Electrical properties of doped conjugated polyelectrolytes with modulated density of the ionic functionalities.
We report the synthesis of a series of water-soluble anionic narrow band-gap conjugated polyelectrolytes with a varied density of the ionic functional groups. The charge density is modulated by incorporating the structural units with tetraethylene glycol (TEG) monomethyl ether side chains. These polymers are readily p-doped during dialysis in water. CPEs with TEG side chains exhibit tighter intermolecular packing in the solid state and higher electrical conductivity.


A new probabilistic seismic hazard assessment for greater Tokyo.
Tokyo and its outlying cities are home to one-quarter of Japan's 127 million people. Highly destructive earthquakes struck the capital in 1703, 1855 and 1923, the last of which took 105,000 lives. Fuelled by greater Tokyo's rich seismological record, but challenged by its magnificent complexity, our joint Japanese-US group carried out a new study of the capital's earthquake hazards. We used the prehistoric record of great earthquakes preserved by uplifted marine terraces and tsunami deposits (17 M approximately 8 shocks in the past 7000 years), a newly digitized dataset of historical shaking (10000 observations in the past 400 years), the dense modern seismic network (300,000 earthquakes in the past 30 years), and Japan's GeoNet array (150 GPS vectors in the past 10 years) to reinterpret the tectonic structure, identify active faults and their slip rates and estimate their earthquake frequency. We propose that a dislodged fragment of the Pacific plate is jammed between the Pacific, Philippine Sea and Eurasian plates beneath the Kanto plain on which Tokyo sits. We suggest that the Kanto fragment controls much of Tokyo's seismic behaviour for large earthquakes, including the damaging 1855 M approximately 7.3 Ansei-Edo shock. On the basis of the frequency of earthquakes beneath greater Tokyo, events with magnitude and location similar to the M approximately 7.3 Ansei-Edo event have a ca 20% likelihood in an average 30 year period. In contrast, our renewal (time-dependent) probability for the great M > or = 7.9 plate boundary shocks such as struck in 1923 and 1703 is 0.5% for the next 30 years, with a time-averaged 30 year probability of ca 10%. The resulting net likelihood for severe shaking (ca 0.9 g peak ground acceleration (PGA)) in Tokyo, Kawasaki and Yokohama for the next 30 years is ca 30%. The long historical record in Kanto also affords a rare opportunity to calculate the probability of shaking in an alternative manner exclusively from intensity observations. This approach permits robust estimates for the spatial distribution of expected shaking, even for sites with few observations. The resulting probability of severe shaking is ca 35% in Tokyo, Kawasaki and Yokohama and ca 10% in Chiba for an average 30 year period, in good agreement with our independent estimate, and thus bolstering our view that Tokyo's hazard looms large. Given 1 trillion US dollars estimates for the cost of an M approximately 7.3 shock beneath Tokyo, our probability implies a 13 billion US dollars annual probable loss.


Peptide-based molecular beacons for cancer imaging and therapy.
Peptide-based molecular beacons are Förster resonance energy transfer-based target-activatable probes. They offer control of fluorescence emission in response to specific cancer targets and thus are useful tools for in vivo cancer imaging. With our increasing knowledge about human genome in health and disease, peptide-based "smart" probes are continually developed for in vivo optical imaging of specific molecular targets, biological pathways and cancer progression and diagnosis. A class of fluorescent photosensitizers further extends the application of peptide beacons to cancer therapeutics. This review highlights the applications of peptide beacons in cancer imaging, the simultaneous treatment and response monitoring and smart therapeutics with a focus on recent improvements in the design of these probes.


The gall bladder cholecystokinin receptor exists in two guanine nucleotide-binding protein-regulated affinity states.
To study proximal events in cholecystokinin (CCK) action on bovine gall bladder smooth muscle, we used the hormone analogue D-Tyr-Gly-[(N1e28,31)CCK-26-32]-phenethyl ester (OPE), which has unique biological properties. This fully efficacious agonist differs from native CCK by not expressing supramaximal inhibition of cell shortening, yet it clearly interacts with the same receptor molecule. This was demonstrated in binding and affinity labeling studies, where both peptides label the same Mr 70,000-85,000 protein and both fully compete for binding of the other ligand. Further, its relatively high affinity for the low affinity CCK receptor permits the clear demonstration of two affinity states of a CCK receptor on a membrane preparation and makes possible evaluation of the molecular basis of these affinity states and their regulation. Analysis of homologous and heterologous binding curves performed with both CCK and OPE peptides and radioligands demonstrated the presence of two affinity states, with CCK being able to distinguish them (Kd1 = 0.48 +/- 0.04 nM and Kd2 = 56.5 +/- 7.4 nM) and OPE recognizing them equally (Kd1 = 0.94 +/- 0.31 nM and Kd2 = 0.96 +/- 0.23 nM). In the presence of nonhydrolyzable GTP analogues, there was a shift in distribution of receptors toward the low affinity state, with the total number of receptors and their absolute affinities for each peptide remaining constant. Thus, the gall bladder CCK receptor is a single molecule capable of assuming two interconvertible affinity states, regulated by a guanine nucleotide-binding protein. Two full agonists are capable of interacting with this molecule to yield different biological responses via different molecular events.


Scattering by an infinite homogenous anisotropic elliptic cylinder in terms of Mathieu functions and Fourier series.
An exact solution to the two-dimensional scattering properties of an anisotropic elliptic cylinder for transverse electric polarization is presented. The internal field in an anisotropic elliptic cylinder is expressed as integral representations of Mathieu functions and Fourier series. The coefficients of the series expansion are obtained by imposing boundary conditions on the anisotropic-free-space interface. A matrix is developed to solve the nonorthogonality properties of Mathieu functions at the interface between two different media. Numerical results are given for the bistatic radar cross section and the amplitude of the total magnetic field along the x and y axes. The result is in agreement with that available as expected when an elliptic cylinder degenerates to a circular one.


Efficiency versus thoroughness in medication review: a qualitative interview study in UK primary care.
Medication reviews may improve the safety of prescribing and the National Institute for Health and Care Excellence (NICE) highlights the importance of involving patients in this process. To explore GP and pharmacist perspectives on how medication reviews were conducted in general practice in the UK. Analysis of semi-structured interviews with GPs and pharmacists working in the South West of England, Northern England, and Scotland, sampled for heterogeneity. Interviews took place between January and October 2017. Interviews focused on experience of medication review. Data saturation was achieved when no new insights arose from later interviews. Interviews were analysed thematically. In total, 13 GPs and 10 pharmacists were interviewed. GPs and pharmacists perceived medication review as an opportunity to improve prescribing safety. Although interviewees thought patients should be involved in decisions about their medicines, high workload pressures meant that most medication reviews were conducted with limited or no patient input. For some GPs, a medication review was done 'in the quickest way possible to say that it was done'. Pharmacists were perceived by both professions as being more thorough but less time efficient than GPs, and few pharmacists were routinely involved in medication reviews even in practices employing a pharmacist. Interviewees argued that it was easier to continue medicines than it was to stop them, particularly because stopping medicines required involving the patient and this generated extra work. Practices tended to prioritise being efficient (getting the work done) rather than being thorough (doing it well), so that most medication reviews were carried out with little or no patient involvement, and medicines were rarely stopped or reduced. Time and resource constraints are an important barrier to implementing NICE guidance.


In-vitro 3D Analysis of Sacroiliac Joint Kinematics: Primary and Coupled Motions.
An in-vitro biomechanical study of human cadaver sacroiliac joints. Our study aimed to develop a more comprehensive understanding of the native motion of the SIJ within the context of spinal kinematics and spinal implant evaluation. Increasing attention has been given to the sacroiliac joint (SIJ) as a source of low back pain, despite its limited range of motion. We sought to characterize the rotational and translational motion in each axis utilizing standard pure moment flexion-extension (FE), lateral bending (LB), and axial rotation (AR) testing. Sixteen sacroiliac joints were evaluated from eight lumbosacral cadaver specimens (six females, two males) from subjects aged 28 to 57 years (mean age 46.8) with body mass index (BMI) 22 to 36 (mean BMI 30). Single leg stance was modeled by clamping the blocks on one ischium in a vise and letting the contralateral ischium hang freely. Pure moment loading was applied in FE, right/left AR, and right/left LB. Relative motions were collected with infrared markers. The on-axis ratio was significantly lower in LB than in FE (P = 0.012) and in AR (P = 0.017). The rotation deviation angle measured 13.9 ± 9.1° in FE, 17.1 ± 8.7° in AR, and 35.7 ± 25.7° in LB. In LB the rotational deviation angle is significantly higher than both FE and AR (P = 0.003 and P = 0.011, respectively). In-plane translation was significantly higher (P = 0.005) in FE loading than in LB loading. A nontrivial amount of rotation and translation occurred out of the expected axis of motion. The largest amount of off-axis rotation was observed in lateral bending. Relative to resultant translation, in-plane translation was lowest in lateral bending. Our results indicate that rotation of the SIJ is not fully described with the in-plane metrics which are normally reported in evaluation of fusion devices. Future studies of the SIJ may need to consider including off-axis rotation measurements when describing SIJ kinematics.Level of Evidence: 5.


A role for CD4+ but not CD8+ T cells in immunity to Schistosoma mansoni induced by 20 krad-irradiated and Ro 11-3128-terminated infections.
The role of CD4+ (L3/T4+) and CD8+ (Lyt-2+) T cells in immunity to Schistosoma mansoni induced by 20 krad-irradiated and Ro 11-terminated infections in mice was investigated directly by in vivo depletion of these subsets with cytotoxic rat monoclonal antibodies (mAb). Effective physical depletion was demonstrated by flow cytometric analysis and immunohistochemical staining. Functional depletion of helper activity following anti-CD4 treatment was indicated by an abrogation of concanavalin A(Con A)-induced colony-stimulating factor (CSF) release, while anti-CD8 treatment had no effect in these assays. Pre-existing S. mansoni-specific antibody levels were unaffected by anti-CD4 and anti-CD8 treatment. In vivo depletion of CD4+ T cells resulted in a dramatic reduction in immunity induced by one (up to 100%) and two (up to 70%) vaccinations with 20 krad-irradiated cercariae and also of resistance induced by Ro 11-attenuated infections (up to 100%). Depletion of CD8+ T cells had no effect on resistance induced by any of the vaccination protocols investigated. A correlation was observed between resistance and T cell-induced, macrophage-mediated killing of schistosomula in vitro, both of which were abrogated following anti-CD4 treatment but were unaffected by CD8+ T-cell depletion. The possible role of CD4+ T cells in vivo and the implications for vaccine development are discussed.


