the circulating blood, leading to septicemia. Septicemia originating in the intestinal tract was frequently associated with the development of vitamin K deficiency. Besides changes in the intestinal flora, a decrease in oral food intake and the presence of a methylthiotetrazole group in the structure of the administered antibiotics were also found to play a crucial role in causing vitamin K deficiency.


Involvement of Bacterial Extracellular Membrane Nanovesicles in Infectious Diseases and Their Application in Medicine.
Bacterial extracellular membrane nanovesicles (EMNs) are attracting the attention of scientists more and more every year. These formations are involved in the pathogenesis of numerous diseases, among which, of course, the leading role is occupied by infectious diseases, the causative agents of which are a range of Gram-positive and Gram-negative bacteria. A separate field for the study of the role of EMN is cancer. Extracellular membrane nanovesicles nowadays have a practical application as vaccine carriers for immunization against many infectious diseases. At present, the most essential point is their role in stimulating immune response to bacterial infections and tumor cells. The possibility of nanovesicles' practical use in several disease treatments is being evaluated. In our review, we listed diseases, focusing on their multitude and diversity, for which EMNs are essential, and also considered in detail the possibilities of using EMNs in the therapy and prevention of various pathologies.


Genomic analysis of penicillin-binding proteins and recombination events in an emerging amoxicillin- and meropenem-resistant PMEN3 (Spain9V-3, ST156) variant in Taiwan and comparison with global descendants of this lineage.
From 2008 to 2020, the Taiwan National Notifiable Disease Surveillance System database demonstrated that the incidence of non-vaccine serotype 23A invasive pneumococcal disease (IPD) approximately doubled. In this study, 276 non-repetitive pneumococcal clinical isolates were collected from two medical centers in Taiwan between 2019 and 2021. Of these 267 pneumococci, 60 were serotype 23A. Among them, 50 (83%) of serotype 23A isolates belonged to the sequence type (ST) 166 variant of the Spain9V-3 clone. Pneumococcal 23A-ST166 isolates were collected to assess their evolutionary relationships using whole-genome sequencing. All 23A-ST166 isolates were resistant to amoxicillin and meropenem, and 96% harbored a novel combination of penicillin-binding proteins (PBPs) (1a:2b:2x):15:11:299, the newly identified PBP2x-299 in Taiwan. Transformation of the pbp1a, pbp2b, and pbp2x alleles into the β-lactam-susceptible R6 strain revealed that PBP2x-299 and PBP2b-11 increased the MIC of ceftriaxone and meropenem by 16-fold, respectively. Prediction analysis of recombination sites in PMEN3 descendants (23A-ST166 in Taiwan, 35B-ST156 in the United States, and 11A-ST838/ST6521 in Europe) showed that adaptive evolution involved repeated, selectively favored convergent recombination in the capsular polysaccharide synthesis region, PBPs, murM, and folP genome sites. In the late 13-valent pneumococcal conjugate vaccine era, PMEN3 continuously displayed an evolutionary capacity for global dissemination and persistence, increasing IPD incidence, leading to an offset in the decrease of pneumococcal conjugate vaccine serotype-related diseases, and contributing to high antibiotic resistance.A clonal shift with a highly β-lactam-resistant non-vaccine serotype 23A, from ST338 to ST166, increased in Taiwan. ST166 is a single-locus variant of the Spain9V-3 clone, which is also called the PMEN3 lineage. All 23A-ST166 isolates, in this study, were resistant to amoxicillin and meropenem, and 96% harbored a novel combination of penicillin-binding proteins (PBPs) (1a:2b:2x):15:11:299. PBP2x-299 and PBP2b-11 contributed to the increasing MIC of ceftriaxone and meropenem, respectively. Prediction analysis of recombination sites in PMEN3 descendants showed that adaptive evolution involved repeated, selectively favored convergent recombination in the capsular polysaccharide synthesis region, PBPs, murM, and folP genome sites. In the late 13-valent pneumococcal conjugate vaccine era, PMEN3 continuously displays the evolutionary capacity for dissemination, leading to an offset in the decrease of pneumococcal conjugate vaccine serotype-related diseases and contributing to high antibiotic resistance.


Risk Factors for Early-Onset Peripheral Neuropathy Caused by Vincristine in Patients With a First Administration of R-CHOP or R-CHOP-Like Chemotherapy.
Peripheral neuropathy is a well-known side effect of vincristine (VCR), a microtubule inhibitor used for R-CHOP or R-CHOP-like (namely R-CVP and R-THP-COP) regimens. Previous studies have shown that both the total dose of VCR and the number of treatment cycles are related to the incidence of VCR-induced peripheral neuropathy (VIPN). However, VIPN will also occur during the first treatment cycle regardless of the total dose of VCR or number of treatment cycles (early-onset VIPN). There is little information about early-onset VIPN, and it is difficult to predict. The present study's goal was to identify risk factors for early-onset VIPN. We analyzed the case records of patients who had their first administration of an R-CHOP or R-CHOP-like regimen between April 2008 and August 2013 at Tokushima University Hospital in Tokushima, Japan. To identify the risk factors for early-onset VIPN, we performed univariate and multivariate logistic regression analyses. Forty-one patients underwent an R-CHOP or R-CHOP-like regimen for the first time at Tokushima University Hospital between April 2008 and August 2013, and 14 patients had grade 1 or higher early-onset VIPN. A univariate analysis revealed that age, the dose of VCR and the concomitant use of aprepitant appeared to be the risk factors of early-onset VIPN. In our calculation using receiver-operator characteristics curves, the cut-off value for patient age was 65 years and that of the dose of VCR was 1.9 mg. A multivariate analysis revealed that VCR dose ≥ 1.9 mg and the concomitant use of the antiemetic aprepitant were independent risk factors for early-onset VIPN. Our present study showed that the patients who had VCR dose ≥ 1.9 mg and the concomitant use of aprepitant had the risk for early-onset VIPN. This suggests that it is important to use aprepitant in light of the risk of early-onset VIPN and the benefit of aprepitant's antiemetic effect in R-CHOP and R-CHOP-like regimens.


Combined detection of mannanaemia and antimannan antibodies as a strategy for the diagnosis of systemic infection caused by pathogenic Candida species.
A novel strategy for the diagnosis of systemic candidosis was evaluated, based on the combination of two enzyme immunoassays that detect a candida oligomannoside repetitive epitope expressed in large amounts by Candida albicans (Platelia Candida Ag), and antibodies against C. albicans mannan, the major cell-wall immunogen in which this epitope is present (Platelia Candida Ab). Sera were selected retrospectively from intensive care and haematology patients with clinically suspected systemic candidosis, and from whom Candida spp. had been isolated from normally sterile sites. Of the 21 patients infected with C. albicans, 13 had positive antigenaemia and 14 had a positive antibody response, including eight patients who were antigenaemia negative. The sensitivity of the combined tests was 100%. In patients infected with C. glabrata (n = 12) or C. tropicalis (n = 10), the sensitivity was 83% and 80%, respectively. For the remaining patients, infected with C. parapsilosis (n = 10), C. krusei (n = 8) or C. kefyr (n = 2), the sensitivity of the combined tests was 40%, 50% and 50%, respectively. At least one of the serological tests was positive before yeast growth occurred in 60% of patients for whom a serum sample was available before blood culture sampling. An increase in serological test positivity to >80% was observed for sera obtained around the date of positive culture, irrespective of the Candida species isolated. These results suggest that regular serological monitoring for both mannanaemia and anti-mannan antibodies in at-risk patients may contribute to the early diagnosis of candidosis.


Diacetylenic lipid tubules: experimental evidence for a chiral molecular architecture.
Molecular self-assembly is of key importance for the rational design of advanced materials. To investigate the causal relation between molecular structure and the consequent self-assembled microstructure, self-assembled tubules of diacetylenic lipids were studied. Circular-dichroism studies give experimental evidence that the formation of tubules is driven by chiral molecular packing, in agreement with recent theories of tubules. On the basis of these results, a molecular mechanism for the formation of tubules is proposed.


[Oral status of drug addicted patients. Study of 66 cases].
We studied 66 heroin-addicted patients from Valencia Community, comparing them with a control group of 30 healthy subjects. In all cases the CAO index and gingival state were analysed, including possible pathology of oral mucous. It was revealed that the major oral deterioration was in the experimental group. This was directly related to oral hygiene deficiency, which was brought about in the period of active addiction. At the same time the difference in oral mucous pathology between the two groups was not significant.


