Role of the Akt pathway in mRNA translation of interferon-stimulated genes.
Multiple signaling pathways are engaged by the type I and II IFN receptors, but their specific roles and possible coordination in the generation of IFN-mediated biological responses remain unknown. We provide evidence that activation of Akt kinases is required for IFN-inducible engagement of the mTOR/p70 S6 kinase pathway. Our data establish that Akt activity is essential for up-regulation of key IFN-alpha- and IFN-gamma-inducible proteins, which have important functional consequences in the induction of IFN responses. Such effects of the Akt pathway are unrelated to regulatory activities on IFN-dependent STAT phosphorylation/activation or transcriptional regulation. By contrast, they reflect regulatory activities on mRNA translation via direct control of the mTOR pathway. In studies using Akt1 and Akt2 double knockout cells, we found that the absence of Akt kinases results in dramatic reduction in IFN-induced antiviral responses, establishing a critical role of the Akt pathway in IFN signaling. Thus, activation of the Akt pathway by the IFN receptors complements the function of IFN-activated JAK-STAT pathways, by allowing mRNA translation of IFN-stimulated genes and, ultimately, the induction of the biological effects of IFNs.


Therapeutic relationships and the problem of containment: Experiences of patients at a psychiatric training hospital.
The biopsychosocial model emphasises the role of human relationships in psychiatric care. Therapeutic relationships that improve treatment outcome and provide containment are desperately needed by patients in distress. Despite the importance of human relationships, they are neglected in an era dominated by biological psychiatry. This qualitative research project explores the experiences, perceptions and subsequent needs of patients. The role of therapeutic relationships, and the factors that patients felt influenced their relationship with their therapists, were examined. A psychiatric training hospital in South Africa. Thirty in-depth semi-structured interviews were conducted with 15 inpatients. A qualitative, explorative-descriptive, collective case study design was used. Purposive sampling ensured maximum variation and richness of information. Grounded theory methods were used to analyse transcribed recordings. Patients valued therapeutic relationships that provide containment and potentially obviate the need for 'measures of control'. A model of containment was developed to demonstrate the various factors that interact in the attempt to provide containment to patients in a psychiatric training hospital system. Training hospitals should emphasise the role of therapeutic relationships in achieving containment and positive treatment outcomes. In developing countries, severe shortcomings in mental healthcare resources hinder the building of personal therapeutic relationships.


Characterization of an Injury Induced Population of Muscle-Derived Stem Cell-Like Cells.
We recently discovered a novel population of stem cells from the injured murine skeletal muscle. These injury induced muscle-derived stem cell-like cells (iMuSCs) are partially reprogrammed from differentiated myogenic cells and display a pluripotent-like state. The iMuSCs exhibit stem cell properties including the ability to differentiate into multiple lineages, such as neurogenic and myogenic differentiations; they also display a superior migration capacity that demonstrating a strong ability of muscle engraftment in vivo. IMuSCs express several pluripotent and myogenic stem cell markers; have the capability to form embryoid bodies and teratomas, and can differentiate into all three germ layers. Moreover, blastocyst microinjection showed that the iMuSCs contributed to chimeric embryos but could not complete germline transmission. Our results indicate that the iMuSCs are in a partially reprogrammed state of pluripotency, which are generated by the microenvironment of injured skeletal muscle.


Production and Preliminary In Vivo Evaluations of a Novel in silico-designed L2-based Potential HPV Vaccine.
L2-based Human Papillomavirus (HPV) prophylactic vaccines, containing epitopes from HPV minor capsid proteins, are under investigation as second-generation HPV vaccines. No such vaccine has passed clinical trials yet, mainly due to the low immunogenicity of peptide vaccines; so efforts are being continued. A candidate vaccine composed of two HPV16 L2 epitopes, flagellin and a Toll-Like Receptor (TLR) 4 agonist (RS09) as adjuvants, and two universal T-helper epitopes was designed in silico in our previous researches. The designed vaccine construct was expressed in E. coli BL21 (DE3) and purified through metal affinity chromatography. Following mice vaccination, blood samples underwent ELISA and flow cytometry analyses for the detection of IgG and seven Th1 and Th2 cytokines. Following immunization, Th1 (IFN-γ, IL-2) and Th2 (IL-4, IL-5, IL-10) type cytokines, as well as IgG, were induced significantly compared with the PBS group. Significant increases in IFN-γ, IL-2, and IL-5 levels were observed in the vaccinated group versus Freund's adjuvant group. The obtained cytokine induction profile implied both cellular and humoral responses, with a more Th-1 favored trend. However, an analysis of specific antibodies against L2 is required to confirm humoral responses. No significant elevation in inflammatory cytokines, (IL-6 and TNF-α), suggested a lack of unwanted inflammatory side effects despite using a combination of two TLR agonists. The designed construct might be capable of inducing adaptive and innate immunity; nevertheless, comprehensive immune tests were not conducted at this stage and will be a matter of future work.


Impact of an accelerated measles-mumps-rubella (MMR) vaccine schedule on vaccine coverage: An ecological study among London children, 2012-2018.
Vaccination coverage of dose two of MMR (MMR2) at 5th birthday has been consistently low in London and measured 76.3% in 2018/19. Since the early 2000s seven boroughs in London started offering dose two earlier, from 15 to 18 months onwards instead of the recommended 3 years 4 months. In this study we investigate whether the accelerated schedule of MMR2 leads to a change in coverage of MMR2 and other childhood vaccines with an ecological study using childhood immunisation data from 2009 to 2018 in London. We modelled coverage used generalized estimating equations (GEE) adjusted for year and DTaP/IPV/Hib3 coverage measured at 2nd birthday as a proxy for baseline local vaccination programme performance to determine the percentage point difference in coverage of MMR2 and other childhood vaccines. Average MMR2 coverage was higher among early implementing boroughs from 2012/13 onwards. Coverage difference was highest in 2017/18 (9.2 percentage points, 95% CI 4.8, 13.5, p < 0.001). On average over the 6 years, compared to London boroughs on the routine schedule, MMR2 coverage among early implementing boroughs was 3.3 percentage points higher (95% CI 1.3, 5.3, p = 0.01) after adjusting for DTaP/IPV/Hib3 coverage, IMD score and year. Earlier vaccination of MMR2 is associated with significantly higher coverage at five years for this vaccine in London. Further research is needed to assess the association at a more granular level, but our findings underline a potential opportunity to increase MMR coverage.


Effect of biodrying of lignocellulosic biomass on humification and microbial diversity.
By optimizing the carbon to nitrogen (C/N) ratio, this study accomplished an improved level of humification and microbial diversity in the biodrying process of lignocellulosic biomass. The results demonstrated that C/N ratio of 20 accelerated the decomposition of refractory lignocellulose, resulting in lower greenhouse gas emissions and the production of highly mature fertilizer with a germination index of 119.0% and a humic index of 3.2. Moreover, C/N ratio of 20 was found to diversify microbial communities, including Pseudogracilibacillus, Sinibacillus, and Georgenia, which contributed to the decomposition of lignocellulosic biomass and the production of humic acid. Hence, it is recommended to regulate the C/N ratio to 20:1 during the biodrying of biogas residue and wood chips to promote the economic feasibility and bioresource recycling.


A comparative study of open surgical and percutaneous renal biopsies.
Percutaneous renal biopsy was performed on 171 patients during the same interval as an open surgical biopsy was done on 100 patients in the same hospital. Patients who underwent an open biopsy had more severe renal dysfunction and hypertension than those who had a percutaneous biopsy. Tissue adequate for diagnosis was obtained in 100 per cent of the open biopsies. The kidney was reached in 88 per cent of all percutaneous biopsies with tissue adequate for diagnosis in 82.5 per cent. Of 87 patients biopsied with image-amplification fluoroscopy renal tissue was obtained in 97 per cent with tissue adequate for diagnosis in 92 per cent. No nephrectomies were done and no deaths occurred in either group. Our complications and yield by percutaneous biopsy compared favorably to those of other series. Further, open biopsy in a high risk population was associated with a high yield of tissue adequate for diagnosis with no increase in complications. Procurement of renal tissue need rarely be denied a patient if a biopsy is clinically indicated.


Identifying rare disease variants in the Genetic Analysis Workshop 17 simulated data: a comparison of several statistical approaches.
Genome-wide association studies have been successful at identifying common disease variants associated with complex diseases, but the common variants identified have small effect sizes and account for only a small fraction of the estimated heritability for common diseases. Theoretical and empirical studies suggest that rare variants, which are much less frequent in populations and are poorly captured by single-nucleotide polymorphism chips, could play a significant role in complex diseases. Several new statistical methods have been developed for the analysis of rare variants, for example, the combined multivariate and collapsing method, the weighted-sum method and a replication-based method. Here, we apply and compare these methods to the simulated data sets of Genetic Analysis Workshop 17 and thereby explore the contribution of rare variants to disease risk. In addition, we investigate the usefulness of extreme phenotypes in identifying rare risk variants when dealing with quantitative traits. Finally, we perform a pathway analysis and show the importance of the vascular endothelial growth factor pathway in explaining different phenotypes.


Effect of green tea catechins in patients with high-grade prostatic intraepithelial neoplasia: Results of a short-term double-blind placebo controlled phase II clinical trial.
Several studies suggest a protective role of green tea catechins against prostate cancer (PCa). In order to evaluate the efficacy of green tea catechins for chemoprevention of PCa in patients with high-grade prostate intraepithelial neoplasia (HG-PIN) we performed a phase II clinical trial. Sixty volunteers with HG-PIN were enrolled to carry out a double-blind randomized placebo-controlled phase II clinical trial. Treated group took daily 600 mg of green tea catechins (Categ Plus®) for 1 year. Patients were screened at 6 and 12 months through prostatic biopsy and measurements of prostate-specific antigen (PSA). Despite the statistically significant reduction of PSA observed in subjects who received green tea catechins for 6 and 12 months, we did not find any statistical difference in PCa incidence between the experimental groups neither after 6 nor after 12 months. However, throughout the one-year follow- up we observed very limited adverse effects induced by green tea catechins and a not significant improvement in lower urinary tract symptoms and quality of life. Although the small number of patients enrolled in our study and the relatively short duration of intervention, our findings seems to deny the efficacy of green tea catechins. However, results of our clinical study, mainly for its low statistical strength, suggest that the effectiveness of green tea catechins should be evaluated in both a larger cohort of men and longer trial.