Homotypic antibody responses to fresh clinical isolates of human immunodeficiency virus.
Human immunodeficiency virus type 1 (HIV-1) exhibits extensive genomic and antigenic diversity, which is thought to contribute to the failure of the host's immune response to control infection and prevent clinical progression. Part of this failure may be due to utilization by the virus of antigenic variation as a means to escape protective immune responses. Antibody-escape variants of HIV-1 were studied here using fresh clinical isolates and autologous plasmas. HIV-1 was isolated from the plasma of seven people who were all seropositive for at least 2 years, and symptomatic sometime during that period. Isolated viruses were confirmed as HIV-1 by the presence of reverse transcriptase activity in infected culture supernatants, and by positive immunofluorescence using human monoclonal antibody to HIV-1 core protein. Plasma from these people were positive by Western immunoblot (DuPont) for most major HIV-1 (strain IIIB) antigens. These plasmas neutralized three laboratory strains of HIV-1 (i.e., IIIB, RF, and MN) but did not neutralize the homotypic strain in five cases, and had greatly reduced neutralizing titers against the homotypic strain in two cases. Homotypic neutralizing antibodies were absent in autologous plasma obtained 3 months later. When antibody titers were measured by fixed-cell indirect immunofluorescence assays (IFAs), high titers of IgG (1:6400 to 1:25,600) were detected against HIV-1 IIIB, while low titers of only 1:20 to 1:160 were detected against homotypic viral antigens at the time of virus isolation, and remained low 12 and 16 weeks later. No class IgA, IgD, IgE, or IgM antibodies to homotypic viral antigens, as possible IgG-blocking antibodies, were detected by fixed-cell IFAs. Cross-reactions with heterologous donor's plasmas were observed in some cases, and in these cases the cross-reactions were unidirectional. Live-cell IFAs detected IgG in patient's plasma to HIV-1 IIIB-infected cells but not to cells infected with homotypic isolates. These results suggest that it is common for neutralization-resistant HIV-1 variants to appear during the course of infection, and that all or most antigens of these variants are capable of escaping antibody recognition.


Catalytic Conversion of Hexagonal Boron Nitride to Graphene for In-Plane Heterostructures.
Heterostructures of hexagonal boron nitride (h-BN) and graphene have attracted a great deal of attention for potential applications in 2D materials. Although several methods have been developed to produce this material through the partial substitution reaction of graphene, the reverse reaction has not been reported. Though the endothermic nature of this reaction might account for the difficulty and previous absence of such a process, we report herein a new chemical route in which the Pt substrate plays a catalytic role. We propose that this reaction proceeds through h-BN hydrogenation; subsequent graphene growth quickly replaces the initially etched region. Importantly, this conversion reaction enables the controlled formation of patterned in-plane graphene/h-BN heterostructures, without needing the commonly employed protecting mask, simply by using a patterned Pt substrate.


Investigating the effectiveness of full-time wrist splinting and education in the treatment of carpal tunnel syndrome: a randomized controlled trial.
This study investigated the effects of wearing a wrist support splint for 8 wk and receiving a formal education program on patients with carpal tunnel syndrome (CTS), as well as factors associated with patients' desire to seek surgical intervention. Participants were recruited from a hospital surgical wait list and randomly assigned to an intervention group (n = 30) or a control group (n = 24). Significant improvements in measures of symptom severity and functional status over the duration of the study appeared in the intervention group but not in the control group. Logistic regression for the intervention group showed that symptom severity (odds ratio [OR] = 1.53, 95% confidence interval [CI] [1.20-1.93]), functional deficits (OR = 1.31, 95% CI [1.08-1.57]), pain score (OR = 1.25, 95% CI [1.11-1.61]), and symptom duration (OR = 1.11, 95% CI [1.01-1.24]) were positively associated with the desire to seek surgical intervention. This conservative CTS treatment program conducted by occupational therapists can improve symptoms and hand function in CTS patients.


Mitochondrial genome sequence analysis: a custom bioinformatics pipeline substantially improves Affymetrix MitoChip v2.0 call rate and accuracy.
Mitochondrial genome sequence analysis is critical to the diagnostic evaluation of mitochondrial disease. Existing methodologies differ widely in throughput, complexity, cost efficiency, and sensitivity of heteroplasmy detection. Affymetrix MitoChip v2.0, which uses a sequencing-by-genotyping technology, allows potentially accurate and high-throughput sequencing of the entire human mitochondrial genome to be completed in a cost-effective fashion. However, the relatively low call rate achieved using existing software tools has limited the wide adoption of this platform for either clinical or research applications. Here, we report the design and development of a custom bioinformatics software pipeline that achieves a much improved call rate and accuracy for the Affymetrix MitoChip v2.0 platform. We used this custom pipeline to analyze MitoChip v2.0 data from 24 DNA samples representing a broad range of tissue types (18 whole blood, 3 skeletal muscle, 3 cell lines), mutations (a 5.8 kilobase pair deletion and 6 known heteroplasmic mutations), and haplogroup origins. All results were compared to those obtained by at least one other mitochondrial DNA sequence analysis method, including Sanger sequencing, denaturing HPLC-based heteroduplex analysis, and/or the Illumina Genome Analyzer II next generation sequencing platform. An average call rate of 99.75% was achieved across all samples with our custom pipeline. Comparison of calls for 15 samples characterized previously by Sanger sequencing revealed a total of 29 discordant calls, which translates to an estimated 0.012% for the base call error rate. We successfully identified 4 known heteroplasmic mutations and 24 other potential heteroplasmic mutations across 20 samples that passed quality control. Affymetrix MitoChip v2.0 analysis using our optimized MitoChip Filtering Protocol (MFP) bioinformatics pipeline now offers the high sensitivity and accuracy needed for reliable, high-throughput and cost-efficient whole mitochondrial genome sequencing. This approach provides a viable alternative of potential utility for both clinical diagnostic and research applications to traditional Sanger and other emerging sequencing technologies for whole mitochondrial genome analysis.


Accuracy of infrared ear thermometry and other temperature methods in adults.
To compare the accuracy of infrared ear-based temperature measurement in relation to thermometer, ear position, and other temperature methods, with pulmonary artery temperature as the reference. Ear-based temperature measurements were made with four infrared thermometers, three in the core mode and two in the unadjusted mode, each with tug and no-tug techniques. Pulmonary artery, bladder (n = 21), and axillary temperatures were read after each ear-based measurement and oral temperature was measured once when possible (n = 32). Subjects consisted of a convenience sample of 50 patients with pulmonary artery catheters who were in adult critical care units of a university teaching hospital. Ear-based measurements correlated well with pulmonary artery temperature (r = .87 to .91), although closeness of agreement differed among thermometer-mode combinations (mean offsets = -0.7 to 0.5 degree C) and had moderately high variability between subjects (SD = +/- 0.5 degree C) with all instruments. Use of an ear tug either made no difference or resulted in slightly lower readings. Bladder temperature was nearly identical to pulmonary artery temperature values (r = .99, offset = 0.0 +/- 0.2 degree C). Oral readings were slightly lower (r = .78, offset = -0.2 degree C) and axillary readings much more so (r = .80 to .82, offset = -0.7 degree C); both were highly variable (SD = +/- 0.6 degree C) and affected by external factors. Infrared ear thermometry is useful for clinical temperature measurement as long as moderately high variability between patients is acceptable. Readings differ among thermometers, although several instruments provide values close to pulmonary artery temperature in adults. Readings are not higher with an ear tug. Bladder temperature substitutes well for pulmonary artery temperature, whereas oral and axillary values may be influenced by external factors in the critical care setting.


Tests and expenditures in the initial evaluation of peripheral neuropathy.
Peripheral neuropathy is a common disorder in which an extensive evaluation is often unrevealing. We sought to define diagnostic practice patterns as an early step in identifying opportunities to improve efficiency of care. The 1996-2007 Health and Retirement Study Medicare claims-linked database was used to identify individuals with an incident diagnosis of peripheral neuropathy using International Classification of Diseases, Ninth Revision, codes and required no previous neuropathy diagnosis during the preceding 30 months. Focusing on 15 relevant tests, we examined the number and patterns of tests and specific test utilization 6 months before and after the incident neuropathy diagnosis. Medicare expenditures were assessed during the baseline, diagnostic, and follow-up periods. Of the 12, 673 patients, 1031 (8.1%) received a new International Classification of Diseases, Ninth Revision, diagnosis of neuropathy and met the study inclusion criteria. Of the 15 tests considered, a median of 4 (interquartile range, 2-5) tests were performed, with more than 400 patterns of testing. Magnetic resonance imaging of the brain or spine was ordered in 23.2% of patients, whereas a glucose tolerance test was rarely obtained (1.0%). Mean Medicare expenditures were significantly higher in the diagnostic period than in the baseline period ($14,362 vs $8067, P < .001). Patients diagnosed as having peripheral neuropathy typically undergo many tests, but testing patterns are highly variable. Almost one-quarter of patients receiving neuropathy diagnoses undergo high-cost, low-yield magnetic resonance imaging, whereas few receive low-cost, high-yield glucose tolerance tests. Expenditures increase substantially in the diagnostic period. More research is needed to define effective and efficient strategies for the diagnostic evaluation of peripheral neuropathy.