Insulin regulation of protein translation repressor 4E-BP1, an eIF4E-binding protein, in renal epithelial cells.
Augmented protein translation by insulin involves activation of eukaryotic initiation factor 4E (eIF4E) that follows release of eIF4E from a heterodimeric complex by phosphorylation of its inhibitory binding protein, 4E-BP1. We examined insulin regulation of 4E-BP1 phosphorylation in murine proximal tubular epithelial cells. Insulin (1 nmol/L) increased de novo protein synthesis by 58 +/- 11% (P < 0.001). Insulin also augmented 4E-BP1 phosphorylation and phosphatidylinositol 3-kinase (PI 3-kinase) activity in antiphosphotyrosine immunoprecipitates. This could be prevented by PI 3-kinase inhibitors, Wortmannin, and LY294002. Insulin also activated Akt that lies downstream of PI 3-kinase. Rapamycin abrogated 4E-BP1 phosphorylation in response to insulin, suggesting involvement of mammalian target of rapamycin (mTOR), a kinase downstream of Akt. Insulin-stimulated phosphorylation of 4E-BP1 was also inhibited by PD098059, implying involvement of Erk-1/-2 mitogen-activated protein (MAP) kinase. An increase in Erk-1/-2 type MAP kinase activity by insulin was directly confirmed in an immunokinase assay and was found to be PI 3-kinase dependent. In proximal tubular epithelial cells, insulin augments 4E-BP1 phosphorylation, which is PI 3-kinase and mTOR dependent. The requirement for Erk-1/-2 MAP kinase activation for 4E-BP1 phosphorylation by insulin suggests a cross-talk between PI 3-kinase and Erk-1/-2-type MAP kinase pathways.


Automatic correction of contaminated images for assessment of reservoir surface area dynamics.
The potential of using Landsat for assessing long-term water surface dynamics of individual reservoirs at a global scale has been significantly hindered by contaminations from clouds, cloud shadows, and terrain shadows. A novel algorithm was developed towards the automatic correction of these contaminated image classifications. By applying this algorithm to the dataset by Pekel et al. (2016), time series of area values for 6817 global reservoirs (with an integrated capacity of 6099 km3) were generated from 1984 to 2015. The number of effective images that can be used in each time series has been improved by 81% on average. The long-term average area for these global reservoirs was corrected from 1.73×105 km2 to 3.94×105 km2. The results were proven to be robust through validation using observations, synthetic data, and visual inspection. This continuous reservoir surface area dataset can provide benefit to various applications (both at continental and local scales).


Haemodynamic correlates of early and delayed responses to sublingual administration of isosorbide dinitrate in migraine patients: a transcranial Doppler study.
In normal subjects or migraine patients, nitrates induce a non-specific early headache caused by vasodilation of intracranial arteries. In migraineurs a delayed headache response to nitrates may have a typical clinical profile of a spontaneous migraine attack. The cerebral vasomotor changes of this delayed response require further study. Isosorbide dinitrate (IDN), an exogenous nitric oxide (NO) donor, was given at a dose of 5 mg sublingually and a bilateral transcranial Doppler device was used to monitor bilateral mean velocity (Vm) changes at the middle cerebral artery (MCA) after IDN administration and until delayed headache occurred. Spontaneous migraine-like headache occurred only in migraine patients during the delayed phase after IDN and was accompanied by a prolonged arterial vasodilation compared to normal subjects. This vasomotor response was more evident on the customary side of the head pain of a spontaneous migraine attack. Our findings suggest a particular vasomotor response to nitrates in migraine patients. This response is associated with the nitrate-induced headache and it is not evident in healthy pain-free controls during the delayed phase after administration of an NO donor. Owing to the short half-life of NO, the neurotransmitter released by IDN, and because of the late onset of headache, we believe the mechanism is unlikely to be vascular in origin, but may have a neurogenic component.


Understanding the Mechanism of the Broad-Spectrum Antiviral Activity of Favipiravir (T-705): Key Role of the F1 Motif of the Viral Polymerase.
Favipiravir (T-705) is a broad-spectrum antiviral agent that has been approved in Japan for the treatment of influenza virus infections. T-705 also inhibits the replication of various RNA viruses, including chikungunya virus (CHIKV). We demonstrated earlier that the K291R mutation in the F1 motif of the RNA-dependent RNA polymerase (RdRp) of CHIKV is responsible for low-level resistance to T-705. Interestingly, this lysine is highly conserved in the RdRp of positive-sense single-stranded RNA (+ssRNA) viruses. To obtain insights into the unique broad-spectrum antiviral activity of T-705, we explored the role of this lysine using another +ssRNA virus, namely, coxsackievirus B3 (CVB3). Introduction of the corresponding K-to-R substitution in the CVB3 RdRp (K159R) resulted in a nonviable virus. Replication competence of the K159R variant was restored by spontaneous acquisition of an A239G substitution in the RdRp. A mutagenesis analysis at position K159 identified the K159M variant as the only other viable variant which had also acquired the A239G substitution. The K159 substitutions markedly decreased the processivity of the purified viral RdRp, which was restored by the introduction of the A239G mutation. The K159R A239G and K159M A239G variants proved, surprisingly, more susceptible than the wild-type virus to T-705 and exhibited lower fidelity in polymerase assays. Furthermore, the K159R A239G variant was found to be highly attenuated in mice. We thus demonstrate that the conserved lysine in the F1 motif of the RdRp of +ssRNA viruses is involved in the broad-spectrum antiviral activity of T-705 and that it is a key amino acid for the proper functioning of the enzyme.IMPORTANCE In this study, we report the key role of a highly conserved lysine residue of the viral polymerase in the broad-spectrum antiviral activity of favipiravir (T-705) against positive-sense single-stranded RNA viruses. Substitutions of this conserved lysine have a major negative impact on the functionality of the RdRp. Furthermore, we show that this lysine is involved in the fidelity of the RdRp and that the RdRp fidelity influences the sensitivity of the virus for the antiviral efficacy of T-705. Consequently, these results provide insights into the mechanism of the antiviral activity of T-705 and may lay the basis for the design of novel chemical scaffolds that may be endowed with a more potent broad-spectrum antiviral activity than that of T-705.


A Closer Look at Warning Cues on the Sustained Attention to Response Task Performance.
We investigated the effects of auditory cues of varying reliability levels on response inhibition performance using a target detection task to determine if external cues offer performance benefits. Further, we examined how the slope of the speed accuracy trade-off changes as a function of auditory cue reliability and used the trade-off to understand where any performance gains may be realized. Researchers have proposed that the sustained attention to response task (SART) can be used to study the mechanisms causing failures of response inhibition. External cues may mitigate the results of motor inhibition failure. The extent to which external cues can effectively aid performance depends on the level of cue reliability. Ninety-one participants performed three SARTs with auditory cue assistance at three different levels of reliability (i.e. 0%, 60% and 100% reliable at cueing imminent No-Go stimuli). We observed fewer errors of commission and faster reaction time in conditions with higher cue reliability. The slope of speed-accuracy trade-off relationship was impacted by cue reliability and was not a simple linear function. Reliable auditory cues aid performance by reducing reaction time and errors of commission. Auditory cues also impact the relationship between speed and accuracy trade-off. Insights of cue effectiveness at different reliability levels help people make informed decisions in developing automation interfaces or sensors based on expected performance. Reliable cues mitigate the risk of impulsive errors; however, the reliability has to be high to have a noticeable impact on the speed-accuracy trade-off.


Tetracycline in the treatment of periodontal disease: review of current literature.
For many years, tetracycline has been prescribed in the treatment of periodontal disease with little research data supporting such use. Current data suggest that most patients with periodontal disease are best treated by conventional techniques of subgingival debridement.


Deregulation of cyclins D1 and E and suppression of cdk2 and cdk4 in senescent human fibroblasts.
The state of cellular senescence is characterised by an irreversible arrest in the G1 phase of the cell cycle. It has previously been shown that three cell cycle genes, cyclin A, cyclin B and cdc2, are not expressed in senescent human fibroblasts. All three gene products have functions after S-phase entry, so that their suppression cannot explain the irreversible G1 arrest. Here, we report that the abundance of transcripts from two other cell cycle genes, cdk2 and cdk4, thought to act during G1-->S progression, is significantly diminished in senescent cells of the diploid human fibroblast line WI-38. Surprisingly, two other cyclins, D1 and E, behave in a completely different way, in that their expression is elevated in senescent cells, especially under conditions of serum starvation. Both the synthesis and the steady-state level of cyclin D1 protein were also found to be markedly higher in senescent cells (3- to 6-fold). Cyclins D1 and E are thus the first genes shown to be overexpressed or deregulated in senescent cells. It is tempting to speculate that this deregulation may be due to the absence, in senescent cells, of a regulatory loop that would normally control their expression. This is supported by our finding that cyclin E-associated kinase activity in senescent cells is reduced approx. 14-fold. Our data also suggest that the deregulated expression of cyclin D1 and E is not sufficient to drive senescent cells into DNA replication.