The Effect of Repeated Botulinum Toxin A Therapy Combined with Intensive Rehabilitation on Lower Limb Spasticity in Post-Stroke Patients.
This study is a retrospective investigation of the effects of repetitive botulinum toxin A therapy (BoNT-A) and intensive rehabilitation (IR) on lower limb spasticity in post-stroke patients. Thirty-five post-stroke patients was included in this study and received BoNT-A for the first time. A 12-day inpatient protocol was with 4 cycles of the treatment protocol. The severity of spasticity, motor function and brace status were evaluated. The modified Ashworth Scale (MAS) score of ankle dorsiflexors, range of motion, walking speed and balancing ability were significantly improved after cycle 1. The improvement of spasticity and motor function was persistent through cycles 2⁻4. One-third of brace users were able to discontinue the use of a brace. All of these brace users showed a forward gait pattern prior to therapy. Repeated BoNT-A combined with IR improved lower limb spasticity in post-stroke patients. Our results suggest that patients who show the forward gait pattern prior to therapy may be able to discontinue the use of their brace after therapy.


Various heterologous cells exhibit interferon induced transfer of viral resistance.
Previously it was shown that cocultivation of mouse L and human WISH or baby hamster kidney cells in the presence of mouse interferon resulted in decreased viral yield from both cell species. We now show that this phenomenon also occurs when rabbit kidney and human WISH cells, with their corresponding interferons, are cocultivated with human WISH and baby hamster kidney cells, respectively. This finding increases the number of donor cell types to three. The related finding that monkey VERO and chick embryo cells can be recipients of transferred resistance expands the number of heterologous recipient cell species capable of receiving transferred resistence to five. Not all cell types tested have been shown to function in this transfer system. The fact that VERO cells, which do not produce interferon, are capable of receiving transferred resistence is significant because it indicates that the mechanism of transfer does not involve production or interferon by the recipient cells.


Biological response of bladders rendered continent by insertion of artificial sphincter.
Artificial sphincters were inserted in 44 children for intractable urinary incontinence. Of the patients 70 per cent achieved satisfactory continence (minimum followup 18 months) with no revisions, and 10 per cent were rendered continent with 1 revision and 10 per cent with multiple revisions. The procedure failed in 10 per cent of the patients primarily because of infection. Extensive urodynamic studies were performed at reoperation and postoperatively. Long-term urodynamic followup was available in 36 continent children. Hydronephrosis occurred in 2 per cent of the upper tracts assessed and 18 per cent of the patients had significant residual urine. Mean bladder capacity tended to increase progressively postoperatively. Mean bladder compliance also was well maintained.


The choleretic effects of N-acetylglucosaminides, major urinary metabolites of ursodeoxycholic acid, in bile fistula rats.
We investigated the effects of three bile acids conjugated with N-acetylglucosamine, ursodeoxycholate N-acetylglucosaminide, tauroursodeoxycholate N-acetylglucosaminide and glycoursodeoxycholate N-acetylglucosaminide, on bile flow and biliary excretion of various markers in comparison with ursodeoxycholic acid, tauroursodeoxycholic acid and glycoursodeoxycholic acid in bile fistula rats. These bile acids were infused intravenously at a constant rate of 0.3 or 0.6 micromol/min/100 g b.w. for 2 h. All bile acids examined increased bile flow in a dose-dependent manner. In particular, ursodeoxycholate N-acetylglucosaminide has a longer-lasting effect after its infusion on bile flow than the other bile acids. Furthermore, these bile acids markedly increased biliary total bile acid excretion. At a higher dose level, the coefficient of determination (r2) between the biliary total bile acid excretion and bile flow for ursodeoxycholate N-acetylglucosaminide (r2 = 0.39) was lower than that for the other bile acids (r2 = 0.75-0.92). The ursodeoxycholate N-acetylglucosaminide, as well as tauroursodeoxycholic acid, glycoursodeoxycholic acid, tauroursodeoxycholate N-acetylglucosaminide and glycoursodeoxycholate N-acetylglucosaminide, was mostly excreted in an unchanged form in bile, whereas ursodeoxycholic acid was excreted as a conjugate with taurine. The three N-acetylglucosaminides as well as ursodeoxycholic acid, tauroursodeoxycholic acid and glycoursodeoxycholic acid significantly increased the biliary excretion of cholesterol, phospholipid, bilirubin and total Ca2+. In contrast, the N-acetylglucosaminides significantly decreased in biliary bicarbonate concentration, whereas ursodeoxycholic acid significantly increased biliary bicarbonate concentration. However, tauroursodeoxycholic acid and glycoursodeoxycholic acid did not significantly change the biliary bicarbonate concentration. The results indicate that N-acetylglucosaminides have a choleretic effect in bile fistula rats. Our present study also demonstrates that N-acetylglucosaminides, but not ursodeoxycholic acid, tauroursodeoxycholic acid or glycoursodeoxycholic acid, can significantly reduce the biliary bicarbonate concentration. Furthermore, our findings suggest that ursodeoxycholate N-acetylglucosaminide may partly exert a choleretic effect via mechanisms different from those of the other bile acids.


Expression of calcitonin receptor-like receptor and receptor-activity-modifying proteins in human cranial arteries.
Recently a new type of proteins modulating the pharmacological profile of the calcitonin receptor-like receptor (CRLR) were identified. The receptor-activity-modifying proteins (RAMPs) were shown to be essential for the expression of a functional CRLR and furthermore the RAMPs seemed to modify ligand selectivity of CRLR: coexpression of CRLR and RAMP1 resulted in a CGRP1 type of receptor while an adrenomedullin receptor resulted when CRLR and RAMP2 were coexpressed. In the present study significant molecular expression of CRLR concomitant with RAMP1, 2 and 3 were demonstrated in human meningeal, cerebral and temporal arteries by use of reverse transcriptase polymerase chain reactions (RT-PCR). These findings support previous studies demonstrating functional CGRP1 receptors in human cranial arteries. Furthermore the present study suggests the potential for functional adrenomedullin receptors in human cranial arteries.


Nanosecond spectroscopy of retinol.
Nanosecond fluorescence spectroscopy was used to study the unique binding site of the retinol-binding protein (RBP) from human serum. At pH 7.4, the binding of retinol to RBP caused the following spectroscopic changes in the ligand: (a) an enhancement of the fluorescence decay time (gamma = 8 ns); and (b) an increase in the emission anisotropy (A = 0.29). Retinol in hexane has a fluorescent decay time of 4.2 ns and a low emission anisotropy (A = 0.02). The increase in the fluorescence decay time of bound retinol is not due to dielectric relaxation effects of polar groups, since nanosecond time-resolved emission spectra of either retinol in glycerol or retinol bound to RBP, failed to show any time-dependent shifts in emission maxima during the time period investigated 0 to 30 ns. The degree of rotational mobility of bound retinol was investigated by time emission anisotropy measurements. The observed rotational correlation time (theta = 7.2 ns) is consistent with a rigid compact macromolecule of 21,000 molecular weight.


Bone marrow involvement in a primary mediastinal extragonadal non-seminomatous germ cell tumour.
A case of primary mediastinal extragonadal germ cell tumour with involvement of bone marrow, a rare finding, is reported with a review of literature.


Now, I think doctors can be heroes … Medical student's attitudes towards the COVID-19 pandemic's impact on key aspects of medical education and how the image of the medical profession has changed due to the COVID-19 pandemic.
The COVID-19 (coronavirus disease 2019) pandemic hit the world in early 2020 and influenced medical education worldwide. Distance learning, risk of infection and patient care, telehealth literacy, medical ethics and research in medical education are key factors of medical education challenged by the pandemic. Additionally, the following question arises: "What do medical students think about their future profession in the face of this crisis?" A cross-sectional survey was conducted among all undergraduate medical students of the Medical University of Vienna. 872 students answered the self-developed questionnaire. Qualitative and quantitative data analyses were conducted. The data show that the COVID-19 pandemic has raised awareness for the key aspects. In all areas of interest, students' feeling of petaredness is limited. Limitations in practical training and distance learning as well as social isolation concern a majority of students. Neutral, positive, and negative themes emerged in qualitative data analysis. Only 8% of the students of the first 3 years of study versus 13.4% of the students in higher years commented negatively. 18.7% of male vs. only 12.5% of all female students' comments were positive. A large proportion of positive comments were dedicated to the relevance and deeper meaning of the medical profession. Infection risk and the demanding nature of the medical profession were predominant in negative comments. The COVID-19 crisis has turned the spotlight on several aspects of medical education in need of reform. In addition, the occupational image of the medical profession seems to shift under the weight of this pandemic.


Intrauterine diagnosis of omphalocele.
Omphalocele can be diagnosed in utero by ultrasonography. Cesarean section can then help reduce neonatal morbidity and mortality.


Establishing research priorities to improve the One Health efficacy of Australian general practitioners and veterinarians with regard to zoonoses: A modified Delphi survey.
While general medical practitioners (GPs) and veterinarians are often the first line responders in the face of a disease outbreak, pathways to improving the One Health efficacy of these clinicians remain unclear. A two-phase modified Delphi survey of professionals with known expertise in One Health ('expert panel') was used to 1) identify key knowledge, attitudes and practices (KAPs) of GPs and veterinarians that would be consistent with a One Health approach to zoonoses; and 2) determine priorities for future surveys with Australian GPs and veterinarians to identify important gaps that impede effective diagnosis and management of zoonoses. A list of 13 topics/sub-topics, as well as a list of 25 specific zoonotic diseases/agents emerged from the first phase of the survey. In the second phase the expert panel identified general knowledge of the clinical aspects and epidemiological aspects of zoonoses, as well as risk management practices, as the most important KAPs and research priorities for both GPs and veterinarians. In terms of diseases, the expert panel regarded knowledge of Hendra virus, Q fever, Australian bat lyssavirus (ABLV), anthrax and Brucella suis most important for veterinarians, whilst for GPs, Q fever, gastrointestinal/foodborne diseases, influenza, ABLV and local vector-borne diseases were found to be most important by the expert panel. Some differences were noted in terms of prioritization of topics/sub-topics and diseases/agents according to expert background (veterinary and non-veterinary). The Delphi survey technique enabled efficient collection of data from a diverse range of One Health 'experts'/specialists and provided clear priorities for proposed future research, and potentially for educational interventions to improve One Health efficacy of clinicians.


p63 gene structure in the phylum mollusca.
Roles of p53 family ancestor (p63) in the organisms' response to stressful environmental conditions (mainly pollution) have been studied among molluscs, especially in the genus Mytilus, within the last 15 years. Nevertheless, information about gene structure of this regulatory gene in molluscs is scarce. Here we report the first complete genomic structure of the p53 family orthologue in the mollusc Mediterranean mussel Mytilus galloprovincialis and confirm its similarity to vertebrate p63 gene. Our searches within the available molluscan genomes (Aplysia californica, Lottia gigantea, Crassostrea gigas and Biomphalaria glabrata), found only one p53 family member present in a single copy per haploid genome. Comparative analysis of those orthologues, additionally confirmed the conserved p63 gene structure. Conserved p63 gene structure can be a helpful tool to complement or/and revise gene annotations of any future p63 genomic sequence records in molluscs, but also in other animal phyla. Knowledge of the correct gene structure will enable better prediction of possible protein isoforms and their functions. Our analyses also pointed out possible mis-annotations of the p63 gene in sequenced molluscan genomes and stressed the value of manual inspection (based on alignments of cDNA and protein onto the genome sequence) for a reliable and complete gene annotation.