Reconfigurable microfluidic device with integrated antibody arrays for capture, multiplexed stimulation, and cytokine profiling of human monocytes.
Monocytes represent a class of immune cells that play a key role in the innate and adaptive immune response against infections. One mechanism employed by monocytes for sensing foreign antigens is via toll-like receptors (TLRs)-transmembrane proteins that distinguish classes of foreign pathogens, for example, bacteria (TLR4, 5, and 9) vs. fungi (TLR2) vs. viruses (TLR3, 7, and 8). Binding of antigens activates a signaling cascade through TLR receptors that culminate in secretion of inflammatory cytokines. Detection of these cytokines can provide valuable clinical data for drug developers and disease investigations, but this usually requires a large sample volume and can be technically inefficient with traditional techniques such as flow cytometry, enzyme-linked immunosorbent assay, or luminex. This paper describes an approach whereby antibody arrays for capturing cells and secreted cytokines are encapsulated within a microfluidic device that can be reconfigured to operate in serial or parallel mode. In serial mode, the device represents one long channel that may be perfused with a small volume of minimally processed blood. Once monocytes are captured onto antibody spots imprinted into the floor of the device, the straight channel is reconfigured to form nine individually perfusable chambers. To prove this concept, the microfluidic platform was used to capture monocytes from minimally processed human blood in serial mode and then to stimulate monocytes with different TLR agonists in parallel mode. Three cytokines, tumor necrosis factor-α, interleukin (IL)-6, and IL-10, were detected using anti-cytokine antibody arrays integrated into each of the six chambers. We foresee further use of this device in applications such as pediatric immunology or drug/vaccine testing where it is important to balance small sample volume with the need for high information content.


Percutaneous transcatheter aortic valve implantation for degenerated surgical bioprostheses: the first case series in Asia with one-year follow-up.
Percutaneous transcatheter aortic valve implantation (TAVI) has become an established therapy for inoperable and high-surgical-risk patients with severe aortic stenosis. Although TAVI in patients with degenerated surgical aortic bioprostheses (i.e. valve-in-valve TAVI) is increasingly reported in Western studies, such data is lacking in Asian patients. We describe the initial experience of valve-in-valve TAVI in Asia. Eight patients who underwent valve-in-valve TAVI due to degenerated aortic bioprostheses were enrolled. The mechanism of bioprosthetic valve failure was stenotic, regurgitation or mixed. All procedures were performed via transfemoral arterial access, using the self-expanding CoreValve prosthesis or balloon-expandable SAPIEN XT prosthesis. The mean age of the patients was 71.6 ± 13.2 years and five were male. Mean duration to surgical bioprosthesis degeneration was 10.2 ± 4.1 years. Valve-in-valve TAVI was successfully performed in all patients. CoreValve and SAPIEN XT prostheses were used in six and two patients, respectively. There were no deaths, strokes or permanent pacemaker requirement at 30 days, with one noncardiac mortality at one year. All patients experienced New York Heart Association functional class improvement. Post-procedure mean pressure gradients were 20 ± 11 mmHg and 22 ± 8 mmHg at 30 days and one year, respectively. Residual aortic regurgitation (AR) of more than mild severity occurred in one patient at 30 days. At one year, only one patient had mild residual AR. In our experience of valve-in-valve TAVI, procedural success was achieved in all patients without adverse events at 30 days. Good clinical and haemodynamic outcomes were sustained at one year.


The South London and Maudsley NHS Foundation Trust Biomedical Research Centre (SLAM BRC) case register: development and descriptive data.
Case registers have been used extensively in mental health research. Recent developments in electronic medical records, and in computer software to search and analyse these in anonymised format, have the potential to revolutionise this research tool. We describe the development of the South London and Maudsley NHS Foundation Trust (SLAM) Biomedical Research Centre (BRC) Case Register Interactive Search tool (CRIS) which allows research-accessible datasets to be derived from SLAM, the largest provider of secondary mental healthcare in Europe. All clinical data, including free text, are available for analysis in the form of anonymised datasets. Development involved both the building of the system and setting in place the necessary security (with both functional and procedural elements). Descriptive data are presented for the Register database as of October 2008. The database at that point included 122,440 cases, 35,396 of whom were receiving active case management under the Care Programme Approach. In terms of gender and ethnicity, the database was reasonably representative of the source population. The most common assigned primary diagnoses were within the ICD mood disorders (n = 12,756) category followed by schizophrenia and related disorders (8158), substance misuse (7749), neuroses (7105) and organic disorders (6414). The SLAM BRC Case Register represents a 'new generation' of this research design, built on a long-running system of fully electronic clinical records and allowing in-depth secondary analysis of both numerical, string and free text data, whilst preserving anonymity through technical and procedural safeguards.


Multiple paired forward and inverse models for motor control.
Humans demonstrate a remarkable ability to generate accurate and appropriate motor behavior under many different and often uncertain environmental conditions. In this paper, we propose a modular approach to such motor learning and control. We review the behavioral evidence and benefits of modularity, and propose a new architecture based on multiple pairs of inverse (controller) and forward (predictor) models. Within each pair, the inverse and forward models are tightly coupled both during their acquisition, through motor learning, and use, during which the forward models determine the contribution of each inverse model's output to the final motor command. This architecture can simultaneously learn the multiple inverse models necessary for control as well as how to select the inverse models appropriate for a given environment. Finally, we describe specific predictions of the model, which can be tested experimentally.


Mismatch repair gene mutation spectrum in the Swedish Lynch syndrome population.
Lynch syndrome caused by constitutional mismatch‑repair defects is one of the most common hereditary cancer syndromes with a high risk for colorectal, endometrial, ovarian and urothelial cancer. Lynch syndrome is caused by mutations in the mismatch repair (MMR) genes i.e., MLH1, MSH2, MSH6 and PMS2. After 20 years of genetic counseling and genetic testing for Lynch syndrome, we have compiled the mutation spectrum in Sweden with the aim to provide a population-based perspective on the contribution from the different MMR genes, the various types of mutations and the influence from founder mutations. Mutation data were collected on a national basis from all laboratories involved in genetic testing. Mutation analyses were performed using mainly Sanger sequencing and multiplex ligation-dependent probe amplification. A total of 201 unique disease-predisposing MMR gene mutations were identified in 369 Lynch syndrome families. These mutations affected MLH1 in 40%, MSH2 in 36%, MSH6 in 18% and PMS2 in 6% of the families. A large variety of mutations were identified with splice site mutations being the most common mutation type in MLH1 and frameshift mutations predominating in MSH2 and MSH6. Large deletions of one or several exons accounted for 21% of the mutations in MLH1 and MSH2 and 22% in PMS2, but were rare (4%) in MSH6. In 66% of the Lynch syndrome families the variants identified were private and the effect from founder mutations was limited and predominantly related to a Finnish founder mutation that accounted for 15% of the families with mutations in MLH1. In conclusion, the Swedish Lynch syndrome mutation spectrum is diverse with private MMR gene mutations in two-thirds of the families, has a significant contribution from internationally recognized mutations and a limited effect from founder mutations.


Methodology of assessment and reporting of safety in anti-malarial treatment efficacy studies of uncomplicated falciparum malaria in pregnancy: a systematic literature review.
Considering the uncertainty of safety of anti-malarial drugs in pregnancy, efficacy studies are one of the few sources of clinical safety data. Complete safety evaluation is not usually incorporated in efficacy studies due to financial and human resource constraints. This review reports the methods used for the assessment of safety of artemisinin-based and quinine-based treatments in efficacy studies in pregnancy. Methodology of assessment and reporting of safety in efficacy studies of artemisinin-based and quinine-based treatment in pregnancy was reviewed using seven databases and two clinical trial registries. The protocol was registered to PROSPERO (CRD42017054808). Of 48 eligible efficacy studies the method of estimation of gestational age was reported in only 32 studies (67%, 32/48) and ultrasound was used in 18 studies (38%, 18/48). Seventeen studies (35%, 17/48) reported parity, 9 (19%, 9/48) reported gravidity and 13 (27%, 13/48) reported both. Thirty-eight studies (79%, 38/48) followed participants through to pregnancy outcome. Fetal loss was assessed in 34 studies (89%, 34/38), but the definition of miscarriage and stillbirth were defined only in 11 (32%, 11/34) and 7 (21%, 7/34) studies, respectively. Preterm birth was assessed in 26 studies (68%, 26/38) but was defined in 16 studies (62%, 16/26). Newborn weight was assessed in 30 studies (79%, 30/38) and length in 10 studies (26%, 10/38). Assessment of birth weight took gestational age into account in four studies (13%, 4/30). Congenital abnormalities were reported in 32 studies (84%, 32/38). Other common risk factors for adverse pregnancy outcomes were not well-reported. Incomplete reporting and varied methodological assessment of pregnancy outcomes in anti-malarial drug efficacy studies limits comparison across studies. A standard list of minimal necessary parameters to assess and report the safety component of efficacy studies of anti-malarials in pregnancy is proposed.


Eye disorders associated with obstructive sleep apnoea.
Obstructive sleep apnoea (OSA) is increasing in prevalence due to rising obesity. Public awareness is also growing. Although OSA is a disorder primarily of the upper airway during sleep, its physiological impact on other parts of the body is now well recognized. There is increasing interest in the association of OSA with various eye disorders. Work in this field has been directed predominantly to OSA prevalence and association studies, but some authors have tried to elucidate the effect of OSA therapies on eye diseases, including continuous positive airway pressure, upper airway surgery or bariatric surgery. This review discusses the publications in this area from the past year. The key ocular disorders featured in the studies and meta-analayses include glaucoma, floppy eyelid syndrome, nonarteritic ischaemic optic neuropathy, keratoconus, age-related macular degeneration and diabetic retinopathy. Associations with OSA were found with all these conditions, but aspects of the studies still leave gaps in our knowledge. This review highlights the need for ophthalmologists to consider OSA in their patients and also makes recommendations for future research studies, especially whether therapies for OSA can be effective for ocular disorders also.


Study Protocol for the Peruvian Registry of Advanced Heart Failure (REPICAV).
Heart failure (HF) is a global problem with a high mortality rate, and advanced HF (AHF) represents the stage with the highest morbidity and mortality. We have no local data on this population and its treatment. The aim of this study will be to determine the epidemiological, clinical, therapeutic, and annual survival characteristics of patients diagnosed with AHF treated in hospitals with HF units in the city of Lima, Peru. An observational, prospective, multicenter study will be conducted with evaluation at baseline and follow-up at 1, 3, 6, and 12 months after study entry. Patients over 18 years of age with AHF seen in referral health facilities in metropolitan Lima will be included. The cumulative mortality during follow-up will be estimated by the Kaplan-Meier method, and Cox regression models will calculate hazard ratios (HRs) and 95% confidence intervals (CI). Likewise, risk ratio (RR) and 95% CI will be estimated using generalized linear models with binomial family and log link function. This study was approved by the Ethics and Research Committee of the National Cardiovascular Institute (Instituto Nacional Cardiovascular "Carlos Alberto Peschiera Carrillo"-INCOR [in Spanish]; Approval report 46/2021-CEI). In Peru, there are no scientific data on the epidemiology of AHF in the population. This means that physicians are not adequately trained in the characteristics of the Peruvian population to identify patients who could be candidates for advanced therapies and to recognize the optimal time to refer these patients to more complex HF units. This study will be the first to examine the clinical-epidemiological characteristics of AHF in Peru with a follow-up of 1 year after the event and will provide relevant information on these observable characteristics for the management of high-complexity patients.