Deprescribing in Older Nursing Home Patients: Focus on Innovative Composite Measures for Dosage Deintensification.
Deprescribing, which includes stopping or reducing the dosage of medications, is designed to improve safety and prevent adverse drug reactions in older patients. To date, there has been limited work on measuring decreases in dosage intensity, or deintensification, across therapeutic classes of medications. Given the ongoing focus on central nervous system (CNS) medications and the frequency with which providers encounter hypertension and diabetes in older nursing home patients, the objective of this expert review is to describe and critique innovative composite dosage intensity measures that have been, or could be, applied to quantify deintensification within three therapeutic medication targets commonly encountered in nursing home patients: CNS agents, antihypertensive therapy, and antidiabetic therapy and the extent to which they are associated with health outcomes. Composite measures for CNS medication intensity considered dividing a patient's daily dose by defined daily dosage (DDD), or the minimum effective adult or geriatric daily dosage. In contrast, composite measures for antihypertensives used either DDD or maximum recommended daily dosage in the denominator. We were not able to identify any composite measure of intensity for antidiabetic classes. There was a paucity of interventional studies that showed reducing the dosage intensity resulted in improved health outcomes. In conclusion, we identified several innovative composite measures of dosage intensity for CNS and antihypertensive medications, and discussed possible approaches for developing an antidiabetic regimen composite measure. It is critical for future research to compare and contrast various measures and to determine their impact on important clinical outcomes.


Dimethyl sulfoxide (DMSO) as the sulfur source for the production of desulfurizing resting cells of Gordonia alkanivorans RIPI90A.
The sulfate repression of desulfurization (Dsz) phenotype represents a major barrier to the mass production of desulfurizing resting cells. This repression can be avoided by replacing sulfate with dibenzothiophene (DBT) as the main substrate for the 4S pathway. However, mass production of biocatalyst using DBT is impractical because of its high price, low water solubility, and growth inhibition by 2-hydroxybiphenyl (2-HBP), which is the end product of the 4S pathway. In this work, the results showed that readily bioavailable sulfur compounds led to repression of the desulfurization activity of Gordonia alkanivorans RIPI90A. However, the Dsz phenotype was expressed through the 4S pathway in the presence of DMSO as the sulfur source for growth. Resting cells grown on DMSO were more active than the resting cells grown on DBT. The growth rate of strain RIPI90A on DMSO was higher than when DBT was used as the sole sulfur source. DMSO concentration significantly influenced the growth pattern of the strain, and the highest growth rate was observed at a concentration of 200 microg ml(-1). Above this concentration, the growth rate gradually decreased. DBT was found to induce the Dsz phenotype, with no observed lag period, in cells grown on DMSO as the sole sulfur source. Prior to induction, the specific activity was detected as 1.4 micromol 2-HBP (g dry cell weight)(-1) h(-1), and following incubation (5 h) the highest specific activity was observed as 5.11 micromol 2-HBP (g dry cell weight)(-1 )h(-1). This study identified that resting cells can be prepared in a two-step process. First, resting cells can be produced using DMSO as the sulfur source for growth; in the second step, improvements to their desulfurizing activity can be made using DBT as an inducer. DMSO is recommended as an appropriate sulfur source for the mass production of G. alkanivorans RIPI90A.


Azacitidine Pre-Treatment Followed by Reduced-Intensity Stem Cell Transplantation in Patients with Higher-Risk Myelodysplastic Syndrome.
Azacitidine (AZA) is commonly used in patients with myelodysplastic syndrome (MDS). To determine the role of AZA before allogeneic stem cell transplantation (allo-SCT), we conducted a prospective study of AZA pre-treatment followed by allo-SCT in patients with higher-risk MDS. Twenty-one patients who were scheduled for their third to sixth cycle of AZA pre-treatment followed by allo-SCT were enrolled. AZA pre-treatment was interrupted early in 3 patients (14.3%) because of leukaemic transformation or death. The overall response rate to AZA pre-treatment was 57.1%. There were 2 cases of complete remission, 1 case of partial remission, and 9 cases of haematologic improvement. Fourteen patients (66.7%) received the planned allo-SCT and 5 patients were alive at the last follow-up. Three-year progression-free survival (PFS) and 3-year overall survival (OS) in the 14 patients who received allo-SCT were 30.0% (95% CI 3.3-56.7) and 42.9% (95% CI 17.1-68.7), respectively. PFS and OS were not influenced by response to AZA pre-treatment (p > 0.05). In this study, AZA had a role as a bridge therapy to prevent leukaemic transformation prior to selection of a donor for allo-SCT and showed low toxicity. It may be considered in patients with higher-risk MDS.


Behaviour of pollinator insects within inflorescences of Scrophularia species from Iberian Peninsula.
Several hypotheses have been proposed to explain the pattern of pollinator visits to vertical inflorescences. These range from a response to a pattern of resources to merely instinctive behaviour. In dichogamous plants, such behaviour has been associated with promoting outcrossing and avoiding geitonogamy. We here analyse behaviour of the principal pollinator groups in five protogynous species of Scrophularia with different flower sizes (S. sambucifolia, S. grandiflora, S. lyrata, S. scorodonia and S. canina), and the distribution of sexual phases along the inflorescences. The results in all cases show that pollinators follow a pattern of ascending visits accompanied by movements between flowers of the same whorl (horizontal movements). The relative frequency of these horizontal movements depends on the flower size, with a higher frequency in species with large flowers. In vertical movements of the three more common pollinator groups to several plant species (bumblebees, wasps and small bees), the behaviour was essentially independent of flower size, with bumblebees having the highest ratio of ascents to descents. Behaviour of the pollinators, together with the absence of a definite pattern of distribution of the sexual phases along the inflorescence, implies that geitonogamy is not avoided in any of the Scrophularia species studied.


When cultures meet in general practice: intercultural differences between GPs and parents of child patients.
Although health care professionals in The Netherlands are increasingly confronted with diverse immigrant groups, medical counselling and treatment of these groups has not been the subject of extensive research yet. From other studies it is well known that intercultural differences can have serious consequences for health care, e.g. in terms of risk of incorrect diagnoses or non-compliance. Eighty-seven autochthonous Dutch and immigrant (mainly from Turkey and Surinam) parents of child patients and their general practitioners (GPs) were recruited to investigate the influence of cultural differences on mutual understanding and patient compliance. Analyses of questionnaires and home interviews revealed that there is a relation between the cultural background of the patient and effectiveness of communication. Communication in consultations between GPs and persons from ethnic minorities is less effective than in consultations with Dutch persons: there is more misunderstanding, and also more non-compliance. In general, mutual understanding between GP and patient proves to be a strong predictor for patient compliance. These findings hold especially true for patients living in two worlds, i.e. a mixture of traditional and western cultures. The results are discussed in terms of methodological issues and practical implications for the health care providers.


Predicting steady-state endoxifen plasma concentrations in breast cancer patients by CYP2D6 genotyping or phenotyping. Which approach is more reliable?
In previous studies, steady-state Z-endoxifen plasma concentrations (ENDOss) correlated with relapse-free survival in women on tamoxifen (TAM) treatment for breast cancer. ENDOss also correlated significantly with CYP2D6 genotype (activity score) and CYP2D6 phenotype (dextromethorphan test). Our aim was to ascertain which method for assessing CYP2D6 activity is more reliable in predicting ENDOss. The study concerned 203 Caucasian women on tamoxifen-adjuvant therapy (20 mg q.d.). Before starting treatment, CYP2D6 was genotyped (and activity scores computed), and the urinary log(dextromethorphan/dextrorphan) ratio [log(DM/DX)] was calculated after 15 mg of oral dextromethorphan. Plasma concentrations of TAM, N-desmethyl-tamoxifen (ND-TAM), Z-4OH-tamoxifen (4OH-TAM) and ENDO were assayed 1, 4, and 8 months after first administering TAM. Multivariable regression analysis was used to identify the clinical and laboratory variables predicting log-transformed ENDOss (log-ENDOss). Genotype-derived CYP2D6 phenotypes (PM, IM, NM, EM) and log(DM/DX) correlated independently with log-ENDOss. Genotype-phenotype concordance was almost complete only for poor metabolizers, whereas it emerged that 34% of intermediate, normal, and ultrarapid metabolizers were classified differently based on log(DM/DX). Multivariable regression analysis selected log(DM/DX) as the best predictor, with patients' age, weak inhibitor use, and CYP2D6 phenotype decreasingly important: log-ENDOss = 0.162 - log(DM/DX) × 0.170 + age × 0.0063 - weak inhibitor use × 0.250 + IM × 0.105 + (NM + UM) × 0.210; (R2 = 0.51). In conclusion, log(DM/DX) seems superior to genotype-derived CYP2D6 phenotype in predicting ENDOss.


[Preoperative stress. Research approach and methods of treatment].
The scientific approach to preoperative stress includes the identification of stressors and the measurement of stress responses. The stress responses include physiological (sympathetic nervous system, hypothalamic--hypophyseal and adrenocortical system), psychological (anxiety, depression etc.) and behavioural ones. The choice of variables to be measured should allow answers for our questions, i.e., they must be adequate and economic, without or only minimal disturbance of the preoperative situation. The value of physiological and biochemical parameters is often overestimated. Their assessment is indicated in special scientific settings. For simple questions such as premedication studies, the measurement of blood pressure and heart rate is sufficient. Anxiety should be self-estimated by the patient, using visual analogue scales, questionnaires or adjective check lists. Depression and other emotional stress responses can be assessed if necessary. For physicians and nurses, an observation of the patient's behavioural stress responses is a useful criterion for assessing patient stress. One important stress-reducing approach is prevention: identification and avoidance of stressors. This includes organisation and a high degree of self-control by the medical personnel. Premedication with benzodiazepines is the most successful pharmacological approach.