Optimising water holding capacity and hydrophobicity of biochar for soil amendment - A review.
Biochar is a product of the thermal treatment of biomass, and it can be used for enhancing soil health and productivity, soil carbon sequestration, absorbance of pollutants from water and soil, and promoting environmental sustainability. Extensive research has been done on applications of biochar to enhance the Water Holding Capacity (WHC) of biochar amended soil. However, a comprehensive road map of biochar optimised for enhanced WHC, and reduced hydrophobicity is not yet published. This review is the first to provide not only quantitative information on the impacts of biochar properties in WHC and hydrophobicity, but also a road map to optimise biochar for enhanced WHC when applied as a soil amendment. The review shows that straw or grass-derived biochar (at 500-600 °C) increases the WHC of soil if applied at 1 to 3 % in the soil. It is clear from the review that soil of varying texture requires different particle sizes of biochar to enhance the WHC and reduce hydrophobicity. Furthermore, the review concludes that ageing biochar for at least a year with enhanced oxidation is recommended for improving the WHC and reducing hydrophobicity compared to using biochar immediately after production. Additionally, while producing biochar a residence time of 1 to 2 h is recommended to reduce the biochar's hydrophobicity. Finally, a road map for optimising biochar is presented as a schematic that can be a resource for making decisions during biochar production for soil amendment.


Antihistamines and Ovarian Cancer Survival: Nationwide Cohort Study and in Vitro Cell Viability Assay.
Antihistamines with cationic amphiphilic drug (CAD) characteristics induce cancer-specific cell death in experimental studies. Epidemiologic evidence is, however, limited. In a Danish nationwide cohort of ovarian cancer patients diagnosed during 2000-2015 (n = 5075), we evaluated the association between filled antihistamine prescriptions and cancer mortality. We used Cox regression models to estimate hazard ratios (HRs) with 95% confidence intervals (CIs) for ovarian cancer mortality. In an in vitro cell viability assay, we evaluated cell death in three ovarian cancer cell lines after treatment with clinically relevant doses of eight antihistamines. In our cohort study, CAD antihistamine use (≥1 prescription; n = 133) was associated with a hazard ratio of 0.63 (95% CI = 0.40 to 0.99) compared to use of non-CAD antihistamines (n = 304), and we found a tendency toward a dose-response association. In our cell viability assay, we found consistent and dose-dependent cytotoxicity for all CAD but not non-CAD antihistamines. In this nationwide cohort study, use of antihistamines with CAD characteristics is associated with a prognostic benefit in ovarian cancer patients.


Reinterpreting mortality statistics: some uses of Gompertzian analysis in epidemiological research.
Gompertzian analyses of mortality data have recently been undertaken for a number of individual conditions (Riggs JE. Mech Ageing Dev 1990; 55: 207-220 [1]; Neilson S et al. Mech Ageing Dev 1992; 64: 201-216 [2]; Neilson S et al. Acta Neurol Scand 1993; 87: 184-191 [3]). Such analyses are in principle of particular epidemiological value in circumstances where demographic change is occurring and where the balance between mortality from different conditions is subject to change. However, the extent to which a Gompertzian relationship between age and mortality holds for particular conditions has been subject to debate. In this analysis it is demonstrated that even some conditions which do not superficially hold to a Gompertz relationship do in fact do so, if such conditions are considered to be restricted to small, inherently susceptible subpopulations. By analysing mortality from a range of neurological conditions within the context of general mortality in England and Wales, conditions with different aetiologies such as Huntington's chorea, amyotrophic lateral sclerosis and multiple sclerosis can be shown to have a Gompertzian mortality rate distribution. Such analyses are of substantial value in indicating how demographic change affects the balance of mortality between conditions, as well as directing interest to revised aetiological possibilities.


A model for communication of sensory quality in the seafood processing chain.
Sensory quality has a key influence of consumer perception of a product. It is therefore of great importance for the processing industry that the sensory quality fulfils the expectations of the consumer. Sensory evaluations are the ultimate tool to measure and communicate sensory quality, but it is generally not fully implemented in the chain from catch to consumer. The importance of communicating sensory demands and results from evaluations in the seafood processing chain is described and a Seafood Sensory Quality Model (SSQM) is suggested as a communication tool.


An improved enzyme linked immunosorbent assay for detection of anti-ranavirus antibodies in the serum of the giant toad (Bufo marinus).
An improved ranavirus antibody ELISA (R Ab ELISA) for the specific detection of anti-ranavirus antibodies in toad sera was developed. Sheep anti-epizootic haematopoietic necrosis virus (EHNV) was used as the antigen-capture antibody. EHNV was used as the antigen and sera from field and challenged toads were used to detect the virus. Rabbit anti-toad IgG and IgM were used to detect bound toad antibody. Pre-absorption of toad sera with a monoclonal antibody, raised against the 50 kDa EHNV protein, improved the specificity of the technique. A blocking ELISA, immunofluorescence and immuno-electron microscopy were used to confirm the validity of the ELISA. The assay has potential use in screening sera from Bufo marinus for the presence of antibodies against ranaviruses and to facilitate understanding of the humoral immunological response in toads during virus infection.


Potential Use of Tannin Extracts as Additives in Semen Destined for Cryopreservation: A Review.
Cryopreservation and storage of semen for artificial insemination (AI) result in excessive accumulation of reactive oxygen species (ROS). This leads to a shortened life span and reduced motility of spermatozoa post-thawing, with consequent impairment of their function. However, certain levels of ROS are essential to facilitate the capacitation of spermatozoa required for successful fertilisation. Tannins, as well-known antioxidant compounds, may act as ROS binders/acceptors/scavengers to inhibit the damaging effects of ROS. This review comprises an analysis of the semen cryopreservation protocol and health functions of tannins, as well as the effects of ROS on fresh and cryopreserved semen's longevity and fertilisation. Additionally, we surveyed available evidence of the effects of tannin extract feed supplementation on male fertility. We furthermore interrogated existing theories on tannin use as a potential additive to semen extenders, its relationship with semen quality, and to what degree existing theories have been investigated to develop testable new hypotheses. Emphasis was placed on the effects of tannins on ROS, their involvement in regulating sperm structure and function during cryopreservation, and on post-thaw sperm motility, capacitation, and fertilising ability. The diverse effects of tannins on the reproductive system as a result of their potential metal ion chelation, protein precipitation, and biological antioxidant abilities have been identified. The current data are the first to support the further investigation of the incorporation of tannin-rich plant extracts into semen extenders to enhance the post-thaw survival, motility, and fertilising ability of cryopreserved spermatozoa.


Impact of policies and performance measurement on development of organizational coordinating strategies for chronic care delivery.
To examine the impact of policy directives and performance feedback on the organization (specifically the coordination) of foot care programs for veterans, as mandated by public law within the Department of Veterans Affairs Health Care System (VA). Case study of 10 VA medical centers performing diabetes-related amputations. Based on expert consensus, we identified 16 recommended foot care delivery coordination strategies. Structured interview protocols developed for primary care, foot care, and surgical providers, as well as administrators, were adapted from a prior study of surgical departments. Although performance measurement results for foot risk screening and referral were high at all study sites over 2 calendar years (average 85%, range 69% to 92%), the number of coordination strategies implemented by any site was relatively low, averaging only 5.4 or 34% (range 1-12 strategies). No facility had systematically collected data to evaluate whether preventive foot care was provided to patients with high-risk foot conditions, or whether these patients had unmet foot care needs. Although foot care policies and data feedback resulted in extremely high rates of adherence to foot-related performance measurement, there remained opportunities for improvement in the development of coordinated, technology-supported, data-driven, patient-centered foot care programs.


Granuloma formation after chevron osteotomy fixation with absorbable copolymer pin: a case report.
After fixation of a chevron osteotomy for hallux valgus of the left foot with a bioabsorbable copolymer, a delayed onset of pain at the surgical site developed. Magnetic resonance imaging showed a cystic lesion that was unresponsive to conservative treatment. Surgical excision determined the lesion to be a giant-cell granuloma formed as a foreign body response to the degrading copolymer.


Bioresponsive Nanoparticles Targeted to Infectious Microenvironments for Sepsis Management.
Sepsis is a life-threatening disease resulted from a dysregulated host immune response to bacterial infections, continuing to cause high morbidity and mortality worldwide. Despite discoveries of many potential therapeutic targets, effective treatments of sepsis are lacking. Here, a strategy is reported to target infectious microenvironments (IMEs) via bioresponsive nanoparticles that simultaneously eliminate bacteria and alleviate the host inflammation response, thus managing sepsis in mice. The nanoparticle is made of copolymers sensitive to pH and bacterial enzymes to self-assemble into a micelle loaded with both an antibiotic (ciprofloxacin) and an anti-inflammatory agent ((2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide). In addition, the nanoparticle is conjugated with intercellular adhesion molecule-1 antibodies to target IMEs. Nanoparticle targeting to IMEs and local cues as triggers to deliver therapeutics in on-demand manners is demonstrated using an acute lung bacterial infection mouse model. In the sepsis mouse model induced by peritonitis at a lethal dose of bacterial invasion, it is shown that concurrently targeting pathogens and excessive inflammation pathways is valuable to manage the sepsis. The study illustrates not only the development of a new delivery system but also the mechanism-based therapy of nanomedicine for infectious diseases.