Lack of benefit with percutaneous intervention for late persistent occlusion after myocardial infarction: summary of the occluded artery trial.
The clinical utility of establishing late patency of the persistently occluded infarct-related artery with percutaneous coronary intervention (PCI) was uncertain. The Occluded Artery Trial was a National Heart, Lung, and Blood Institute-supported, international, multicenter, randomized controlled trial comparing a test strategy of late PCI (3-28 days) of the occluded infarct-related artery and optimal medical therapy to optimal medical therapy alone. The primary end point of the trial was a centrally adjudicated composite of death, reinfarction, and New York Heart Association class IV heart failure over 4 years (mean follow-up, 1,059 +/- 11 days). The final study population of 2,166 patients gave the trial 94% power to detect the anticipated 25% reduction in event rate with PCI. The combined primary outcome occurred in 161 patients in the PCI group and in 140 subjects receiving medical therapy alone. The 4-year cumulative primary event rate was 17.2% in the PCI group and 15.6% in the medical therapy group (hazard ratio, 1.16; 95% confidence interval, 0.92-1.45; P = .20; covariate-adjusted hazard ratio, 1.17; 95% confidence interval, 0.93 to 1.47; P = .18). Rates of New York Heart Association class IV heart failure and death were similar in both groups. A trend toward increased nonfatal myocardial infarction in the PCI group (hazard ratio, 1.44; 95% confidence interval, 0.96-2.16; P = .08) unrelated to periprocedural events was apparent. No significant interaction between treatment effect and prespecified subgroups was observed. This lack of clinical benefit supports optimal medical therapy alone for Occluded Artery Trial-eligible patients in current clinical practice.


Quantification of the optical properties of two-layer turbid materials using a hyperspectral imaging-based spatially-resolved technique.
Recent research has shown that a hyperspectral imaging-based spatially-resolved technique is useful for determining the optical properties of homogenous fruits and food products. To better characterize fruit properties and quality attributes, it is desirable to consider fruit to be composed of two homogeneous layers of skin and flesh. This research was aimed at developing a nondestructive method to determine the absorption and scattering properties of two-layer turbid materials with the characteristics of fruit. An inverse algorithm along with the sensitivity coefficient analysis for a two-layer diffusion model was developed for the extraction of optical properties from the spatially-resolved diffuse reflectance data acquired using a hyperspectral imaging system. The diffusion model and the inverse algorithm were validated with Monte Carlo simulations and experimental measurements from solid model samples of known optical properties. The average errors of determining two and four optical parameters were 6.8% and 15.3%, respectively, for Monte Carlo reflectance data. The optical properties of the first or top layer of the model samples were determined with errors of less than 23.0% for the absorption coefficient and 18.4% for the reduced scattering coefficient. The inverse algorithm did not give acceptable estimations for the second or lower layer of the model samples. While the hyperspectral imaging-based spatially-resolved technique has the potential to measure the optical properties of two-layer turbid materials like fruits and food products, further improvements are needed in determining the optical properties of the second layer.


Effects of a few food diet in attention deficit disorder.
Seventy-eight children, referred to a diet clinic because of hyperactive behaviour, were placed on a 'few foods' elimination diet. Fifty nine improved in behaviour during this open trial. For 19 of these children it was possible to disguise foods or additives, or both, that reliably provoked behavioural problems by mixing them with other tolerated foods and to test their effect in a placebo controlled double blind challenge protocol. The results of a crossover trial on these 19 children showed a significant effect for the provoking foods to worsen ratings of behaviour and to impair psychological test performance. This study shows that observations of change in behaviour associated with diet made by parents and other people with a role in the child's care can be reproduced using double blind methodology and objective assessments. Clinicians should give weight to the accounts of parents and consider this treatment in selected children with a suggestive medical history.


Latest in vitro and in vivo models of celiac disease.
Currently, the only treatment for celiac disease is a gluten-free diet, and there is an increased desire for alternative therapies. In vitro and in vivo models of celiac disease have been generated in order to better understand the pathogenesis of celiac disease, and this review will discuss these models as well as the testing of alternative therapies using these models. The research discussed describes the different in vitro and in vivo models of celiac disease that currently exist and how they have contributed to our understanding of how gluten can stimulate both innate and adaptive immune responses in celiac patients. We also provide a summary on the alternative therapies that have been tested with these models and discuss whether subsequent clinical trials were done based on these tests done with these models of celiac disease. Only a few of the alternative therapies that have been tested with animal models have gone on to clinical trials; however, those that did go on to clinical trial have provided promising results from a safety standpoint. Further trials are required to determine if some of these therapies may serve as an effective adjunct to a gluten-free diet to alleviate the adverse affects associated with accidental gluten exposure. A "magic-bullet" approach may not be the answer to celiac disease, but possibly a future cocktail of these different therapeutics may allow celiac patients to consume an unrestricted diet.


Spanish dermatological vocabulary used by Mexican workers.
Spanish is the second most common language spoken in the United States. Physicians can expect a growing population of patients who speak Spanish as their primary language. The purpose of this study was to develop a glossary of Spanish dermatological vocabulary commonly used by Mexican workers. Dermatologic terms were gathered from transcripts of in-depth interviews with 31 Latino farmworkers in North Carolina. Participants were asked to name and define words related to skin irritations. Farmworkers provided several definitions of some terms, and several Spanish equivalents exist for specific English words. This glossary is a supplement to other resources for learning medical Spanish and expands health care professionals' knowledge of dermatology-related Spanish vocabulary.


Time-gated luminescence imaging of singlet oxygen photoinduced by fluoroquinolones and functionalized graphenes in Daphnia magna.
Singlet oxygen (1O2) can be photogenerated by photoactive xenobiotics and is capable of causing adverse effects due to its electrophilicity and its high reactivity with biological molecules. Detection of the production and distribution of 1O2 in living organisms is therefore of great importance. In this study, a luminescent probe ATTA-Eu3+ combined with time-gated luminescence imaging was adopted to detect the distribution and temporal variation of 1O2 photoinduced by fluoroquinolone antibiotics and carboxylated/aminated graphenes in Daphnia magna. Results show that the xenobiotics generate 1O2 in living daphnids under simulated sunlight irradiation (SSR). The photogeneration of 1O2 by carboxylated/aminated graphenes was also confirmed by electron paramagnetic resonance spectroscopy. The strongest luminescence signals of 1O2 were observed in the hindgut of daphnids, and the signals in different areas of the daphnids (gut, thoracic legs and post-abdominal claw) displayed a similar trend of enhancement over irradiation time. Mean 1O2 concentrations at different regions of daphnids within one hour of SSR irradiation were estimated to be in the range of 0.5∼4.8μM. This study presented an efficient method for visualizing and quantifying the temporal and spatial distribution of 1O2 photogenerated by xenobiotics in living organisms, which can be employed for phototoxicity evaluation of xenobiotics.


On optimizing distance-based similarity search for biological databases.
Similarity search leveraging distance-based index structures is increasingly being used for both multimedia and biological database applications. We consider distance-based indexing for three important biological data types, protein k-mers with the metric PAM model, DNA k-mers with Hamming distance and peptide fragmentation spectra with a pseudo-metric derived from cosine distance. To date, the primary driver of this research has been multimedia applications, where similarity functions are often Euclidean norms on high dimensional feature vectors. We develop results showing that the character of these biological workloads is different from multimedia workloads. In particular, they are not intrinsically very high dimensional, and deserving different optimization heuristics. Based on MVP-trees, we develop a pivot selection heuristic seeking centers and show it outperforms the most widely used corner seeking heuristic. Similarly, we develop a data partitioning approach sensitive to the actual data distribution in lieu of median splits.


A comprehensive approach to reablement in dementia.
As society grapples with an aging population and increasing prevalence of disability, "reablement" as a means of maximizing functional ability in older people is emerging as a potential strategy to help promote independence. Reablement offers an approach to mitigate the impact of dementia on function and independence. This article presents a comprehensive reablement approach across seven domains for the person living with mild-to-moderate dementia. Domains include assessment and medical management, cognitive disability, physical function, acute injury or illness, assistive technology, supportive care, and caregiver support. In the absence of a cure or ability to significantly modify the course of the disease, the message for policy makers, practitioners, families, and persons with dementia needs to be "living well with dementia", with a focus on maintaining function for as long as possible, regaining lost function when there is the potential to do so, and adapting to lost function that cannot be regained. Service delivery and care of persons with dementia must be reoriented such that evidence-based reablement approaches are integrated into routine care across all sectors.


Lower extremity performance in nondisabled older persons as a predictor of subsequent hospitalization.
This study examines, in initially nondisabled older persons, the impact of reduced lower extremity performance on subsequent hospitalizations. A 4-year prospective cohort study was conducted among 3381 persons, aged 71 years and older, who initially reported no disability. At baseline, lower extremity performance was measured by an assessment of standing balance, a timed 2.4-m walk, and a timed test of rising from a chair five times. Data on subsequent hospital admissions and discharge diagnoses over 4 years were obtained from the Medicare database. During the follow-up period, nondisabled persons with poor lower extremity performance spent significantly more days in the hospital (17.7 days) than those with intermediate and high performance (11.6 and 9.7 days, respectively). Poor lower extremity performance in nondisabled persons significantly predicted subsequent hospitalization over 4 years (relative risk for hospitalization in those with poor vs high performance: 1.78; 95% confidence interval, 1.45-2.17). This increased hospitalization risk could not be explained by several indicators of baseline health status. Increased hospitalization risks were especially found for geriatric conditions, such as dementia, decubitus ulcer, hip fractures, other fractures, pneumonia, dehydration, and acute infections. Even in persons who are currently nondisabled, a simple measure of lower extremity performance is predictive of subsequent hospitalization, especially for geriatric conditions.