Influenza virus-specific neutralizing IgM antibodies persist for a lifetime.
Detection of immunoglobulin M (IgM) antibodies has long been used as an important diagnostic tool for identifying active viral infections, but their relevance in later stages has not been clearly defined in vivo. In this study, we followed the kinetics, longevity, and function of influenza virus-specific IgM antibodies for 2 years following sublethal infection of mice with live mouse-adapted A/PR/8/34 virus or immunization with formalin-inactivated virus. These groups mounted robust protective immune responses and survived lethal challenges with 50 × 50% lethal dose (LD50) mouse-adapted A/PR/8/34 virus 600 days after the primary exposure. Surprisingly, the virus-specific IgM antibodies persisted along with IgG antibodies, and we found a significantly higher number of IgM-positive (IgM(+)) virus-specific plasma cells than IgG(+) plasma cells that persisted for at least 9 months postexposure. The IgM antibodies were functional as they neutralized influenza virus in the presence of complement just as well as IgG antibodies did.


[Relationship between the efficacy of mebendazole and immune level of mice infected with secondary cysts of Echinococcus granulosus].
The proliferative response of lymphocytes to Con A and enzyme-linked absorbent assay were used to determine the levels of cellular and humoral immunity of mice infected with ip 2000 protoscolices of Echinococcus granulosus for 2, 4 or 8 months and treated twice with ip BCG 0.2 mg/mouse at an interval of 10 d, or with ip cyclophosphamide (Cy) 10 mg/(kg.d) x 5 d. The results showed that the immune levels of the host were stimulated by BCG, but depressed by Cy significantly. When the mice were treated with ip mebendazole (Meb) 25 mg/(kg.d) x 10 d in combination with ip BCG 0.2 mg/mouse on d 3 before Meb treatment and on d 7 after the beginning of Meb treatment, or with ip Cy 10 mg/(kg.d) x 5 d before Meb treatment, the inhibition rates of cyst weight and the alterations of germinal layers induced by the drugs were similar to those of corresponding groups treated with Meb alone. Cy also exhibited an apparent effect on mice infected with protoscolices for 2 months. Even so, no apparent synergic effect was seen after combined treatment with Cy and Meb. The results suggest that the effect of Meb on secondary cysts of E granulosus was not affected by the host immune level.


Online Oncology Careers Events to Attract the Future Oncology Medical Workforce: An Exemplar Event and Review of the Literature.
There are too few oncologists to meet the increasing burden imposed by the rising incidence of cancer. This results from issues with the retention of established oncologists and longstanding challenges to the recruitment of adequate numbers of trainees. To counter this, the British Oncology Network for Undergraduate Societies (BONUS) devised an online oncology careers event for medical students and junior doctors who are yet to select a specialty. An online careers event was devised with a focus on oncology practice and related subspecialties, as well as research. Event attendees were asked to respond to piloted pre- and post-event surveys. Knowledge and attitudes towards a career in oncology were evaluated using Likert scale and multiple-choice questions. A systematic literature search was carried out to contextualise these data. Of the 73 attendees, 44 (60%) participants completed both the pre- and post-event surveys; 79.5% of attendees believed that information on a career in oncology is lacking in medical training. This viewpoint was supported by the systematic review, which highlighted a need for relevant focussed interventions targeted at medical students and fledgling doctors. The education event led to an increase in the median reported understanding of the oncology career pathway from 6.0 to 8.0 (P < 0.05 and P < 0.001), as well as the likelihood of pursuing a career in oncology (8.0-9.0; P < 0.05). It was also associated with a proportional increase in medical and surgical oncology interest, albeit with a fall in interest in clinical and interventional oncology as well as academia. A targeted online careers event increases knowledge of and interest in a career in oncology, albeit predominantly for medical and surgical subspecialties. Broader initiatives based on our model should be developed and careers in academia as well as clinical and interventional oncology emphasised.


A tale of three cities--where RHIOS meet the NHIN.
Regional health information exchanges in California, Indiana, and Massachusetts have been collaborating on a prototype for a nationwide health information network, first under the auspices of the Markle Foundation's Connecting for Health program and now under contract to the Department of Health and Human Services' Office of the National Coordinator for Health Information Technology. Since mid-2004, this collaboration has evolved from a collection of regional efforts to a standards-driven cooperative and now to one of four prototype national networks fostered by federal efforts. This development reflects a maturing market for interoperability and integration in healthcare information technology, starting with RHIOs, and suggests one response to the industry's need for the type of plug-and-play information exchange available in other industries. The authors share their experiences and their views of how RHIOs and a Nationwide Health Information Network will further develop to make interoperable electronic health records a reality in coming years. The content of this article is solely the responsibility of the authors and does not necessarily represent the official view of the Office of the National Coordinator for Health Information Technology.


Midterm outcomes of total hip arthroplasty with a modular Birmingham head.
Birmingham Hip Resurfacing (BHR) implants may be combined with a conventional femoral stem to create a modular metal-on-metal total hip arthroplasty (BHR MoM THA). There is little outcome data regarding this construct. This study examines midterm outcomes of BHR MoM THA compared to oxidised zirconium total hip arthroplasty (THA). A retrospective institutional review identified all patients receiving BHR MoM THA between April 2005 and February 2011 and a matched control cohort of zirconium THA patients. Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Harris Hip Score (HHS), and SF-12 Health status scores were obtained. Revisions and complications were collected from clinical records. Radiographs were assessed for evidence of component malposition, loosening, osteolysis, or heterotopic ossification. 63 modular BHR MoM THA were identified in 61 patients (36 with BHR cups, 27 with R3 cups) and 63 zirconium THA in 58 matched controls. Mean follow-up was 58 months. 14 BHR MoM THA hips (22.2%) were revised (4 infections, 1 dislocation, 9 soft tissue reactions) compared to 3 (4.8%) zirconium THA (all infections). At latest follow-up, 18.4% of surviving BHR MoM THA hips were painful compared to 0.5% of zirconium THA controls (p < 0.001). WOMAC, HHS, and SF-12 did not differ significantly between surviving members of the 2 groups. BHR MoM THA demonstrated a high revision rate, largely for adverse local soft tissue reaction and pain. Among those not revised, many reported some residual pain despite similar quality of life measures to those who received zirconium THA.


[Cerebral angiography: a study of complications in 450 consecutive procedures].
To evaluate all types of complications, both minor and major, associated with modern cerebral angiography. A prospective study of 450 consecutive cerebral angiographic procedures is reported. One patient (0.2%) died from a cholesterol embolus. In seven patients (1.6%), thromboembolic events occurred within 24 hours after the procedure, leading to transient ischemic symptoms in six and permanent hemiplegia in one. Two patients suffered from acute renal failure (0.4%). Transient cardiac arrhythmias were observed in three patients without consequence on the clinical outcome. Most complications of angiography occurred in patients referred from the neurology department for work-up of stroke syndrome. Our results show that morbidity and mortality rates related to the angiographic procedure did not decrease in spite of major improvement of angiographic materiel. Atherosclerosis is the main risk factor for complication. Most of the complications could be avoided by appropriate selection of indications and by using non-invasive techniques such as magnetic resonance angiography or helical CT angiography.


Structure of the human desmoplakins. Implications for function in the desmosomal plaque.
Desmoplakins (DPs) I and II are two major related proteins located in the innermost portion of the desmosomal plaque where it is thought they may play a role in attaching intermediate filaments (IF) to the cell surface. We have isolated and sequenced human cDNA clones encoding two major DP domains and a portion of a third. These clones can be divided into two classes that we believe to represent DPI and DPII cDNAs; our evidence suggests that the DPII message is derived at least in part from the processing of a larger transcript encoded by a single gene. Computer-aided analysis of the DPI-predicted amino acid sequence indicates that the central domain, which contains the heptad repeat characteristic of many alpha-fibrous proteins, will participate in the formation of a coiled coil dimer approximately 130 nm in length. The periodicity of acidic and basic residues in the rod suggests that DPI will aggregate with itself or similar molecules into higher order filamentous structures. The carboxyl terminus contains three regions with significant homology, each of which comprises almost five repeats of a 38-residue motif. It is likely that these regions each fold into a compact globular conformation stabilized by intrachain ionic interactions. Comparison of the predicted amino acid sequence of a cDNA encoding a portion of the 230-kDa bullous pemphigoid antigen (Stanley, J. R., Tanaka, T., Mueller, S., Klaus-Kovtun, V., and Roop, D. (1988) J. Clin. Invest. 82, 1864-1870) with DP revealed the presence of a 38-residue repeat with striking similarity to that of the DPs. Significantly, the periodicity in acidic and basic residues of these domains is the same as that found in the 1B rod domain of IF proteins. This suggests the possibility that the DPs might interact with IF via their common periodicity of charged residues.