Selenium metabolism in Escherichia coli.
Escherichia coli will reduce selenite (SeO3(2-)) and selenate (SeO4(2-)) to elemental selenium Se0. Selenium will also become incorporated into proteins as part of the amino acids selenocysteine or selenomethionine. The reaction of selenite with glutathione produces selenodiglutathione (GS-Se-GS). Selenodiglutathione and its subsequent reduction to glutathioselenol (GS-SeH) are likely the key intermediates in the possible metabolic fates of selenium. This review presents the possible pathways involving selenium in E. coli. Identification of intermediates and potential processes from uptake of the toxic oxyanions through to their detoxification will assist us in understanding the complexities of metalloid oxyanion metabolism in these bacteria.


Oesophageal varices predict complications in compensated advanced non-alcoholic fatty liver disease.
We aimed to evaluate the impact of oesophageal varices (OV) and their evolution on the risk of complications of compensated advanced chronic liver disease (cACLD) caused by non-alcoholic fatty liver disease (NAFLD). We also assessed the accuracy of non-invasive scores for predicting the development of complications and for identifying patients at low risk of high-risk OV. We performed a retrospective assessment of 629 patients with NAFLD-related cACLD who had baseline and follow-up oesophagogastroduodenoscopy and clinical follow-up to record decompensation, portal vein thrombosis (PVT), and hepatocellular carcinoma. Small and large OV were observed at baseline in 30 and 15.9% of patients, respectively. The 4-year incidence of OV from absence at baseline, and that of progression from small to large OV were 16.3 and 22.4%, respectively. Diabetes and a ≥5% increase in BMI were associated with OV progression. Multivariate Cox regression revealed that small (hazard ratio [HR] 2.24, 95% CI 1.47-3.41) and large (HR 3.86, 95% CI 2.34-6.39) OV were independently associated with decompensation. When considering OV status and trajectories, small (HR 2.65, 95% CI 1.39-5.05) and large (HR 4.90, 95% CI 2.49-9.63) OV at baseline and/or follow-up were independently associated with decompensation compared with the absence of OV at baseline and/or follow-up. The presence of either small (HR 2.8, 95% CI 1.16-6.74) or large (HR 5.29, 95% CI 1.96-14.2) OV was also independently associated with incident PVT. In NAFLD-related cACLD, the presence, severity, and evolution of OV stratify the risk of developing decompensation and PVT. Portal hypertension is the main driver of liver decompensation in chronic liver diseases, and its non-invasive markers can help risk prediction. The presence, severity, and progression of oesophageal varices stratify the risk of complications of non-alcoholic fatty liver disease. Easily obtainable laboratory values and liver stiffness measurement can identify patients at low risk for whom endoscopy may be withheld, and can also stratify the risk of liver-related complications.


Psychosocial and Health-Related Factors Associated With Discordance Between 13-Year Trajectories of Self-Reported Functional Limitations and Performance-Based Physical Functioning in Old Age.
Objective: The objective of this study was to examine correlates of discordance between 13-year trajectories of self-reported functional limitations and performance-based physical functioning in older adults. Method: We included 2,135 participants from the population-based Longitudinal Aging Study Amsterdam, the Netherlands, followed across 1995-2008. Self-reported functional limitations included six (instrumental) activities of daily living. Performance-based functioning was a composite of four tests. We used latent class growth analysis and multinomial logistic regression to examine discordance and its correlates. Results: Patterns of discordance and concordance (41% concordance, 46% "overestimation" of daily functioning, 13% "underestimation") appeared to be persistent over 13 years. Older age, male sex, cognitive impairment, absence of pain, and light alcohol use were associated with overestimation. Younger age, female sex, and lower self-rated health were associated with underestimation. Discussion: Factors associated with overestimation partly differ from those associated with underestimation. Factors that are highly stable over time are particularly good indicators of persistent discordance.


Single tonic-clonic seizure after energy drink abuse.
Energy drinks are soft beverages especially marketed for adolescents in order to obtain a heightened sense of awareness. Concerns about the safety of these drinks are raised based on our observation of potentially serious adverse effects. Caffeine and taurine are psychoactive agents highly present in energy drinks, which may lead to modification of neurotransmission. We herein report the case of a 20-year-old man presenting with a generalized epileptic seizure after energy drink consumption.


Bilirubin Induces A1-Like Reactivity of Astrocyte.
Astrocytes play an important role in the pathogenesis of bilirubin neurotoxicity, and activated astrocytes might be potential mediators of neuroinflammation processes contributing to neuronal cell death and tissue injury. Recent studies have reported that activated microglia induce two types of reactive astrocytes. A1 astrocytes could cause neuronal death and synaptic damage, as well as impaired phagocytosis. Therefore, the purpose of this study was to investigate whether unconjugated bilirubin (UCB)-induced A1-like astrocytes take on a neuroinflammation type and the underlying regulatory mechanisms. In this study, primary cortical astrocytes were treated with UCB in vitro. We detected the expression of complement component 3 (C3), S100 calcium binding protein A10 (S100A10), nuclear factor kappa B (NF-κB), NLR family pyrin domain containing 3 (NLRP3), activated caspase-1, gasdermin D N-terminal (GSDMD-N), PSD95, synaptophysin (SYP), the transcription levels of interleukin (IL)-1β and IL-18, and the survival rate of astrocytes after UCB treatment. The results showed that an increase in C3 was accompanied by a decrease in S100A10, and that A1-like astrocytes were functionally expressed after UCB stimulation. Meanwhile, the NF-κB and caspase-1 pathways were activated after UCB stimulation. After adding the NF-κB-specific inhibitor trans-activator of transcriptional-NEMO-binding domain (TAT-NBD) and caspase-1 specific inhibitor VX-765, the survival rate of astrocytes and neurons increased, whereas the protein expression of C3, NF-κB, NLRP3, activated caspase-1, and GSDMD-N decreased, and the mRNA levels of IL-1β and IL-18 reduced. Thus, we concluded that UCB stimulates the activation of A1-like astrocytes. Inhibition of NF-κB and caspase-1 alleviated A1-like astrocytes and exerted anti-inflammatory protective effects.


Characterization of allosteric modulators that disrupt androgen receptor co-activator protein-protein interactions to alter transactivation-Drug leads for metastatic castration resistant prostate cancer.
Three series of compounds were prioritized from a high content screening campaign that identified molecules that blocked dihydrotestosterone (DHT) induced formation of Androgen Receptor (AR) protein-protein interactions (PPIs) with the Transcriptional Intermediary Factor 2 (TIF2) coactivator and also disrupted preformed AR-TIF2 PPI complexes; the hydrobenzo-oxazepins (S1), thiadiazol-5-piperidine-carboxamides (S2), and phenyl-methyl-indoles (S3). Compounds from these series inhibited AR PPIs with TIF2 and SRC-1, another p160 coactivator, in mammalian 2-hybrid assays and blocked transcriptional activation in reporter assays driven by full length AR or AR-V7 splice variants. Compounds inhibited the growth of five prostate cancer cell lines, with many exhibiting differential cytotoxicity towards AR positive cell lines. Representative compounds from the 3 series substantially reduced both endogenous and DHT-enhanced expression and secretion of the prostate specific antigen (PSA) cancer biomarker in the C4-2 castration resistant prostate cancer (CRPC) cell line. The comparatively weak activities of series compounds in the H3-DHT and/or TIF2 box 3 LXXLL-peptide binding assays to the recombinant ligand binding domain of AR suggest that direct antagonism at the orthosteric ligand binding site or AF-2 surface respectively are unlikely mechanisms of action. Cellular enhanced thermal stability assays (CETSA) indicated that compounds engaged AR and reduced the maximum efficacy and right shifted the EC50 of DHT-enhanced AR thermal stabilization consistent with the effects of negative allosteric modulators. Molecular docking of potent representative hits from each series to AR structures suggest that S1-1 and S2-6 engage a novel binding pocket (BP-1) adjacent to the orthosteric ligand binding site, while S3-11 occupies the AR binding function 3 (BF-3) allosteric pocket. Hit binding poses indicate spaces and residues adjacent to the BP-1 and BF-3 pockets that will be exploited in future medicinal chemistry optimization studies. Small molecule allosteric modulators that prevent/disrupt AR PPIs with coactivators like TIF2 to alter transcriptional activation in the presence of orthosteric agonists might evade the resistance mechanisms to existing prostate cancer drugs and provide novel starting points for medicinal chemistry lead optimization and future development into therapies for metastatic CRPC.


The benefits of early androgen blockade.
Altering the effects of testosterone on prostate cancer cells by blocking androgen production or action is a critical part of treating this malignancy. The appropriate timing of androgen blockade for advanced prostate cancer is controversial. Despite many phase-III studies, gaps in clinical information still remain, and some questions are still not conclusively answered. This review concentrates on high-level evidence to address common clinical situations in advanced and metastatic prostate cancer. Additionally, the side-effects of androgen blockade have become increasingly visible and relevant.


[Panorama of self-management initiatives in Spain].
To describe the current situation of self-management initiatives in Spain. We performed a descriptive study of self-management support initiatives in Spain from the perspective of the patient as expert. Three databases were searched in October 2010 (Pubmed, Scientific Electronic Library Online [SCIELO] and Indice Médico Español [IME]), using the following Keywords «paciente experto» (expert patient), «paciente activo» (active patient) and «apoyo al autocuidado» (self-management support). Web sites were also consulted, using the same key words. Of the initiatives found, we selected those with the most advanced development and continuity, using the perspective of the expert patient (in which patients have an active role) and with a systematic format and methodology. A questionnaire was designed and was sent to the heads of the selected initiatives in the last quarter of 2010. To update the information, the questionnaire was sent again between August and September, 2012. Subsequently, the web sites were visited to review their contents and presence in social networks. Seven initiatives were identified in the autonomous regions of Murcia, Andalusia, Galicia, Castile-La Mancha, Basque Country, and Catalonia. These initiatives used distinct methodologies, formats and assessment systems. In Spain, there is increasing interest in the development of self-management support programs, although their scope is limited and their impact is mostly unknown, except for patient satisfaction. There is a need for studies on results assessment to identify the impact of these initiatives in our setting, as well as for studies on their implementation to encourage the introduction of patient activation initiatives in routine clinical practice.