The evolution of Mercury's crust: a global perspective from MESSENGER.
Mapping the distribution and extent of major terrain types on a planet's surface helps to constrain the origin and evolution of its crust. Together, MESSENGER and Mariner 10 observations of Mercury now provide a near-global look at the planet, revealing lateral and vertical heterogeneities in the color and thus composition of Mercury's crust. Smooth plains cover approximately 40% of the surface, and evidence for the volcanic origin of large expanses of plains suggests that a substantial portion of the crust originated volcanically. A low-reflectance, relatively blue component affects at least 15% of the surface and is concentrated in crater and basin ejecta. Its spectral characteristics and likely origin at depth are consistent with its apparent excavation from a lower crust or upper mantle enriched in iron- and titanium-bearing oxides.


Removal of human enteric viruses by a full-scale membrane bioreactor during municipal wastewater processing.
In the US, human enteric viruses are the main etiologic agents of childhood gastroenteritis, resulting in several hospitalizations and deaths each year. These viruses have been linked to several waterborne diseases, such as acute gastroenteritis, conjunctivitis and respiratory illness. The removal of human enterovirus (EV) and norovirus genogroup II (NoV GGII) was studied in a full-scale membrane bioreactor (MBR) wastewater treatment plant (WWTP) and compared with the removal of human adenovirus (HAdV). In total, 32 samples were quantified using real-time reverse transcription-PCR (RT-PCR) from four separate locations throughout the treatment process; influent, primary settling effluent, membrane influent (which includes the MLSS) and membrane effluent. EV was detected in all 32 samples (100%) with an average concentration of 1.1 × 10(7) and 7.8 × 10(1) viruses/L for the membrane influent and membrane effluent, respectively. NoV GGII was detected in 20 of 32 samples (63%) with an average membrane influent and membrane effluent concentration of 2.8 × 10(5) and 1.2 × 10(1) viruses/L, respectively. HAdV was detected in all 32 samples with an average membrane influent concentration of 5.2 × 10(8) and 2.7 × 10(3) viruses/L in the membrane effluent. Our findings indicate that this particular full-scale MBR treatment was able to reduce the viral loads by approximately 5.1 and 3.9 log units for EV and NoV GGII as compared to 5.5 log units for HAdV. This full-scale MBR system outperformed the removal observed in previous pilot and bench scale studies by 1 to 2 log units. To the best of our knowledge, this is the first study focusing on the removal of EV in a full-scale MBR WWTP using real-time RT-PCR, and on the solid-liquid distribution of EV and NoV GII in secondary biological treatment.


The human Transmembrane Protease Serine 2 is necessary for the production of Group 2 influenza A virus pseudotypes.
The monomer of influenza haemagglutinin is synthesized as a single polypeptide precursor that during maturation is cleaved by proteases into two active subunits. Other studies have demonstrated that the human Transmembrane Protease Serine 2 (TMPRSS2) can cleave the HA of human seasonal influenza viruses. Consequently, we have investigated the use of human Transmembrane Protease Serine 2 to produce high titre influenza haemmagglutinin (HA) lentiviral pseudotypes from Group 2 influenza viruses. Such pseudotypes represent powerful and safe tools to study viral entry and immune responses. Influenza pseudotype particles are obtained by co-transfecting human embryonic kidney HEK293T/17 cells using plasmids coding for the influenza HA, HIV gag-pol and a lentiviral vector incorporating firefly luciferase. However, in order to produce Group 2 pseudotypes, it was necessary to co-transfect a plasmid expressing the TMPRSS2 endoprotease, to achieve the necessary HA cleavage for infective particle generation. These lentiviral pseudotypes were shown to transduce HEK293T/17 cells with high efficiency. This demonstrates that TMPRSS2 is necessary for the functional activation, in vitro, of both the HA of human seasonal influenza and other Group 2 HA influenza strains. Additionally, we show that the Group 2 influenza pseudotype particles can be used as surrogate antigens in neutralization assays and are efficiently neutralized by corresponding influenza virus reference sera. These data demonstrate that the viral pseudotype system is a powerful method for serological surveillance of a wide range of influenza viruses.


Comparison of 3MP medical-grade to 1MP office-grade LCD monitors in mammographic diagnostic and perceptual performance.
Picture archiving and communication systems images designed to be viewed on high-resolution medical-grade monitors are routinely viewed on office-grade monitors on the wards or at home. This study aimed to determine whether a statistically significant difference in diagnostic (cancer detection) and perceptual (microcalcification detection) performance exists between 3MP grade and 1MP office-grade monitors. 3MP Dome medical-grade liquid crystal display (LCD) monitors (Planar, Beaverton, OR, USA) were compared to 1MP Dell office-grade LCD monitors (Dell Inc, Round Rock, TX, USA). Eight radiologists (reader experience 8-30 years) read the same set of 100 mammograms (23/100 with proven cancers and 52/100 with microcalcifications) presented in random order on three occasions separated by two time intervals of 12 weeks. Reads 1 and 3 utilised 3MP monitors and formed the baseline read. Read 2 utilised 1MP monitors and constituted the experimental read. Reading conditions were standardised. Readers were aware of which monitors they were using. Multivariate logistic regression analysis (to account for reader variability and monitor impact) was performed to assess for statistical significance. At α = 5%, confidence intervals analysis comparing the measured parameters between 1MP to 3MP monitors demonstrated no statistically significant difference in diagnostic and perceptual performance for the reader group. In cancer detection (the diagnostic task), reader accuracy remained high irrespective of monitor type. Regression analysis comparing performance with 1MP against 3MP monitors found P values of 0.693 and 0.324 for diagnostic and perceptual performance, respectively. There were no statistically and clinically significant differences between 3MP and 1MP monitors in mammographic diagnostic and perceptual performance. Comparable performance may be due to compensatory behaviour by readers.


Evaluation of VEGF-mediated signaling in primary human cells reveals a paracrine action for VEGF in osteoblast-mediated crosstalk to endothelial cells.
Communication between endothelial and bone cells is crucial for controlling vascular supply during bone growth, remodeling, and repair but the molecular mechanisms coordinating this intercellular crosstalk remain ill-defined. We have used primary human and rat long bone-derived osteoblast-like cells (HOB and LOB) and human umbilical vein endothelial cells (HUVEC) to interrogate the potential autocrine/paracrine role of vascular endothelial cell growth factor (VEGF) in osteoblast:endothelial cell (OB:EC) communication and examined whether prostaglandins (PG), known modulators of both OB and EC behavior, modify VEGF production. We found that the stable metabolite of PGI2, 6-keto-PGF(1alpha) and PGE2, induced a concentration-dependent increase in VEGF release by HOBs but not ECs. In ECs, VEGF promoted early ERK1/2 activation, late cyclooxygenase-2 (COX-2) protein induction, and release of 6-keto-PGF1alpha. In marked contrast, no significant modulation of these events was observed in HOBs exposed to VEGF, but LOBs clearly exhibited COX-dependent prostanoid release (10-fold less than EC) following VEGF treatment. A low level of osteoblast-like cell responsiveness to exogenous VEGF was supported by VEGFR2/Flk-1 immunolabelling and by blockade of VEGF-mediated prostanoid generation by a VEGFR tyrosine kinase inhibitor (TKI). HOB alkaline phosphatase (ALP) activity was increased following long-term non-contact co-culture with ECs and exposure of ECs to VEGF in this system further increased OB-like cell differentiation and markedly enhanced prostanoid release. Our studies confirm a paracrine EC-mediated effect of VEGF on OB-like cell behavior and are the first supporting a model in which prostanoids may facilitate this unidirectional VEGF-driven OB:EC communication. These findings may offer novel regimes for modulating pathological bone remodeling anomalies through the control of the closely coupled vascular supply.


Towards a genuinely medical model for psychiatric nosology.
Psychiatric nosology is widely criticized, but solutions are proving elusive. Planned revisions of diagnostic criteria will not resolve heterogeneity, comorbidity, fuzzy boundaries between normal and pathological, and lack of specific biomarkers. Concern about these difficulties reflects a narrow model that assumes most mental disorders should be defined by their etiologies. A more genuinely medical model uses understanding of normal function to categorize pathologies. For instance, understanding the function of a cough guides the search for problems causing it, and decisions about when it is expressed abnormally. Understanding the functions of emotions is a foundation missing from decisions about emotional disorders. The broader medical model used by the rest of medicine also recognizes syndromes defined by failures of functional systems or failures of feedback control. Such medical syndromes are similar to many mental diagnoses in their multiple causes, blurry boundaries, and nonspecific biomarkers. Dissatisfaction with psychiatric nosology may best be alleviated, not by new diagnostic criteria and categories, but by more realistic acknowledgment of the untidy landscape of mental and other medical disorders.


A glutathione-depleting chemodynamic therapy agent with photothermal and photoacoustic properties for tumor theranostics.
Nowadays, Fenton reaction-based chemodynamic therapy (CDT) strategies have drawn extensive attention as tumor-specific nanomedicine-based therapy. Nevertheless, current existing CDTs normally suffer from therapeutic bottlenecks such as the scavenging of hydroxyl radical (˙OH) by intracellular antioxidants and unideal therapeutic outcome of single treatment modality. Herein, we constructed novel all-in-one AFP nanoparticles (NPs) as CDT agents through a one-pot process for multifunctional nanotheranostics. The as-constructed AFP NPs could simultaneously produce ˙OH through the Fenton reaction and scavenge intracellular glutathione, functioning as self-reinforced CDT agents to achieve tumor-triggered enhanced CDT (ECDT). In addition, the AFP NPs possessed the capability of H2O2 and acid-boosted photoacoustic imaging and photothermal therapy, enabling a precise and effective tumor therapeutic outcome with minimal nonspecific damage in combination with ECDT. Our novel nanoplatform would open new perspectives on multi-functional CDT agents for accurate and non-invasive tumor theranostics.