Capsaicin mediates apoptosis in human nasopharyngeal carcinoma NPC-TW 039 cells through mitochondrial depolarization and endoplasmic reticulum stress.
Capsaicin, a pungent compound found in hot chili peppers, has been reported to have antitumor activities in many human cancer cell lines, but the induction of precise apoptosis signaling pathway in human nasopharyngeal carcinoma (NPC) cells is unclear. Here, we investigated the molecular mechanisms of capsaicin-induced apoptosis in human NPC, NPC-TW 039, cells. Effects of capsaicin involved endoplasmic reticulum (ER) stress, caspase-3 activation and mitochondrial depolarization. Capsaicin-induced cytotoxic effects (cell death) through G0/G1 phase arrest and induction of apoptosis of NPC-TW 039 cells in a dose-dependent manner. Capsaicin treatment triggered ER stress by promoting the production of reactive oxygen species (ROS), increasing levels of inositol-requiring 1 enzyme (IRE1), growth arrest and DNA-damage-inducible 153 (GADD153) and glucose-regulated protein 78 (GRP78). Other effects included an increase in cytosolic Ca(2+), loss of the mitochondrial transmembrane potential (ΔΨ(m)), releases of cytochrome c and apoptosis-inducing factor (AIF), and activation of caspase-9 and -3. Furthermore, capsaicin induced increases in the ratio of Bax/Bcl-2 and abundance of apoptosis-related protein levels. These results suggest that ER stress- and mitochondria-mediated cell death is involved in capsaicin-induced apoptosis in NPC-TW 039 cells.


Cloning and nucleotide sequence analysis of human embryonic zeta-globin cDNA.
Clones of human embryonic alpha-like zeta-globin cDNA were isolated, by detection using cross-hybridization to human alpha-globin cDNA probes, from a cDNA library derived from the mRNA of the human erythroleukemia cell line K562. Nucleotide sequence analysis of these cDNA clones revealed a coding sequence that corresponds perfectly to the independently derived amino acid sequence of the human zeta-globin chain. Comparison of the nucleotide sequence of human zeta-globin cDNA with that of human alpha-globin cDNA confirmed previous estimates of very distant evolutionary divergence between the human zeta- and alpha-globin genes. Nevertheless, the human zeta-globin cDNA sequence shares a remarkable similarity to that of the alpha-globin gene in its codon usage, high G + C base composition, and lack of bias against usage of CG dinucleotides.


Lyme disease presenting as subacute transverse myelitis.
Lyme disease (borreliosis) is a systemic illness resulting from infection with the spirochete Borrelia burgdorferi. It is transmitted to humans by the bites of infected ticks belonging to several species of the genus Ixodes. After the bacteria enter the body via the dermis, most patients develop the early, localised form of Lyme disease, which is characterised by erythema migrans and influenza-like symptoms. This disease may also affect the heart, nervous system and joints. The neurological findings of this disease may include peripheral and central nervous system signs. A 21-year-old woman attended a family medicine outpatient clinic complaining of unexplained pain and muscle power loss in her lower extremities. The problem had started in her right leg 3 months earlier and worsened in the last week. She had a neurology consultation and was hospitalised. Her neurological examination revealed bilateral facial paralysis and sensory impairment. Immunoglobulin M antibody to B. burgdorferi was positive on Western blotting in both serum and cerebrospinal fluid. The patient was diagnosed with subacute neuroborreliosis and treated.


Functional characterization of MexXY and OpmG in aminoglycoside efflux in Pseudomonas aeruginosa.
MexXY is an active efflux system that contributes to intrinsic resistance to aminoglycosides in Pseudomonas aeruginosa. MexXY can function in combination with OprM in aminoglycoside efflux but may also functionally associate with another as yet unidentified outer membrane channel. The possible role of OpmG as a third component of MexXY in aminoglycoside efflux was investigated by construction of unmarked opmG mutants. Loss of OpmG did not have any impact on minimum inhibitory concentrations for aminoglycosides regardless of the presence of oprM, indicating that MexXY does not interact with OpmG in aminoglycoside efflux. In a clinical isolate PAJ010, (mexXY) enhanced streptomycin susceptibility but neither oprM nor opmG could, suggesting that MexXY functionally associates with an unidentified outer membrane protein for aminoglycoside efflux. Expression of an opmG-lacZ transcriptional fusion revealed that OpmG expression was neither constitutive nor inducible by gentamicin. Growth rates of wildtype P. aeruginosa and opmG mutant derivatives were not different, indicating that expression of opmG is not essential for P. aeruginosa growth.


[Mucopolysaccharidosis type VI: clinical aspects, diagnosis and treatment with enzyme replacement therapy].
Mucopolysaccharidosis VI (MPS VI) is a lysosomal storage disease associated with a deficiency or absence of arylsulfatase B leading to the abnormal accumulation of dermatan sulfate. MPS VI shows a wide spectrum of symptoms from slowly to rapidly progressing forms. The characteristic spectrum includes skeletal displasia, coarse facies, cardiomyopathy, pulmonary complications and spinal compression. Diagnosis generally requires measurement ofurinary glycosaminoglycans and arylsulfatase B enzyme activity in dried blood spot, leukocytes or cultured fibroblasts. Enzyme replacement therapy (ERT) with galsulfase is now widely available providing improvement in skeletal performance and stabilization in pulmonary and cardiac functioning. Spinal involvement does not respond to ERT when is present, surgical decompression should be indicated early. Prognosis is variable depending on the age of onset and age at initiation of ERT.


MLCK210 gene knockout or kinase inhibition preserves lung function following endotoxin-induced lung injury in mice.
Barrier dysfunction, involving the endothelium or epithelium, is implicated in the pathophysiology of many disease states, including acute and ventilator-associated lung injury. Evidence from cell culture, in vivo and clinical studies, has identified myosin light chain kinase as a drug discovery target for such diseases. Here, we measured disease-relevant end points to test the hypothesis that inhibition of myosin light chain kinase is a potential therapeutic target for treatment of barrier dysfunction resulting from acute lung injury. We used a combined gene knockout and chemical biology approach with an in vivo intact lung injury model. We showed that inhibition of myosin light chain kinase protects lung function, preserves oxygenation, prevents acidosis, and enhances survival after endotoxin exposure with subsequent mechanical ventilation. This protective effect provided by the small molecule inhibitor of myosin light chain kinase is present when the inhibitor is administered during a clinically relevant injury paradigm after endotoxin exposure. Treatment with inhibitor confers additional protection against acute lung injury to that provided by a standard protective mode of ventilation. These results support the hypothesis that myosin light chain kinase is a potential therapeutic target for acute lung injury and provide clinical end points of arterial blood gases and pulmonary compliance that facilitate the direct extrapolation of these studies to measures used in critical care medicine.


Waste Informatics: Establishing Characteristics of Contemporary U.S. Landfill Quantities and Practices.
Waste generation is expected to increase in most countries for many decades with landfill disposal still the dominant solid waste management method1-3. Yet, operational characteristics of landfills are often poorly understood with comparative statistics substantially lacking. Here, we call for a more formal waste informatics to organize and standardize waste management knowledge at multiple spatial scales through analysis of recently reported data from 1232 U.S. landfills and other high resolution data sets. We create the first known estimate of available U.S. municipal waste stocks (8.5 billion tonnes) and go on to resolve these stocks at the county level, reflecting prospective urban mining opportunities. Our analysis of disposal rates and landfill capacities reveals that more than half of U.S. states have more than 25 years of life remaining. We also estimate the gross energy potential of landfill gas in the U.S. (338 billion MJ/yr) by examining 922 operational methane collection systems and demonstrate that the greatest energy recovery opportunities lie at landfills with existing collection systems and energy conversion infrastructure. Finally, we found that the number of landfills reaching the federally defined 30-year postclosure care period will more than triple in the coming two decades, with 264 sites expected by the year 2044, highlighting the need to develop and standardize metrics carefully to define and standardize when it is appropriate to end or scale back long-term landfill monitoring.