Diminution of ethanol-induced hyperglycaemia in the rat by administration of beta-lactam antibiotics with a tetrazole thiol group.
The acute-onset hyperglycaemia produced by i.p. injection of 2 g of ethanol/kg body weight in adult female SPF Sprague-Dawley rats (200-220 g) was considerably less pronounced when 100 mumol/kg b.w. of one of the following tetrazole thiol-containing beta-lactam antibiotics (BLAs) was given i.p. 3 hours in advance: cephamandole (CMD), moxalactam (MOX), cephoperazone (CPZ) or cephothiam (CTM). An equimolar dose of cephazolin (CEZ), a thiadiazole thiol-containing cephalosporin, did not affect the ethanol-induced hyperglycaemia. Administration of an equimolar oral dose of disulfiram, which has an NCS structure similar to that of the tetrazole thiol-containing BLAs, produced an additional increase of the ethanol-induced hyperglycaemia. The diminution of the ethanol-induced hyperglycaemia by BLAs with a tetrazole thiol group appears to be linked to their NCS structure. It is conceivable that this effect which might be important for human therapy, is causally related to an inhibition of the glycolytic or gluconeogenetic enzyme system.


Antifouling paints leach copper in excess - study of metal release rates and efficacy along a salinity gradient.
Antifouling paints are biocidal products applied to ship and boat hulls in order to prevent the growth and settlement of marine organisms, i.e. fouling. The release of biocides from the surface of the paint film act to repel or poison potential settling organisms. Currently, the most commonly used biocide in antifouling paints is cuprous oxide. In the EU, antifouling products are regulated under the Biocidal Products Regulation (BPR), which states that the recommended dose should be the minimum necessary to achieve the desired effect. For antifouling products, the dose is measured as the release rate of biocide(s) from coating. In this study, the release rates of copper and zinc from eight different coatings for leisure boats were determined through static exposure of coated panels in four different harbors located in Swedish waters along a salinity gradient ranging from 0 to 27 PSU. The results showed the release rate of copper to increase with increasing salinity. Paints with a higher content of cuprous oxide were also found to release larger amounts of copper. The coatings' ability to prevent biofouling was also evaluated and no significant difference in efficacy between the eight tested products was observed at the brackish and marine sites. Hence, the products with high release rates of copper were equally efficient as those with 4 - 6 times lower releases. These findings suggest that current antifouling paints on the market are leaching copper in excess of the effective dose in brackish and marine waters. Additionally, the results from the freshwater site showed no benefit in applying a copper-containing paint for the purpose of fouling prevention. This indicates that the use of biocidal paints in freshwater bodies potentially results in an unnecessary release of copper. By reducing the release rates of copper from antifouling paints in marine waters and restricting the use of biocidal paints in freshwater, the load of copper to the environment could be substantially reduced.


Contamination evaluation and source identification of heavy metals in sediments near outlet of Shekou industrial district of Shenzhen City.
Sediment samples in this study were taken from five typical sites near the outlet of Shekou industrial district of Shenzhen City, China. The concentrations of seven elements including Cr, Cu, Cd, Pb, Hg, Zn, and As were determined respectively by atomic absorption spectrometry or atomic fluorescence spectrometry. The pollution degrees of the seven elements were assessed with the contamination factor and potential ecological risk index respectively, and their different sources were identified using multivariate statistical methods. The calculated contamination factors of these elements indicated that the sediments were at least moderately polluted by all the surveyed elements except As. The values of potential ecological risk indexes obtained decreased as the following order: Cd > Hg > Cu > As > Pb > Zn > Cr, and suggested that Cd and Hg were respectively at high and considerable environmental health risks. In addition, multivariate statistical analyses indicated that Cd, Hg, Cu, and Zn were most likely from the waste of electroplating, metal, and battery industries at Shekou industrial district, while Pb and As originated from both natural processes and anthropogenic activities along the bank of Pear River such as coal transportation and combustion, glass manufacturing, and painting, and Cr came mostly from Cr-related industries especially leather tanning within Shenzhen City. This study provided useful reference information about heavy metal contamination in the sediments in the estuarine and coastal areas with rapid urbanization and industrialization, and should be very helpful for the local governments to make relevant policies and strategies of heavy metal contamination control and management in developing countries.


Efficacy evaluation of Der p 1 DNA vaccine for allergic asthma in an experimental mouse model.
Der p 1 is a major allergen that reacts with IgE in more than 70% allergic sera, hence a good candidate for DNA vaccine development. Previous study demonstrated that intramuscular injection of plasmid DNA containing wild type Der p 1 gene induced specific Th1 skewed immune response. The aim of this study is to further optimize the DNA construct to achieve preventive and therapeutic efficacies. An optimized construct, containing mouse Igkappa light chain leader sequence followed by codon optimized mature Der p 1 gene, was generated and used to prevent or treat Der p1 induced allergic asthma in an experimental model. Results showed that this optimized construct was effective in the inhibition of Der p 1 specific IgE levels, attenuation of Th2 response and reduction of airway hyperresponsiveness in terms of prophylactic efficacy. It also showed therapeutic efficacy in IgE inhibition and up-regulation of IgG2a and IFN-gamma. The information obtained here could facilitate the design of DNA vaccines for allergy in general.


Hip arthroscopy and hypothermia.
To determine the incidence of and factors that contribute to the development of hypothermia during hip arthroscopic surgery. An analytic observational study was carried out in a cohort of 73 consecutive patients. All patients underwent hip arthroscopy for the treatment of femoroacetabular impingement. The patients' core temperature (esophageal) was measured throughout the surgery. Relevant information was collected on the patients (age, gender, body mass index, blood pressure) and on the procedure (volume and temperature of saline solution, pressure of fluid pump, surgery time, room temperature). The corresponding statistical analysis was performed with Stata 10.0 (StataCorp, College Station, TX), by use of a repeated-measures generalized estimating equations model. The patients' mean age was 33 years, and there were 39 female and 34 male patients. The mean body mass index was 23.9; systolic blood pressure, 97.5 mm Hg; and diastolic blood pressure, 52.2 mm Hg. The incidence of hypothermia below 35°C (95°F) was 2.7%. The multivariate statistical analysis of the results showed a direct relation between hypothermia and surgery time of more than 120 minutes (P < .001). There was an inverse relation between core body temperature and surgery time (P < .001), with a drop of 0.19°C/h (32.342°F/h). Of the patients, 68.22% had a decrease in temperature of more than 0.5°C (32.9°F) until the end of surgery. There was also a direct relation between core body temperature and saline solution temperature (P < .001), body mass index (P < .01), and diastolic blood pressure (P < .03). The incidence of hypothermia below 35°C (95°F) in patients who underwent hip arthroscopy for the treatment of femoroacetabular impingement is 2.7%. The factors that contribute toward the development of hypothermia during hip arthroscopic surgery are prolonged surgery time, low body mass index, low blood pressure during the procedure, and low temperature of the arthroscopic irrigation fluid.


A practical method of modeling a treatment couch using cone-beam computed tomography for intensity-modulated radiation therapy and RapidArc treatment delivery.
The effect of a treatment couch on dose perturbation is not always fully considered in intensity-modulated radiation therapy (IMRT) and volumetric modulated arc therapy (VMAT). In the course of inverse planning radiotherapy techniques, beam parameter optimization may change in the absence of the couch, causing errors in the calculated dose distributions. Although modern treatment planning systems (TPS) include data for the treatment couch components, they are not manufactured identically. Thus, variations in their Hounsfield unit (HU) values may exist. Moreover, a radiotherapy facility may wish to have a third-party custom tabletop installed that is not included by the TPS vendor. This study demonstrates a practical and simple method of acquiring reliable computed tomography (CT) data for the treatment couch and shows how the absorbed dose calculated with the modeled treatment couch can differ from that with the default treatment couch found in the TPS. We also experimentally verified that neglecting to incorporate the treatment couch completely in the treatment planning process might result in dose differences of up to 9.5% and 7.3% for 4-MV and 10-MV photon beams, respectively. Furthermore, 20 RapidArc and IMRT cases were used to quantify the change in calculated dose distributions caused by using either the default or modeled couch. From 2-dimensional (2D) ionization chamber array measurements, we observed large dose distribution differences between the measurements and calculations when the couch was omitted that varied according to the planning technique and anatomic site. Thus, incorporating the treatment couch in the dose calculation phase of treatment planning significantly decreases dose calculation errors.


Comparative study of binding of ovine complement factor H with different Borrelia genospecies.
This study presents the binding of ovine factor H (fH) by various serotypes of Borrelia and simultaneously correlates their complement resistance to sheep serum. Affinity ligand binding assay was employed to study the binding of borrelial proteins to ovine recombinant fH and its truncated forms (short consensus repeat, SCR 7 and SCRs 19-20). From a repertoire of 17 borrelial strains, only two strains showed affinity to sheep fH. A ~28-kDa protein of Borrelia burgdorferi sensu stricto (B. burgdorferi s.s., strain SKT-2) bound full-length fH as well as SCRs 19-20. This fH-binding protein was further identified as complement regulator-acquiring surface protein of B. burgdorferi (BbCRASP-1) by MALDI-TOF analysis. Surprisingly, a ~26-kDa protein of Borrelia bissettii (DN127) showed affinity to full-length fH but not to SCR 7 and SCRs19-20. In complement sensitivity assay, both strains-SKT-2 and DN127-were resistant to normal sheep serum. Significant complement resistance of two Borrelia garinii strains (G117 and T25) was also observed; however, none of those strains was able to bind sheep fH. Our study underscores the need of further exploration of fH-mediated evasion of complement system by Borrelia in domestic animals.


Theoretical considerations of the sensitivity of quantitative subunit hybridization.
The theoretical basis of the quantitative subunit hybridization technique and its ability to measure evolutionary amino acid substitutions is examined. Homospecific:heterospecific enzyme ratios found after subunit reassociation depend upon K1.2, the equilibrium constant for the dissociation of the heterospecific enzyme. It is shown that if this constant is near the geometric mean of the two homospecific enzyme dissociation constants, as it should be in enzymes whose subunits pair isologously, the quantitative subunit hybridization method will not detect most changes in the subunit contact regions of homologous proteins.


How much practice makes perfect? A quantitative measure of the experience needed to achieve procedural competence.
Based on the general learning curve formula, the authors have devised a method to estimate the experience needed to achieve competence in the performance of medical and surgical procedures. The method quantitates the learning ability of the trainee, the technical difficulty of the procedure, and the expected level of competence. Two retrospective applications demonstrate the ease of use and reliability of the method.