A novel method of preserving cardiac grafts using a hydrogen-rich water bath.
Exogenously administered hydrogen exerts cytoprotective effects through anti-oxidant, anti-inflammatory, and anti-apoptotic mechanisms in various disease settings, including organ transplantation. Our objective in this study was to evaluate the efficacy of a novel cold storage device equipped with a hydrogen-rich water bath. The study used an established rat heterotopic transplantation model. Syngeneic heart grafts from elderly donors (60- to 70-week-old Lewis rats) or allografts from adult donors (12-week-old Brown Norway rats) were exposed to prolonged cold preservation. The cardiac grafts were stored in plastic bags containing Celsior, which were immersed in the cold water bath equipped with an electrolyzer to saturate the water with hydrogen. The cardiac grafts then were heterotopically engrafted into Lewis rat recipients. In both experimental settings, serum troponin I and creatine phosphokinase were markedly elevated 3 hours after reperfusion in the control grafts without hydrogen treatment. The grafts exhibited prominent inflammatory responses, including neutrophil infiltration and the upregulation of messenger RNAs for pro-inflammatory cytokines and chemokines. Myocardial injury and inflammatory events were significantly attenuated by organ storage in the hydrogen-rich water bath. The grafts stored using the hydrogen-rich water bath also exhibited less mitochondrial damage and a higher adenosine triphosphate content. Hydrogen delivery to cardiac grafts during cold preservation using a novel hydrogen-supplemented water bath efficiently ameliorated myocardial injury due to cold ischemia and reperfusion. This new device to saturate organs with hydrogen during cold storage merits further investigation for possible therapeutic and preventative use during transplantation.


Hot and Cool Executive Functions in Adolescence: Development and Contributions to Important Developmental Outcomes.
Despite significant theoretical advancement in the area of child neuropsychology, limited attention has been paid to the developmental features of adolescence. The present study intends to address this issue in relation to executive function (EF). EF refers to the psychological processes that underlie goal-directed behavior; recent studies separate cool EF (psychological process involves pure logic and critical analysis) and hot EF (psychological process driven by emotion). Although neurological findings suggest that adolescence is a sensitive period for EF development, data on comparing the developmental progression in hot or cool EFs is highly missing. Moreover, while evidence has confirmed the relationships between EF and day-to-day functioning, whether and how hot and cool EFs contribute to core developmental outcomes in adolescence is still remained unknown. The current study aims to enhance our understanding of the development and impacts of hot and cool EFs in adolescence. A total of 136 typically developing adolescents from age 12 to 17 completed four cool EF tasks including Backward digit span, Contingency naming test, Stockings of Cambridge, and Stroop Color and Word test, and one hot task on Cambridge gambling task. Data on academic performance and psychological adjustment was also collected. Results showed that cool and hot EF exhibited different patterns of age-related growth in adolescence. Specifically, cool EF ascended with age while hot EF showed a bell-shaped development. Moreover, there were correlations among cool EF measures but no association between cool and hot EFs. Further, cool EF was a better predictor of academic performance, while hot EF uniquely related to emotional problems. The results provide evidence for the association among cool EF tests and the differentiation of hot and cool EFs. The bell-shaped development of hot EF might suggest a period of heightened risk-taking propensity in middle adolescence. Given the plastic nature of EF, especially over adolescence, the current findings may have practical implications for future EF identification and training.


Experimental test of genetic rescue in isolated populations of brook trout.
Genetic rescue is an increasingly considered conservation measure to address genetic erosion associated with habitat loss and fragmentation. The resulting gene flow from facilitating migration may improve fitness and adaptive potential, but is not without risks (e.g., outbreeding depression). Here, we conducted a test of genetic rescue by translocating ten (five of each sex) brook trout (Salvelinus fontinalis) from a single source to four nearby and isolated stream populations. To control for the demographic contribution of translocated individuals, ten resident individuals (five of each sex) were removed from each recipient population. Prior to the introduction of translocated individuals, the two smallest above-barrier populations had substantially lower genetic diversity, and all populations had reduced effective number of breeders relative to adjacent below-barrier populations. In the first reproductive bout following translocation, 31 of 40 (78%) translocated individuals reproduced successfully. Translocated individuals contributed to more families than expected under random mating and generally produced larger full-sibling families. We observed relatively high (>20%) introgression in three of the four recipient populations. The translocations increased genetic diversity of recipient populations by 45% in allelic richness and 25% in expected heterozygosity. Additionally, strong evidence of hybrid vigour was observed through significantly larger body sizes of hybrid offspring relative to resident offspring in all recipient populations. Continued monitoring of these populations will test for negative fitness effects beyond the first generation. However, these results provide much-needed experimental data to inform the potential effectiveness of genetic rescue-motivated translocations.


Prediction of disease-free survival using relative change in FDG-uptake early during neoadjuvant chemoradiotherapy for potentially curable esophageal cancer: A prospective cohort study.
18F-Fluorodeoxyglucose positron emission tomography (FDG-PET) has been investigated as a tool for monitoring response to neoadjuvant chemo- and chemoradiotherapy (CT and CRT, respectively) and as a predictor for survival in patients with esophageal cancer. In contrast to patients who undergo neoadjuvant CT, it is not known whether patients who are clinically identified as responders after neoadjuvant CRT show better disease-free survival (DFS) than patients identified as nonresponders. The aim of the study was to determine the predictive value of FDG-uptake measured prior to and early during neoadjuvant CRT. Patients treated with neoadjuvant CRT between 2004 and 2009 within a randomized trial were included. FDG-uptake was measured at baseline and after 14 days of CRT. According to the PERCIST-criteria, patients were allocated to have metabolic response, stable disease, or progression. Patients were followed until recurrence of disease or death. The predictive value of FDG-PET was determined with univariable and multivariable analysis in patients who underwent potentially curative surgery. One-hundred and six patients were included in the analysis. Minimal follow-up for surviving patients was 60 months. No significant differences in DFS were found between patients with metabolic response, stable disease, or progression, with 5-year DFS rates of 66%, 53%, and 67%, respectively (P = 0.39). Relative change in FDG uptake after 14 days of CRT is not associated with DFS in patients with esophageal cancer undergoing neoadjuvant chemoradiotherapy followed by surgery. These measurements should not be used for prognostication in this specific group of patients.


The correlation between osteopontin adsorption and cell adhesion to mixed self-assembled monolayers of varying charges and wettability.
Osteopontin (OPN) is a key mediator of cell interactions with biomaterials. However, few studies have been dedicated to studying cell adhesion on OPN-adsorbed substrates with controlled charge and wettability. Here, amino-carboxyl (NH2/COOH) and hydroxyl-methyl (OH/CH3) mixed self-assembled monolayers (SAMs) of varying charges and wettability, respectively, were used as controllable model surfaces to study OPN adsorption and subsequent mesenchymal stem cell (MSC) adhesion. The amount of OPN adsorbed onto the NH2/COOH mixed SAMs appeared to monotonically depend on the surface charge, whereas only a moderately hydrophilic surface was conducive to OPN adsorption on OH/CH3 mixed SAMs. The results correlated well with cell spreading on OPN-coated surfaces in a serum-free medium culture. In addition, the OH/CH3 mixed SAMs with moderate wettability tended to promote β1, β3, αv and α5 integrins, indicating that wettability may guide cell adhesion by mediating the integrins signaling pathway. This work will have reference value for designing biologically responsive substrate surfaces.


Rapid identification of cyst (Heterodera spp., Globodera spp.) and root-knot (Meloidogyne spp.) nematodes on the basis of ITS2 sequence variation detected by PCR-single-strand conformational polymorphism (PCR-SSCP) in cultures and field samples.
Cyst and root-knot nematodes show high levels of gross morphological similarity. This presents difficulties for the study of their ecology in natural ecosystems. In this study, cyst and root-knot nematode species, as well as some ectoparasitic nematode species, were identified using the second internal transcribed spacer (ITS2) sequence variation detected by polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP). The ITS2 region was sufficiently variable within the taxa investigated to allow species to be separated on the basis of minor sequence variation. The PCR primers used in this study were effective for 12 species with three genera within the Heteroderinae (Globodera pallida, G. rostochiensis, Heterodera arenaria/avenae, H. ciceri, H. daverti, H. hordecalis, H. mani, H. schachtii, H. trifolii, Meloidogyne ardenensis, M. duytsi and M. maritima). However, pathotypes of Globodera pallida and G. rostochiensis could not be distinguished. The method was tested at two coastal dune locations in The Netherlands (one in the lime-poor dunes of the north and one in calcareous dunes of the south) to determine the population structure of cyst nematodes. At each site, cyst nematodes were associated with three plant species: two plant species on the foredune (Elymus farctus and Ammophila arenaria) and one plant species occurring further inland (Calamagrostis epigejos). Two species of cyst nematodes, H. arenaria and H. hordecalis, were found. H. arenaria associated with vigorous A. arenaria and H. hordecalis in association with degenerating A. arenaria and C. epigejos. The field survey demonstrated that in coastal dunes abiotic factors may be the important affecting the distribution of cyst nematodes.


Efficacy of a guided internet-based intervention (iSOMA) for somatic symptoms and related distress in university students: study protocol of a randomised controlled trial.
Persistent and distressing somatic symptoms are common in younger age cohorts such as university students. However, the majority does not receive adequate psychosocial care. Internet-based and mobile-based interventions may represent low threshold and effective extensions to reduce somatic and associated mental symptom severity. The planned study aims to investigate the feasibility and efficacy of an internet-based intervention in reducing somatic and psychological symptoms in an international population of university students with somatic symptom burden. This parallel two-armed randomised controlled trial evaluates an 8-week guided intervention, including web-based consecutive modules based on cognitive behavioural therapy (CBT) principles against a waitlist control group. Guidance will be provided by trained psychologists with weekly written supportive feedback. As part of the 'Studicare' project, the present study aims to recruit n=154 university students indicating somatic symptom burden at baseline in German-speaking universities. Self-report assessments will take place at baseline and after intervention completion (8, 16 weeks after randomisation). The primary outcome will be the severity of somatic symptoms and associated mental distress. Secondary outcomes include depression, (health) anxiety, disability, intervention satisfaction and adherence. Ethics approval has been granted. Results from this study will be published in peer-reviewed journals and presented at international conferences. DRKS00014375; Pre-results.