Elevated expression of AhR and NLRP3 link polycyclic aromatic hydrocarbon exposure to cytokine storm in preschool children.
Polycyclic aromatic hydrocarbons (PAHs), as a group of persistent organic pollutants, are linked to impaired immune function and low-grade inflammation in adults and children. However, the potential of PAHs to lead to a cytokine storm associated with AhR (aryl hydrocarbon receptor) and NLRP3 (NLR family pyrin domain containing 3) in humans has been poorly studied. We aimed to investigate the associations between PAH exposure, AhR and NLRP3 expression, and cytokines associated with a cytokine storm in healthy preschoolers. Basic demographic surveys and physical examinations were conducted on 248 preschoolers from an electronic waste (e-waste) recycling area (Guiyu, n = 121) and a reference area (Haojiang, n = 127). Ten urinary PAH metabolite (OH-PAH) concentrations were measured. We also measured the expression levels of AhR and NLRP3 and seventeen serum cytokine levels. The concentrations of multiple OH-PAHs were significantly higher in the exposed group than those in the reference group, especially 1-hydroxynaphthalene (1-OH-Nap) and 2-hydroxynaphthalene (2-OH-Nap). PAH exposure was closely related to a child's living environment and hygiene habits. Expression levels of AhR and NLRP3 were significantly higher in the exposed group than in the reference group. Similarly, serum IL-1β, IL-4, IL-5, IL-10, IL-12p70, IL-13, IL-17A, IL-18, IL-22, IL-23, and IFN-γ levels were notably higher in the e-waste-exposed children than in the reference children. After adjusting for age, gender, BMI, family income, parental education level, and second-hand smoke exposure, we found that increased PAH exposure was associated with higher AhR and NLRP3 expression and elevated IL-4, IL-10, IL-12p70, IL-18, IL-22, IL-23, TNF-α, and IFN-γ levels. The associations between PAH exposure and IL-1β, IL-18, IFN-γ, and TNF-β were mediated by NLRP3 expression, and the relationships between PAH exposure and IL-4, IL-10, IL-12p70, IL-22, IL-23, and TNF-α were mediated by AhR expression. Our findings suggest that the association between PAH exposure and a cytokine storm may be mediated by AhR and NLRP3 expression among preschoolers.


Elevated chromosome translocation frequencies in New Zealand nuclear test veterans.
In 1957/58 the British Government conducted a series of nuclear tests in the mid-Pacific codenamed Operation Grapple, which involved several naval vessels from Britain and New Zealand. Two New Zealand frigates with 551 personnel onboard were stationed at various distances between 20 and 150 nautical miles from ground zero. In the present study we applied the cytomolecular technique mFISH (multicolour fluorescent in situ hybridisation) to investigate a potential link between chromosome abnormalities and possible past radiation exposure in New Zealand nuclear test veterans who participated in Operation Grapple. Compared to age matched controls, the veterans showed significantly higher (P < 0.0001) frequencies of chromosomal abnormalities (275 translocations and 12 dicentrics in 9,360 cells vs. 96 translocations and 1 dicentric in 9,548 cells in the controls), in addition to a significant excess of CCRs (complex chromosomal rearrangements) in the veterans. A Kolmogorov-Smirnoff test showed that the distributions of translocations for the two groups were significantly different.


Alchemical Free Energy Calculations and Isothermal Titration Calorimetry Measurements of Aminoadamantanes Bound to the Closed State of Influenza A/M2TM.
Adamantane derivatives, such as amantadine and rimantadine, have been reported to block the transmembrane domain (TM) of the M2 protein of influenza A virus (A/M2) but their clinical use has been discontinued due to evolved resistance in humans. Although experiments and simulations have provided adequate information about the binding interaction of amantadine or rimantadine to the M2 protein, methods for predicting binding affinities of whole series of M2 inhibitors have so far been scarcely applied. Such methods could assist in the development of novel potent inhibitors that overcome A/M2 resistance. Here we show that alchemical free energy calculations of ligand binding using the Bennett acceptance ratio (BAR) method are valuable for determining the relative binding potency of A/M2 inhibitors of the aminoadamantane type covering a binding affinity range of only ∼2 kcal mol(-1). Their binding affinities measured by isothermal titration calorimetry (ITC) against the A/M2TM tetramer from the Udorn strain in its closed form at pH 8 were used as experimental probes. The binding constants of rimantadine enantiomers against M2TMUdorn were measured for the first time and found to be equal. Two series of alchemical free energy calculations were performed using 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipids to mimic the membrane environment. A fair correlation was found for DPPC that was significantly improved using DMPC, which resembles more closely the DPC lipids used in the ITC experiments. This demonstrates that binding free energy calculations by the BAR approach can be used to predict relative binding affinities of aminoadamantane derivatives toward M2TM with good accuracy.


Nasal Application of the Galantamine Pro-drug Memogain Slows Down Plaque Deposition and Ameliorates Behavior in 5X Familial Alzheimer's Disease Mice.
The plant alkaloid galantamine is an established symptomatic drug treatment for Alzheimer's disease (AD), providing cognitive and global relief in human patients. However, as an acetylcholinesterase inhibitor, gastrointestinal side effects limit the dosage and duration of treatment. Memogain (Gln-1062), a pro-drug, liberates galantamine on cleavage by a carboxyesterase in the brain. The possibility to deliver Memogain intranasally may further circumvent side effects, allowing higher dosing compared to galantamine. In this study, the 5X Familial Alzheimer's Disease (5XFAD) mouse model was used to investigate the effect of chronic Memogain treatment on behavior and amyloid-β (Aβ) plaque deposition in the brain. Chronic intranasal dosage of 6 mg/kg body weight twice daily was tolerated well, whereas the double dose caused body weight loss in males and was less effective in some behavioral tests. 8 weeks of chronic treatment resulted in improved performance in behavioral tests, such as open field and light-dark avoidance, and in fear conditioning already at mildly affected stages at the age of 18 weeks compared to untreated controls. Furthermore, after treatment a significantly lower plaque density in the brain, i.e., in the entorhinal cortex (reduction 20% females, 40% males) and the hippocampus (19% females, 31% males) at the age of 18 weeks was observed. These results show that nasal application of Memogain effectively delivers the drug to the brain with the potential to retard plaque deposition and improve behavioral symptoms in AD similar to the approved galantamine.


Resveratrol inhibits angiogenic response of cultured endothelial F-2 cells to vascular endothelial growth factor, but not to basic fibroblast growth factor.
Resveratrol, a natural polyphenol in grapes, is known to prevent the cardiovascular diseases and to exert the antiangiogenic effect in in vivo models with vascular endothelial growth factor (VEGF) or basic fibroblast growth factor (bFGF). We examined the effect of resveratrol on tubule formation of cultured endothelial F-2 cells. In collagen gel matrix, F-2 cells formed an extended network of tubular structures in response to VEGF or bFGF. Resveratrol dose-dependently prevented the VEGF-induced tubule formation, but failed to inhibit the angiogenic response to bFGF. We next examined whether the inhibition of nitric oxide (NO) production is linked to the antiangiogenic effect of resveratrol on VEGF-stimulated F-2 cells, because NO plays a crucial role in VEGF-induced tubular network formation. NO production was increased by VEGF, but not by bFGF, and resveratrol inhibited VEGF-stimulated NO production. N(G)-nitro-L-arginine methyl ester (L-NAME) potently inhibited NO production under all conditions, including VEGF stimulation, and abrogated VEGF-induced tubule formation. However, L-NAME did not inhibit bFGF-induced tubule formation. To investigate the bFGF-induced in vivo antiangiogenic effect of resveratrol, we examined the effect of resveratrol on prostaglandin E(2) (PGE(2)) production and cyclooxygenase (COX) expression in NRK-F fibroblasts. COX-2 and its derived PGE(2) are important factors for bFGF-induced in vivo angiogenesis. Resveratrol dose-dependently prevented both COX-2 induction and PGE(2) production in bFGF-stimulated fibroblasts. These results suggest that resveratrol exerts the inhibitory effects on VEGF- and bFGF-induced angiogenesis through different mechanisms including inhibition of NO production in VEGF-stimulated endothelial cells and inhibition of COX-2 induction in bFGF-stimulated fibroblasts.


ADAM17 mediates epidermal growth factor receptor transactivation and vascular smooth muscle cell hypertrophy induced by angiotensin II.
Angiotensin II (Ang II) promotes growth of vascular smooth muscle cells (VSMCs) via epidermal growth factor (EGF) receptor (EGFR) transactivation mediated through a metalloprotease-dependent shedding of heparin-binding EGF-like growth factor (HB-EGF). However, the identity of the metalloprotease responsible for this process remains unknown. To identify the metalloprotease required for Ang II-induced EGFR transactivation, primary cultured aortic VSMCs were infected with retrovirus encoding dominant negative (dn) mutant of ADAM10 or ADAM17. EGFR transactivation induced by Ang II was inhibited in VSMCs infected with dnADAM17 retrovirus but not with dnADAM10 retrovirus. However, Ang II comparably stimulated intracellular Ca2+ elevation and JAK2 tyrosine phosphorylation in these VSMCs. In addition, dnADAM17 inhibited HB-EGF shedding induced by Ang II in A10 VSMCs expressing the AT1 receptor. Moreover, Ang II enhanced protein synthesis and cell volume in VSMCs infected with control retrovirus, but not in VSMCs infected with dnADAM17 retrovirus. ADAM17 activated by the AT1 receptor is responsible for EGFR transactivation and subsequent protein synthesis in VSMCs. These findings demonstrate a previously missing molecular mechanism by which Ang II promotes vascular remodeling.