Sleep problems and recall memory in children with Down syndrome and typically developing controls.
Research conducted with typically developing (TD) infants and children generally indicates that better habitual sleep and sleep after learning are related to enhanced memory. Less is known, however, about associations between sleep and recall memory in children with Down syndrome (DS). The present study was conducted to determine whether parent-reported sleep problems were differentially associated with encoding, 1-month delayed recall memory, and forgetting over time in children with DS and those who were TD. Ten children with DS (mean age = 33 months, 5 days) and 10 TD children (mean age = 21 months, 6 days) participated in a two-session study. At each session, recall memory was assessed using an elicited imitation paradigm. Immediate imitation was permitted at the first session as an index of encoding, and delayed recall was assessed 1 month later. In addition, parents provided demographic information and reported on child sleep problems. Although parents did not report more frequent sleep problems for children with DS relative to TD children, regression-based moderation analyses revealed that more frequent sleep problems were associated with increased forgetting of individual target actions and their order by children with DS. Evidence of moderation was not found when examining encoding or delayed recall. Although group differences were not found when considering parent-reported sleep problems, more frequent sleep problems were positively associated with increased forgetting by children with DS relative to those who were TD. Although future experimental work is needed to determine causality, these results suggest that improved sleep in children with DS might reduce forgetting, ultimately improving long-term recall memory.


Experimental (13C NMR, 1H NMR, FT-IR, single-crystal X-ray diffraction) and DFT studies on 3,4-bis(isoproylamino)cyclobut-3-ene-1,2-dione.
In this work, 3,4-bis(isoproylamino)cyclobut-3-ene-1,2-dione C(10)H(16)N(2)O(2) (I), was synthesized and characterized by (13)C NMR, (1)H NMR, FT-IR, UV-vis spectroscopy and single-crystal X-ray diffraction. DFT method with 6-31G(d,p) basis set has been used to calculate the optimized geometrical parameters, atomic charges, vibrational frequencies and chemical shift values. The calculated vibrational frequencies and chemical shift values are compared with experimental FT-IR and NMR spectra. The results of the calculation shows good agreement between experimental and calculated values of the compound I. The existence of N-H⋯O type intermolecular ve C-H⋯O type intramolecular hydrogen bonds can be deduced from differences between experimental and calculated results of FT-IR and NMR. In addition, the molecular electrostatic potential map and frontier molecular orbitals and electronic absorption spectra were performed at B3LYP/6-31G(d,p) level of theory. HOMO-LUMO electronic transition of 4.90 eV are derived from the contribution of the bands π→π* and n→π* The spectral results obtained from FT-IR, NMR and X-ray of I revealed that the compound I is in predominantly enamine tautomeric form, which was supported by DFT calculations.


Duplicate testicular veins accompanied by anomalies of the testicular arteries.
Duplicate testicular veins associated with other anomalies of the testicular arteries were observed during dissection of the posterior abdominal wall in a 90-year-old Japanese male cadaver. The right testicular vein was composed of the medial and lateral testicular veins. The medial testicular vein drained into the inferior vena cava, whereas the lateral testicular vein drained into the confluence of the inferior vena cava and right renal vein. Several anastomosing branches were seen between the medial and lateral testicular veins. The left testicular vein was formed after the medial and lateral venous trunks joined and drained into the ipsilateral renal vein. The right testicular artery originated from the anterior surface of the abdominal aorta at the level of the left renal artery, passed posterior to the inferior vena cava, and accompanied the right lateral testicular vein running downwards. The left testicular artery arose from the abdominal aorta at a level of 5 cm below the origin of the right testicular artery, and then ran downwards accompanied by the medial trunk of the left testicular vein.


Lifelong changes of neurotransmitter receptor expression and debilitation of hippocampal synaptic plasticity following early postnatal blindness.
In the weeks immediately after onset of sensory loss, extensive reorganization of both the cortex and hippocampus occurs. Two fundamental characteristics comprise widespread changes in the relative expression of GABA and glutamate receptors and debilitation of hippocampal synaptic plasticity. Here, we explored whether recovery from adaptive changes in the expression of plasticity-related neurotransmitter receptors and hippocampal synaptic plasticity occurs in the time-period of up to 12 months after onset of sensory loss. We compared receptor expression in CBA/J mice that develop hereditary blindness, with CBA/CaOlaHsd mice that have intact vision and no deficits in other sensory modalities throughout adulthood. GluN1-subunit expression was reduced and the GluN2A:GluN2B ratio was persistently altered in cortex and hippocampus. GABA-receptor expression was decreased and metabotropic glutamate receptor expression was altered. Hippocampal synaptic plasticity was persistently compromised in vivo. But although LTP in blind mice was chronically impaired throughout adulthood, a recovery of the early phase of LTP became apparent when the animals reached 12 months of age. These data show that cortical and hippocampal adaptation to early postnatal blindness progresses into advanced adulthood and is a process that compromises hippocampal function. A partial recovery of hippocampal synaptic plasticity emerges in advanced adulthood, however.


A neuronal death model: overexpression of neuronal intermediate filament protein peripherin in PC12 cells.
Abnormal accumulation of neuronal intermediate filament (IF) is a pathological indicator of some neurodegenerative disorders. However, the underlying neuropathological mechanisms of neuronal IF accumulation remain unclear. A stable clone established from PC12 cells overexpressing a GFP-Peripherin fusion protein (pEGFP-Peripherin) was constructed for determining the pathway involved in neurodegeneration by biochemical, cell biology, and electronic microscopy approaches. In addition, pharmacological approaches to preventing neuronal death were also examined. Results of this study showed that TUNEL positive reaction could be detected in pEGFP-Peripherin cells. Swollen mitochondria and endoplasmic reticulum (ER) were seen by electron microscopy in pEGFP-Peripherin cells on day 8 of nerve growth factor (NGF) treatment. Peripherin overexpression not only led to the formation of neuronal IF aggregate but also causes aberrant neuronal IF phosphorylation and mislocation. Western blots showed that calpain, caspase-12, caspase-9, and caspase-3 activity was upregulated. Furthermore, treatment with calpain inhibitor significantly inhibited cell death. These results suggested that the cytoplasmic neuronal IF aggregate caused by peripherin overexpression may induce aberrant neuronal IF phosphorylation and mislocation subsequently trapped and indirectly damaged mitochondria and ER. We suggested that the activation of calpain, caspase-12, caspase-9, and caspase-3 were correlated to the dysfunction of the ER and mitochondria in our pEGFP-Peripherin cell model. The present study suggested that pEGFP-Peripherin cell clones could be a neuronal death model for future studies in neuronal IFs aggregate associated neurodegeneration.


The Chemistry and Biological Activities of Natural Products from Northern African Plant Families: From Taccaceae to Zygophyllaceae.
Traditional medicinal practices have a profound influence on the daily lives of people living in developing countries, particularly in Africa, since the populations cannot generally afford the cost of Western medicines. We have undertaken to investigate the correlation between the uses of plants in Traditional African medicine and the biological activities of the derived natural products, with the aim to validate the use of traditional medicine in Northern African communities. The literature is covered for the period 1959-2015 and part III of this review series focuses on plant families with names beginning with letters T to Z. The authors have focused on curating data from journals in natural products and phytomedicine. Within each journal home page, a query search based on country name was conducted. All articles "hits" were then verified, one at a time, that the species was harvested within the Northern African geographical regions. The current data partly constitutes the bases for the development of the Northern African natural compounds database. The review discusses 284 plant-based natural compounds from 34 species and 11 families. It was observed that the ethnobotanical uses of less than 40 % of the plant species surveyed correlated with the bioactivities of compounds identified.


Discovery and structural characterization of chicoric acid as a SARS-CoV-2 nucleocapsid protein ligand and RNA binding disruptor.
The nucleocapsid (N) protein plays critical roles in coronavirus genome transcription and packaging, representing a key target for the development of novel antivirals, and for which structural information on ligand binding is scarce. We used a novel fluorescence polarization assay to identify small molecules that disrupt the binding of the N protein to a target RNA derived from the SARS-CoV-2 genome packaging signal. Several phenolic compounds, including L-chicoric acid (CA), were identified as high-affinity N-protein ligands. The binding of CA to the N protein was confirmed by isothermal titration calorimetry, 1H-STD and 15N-HSQC NMR, and by the crystal structure of CA bound to the N protein C-terminal domain (CTD), further revealing a new modulatory site in the SARS-CoV-2 N protein. Moreover, CA reduced SARS-CoV-2 replication in cell cultures. These data thus open venues for the development of new antivirals targeting the N protein, an essential and yet underexplored coronavirus target.


Comparable long-term outcome of unrelated cord blood transplantation with related bone marrow or peripheral blood stem cell transplantation in patients aged 45 years or older with hematologic malignancies after myeloablative conditioning.
We investigated whether bone marrow or peripheral blood stem cells from older sibling donors or cord blood from unrelated donors provided a better outcome in allogeneic hematopoietic stem cell transplantation for relatively older patients who were candidates for myeloablative conditioning. Clinical outcomes of 97 patients aged 45 years or older with hematologic malignancies who received unrelated cord blood transplantation (CBT) (n = 66) or bone marrow transplantation (BMT) or peripheral blood stem cell transplantation (PBSCT) from related donors (n = 31) were compared. The cumulative incidences of grades III to IV acute and extensive chronic graft-versus-host diseases were similar between both groups. Although transplant-related mortality was significantly lower after CBT compared with BMT/PBSCT from related donors (hazard ratio [HR], .29, P = .04), overall mortality (HR, .72, P = .47) and relapse (HR, 2.02, P = .23) were not significantly different after CBT and BMT/PBSCT from related donors. These data suggest that CBT could be as safe and effective as BMT/PBSCT from older related donors for relatively older patients when it is used as a primary unrelated stem cell source.


Temporal relationships between awakening cortisol and psychosocial variables in inpatients with anorexia nervosa - A time series approach.
The aim of the study was to investigate the characteristics of the awakening salivary cortisol in patients with anorexia nervosa (AN) using a time series design. We included ten AN inpatients, six with a very low BMI (high symptom severity, HSS group) and four patients with less severe symptoms (low symptom severity, LSS group). Patients collected salivary cortisol daily upon awakening. The number of collected saliva samples varied across patients between n=65 and n=229 (due to the different lengths of their inpatient stay). In addition, before retiring, the patients answered questions daily on the handheld regarding disorder-related psychosocial variables. The analysis of cortisol and diary data was conducted by using a time series approach. Time series showed that the awakening cortisol of the AN patients was elevated as compared to a control group. Cortisol measurements of patients with LSS essentially fluctuated in a stationary manner around a constant mean. The series of patients with HSS were generally less stable; four HSS patients showed a non-stationary cortisol awakening series. Antipsychotic medication did not change awakening cortisol in a specific way. The lagged dependencies between cortisol and depressive feelings became significant for four patients. Here, higher cortisol values were temporally associated with higher values of depressive feelings. Upon awakening, the cortisol of all AN patients was in the standard range but elevated as compared to healthy controls. Patients with HSS appeared to show less stable awakening cortisol time series compared to patients with LSS.