Optimization of subconjunctival biodegradable microfilms for sustained drug delivery to the anterior segment in a small animal model.
We evaluated a biodegradable, sustained-release, prednisolone acetate (PA)-loaded poly[d,l-lactide-co-ε-caprolactone] (PLC) drug delivery system on its biocompatibility, feasibility and release characteristics in vitro and in vivo. Blank and 40% PA-loaded PLC microfilms with a diameter of 2 mm were fabricated, and the degradation and drug release profiles of the microfilms were characterized in vitro and in vivo. The microfilms were implanted into the subconjunctival space of Lewis rats (n = 48). All eyes were assessed clinically using slit-lamp biomicroscopy, and graded with Hackett-McDonald ocular scoring system and anterior segment optical coherence tomography. Histologic and immunohistochemical analyses were performed comparing blank and PA-loaded microfilm groups. PA concentrations in the aqueous humor were determined by HPLC. Subconjunctivally-implanted PLC microfilms were able to deliver PA in a sustained manner over 3 months, with a steady rate of 0.002 mg/d in vivo. Eyes with either blank or PA-loaded implanted microfilms showed a very minimal inflammatory response at the insertion sites and mild degree of collagen encapsulation around the microfilms, with significantly less CD11c cells at 2 and 4 weeks (P = 0.001 and P = 0.002), and collagen formation at 2 weeks (P = 0.001) in the PA-loaded microfilm group. Anterior chamber PA levels were achieved, with concentrations at 76.7 ± 5.9, 70.3 ± 2.3, and 42.7 ± 4.1 ng/mL at 2, 4, and 12 weeks, respectively. PA-loaded PLC microfilms display good biocompatibility, feasibility, and desirable sustained drug release profiles, and have the potential to exhibit antifibrotic and anti-inflammatory effects. This device is applicable to use in small animal models of anterior segment inflammation.


Comparison of miRNA expression patterns using total RNA extracted from matched samples of formalin-fixed paraffin-embedded (FFPE) cells and snap frozen cells.
Archival formalin-fixed paraffin-embedded (FFPE) tissues have limited utility in applications involving analysis of gene expression due to mRNA degradation and modification during fixation and processing. This study analyzed 160 miRNAs in paired snap frozen and FFPE cells to investigate if miRNAs may be successfully detected in archival specimens. Our results show that miRNA extracted from FFPE blocks was successfully amplified using Q-RT-PCR. The levels of expression of miRNA detected in total RNA extracted from FFPE were higher than that extracted from snap frozen cells when the quantity of total RNA was identical. This phenomenon is most likely explained by the fact that larger numbers of FFPE cells were required to generate equivalent quantities of total RNA than their snap frozen counterparts. We hypothesise that methylol cross-links between RNA and protein which occur during tissue processing inhibit the yield of total RNA. However, small RNA molecules appear to be less affected by this process and are recovered more easily in the extraction process. In general miRNAs demonstrated reliable expression levels in FFPE compared with snap frozen paired samples, suggesting these molecules might prove to be robust targets amenable to detection in archival material in the molecular pathology setting.


Effect of delta-9-tetrahydrocannabinol on behavioral despair and on pre- and postsynaptic serotonergic transmission.
Preclinical and clinical studies suggest that direct and indirect cannabinoid agonists, including enhancers of endocannabinoids, engender stress-relieving, anxiolytic and antidepressant effects, mediated by central CB(1) receptors (CB(1)Rs). The effect of the main pharmacologically active principle in cannabis, (-)-trans-Δ(9)-tetrahydrocannabinol (delta-9-THC), on depressive behavior and on the serotonin (5-HT) system, which is implicated in the mechanism of action of antidepressants, has not been extensively clarified. Here, we showed that repeated (5 days), but not single (acute) intraperitoneal (ip) treatment with delta-9-THC (1mg/kg) exerts antidepressant-like properties in the rat forced swim test (FST). This effect was CB(1)R-dependent because it was blocked by the CB(1)R antagonist rimonabant (1mg/kg, ip). Using in vivo electrophysiology, we demonstrated that delta-9-THC modulated dorsal raphe (DR) 5-HT neuronal activity through a CB(1)R-dependent mechanism. Acute intravenous delta-9-THC administration (0.1-1.5mg/kg) elicited a complex response profile, producing excitatory, inhibitory and inert responses of 5-HT neurons. Only excitatory responses were blocked by rimonabant. Finally, repeated but not single delta-9-THC administration (1mg/kg, ip) enhanced tonic 5-HT(1A) receptor activity in the hippocampus, a postsynaptic event commonly elicited by standard antidepressants. These results suggest that delta-9-THC, like other CB(1)R agonists and endocannabinoid enhancers, may possess antidepressant properties at low doses, and could modulate 5-HT transmission in the DR and hippocampus as standard antidepressants such as selective serotonin reuptake inhibitors.


Sero-epidemiology of Toxocara canis infection among aboriginal schoolchildren in the mountainous areas of north-eastern Taiwan.
A sero-epidemiological study of Toxocara canis infection was conducted among Atayal schoolchildren (aged 7-12 years) residing in the mountainous areas of north-eastern Taiwan. The 73 children investigated were each checked for anti-Toxocara IgG, in ELISA based on the larval excretory-secretory antigens of T. canis larvae. A short, self-administered questionnaire was then used to collect relevant information from each subject, including data on the keeping of dogs, playing in soil, eating raw vegetables, and whether the subjects normally washed their hands before eating. Once the seropositive children had been identified, odds ratios (OR), with their corresponding 95% confidence intervals (CI) and P-values, were calculated for each potential risk factor. When diluted 1:64, sera from 42 (57.5%) of the children gave a positive result in the ELISA, indicating that these 42 children were seropositive for T. canis infection. Seropositivity did not appear to be associated with the age or gender of the subject, the eating of raw vegetables, or the regular failure to wash hands prior to a meal. Compared with the other subjects, however, those who admitted living in a household where dogs were kept (OR = 3.79; CI = 1.23-11.69; P = 0.02) or playing in soil (OR = 3.00; CI = 1.10-8.16; P = 0.03) appeared at increased risk of seropositivity.


Biocompatible gelatin nanoparticles for tumor-targeted delivery of polymerized siRNA in tumor-bearing mice.
Structural modifications of the siRNA backbone improved its physiochemical properties for incorporating in gene carriers without loss of gene-silencing efficacy. These modifications provide a wider variety of choice of vector systems for siRNA delivery. We developed a tumor-targeted siRNA delivery system using polymerized siRNA (poly-siRNA) and natural polymer gelatin. The polymerized siRNA (poly-siRNA) was prepared through self-polymerization of thiol groups at the 5'-end of sense and anti-sense strands of siRNA and was encapsulated in the self-assembled thiolated gelatin (tGel) nanoparticles (NPs) with chemical cross-linking. The resulting poly-siRNA-tGel (psi-tGel) nanoparticles (average of 145 nm in diameter) protect siRNA molecules from enzymatic degradation, and can be reversibly reduced to release functional siRNA molecules in reductive conditions. The psi-tGel NPs presented efficient siRNA delivery in red fluorescence protein expressing melanoma cells (RFP/B16F10) to down-regulate target gene expression. In addition, the NPs showed low toxicity at a high transfection dose of 125 μg/ml psi-tGel NPs, which included 1 μM of siRNA molecules. In tumor-bearing mice, the psi-tGel NPs showed 2.8 times higher tumor accumulation than the naked poly-siRNA, suggesting tumor-targeted siRNA delivery of psi-tGel NPs. Importantly, the psi-tGel NPs induced effective tumor RFP gene silencing in vivo without remarkable toxicity. The psi-tGel NPs have great potential for a systemic siRNA delivery system for cancer therapy, based on their characteristics of low toxicity, tumor accumulation, and effective siRNA delivery.


Effects of partial tonsillectomy on the immune functions of children with obstructive sleep apnea-hypopnea syndrome at early stage.
The purpose of this study was to investigate the changes in the humoral and cellular immunity of children with obstructive sleep apnea-hypopnea syndrome and hypertrophy of tonsils before and after plasma-mediated temperature-controlled radiofrequency ablation treatment. Fifty-seven children suffering from obstructive sleep apnea-hypopnea syndrome and with hypertrophy of tonsils were enrolled in this study. Thirty-seven children were grouped in the partial tonsillectomy group and 20, in the tonsillectomy group. The levels of CD3(+), CD4(+), CD8(+), and CD4(+)/CD8(+) were measured for cellular immunity, and the levels of IgG, IgA, and IgM were measured for humoral immunity. Blood samples were collected before and 1 and 3 months after the operation. The IgG, IgA, and IgM levels in the tonsillectomy group were significantly decreased 1 month after the operation, and recovered to the normal levels within 3 months of the operation (P < 0.05). However, the levels of IgG, IgA, and IgM in the partial tonsillectomy group decreased slightly, without a significant difference (P > 0.05). The cellular immunity of the 2 groups was not statistically different pre- and post-operation (P > 0.05). The results from the present study indicate that partial tonsillectomy by plasma-mediated temperature-controlled radiofrequency ablation did not impact on the humoral and cellular immunity of children.


Rescue of secretion of a rare-disease associated mis-folded mutant glycoprotein in UGGT1 knock-out mammalian cells.
Endoplasmic reticulum (ER) retention of mis-folded glycoproteins is mediated by the ERlocalised eukaryotic glycoprotein secretion checkpoint, UDP-glucose glycoprotein glucosyl-transferase (UGGT). The enzyme recognises a mis-folded glycoprotein and flags it for ER retention by reglucosylating one of its N-linked glycans. In the background of a congenital mutation in a secreted glycoprotein gene, UGGT-mediated ER retention can cause rare disease even if the mutant glycoprotein retains activity ("responsive mutant"). Here, we investigated the subcellular localisation of the human Trop-2 Q118E variant, which causes gelatinous droplike corneal dystrophy (GDLD). Compared with the wild type Trop-2, which is correctly localised at the plasma membrane, the Trop-2-Q118E variant is found to be heavily retained in the ER. Using Trop-2-Q118E, we tested UGGT modulation as a rescue-of-secretion therapeutic strategy for congenital rare disease caused by responsive mutations in genes encoding secreted glycoproteins. We investigated secretion of a EYFP-fusion of Trop-2-Q118E by confocal laser scanning microscopy. As a limiting case of UGGT inhibition, mammalian cells harbouring CRISPR/Cas9-mediated inhibition of the UGGT1 and/or UGGT2 gene expressions were used. The membrane localisation of the Trop-2-Q118E-EYFP mutant was successfully rescued in UGGT1-/- and UGGT1/2-/- cells. UGGT1 also efficiently reglucosylated Trop-2-Q118E-EYFP in cellula. The study supports the hypothesis that UGGT1 modulation constitutes a novel therapeutic strategy for the treatment of Trop-2-Q118E associated GDLD, and it encourages the testing of modulators of ER glycoprotein folding Quality Control (ERQC) as broad-spectrum rescueof-secretion drugs in rare diseases caused by responsive secreted glycoprotein mutants.