Inhibition of mRNA turnover in yeast by an xrn1 mutation enhances the requirement for eIF4E binding to eIF4G and for proper capping of transcripts by Ceg1p.
Null mutants of XRN1, encoding the major cytoplasmic exoribonuclease in yeast, are viable but accumulate decapped, deadenylated transcripts. A screen for mutations synthetic lethal with xrn1Delta identified a mutation in CDC33, encoding eIF4E. This mutation (glutamate to glycine at position 72) affected a highly conserved residue involved in interaction with eIF4G. Synthetic lethality between xrn1 and cdc33 was not relieved by high-copy expression of eIF4G or by disruption of the yeast eIF4E binding protein Caf20p. High-copy expression of a mutant eIF4G defective for eIF4E binding resulted in a dominant negative phenotype in an xrn1 mutant, indicating the importance of this interaction in an xrn1 mutant. Another allele of CDC33, cdc33-1, along with mutations in CEG1, encoding the nuclear guanylyltransferase, were also synthetic lethal with xrn1Delta, whereas mutations in PRT1, encoding a subunit of eIF3, were not. Mutations in CDC33, CEG1, PRT1, PAB1, and TIF4631, encoding eIF4G1, have been shown to lead to destabilization of mRNAs. Although such destabilization in cdc33, ceg1, and pab1 mutants can be partially suppressed by an xrn1 mutation, we observed synthetic lethality between xrn1 and either cdc33 or ceg1 and no suppression of the inviability of a pab1 null mutation by xrn1Delta. Thus, the inhibition of mRNA turnover by blocking Xrn1p function does not suppress the lethality of defects upstream in the turnover pathway but it does enhance the requirement for (7)mG caps and for proper formation of the eIF4E/eIF4G cap recognition complex.


US Smokers' Beliefs, Experiences and Perceptions of Different Cigarette Variants Before and After the FSPTCA Ban on Misleading Descriptors Such as "Light," "Mild," or "Low".
In December 2008, the Federal Trade Commission (FTC) took action that prompted the removal of nicotine and tar listings from cigarette packs and ads. As of June 2010, the US Family Smoking Prevention and Tobacco Control Act prohibited the use of explicit or implicit descriptors on tobacco packaging or in advertising that convey messages of reduced risk or exposure, specifically including terms like "light," "mild," and "low" and similar descriptors. This study evaluates the effect of these two policy changes on smokers' beliefs, experiences and perceptions of different cigarettes. Using generalized estimating equations models, this study analyzed survey data collected between 2002 and 2013 by the International Tobacco Control Policy Evaluation Study regarding US smokers' beliefs, experiences, and perceptions of different cigarettes. Between 2002 and 2013, smoker misperceptions about "light" cigarettes being less harmful did not change significantly and remained substantial, especially among those who reported using lower-strength cigarettes. After the two policy changes, reported reliance on pack colors, color terms, and other product descriptors like "smooth" to determine cigarette strength style trended upward. Policies implemented to reduce smoker misperceptions that some cigarettes are safer than others appear to have had little impact. Because of pack colors, color terms, descriptors such as "smooth," cigarette taste or feel, and possibly other characteristics, millions of smokers continue to believe, inaccurately, that they can reduce their harms and risks by smoking one cigarette brand or sub-brand instead of another, which may be delaying or reducing smoking cessation. What this study adds: This study confirms that US policies to reduce smoker misperceptions that some cigarettes are less harmful than others have not been successful. Following the removal of light/low descriptors and tar and nicotine numbers from cigarette packs and ads, pack colors, color words, other descriptors (eg, smooth), and sensory experiences of smoother or lighter taste have helped smokers to continue to identify their preferred cigarette brand styles and otherwise distinguish between which brands and styles they consider "lighter" or lower in tar and, mistakenly, less harmful than others. These findings provide additional evidence to support new enforcement or regulatory action to stop cigarettes and their packaging from misleading smokers about relative risk, which may be reducing or delaying quit attempts.


Sentence comprehension in autism: thinking in pictures with decreased functional connectivity.
Comprehending high-imagery sentences like The number eight when rotated 90 degrees looks like a pair of eyeglasses involves the participation and integration of several cortical regions. The linguistic content must be processed to determine what is to be mentally imaged, and then the mental image must be evaluated and related to the sentence. A theory of cortical underconnectivity in autism predicts that the interregional collaboration required between linguistic and imaginal processing in this task would be underserved in autism. This functional MRI study examined brain activation in 12 participants with autism and 13 age- and IQ-matched control participants while they processed sentences with either high- or low-imagery content. The analysis of functional connectivity among cortical regions showed that the language and spatial centres in the participants with autism were not as well synchronized as in controls. In addition to the functional connectivity differences, there was also a group difference in activation. In the processing of low-imagery sentences (e.g. Addition, subtraction and multiplication are all math skills), the use of imagery is not essential to comprehension. Nevertheless, the autism group activated parietal and occipital brain regions associated with imagery for comprehending both the low and high-imagery sentences, suggesting that they were using mental imagery in both conditions. In contrast, the control group showed imagery-related activation primarily in the high-imagery condition. The findings provide further evidence of underintegration of language and imagery in autism (and hence expand the understanding of underconnectivity) but also show that people with autism are more reliant on visualization to support language comprehension.


Low order modes in microcavities based on silicon colloids.
Silicon colloids based microcavities, with sphere size between 1 and 3 micrometers, have been synthesized and optically characterized. Due to both the small cavity volume and the high refractive index of silicon we are able to tune resonances with extremely low mode index, whose electric field distribution resembles those of electronic orbitals. The value of some parameters such as quality factor Q, effective mode volume, and evanescent field have been calculated for several modes. This calculation indicates silicon colloids can be a serious strategy for developing optical microcavities where may coexist both optical modes with large evanescent fields useful for sensing applications, as well as modes with high Q/V ratio values, of the order of 10(9)(λ/n)(-3).


Fiber type composition of rostral and caudal portions of the digastric muscle in the dog.
The distribution of muscle fiber types in rostral and caudal portions of the musculus digastricus (digastric muscle) was studied in 6 dogs. Staining procedures which stain specifically for type IIM fibers, a fiber type found in other muscles supplied by the trigeminal nerve, were used. Rostral and caudal portions of the muscle were compared because the rostral portion is innervated by the trigeminal nerve, and the caudal portion is innervated by the facial nerve. The musculus triceps brachii (triceps muscle), which contains fiber types I and IIA, and the musculus masseter (masseter muscle), which contains type IIM, were used as controls. Mean fiber diameters were calculated for each of the muscles. Both portions of the digastric muscle exhibited the same histochemical behavior, possessing types I and IIA myofibers. Neither portion contained type IIM fibers. Type I fibers in the masseter muscle were histochemically different from type I fibers in the other muscles studied. Type II fibers predominated in all 3 muscles, but there were significantly (P less than 0.001) more type I fibers in the triceps muscle than in either portion of the digastric muscle or in the masseter muscle. Type II fibers were significantly larger than type I fibers in the caudal digastric (P less than 0.01) and masseter (P less than 0.05) muscles. There was no difference in the size of type I or type II fibers between any of the muscles studied (P greater than 0.20).


Self-template synthesis of defect-rich NiO nanotubes as efficient electrocatalysts for methanol oxidation reaction.
Developing robust and inexpensive non-noble metal based anode electrocatalysts is highly desirable for alkaline direct methanol fuel cells (ADMFCs). Herein, we successfully develop a facile self-template synthetic strategy for gram-grade porous NiO nanotubes (NTs) by pyrolyzing a nanorod-like Ni-dimethylglyoxime complex. The pyrolysis temperature highly correlates with the morphology and crystallinity of NiO NTs. The optimal NiO NTs exhibit a large electrochemically active surface area, a fast catalytic kinetics, and a small charge transfer resistance, which induce an outstanding electrocatalytic activity for the methanol oxidation reaction (MOR). Compared with conventional NiO nanoparticles, NiO NTs achieve a 11.5-fold increase in mass activity at 1.5 V for the MOR due to nanotubal morphology and abundant non-vacancy defects on the NiO NT surface. Moreover, NiO NTs have a higher electrocatalytic activity for the intermediates of the MOR (such as formaldehyde and formate) than conventional NiO nanoparticles, which also contribute to MOR activity enhancement. Given the facile synthesis and enhanced electrocatalytic performance, NiO NTs may be promising anode electrocatalysts for ADMFCs.


Monoclonal antibody-based enzyme linked immunosorbent assay for the analysis of jasmonates in plants.
Methyl jasmonate (MeJA) and its free-acid form, jasmonic acid (JA) are naturally occurring plant growth regulators widely distributed in higher plants. In order to improve the sensitivity for the analysis of MeJA at low levels in small amounts of plant samples, a monoclonal antibody (MAb) (designated as MAb 3E(5)D(7)C(4)B(6)) against MeJA was derived from a JA-bovine serum albumin (BSA) conjugate as an immunogen. The antibody belongs to the IgG(1) subclass with a kappa type light chain and has a dissociation constant of approximately 6.07 x 10(-9) M. MAb3E(5)D(7)C(4)B(6) is very specific to MeJA. It was used to develop a direct competitive enzyme-linked immunosorbent assay (dcELISA), conventional and simplified indirect competitive ELISAs (icELISA). JA was derivatized into MeJA for the ELISA analysis. The IC(50) value and detection range for MeJA were, respectively, 34 and 4-257 ng/mL by the conventional icELISA, 21 and 3-226 ng/mL by the simplified icELISA and 5.0 and 0.7-97.0 ng/mL by the dcELISA. The dcELISA was more sensitive than either the conventional or simplified icELISA. The assays were used to measure the content of jasmonates as MeJA in tobacco leaves under drought stress or inoculated with tobacco mosaic virus and tomato leaves inoculated with tomato mosaic virus or Lirioinyza sativae Blanchard as compared with the corresponding healthy leaves. The increased jasmonates content indicated its role in response to the drought stress and pathogens.