Myeloma bone disease.
The major clinical manifestation of multiple myeloma is related to the osteolytic bone destruction. The bone disease can lead to pathologic fractures, spinal cord compression, hypercalcemia, and pain. It is also a major cause of morbidity and mortality in these patients. These patients frequently require radiation therapy, surgery and analgesic medications. Bisphosphonates are specific inhibitors of osteoclastic activity, and these agents have been evaluated in myeloma patients with bone disease during the past 15 years. Several large randomized trials have been conducted in myeloma patients also receiving chemotherapy. Orally administered bisphosphonates have shown little ability to slow the development of skeletal complications in these patients. In contrast, more potent intravenous monthly infusions of either pamidronate or zoledronic acid have reduced the skeletal complications among these patients and are now a mainstay of myeloma therapy. A number of other types of new anti-bone-resorptive agents are also in early clinical development.


Bisindoylmaleimide I enhances osteogenic differentiation.
The Wnt/β-catenin and bone morphogenetic proteins (BMPs) pathways play important roles in controlling osteogenesis. Using a cell-based kinase inhibitor screening assay, we identified the compound bisindoylmaleimide I (BIM) as a potent agonist of the cytosolic β-catenin accumulation in preosteoblast cells. Through suppressing glycogen synthase kinase 3β enzyme activities, BIM upregulated β-catenin responsive transcription and extended duration of BMP initiated signal. Functional analysis revealed that BIM promoted osteoblast differentiation and bone formation. The treatment of human mesenchymal stem cells with BIM promoted osteoblastogenesis. Our findings provide a new strategy to regulate mesenchymal stem cell differentiation by integration of the cellular signaling pathways.


Inhibition of bacterial transglutaminase by its heat-treated pro-enzyme.
Transglutaminases form a unique family of cross-linking enzymes which may be interesting for pharmaceutical and technical purposes. Bacterial transglutaminase, differing from the eucaryotic counterparts in being independent from Ca2+ ions, is excreted by several Streptomyces species. Until now an endogenous factor regulating activated transglutaminase could not be detected. Here, we investigated whether an inhibitor of transglutaminase is excreted into the culture fluid of Streptomyces mobaraensis. We could demonstrate that heat-resistant inhibitory activity is produced after 24h of growth reaching a maximum after 72h. A two-step ion exchange chromatography purification procedure revealed co-elution of the heat-treated inhibitor with pro-transglutaminase. Experiments with wild-type and recombinant pro-transglutaminase confirmed that the precursor protein indeed inhibits the activity of the mature enzyme.


Long-term outcomes of sentinel node identification using indocyanine green in patients with lung cancer.
Sentinel node (SN) biopsy is used in the management of numerous cancers to avoid unnecessary lymphadenectomy. This was a clinical exploration/feasibility study of a novel identification technique for SN biopsy using indocyanine green (ICG) fluorescence imaging during lung cancer surgery. SN biopsy using ICG was performed on 22 patients who had cT1 or T2N0M0 lung cancer. ICG was injected just around the primary tumor. The fluorescence imaging system enabled visualization of the lymphatic vessels draining from the primary tumor toward the lymph nodes. Fluorescently labeled nodes were dissected, and patients were followed-up for prognosis and recurrence to confirm the pattern of lymph node metastasis after surgery. SNs were successfully identified in 16 (72.7%) of 22 patients. A total of 13 of 16 patients had pathological N0 and three had SN metastasis. The median follow-up time was 92.7 months. Only one patient had no SN metastasis at the postoperative pathological examination and lymph node metastasis during the follow-up period. The accuracy rate was 93.8% (15/16) and the false-negative rate was 7.7% (1/13). SNs were identified by ICG fluorescence imaging, and this technique during lung cancer surgery had good identification and accuracy rates throughout the follow-up period. SIGNIFICANT FINDINGS OF THE STUDY: We attempted to identify sentinel lymph nodes by indocyanine green in lung cancer surgery. The identification rate was 72.7%. The accuracy rate was 100% immediately after surgery, and 93.8% after follow-up. Sentinel node biopsy by indocyanine green may be useful for lymph node dissection during lung cancer surgery to avoid unnecessary lymphadenectomy.


Molecular mechanisms underlying delayed apoptosis in neutrophils from multiple trauma patients with and without sepsis.
Delayed neutrophil apoptosis and overshooting neutrophil activity contribute to organ dysfunction and subsequent organ failure in sepsis. Here, we investigated apoptotic signaling pathways that are involved in the inhibition of spontaneous apoptosis in neutrophils isolated from major trauma patients with uneventful outcome as well as in those with sepsis development. DNA fragmentation in peripheral blood neutrophils showed an inverse correlation with the organ dysfunction at d 10 after trauma in all patients, supporting the important role of neutrophil apoptosis regulation for patient's outcome. The expression of the antiapoptotic Bcl-2 protein members A1 and Mcl-1 were found to be diminished in the septic patients at d 5 and d 10 after trauma. This decrease was also linked to an impaired intrinsic apoptosis resistance, which has been previously shown to occur in neutrophils during systemic inflammation. In patients with sepsis development, delayed neutrophil apoptosis was found to be associated with a disturbed extrinsic pathway, as demonstrated by reduced caspase-8 activity and Bid truncation. Notably, the expression of Dad1 protein, which is involved in protein N-glycosylation, was significantly increased in septic patients at d 10 after trauma. Taken together, our data demonstrate that neutrophil apoptosis is regulated by both the intrinsic and extrinsic pathway, depending on patient's outcome. These findings might provide a molecular basis for new strategies targeting cell death pathways in apoptosis-resistant neutrophils during systemic inflammation.


Quality of life and body image after circumferential body lifting of the lower trunk: a prospective clinical trial.
Plastic surgery after massive weight loss remains a fast growing subspecialty. However, very few studies exist that investigate the quality of life and psychological outcome after body lift surgery. Twenty-seven patients (25 females and 2 males) who had undergone bariatric surgery were scheduled for circumferential body lifting of the lower trunk. The patients, aged 21-58 years (mean = 39.9, SD = 10.9), underwent surgery in our department of plastic and reconstructive surgery between 2008 and 2010. All participants took part in two psychological examinations on a voluntary basis and completed the prospective pre-post study. Post-weight loss surgery improves the quality of life and body image. It is therefore an important part in the interdisciplinary treatment of the massive weight loss patient.


Patient Satisfaction and Clinical Efficacy of Flash Glucose Monitoring in Patients with Type 1 Diabetes: A Prospective, Single-Center, Single-Arm Study.
Frequent self-monitoring of blood glucose (SMBG) is usually required in patients with type 1 diabetes (T1D). However, the fear of self-testing, hygiene, and social obstacles may deter some patients from SMBG. Flash glucose monitoring (FGM), a less-invasive glucose monitoring method, was launched in Japan in September 2017. The aim of this study was to evaluate patient satisfaction and the clinical efficacy of FGM in Japanese patients with T1D. Patient satisfaction with FGM was assessed using the Diabetes Treatment Satisfaction Questionnaire (DTSQ) and Diabetes Therapy-Related Quality of Life (DTR-QOL) questionnaire before (baseline) and 4 and 12 weeks after initiating FGM use in 20 Japanese patients with T1D. Clinical parameters related to glucose metabolism, such as glycated hemoglobin (HbA1c) levels, were assessed, and glucose fluctuations were evaluated using the FGM data. Values at 4 and 12 weeks after initiating FGM were compared with baseline data using the Wilcoxon signed-rank test. The mean absolute relative difference (MARD) between glucose values detected using the FGM device and by SMBG was also calculated. The DTSQ scores significantly improved 12 weeks after initiating FGM (P < 0.001). The DTR-QOL scores related to "burden in social activities" and "treatment satisfaction" also significantly improved 12 weeks after initiating FGM (P = 0.024 and 0.007, respectively). The HbA1c values and the percentage of time within the target glucose range (3.9-7.8 mmol/L) at the 12-week time point also improved, from 58 ± 5 (baseline) to 54 ± 4 mmol/mol (P = 0.002) and from 36% (interquartile range [IQR] 25-44.5%) (baseline) to 43% (IQR 39-50%) (P = 0.016), respectively. The MARD between glucose values detected using FGM and those detected using SMBG was 12.2% throughout the study period. Flash glucose monitoring contributed to improved patient satisfaction and the adjustment of blood glucose levels in patients with T1D. University Hospital Medical Information Network Clinical Trials Registry (UMIN-CTR): UMIN000029673.


Correlations between central and peripheral changes in anterior corneal topography after myopic LASIK and their implications in postsurgical contact lens fitting.
To investigate the changes in central and peripheral anterior corneal curvatures after myopic laser in situ keratomileusis (LASIK) and to correlate them with the manifest refractive change to discuss how such results could affect post-LASIK corneal topography profiles and midterm stability and their implications in postsurgical contact lens fitting. Topographic and refractive data from 18 eyes of 11 patients that had undergone myopic LASIK were collected for 6 months after surgery. Short-term and midterm topographic responses were investigated and correlated with spherical equivalent manifest refractive changes. There was a strong correlation between eccentricity changes and manifest refractive change 15 days after surgery (r = 0.753, P < 0.001), with no significant changes thereafter for the following 6 months (r = 0.148, P = 0.114). A strong linear relationship was found between baseline manifest refraction and changes in corneal curvature at the center (r = 0.810, P < 0.001), 4-mm chord (r = 0.895, P < 0.001), and 6-mm chord (r = 0.696, P < 0.001). Statistically significant changes were also found after the first 15 days (P < 0.005) and showed a regression effect that affects the three zones. In this case, a weaker relationship was found between curvature regression and the final refractive change for the central location (r = 0.412, P = 0.004), 4-mm chord area (r = 0.430, P = 0.003), and 6-mm chord area (r = 0.283, P = 0.023). CONCLUSIONS.: After myopic LASIK, the anterior corneal dioptric power is expected to change, on average, approximately 77% of the attempted spherical equivalent correction at the center; 60% at the 4-mm chord region, where the stronger correlation between topographic and refractive change is found; and 30% at the 6-mm chord area. The paracentral area 4 mm from the center seems to be more likely to predict baseline corneal curvature from manifest refractive change. Some degree of regression in the midterm period is expected to occur after myopic LASIK, which shows a significant correlation with the manifest refractive change. Again, this effect is more evident and more accurately predicted at the 4-mm chord area. The results of the current study are of interest for those fitting contact lenses after myopic LASIK.