Replacement, Reduction, Refinement - Animal welfare progress in European Pharmacopoeia monographs: activities of the European Pharmacopoeia Commission from 2007 to 2017.
Since the opening for signature of the European Convention for the Protection of Animals Used for Experimental and Other Scientific Purposes in 1986, the European Pharmacopoeia Commission and its experts have carried out a programme of work committed to Replacing, Reducing and Refining (3Rs) the use of animals for test purposes. While updates on achievements in the field of the 3Rs are regularly provided, this article summarises the activities of the Ph. Eur. Commission in this field within the last decade.


Effectiveness of mandibular advancement devices in the treatment of obstructive sleep apnea and the impact of different body positions on treatment: A systematic review and meta-analysis.
The meta-analysis aimed to evaluate the efficacy of mandibular advancement device (MAD) for the treatment of obstructive sleep apnea (OSA) and explore the effect of different positions on MAD for OSA. The Embase, PubMed, Medline, and Cochrane Library databases were searched for relevant studies evaluating the effect of MAD on the treatment of OSA from database inception to November 2022. The Bayesian random-effects mode was used to calculate the pooled outcome. Subgroup analysis and sensitivity analysis were applied to investigate the heterogeneity. A total of 6 studies enrolling 643 patients were eligible for further analysis. MAD treatment led to improvements in total apnea-hypopnea index (AHI) for both positional OSA(POSA) and Non-POSA groups, but there was no significant difference in the effect of MAD on Non-POSA and POSA (MD = -1.46,95%CI [-4.89,1.97], P = 0.40). In the supine position, AHI improvement after MAD treatment in POSA group was more than that in Non-POSA group by 15 events/hour in average (MD = 14.82, 95%CI [11.43,18.22], P<0.00001), while in the non-supine position, the change of AHI in Non-POSA group was significantly better than that in POSA group by approximately 8 events/hour (MD = -7.55,95%CI[-10.73,-4.38],p < 0.00001). MAD is more suitable for POSA compared to Non-POSA in patients with habitual sleep in the supine or supine predominant position. While for patients with habitual sleep in the non-supine position, MAD is an effective treatment option for Non-POSA.


Meningitis and encephalitis.
: Neurologists have a vital role in the recognition of meningitis and encephalitis, the accurate evaluation and interpretation of CSF studies, and the management and prevention of the neurologic complications of CNS infectious diseases. ;: Although the tetravalent meningococcal glycoconjugate vaccine has decreased the incidence of meningococcal meningitis, the vaccine does not contain serogroup B, which is responsible for one-third of cases of meningococcal disease. Thus, meningitis due to Neisseria meningitidis is still a concern in a vaccinated individual. Empiric therapy for meningitis associated with sinusitis, otitis, or mastoiditis should include antibiotic therapy for anaerobes. An organism that classically causes a subacute or chronic meningitis, such as Mycobacterium tuberculosis, may on occasion present with an acute onset of symptoms. ;: Unlike most other diseases, the management of patients with suspected meningitis or encephalitis begins with empiric therapy. The etiologic organism cannot always be identified. The goal is to identify those that are treatable, provide supportive care for those that are not, and, when possible, prevent the neurologic complications of these infections.


The need to reform our assessment of evidence from clinical trials: a commentary.
The ideology of evidence-based medicine (EBM) has dramatically altered the way we think, conceptualize, philosophize and practice medicine. One of its major pillars is the appraisal and classification of evidence. Although important and beneficial, this process currently lacks detail and is in need of reform. In particular, it largely focuses on three key dimensions (design, [type I] alpha error and beta [type II] error) to grade the quality of evidence and often omits other crucial aspects of evidence such as biological plausibility, reproducibility, generalizability, temporality, consistency and coherence. It also over-values the randomized trial and meta-analytical techniques, discounts the biasing effect of single centre execution and gives insufficient weight to large and detailed observational studies. Unless these aspects are progressively included into systems for grading, evaluating and classifying evidence and duly empirically assessed (according to the EBM paradigm), the EBM process and movement will remain open to criticism of being more evidence-biased than evidence-based."All scientific work is incomplete--whether it be observational or experimental. All scientific work is liable to be upset or modified by advancing knowledge. That does not confer upon us a freedom to ignore the knowledge we already have, or to postpone the action that it appears to demand at a given time". (Sir Bradford Austin Hill 1).


T cell-dependent immune response in C1q-deficient mice: defective interferon gamma production by antigen-specific T cells.
The role of the classical complement pathway in humoral immune responses was investigated in gene-targeted C1q-deficient mice (C1qA-/-). Production of antigen-specific immunoglobulin (Ig)G2a and IgG3 in primary and secondary responses to T cell-dependent antigen was significantly reduced, whereas IgM, IgG1, and IgG2b responses were similar in control and C1qA-/- mice. Despite abnormal humoral responses, B cells from C1qA-/- mice proliferated normally to a number of stimuli in vitro. Immune complex localization to follicular dendritic cells within splenic follicles was lacking in C1qA-/- mice. The precursor frequency of antigen-specific T cells was similar in C1qA-/- and wild-type mice. However, analysis of cytokine production by primed T cells in response to keyhole limpet hemocyanin revealed a significant reduction in interferon-gamma production in C1qA-/- mice compared with control mice, whereas interleukin 4 secretion was equivalent. These data suggest that the classical pathway of complement may influence the cytokine profile of antigen-specific T lymphocytes and the subsequent immune response.


Endothelial gaps: time course of formation and closure in inflamed venules of rats.
In the rat trachea, substance P causes rapid but transient plasma leakage. We sought to determine how closely the number, morphology, and size of endothelial gaps correspond to the time course of this leakage. Endothelial gaps were examined by scanning electron microscopy (EM), by transmission EM, or by light microscopy after silver nitrate staining. Substance P-induced leakage of the particulate tracer Monastral blue peaked at 1 min but decreased with a half-life of 0.3 min. The number of silver-stained gaps also peaked at 1 min then decreased significantly more slowly (half-life 1.9 min) than the leakage. Scanning EM revealed two types of endothelial gaps, designated vertical gaps and oblique slits. Vertical gaps predominated at peak leakage, whereas oblique slits became more common as the leakage diminished. Measurements of the mean diameter of vertical gaps made by light microscopy, scanning EM, and transmission EM were all in the range of 0.36-0.47 micron. Fingerlike endothelial cell processes that appeared during gap formation became shorter as the leakage diminished (mean length: 1.44 microns at 1 min compared with 1.06 microns at 3 min after substance P), suggesting a role in gap closure. We conclude that the plasma leakage occurring immediately after an inflammatory stimulus results from the rapid formation of endothelial gaps. Multiple factors, including alterations in gap morphology, gap closure, and changes in driving force, are likely to participate in the rapid decrease in the leakage.


Optimization of Long-Term Human iPSC-Derived Spinal Motor Neuron Culture Using a Dendritic Polyglycerol Amine-Based Substrate.
Human induced pluripotent stem cells (hiPSCs) derived from healthy and diseased individuals can give rise to many cell types, facilitating the study of mechanisms of development, human disease modeling, and early drug target validation. In this context, experimental model systems based on hiPSC-derived motor neurons (MNs) have been used to study MN diseases such as spinal muscular atrophy and amyotrophic lateral sclerosis. Modeling MN disease using hiPSC-based approaches requires culture conditions that can recapitulate in a dish the events underlying differentiation, maturation, aging, and death of MNs. Current hiPSC-derived MN-based applications are often hampered by limitations in our ability to monitor MN morphology, survival, and other functional properties over a prolonged timeframe, underscoring the need for improved long-term culture conditions. Here we describe a cytocompatible dendritic polyglycerol amine (dPGA) substrate-based method for prolonged culture of hiPSC-derived MNs. We provide evidence that MNs cultured on dPGA-coated dishes are more amenable to long-term study of cell viability, molecular identity, and spontaneous network electrophysiological activity. The present study has the potential to improve hiPSC-based studies of human MN biology and disease.We describe the use of a new coating substrate providing improved conditions for long-term cultures of human iPSC-derived motor neurons, thus allowing evaluation of cell viability, molecular identity, spontaneous network electrophysiological activity, and single-cell RNA sequencing of mature motor neurons.