Axillary management in breast cancer: what's new for 2012?
Sentinel lymph node (SLN) biopsy has become standard care for management of the axilla in invasive breast cancer, replacing axillary lymph node dissection (ALND) in most subjects, with a progressively diminishing role of ALND. Advances in preoperative imaging have also changed the algorithm for axillary management, and ultrasound-guided needle biopsy has been shown to triage >50% of subjects with node metastases to ALND. However, the past two years have witnessed remarkable and practice-changing advances in our knowledge and approach to management of the axilla, with availability of high-level evidence that demands reappraisal of practice and challenges the role of routine ALND for SLN-positive patients. In particular, for the group of patients defined by eligibility criteria in the Z0011 trial, it appears that ALND has little or no effect on local recurrence and survival, or on the choice of local or systemic therapies. We review the available evidence on staging and management of the axilla in breast cancer, and outline our interpretation of its implications for clinical practice.


Molecular characterization and expression analysis of IκB from Haliotis discus discus.
Innate immune system relies on the recognition of pathogen associated molecular patterns present in the microbes by the pattern recognition receptors leading to the activation of signaling cascade and subsequent synthesis of cytokines. NF-κB is a major stimulus activated transcription factor, which regulates the expression of a diverse array of genes. IκB is an inhibitor of NF-κB, retaining NF-κB in an inactive state in the cytoplasm. In this study, we have reported the characterization of first abalone IκB (HdIκB). The cDNA possessed an ORF of 1200 bp coding for a protein of 400 amino acids with molecular mass of 45 kDa and isoelectric point of 4.7. HdIκB protein possessed a conserved phosphorylation site (58)DSGIFS(63) in the N-terminal region, six ankyrin repeats, and a PEST sequence in the C-terminal region. A casein kinase II phosphorylation site could also be observed in the PEST sequence. Constitutive expression of HdIκB revealed its physiological significance since NF-κB is known to be activated by various stimuli. Elevated expression of HdIκB transcripts could be observed in abalones challenged with various mitogens and live microbes. This novel characterization of abalone IκB would further be a positive approach in the affirmation of evolutionary conservation and significance of this protein as a repressor/inhibitor of a pleiotropic transcription factor like NF-κB.


Mutational analysis of a satellite phage activator.
The late gene activator, Delta, of satellite phage P4 is more efficient than the Delta of satellite phage phiR73 in utilizing a P2 helper prophage that lacks an activator (ogr) gene. Analysis of P4 Delta is complicated by the fact that this protein contains two tandem phiR73 Delta-like domains. We performed a mutational analysis of phiR73 Delta, in order to select mutations that might not be found using P4 Delta. The host RNA polymerase alpha subunit mutation rpoA155 (L289F) blocks the growth of P2, P4, and P4 carrying the delta gene of phiR73. A mutant of this latter phage that can grow in the presence of rpoA155 carries a V19M mutation in phiR73 Delta. This suggests that aa 19 contacts RNA polymerase, in addition to the aa residues 13, 42 and 44, that have been implicated in interactions with RNA polymerase by previous mutational analyses of P2 ogr and P4 delta. In corroboration of the proposed role of the regions at aa residues 19, 42, and 44, we found phiR73 Delta mutations in these regions that showed a reduced activation of late gene expression, but a normal ability to bind to late gene promoters. All activators in the Delta class contain four Cys residues that bind Zn2+. Mutation of these aa residues in phiR73 Delta eliminated late gene activation. Spectroscopic analysis of these mutant proteins revealed that they were unable to bind Zn2+. Histidine residues were substituted for two of the Cys residues (C30 and C35), changing a C2C2 type Zn-binding motif to a C2H2 motif. Although His residues are used in coordinating Zn2+ in other proteins, these His substitutions resulted in complete loss of activity and the inability to bind Zn2+.


Adalimumab with Methotrexate in Treatment-Naïve Japanese Patients with Rheumatoid Arthritis at Risk of Progressive Structural Joint Damage: A Postmarketing Observational Study.
The objective of this study was to evaluate the real-world safety and effectiveness of adalimumab with methotrexate (MTX) in disease-modifying antirheumatic drug (DMARD)- and biologic-naïve Japanese patients with rheumatoid arthritis (RA) at risk of progressive structural joint damage. This multicenter, prospective, observational, postmarketing surveillance study was conducted between February 2013 and April 2015 at 84 centers in Japan. Patients with RA at risk of progressive structural joint damage were enrolled and initiated treatment with adalimumab and MTX. Adverse events were recorded up to week 28. Effectiveness/disease activity was assessed using the Disease Activity Score based on a 28-joint count with erythrocyte sedimentation rate and C-reactive protein (DAS28-4ESR and DAS28-4CRP), Clinical Disease Activity Index, and Simplified Disease Activity Index at 0, 4, 12, and 24 weeks. DAS28-4CRP response was evaluated in the low-dose (<8 mg/week) and high-dose (≥8 mg to ≤16 mg/week) MTX groups at week 24. One hundred fifty-seven of 163 patients comprised the safety cohort: mean (SD) age, 56.5 (13.9) years; females, 65.6%; rheumatoid factor positive, 73.2%; anti-cyclic citrullinated peptide antibody positive, 66.9%; bone erosions, 51.6%; mean disease duration, 9.5 months. The majority of patients (≥80%) had moderate or high disease activity at baseline, and ≥50% with available data achieved remission or low disease activity at week 24 (DAS28-4CRP <3.2). Five serious adverse drug reactions occurred in four patients, including pyelonephritis, Pneumocystis jiroveci pneumonia, interstitial lung disease, pleurisy, and pericarditis; the outcomes were either recovered or recovering. Significant improvements/reductions in disease activity over 24 weeks were noted in all effectiveness measures (P < 0.0001). Most of the population achieved DAS28-4CRP remission (<2.6) at week 24 regardless of the MTX dose. Adalimumab in combination with MTX could be a beneficial treatment option for DMARD- and biologic-naïve Japanese patients with RA at risk of progressive structural joint damage. AbbVie GK and Eisai. ClinicalTrials.gov identifier, NCT01783730.


Pathogenic Specialization and Pathotype Distribution of Puccinia hordei in Australia, 1992 to 2001.
Annual surveys of pathogenic variability in the leaf rust pathogen of barley, Puccinia hordei, from 1992 to 2001 revealed a significant shift in the composition of populations across Austra-lia. Virulence for the resistance gene Rph12, first detected in a single pathotype, 4610P+, in Tasmania in 1991, was subsequently detected in 1993 in South Australia, Victoria, and southern New South Wales. By the end of 2001, eight pathotypes with virulence for Rph12 had been isolated, and virulence for this gene was present in all Australian barley growing regions. Virulence was not detected for the resistance genes Rph3, Rph7, Rph11, or Rph14. The distribution and spread of the pathotypes detected, their possible origins, pathogenicity on several uncharacterized seedling resistance sources, and implications for resistance breeding are discussed.


Effect of post-anthesis waterlogging on biosynthesis and granule size distribution of starch in wheat grains.
Waterlogging is one of the most common abiotic stress types in wheat production in many rainy areas of the world. Two locally widely grown winter wheat (Triticum aestivum L. cv Yumai 34 and Yangmai 9) were subjected to post-anthesis waterlogging in a pot experiment to investigate the impacts of waterlogging on the starch synthesis and the physiochemical properties. Post-anthesis waterlogging significantly decreased grain weight and affected the content of starch components. Waterlogging down-regulated the activity and expression of genes encoding soluble starch synthase [SSS (EC 18.104.22.168)], while up-regulated those of the granule bound starch synthase I [GBSSI (EC:18.104.22.168)]. This further resulted in decreased amylopectin content and increased amylose content. Waterlogging also caused a reduction in the number of starch granules, while increased the mean diameter of starch granules in mature grains, which was mainly due to an increase in the volume frequency percent of the A-type starch granules. Waterlogging also lowered the peak viscosity and trough viscosity of starch, but did not affect the breakdown viscosity and peak time. We concluded that the modified expressions of the starch synthase encoding genes were responsible for the changed size distribution of starch granules, which finally affected the starch pasting properties of wheat growing under post-anthesis waterlogging conditions.


ADHD diagnosis using structural brain MRI and personal characteristic data with machine learning framework.
An essential yet challenging task is an automatic diagnosis of attention-deficit/hyperactivity disorder (ADHD) without manual intervention. The present study emphasises utilizing structural MRI and personal characteristic (PC) data for developing an automated diagnostic system for ADHD classification. Here, an age-balanced dataset of 316 ADHD and 316 Typically Developing Children (TDC) was prepared from the publicly available dataset. We extracted volumetric features from gray matter (GM) volumes from brain regions defined by Automated Anatomical Labelling (AAL3) atlas and cortical thickness-based (CT) features using the Destrieux atlas. A set of salient features were selected independently using minimum redundancy and maximum relevance (mRMR) and ensemble feature selection (EFS) methods. Decision models were trained using five well-known classifiers: K-nearest neighbours, logistic regression, linear Support Vector Machine (SVM), radial-based SVM (RBSVM), and Random Forest. The performance of the proposed system was evaluated using accuracy, recall, and specificity with ten runs of a ten-fold cross-validation scheme. We run seven experiments by considering different combinations of features. The maximum classification accuracy of 75% was obtained with CT and PC features with RBSVM and SVM with the EFS. An increase in GM volume in fifteen brain regions and loss of cortical thickness in twenty-seven brain regions were observed.


Superficial hemosiderosis in a second trimester fetus: pathological and clinical manifestations.
Prenatal brain hemorrhages are associated with considerable morbidity and mortality in neonates. They appear predominantly as bleeding into periventricular germinal matrix with subsequent hemorrhages into lateral ventricles and subarachnoid space. Other patterns of brain hemorrhage are not widely documented in second trimester fetuses. We report a case of hemorrhage presenting as extensive hemosiderin deposits in leptomeninges and adjacent brain parenchyma, as observed in superficial hemosiderosis. The lesion was associated with substantial tissue damage. Clinically, it was diagnosed during the second half of second trimester by successive ultrasound examinations. If differs from germinal matrix hemorrhage and other forms of hemorrhage in origin, pathogenesis, morphology, and probably in clinical manifestations.