Treatment of primary CNS lymphoma.
Primary central nervous system lymphoma is a particular challenge in clinical neuro-oncology. In contrast to most other malignant brain tumors, it may be considered a curable disease at least in younger patients who can tolerate intensive treatment regimens. Yet, therapeutic progress has been limited with little measurable improvement in outcome over the last two decades, mainly due to the low incidence of this tumor, which impedes the execution of large randomized clinical trials, and the failure of most large cooperative groups to conduct such trials. Whenever possible, high-dose methotrexate (HD-MTX) is the backbone of the therapeutic regimen. Response rates can be increased by the addition of second agents like ifosfamide or cytarabine, however, their impact on overall survival is less clear. Similarly, the use of the anti-CD20 antibody rituximab, commonly used in the treatment of B cell lymphomas outside the CNS, remains controversial and has not been examined in adequate clinical trials. The prognosis of patients, who do not qualify for HD-MTX-based chemotherapy, is considerably poorer. Radiation therapy is an active treatment with high response rates but does typically not result in long-lasting remissions. It remains an important therapeutic option as a salvage therapy in patients progressing on or no longer responding to HD-MTX-based treatment. The combination of HD-MTX and radiation therapy does not prolong overall survival. It is associated with significant neurotoxicity, and it should be avoided. Another matter of debate is whether consolidation therapy by other means, such as high-dose chemotherapy followed by stem cell support, is the most promising regimen. Given these numerous uncertainties, neuro-oncologists should strive for a treatment of PCNSL patients within clinical trials to allow for the development of improved therapeutic regimens.


Consequences of occupational asthma on employment and financial status: a follow-up study.
The aim of this study was to describe changes in employment and income following a diagnosis of occupational asthma, and to determine what factors might affect these changes. Two hundred and nine patients with occupational asthma were reviewed on average 3.1 yrs after the diagnosis had been made. They were contacted by telephone or were sent a self-administered questionnaire by post. Multiple logistic regression models were constructed to determine which variables were associated with loss of employment after the diagnosis. At the time of review, 44% of patients had left their previous job and 25% were currently unemployed. Remarkably, 32% remained exposed to the offending agents in the same job. Forty six percent of the patients had suffered a reduction of income (84% of those who had left their employer versus 19% of those still employed in the same company (p < 0.001)). Claims for compensation, size of the company, level of education, and age at the time of diagnosis were significantly associated with a risk for becoming unemployed or having a new employer after the diagnosis of occupational asthma. Occupational asthma results in severe socioeconomic consequences. The French compensation system for occupational asthma should be revised, as the criteria currently used to determine compensation for this disease largely underestimate the social and occupational damages.


Cognitive mapping tobacco control advice for dentistry: a dental PBRN study.
To identify facilitative strategies that could be used in developing a tobacco cessation program for community dental practices. Nominal group technique (NGT) meetings and a card-sort task were used to obtain formative data. A cognitive mapping approach involving multidimensional scaling and hierarchical cluster analysis was used for data analysis. Three NGT meetings conducted with 23 dental professionals yielded 27 nonredundant facilitative strategies. A 2-dimensional 4-cluster cognitive map provided an organizational framework for understanding these strategies. Views of the target population solicited in a structured format provided clear direction for designing a tobacco cessation intervention.


Friedreich's ataxia-associated GAA repeats induce replication-fork reversal and unusual molecular junctions.
Expansion of GAA/TTC repeats is the causative event in Friedreich's ataxia. GAA repeats have been shown to hinder replication in model systems, but the mechanisms of replication interference and expansion in human cells remained elusive. To study in vivo replication structures at GAA repeats, we designed a new plasmid-based system that permits the analysis of human replication intermediates by two-dimensional gel electrophoresis and EM. We found that replication forks transiently pause and reverse at long GAA/TTC tracts in both orientations. Furthermore, we identified replication-associated intramolecular junctions, located between GAA/TTC repeats and other homopurine-homopyrimidine tracts, that were associated with breakage of the plasmid fork not traversing the repeats. Finally, we detected postreplicative, sister-chromatid hemicatenanes on control plasmids, which were replaced by persistent homology-driven junctions at GAA/TTC repeats. These data prove that GAA/TTC tracts interfere with replication in humans and implicate postreplicative mechanisms in trinucleotide repeat expansion.


A review of low and reduced carbohydrate diets and weight loss in type 2 diabetes.
Recent evidence from randomized controlled trials of hypocaloric low carbohydrate diets in people without diabetes has shown that they promote significant weight loss over the short term. There is very little evidence for any effects of reduced carbohydrate intakes on body weight, glycaemia and cardiovascular risk in people with type 2 diabetes. An electronic search was performed using MEDLINE (1966 to March 2007), EMBASE (1988 to March 2007) and Cochrane Central Register of Controlled Trials (1991 to March 2007) using the keywords low carbohydrate, type 2 diabetes and weight loss. Studies including subjects with type 2 diabetes who adopted a reduced carbohydrate weight loss diet were identified. Data were extracted on study design, weight loss, effects on glycaemia and cardiovascular risk and potential adverse effects. Six studies investigating the effects of hypocaloric reduced carbohydrate diets in people with type 2 diabetes were identified. The studies were heterogenous and most included small numbers, were short-term and provided varying amounts of carbohydrate. No studies were identified that were both low carbohydrate (<50 g day(-1)) and also designed as randomized controlled trials. All studies reported reductions in both body weight and glycated haemoglobin, with no deleterious effects on cardiovascular risk, renal function or nutritional intake. Conclusions are limited by study design and small numbers, but it appears that reduced carbohydrate diets are safe and effective over the short term for people with type 2 diabetes.


A Genetically Modified Adenoviral Vector with a Phage Display-Derived Peptide Incorporated into Fiber Fibritin Chimera Prolongs Survival in Experimental Glioma.
The dismal clinical context of advanced-grade glioma demands the development of novel therapeutic strategies with direct patient impact. Adenovirus-mediated virotherapy represents a potentially effective approach for glioma therapy. In this research, we generated a novel glioma-specific adenovirus by instituting more advanced genetic modifications that can maximize the efficiency and safety of therapeutic adenoviral vectors. In this regard, a glioma-specific targeted fiber was developed through the incorporation of previously published glioma-specific, phage-panned peptide (VWT peptide) on a fiber fibritin-based chimeric fiber, designated as "GliomaFF." We showed that the entry of this virus was highly restricted to glioma cells, supporting the specificity imparted by the phage-panned peptide. In addition, the stability of the targeting moiety presented by fiber fibritin structure permitted greatly enhanced infectivity. Furthermore, the replication of this virus was restricted in glioma cells by controlling expression of the E1 gene under the activity of the tumor-specific survivin promoter. Using this approach, we were able to explore the combinatorial efficacy of various adenoviral modifications that could amplify the specificity, infectivity, and exclusive replication of this therapeutic adenovirus in glioma. Finally, virotherapy with this modified virus resulted in up to 70% extended survival in an in vivo murine glioma model. These data demonstrate that this novel adenoviral vector is a safe and efficient treatment for this difficult malignancy.


Association of late-onset Alzheimer disease with a genotype of PLAU, the gene encoding urokinase-type plasminogen activator on chromosome 10q22.2.
Urokinase-type plasminogen activator (uPA) converts plasminogen to plasmin. Plasmin is involved in processing of amyloid precursor protein and degrades secreted and aggregated amyloid-beta, a hallmark of Alzheimer disease (AD). PLAU, the gene encoding uPA, maps to chromosome 10q22.2 between two regions showing linkage to late-onset AD (LOAD). We genotyped a frequent C/T single nucleotide polymorphism in codon 141 of PLAU (P141L) in 347 patients with LOAD and 291 control subjects. LOAD was associated with homozygous C/C PLAU genotype in the whole sample (chi2=15.7, P=0.00039, df 2), as well as in all sub-samples stratified by gender or APOE epsilon4 carrier status (chi2> or = 6.84, P< or =0.033, df 2). Odds ratio for LOAD due to homozygosity C/C was 1.89 (95% confidence interval 1.37-2.61). PLAU is a promising new candidate gene for LOAD, with allele C (P141) being a recessive risk allele or allele T (L141) conferring protection.


Interleukin-1 induction of collagenase 3 (matrix metalloproteinase 13) gene expression in chondrocytes requires p38, c-Jun N-terminal kinase, and nuclear factor kappaB: differential regulation of collagenase 1 and collagenase 3.
To examine the mechanism of interleukin-1 (IL-1)-induced collagenase 3 (matrix metalloproteinase 13 [MMP-13]) gene expression in cultured chondrocytes for the purpose of better understanding how the gene is induced in these cells, and how it contributes to cartilage degradation in osteoarthritis. The transcriptional and posttranscriptional responses of the MMP-13 gene to IL-1 were assessed first. Then, direct inhibitors of mitogen-activated protein kinase (MAPK) signaling pathways and a constitutive repressor of nuclear factor kappaB (NF-kappaB) were used to assess the role of each pathway in IL-1-mediated induction of MMP-13. We found that IL-1 induction of MMP-13 requires p38 activity, c-Jun N-terminal kinase (JNK) activity and NF-kappaB translocation. These results suggest that both NF-kappaB and activator protein 1 transcription factors are necessary for IL-1 induction of MMP-13. We also compared the signaling pathways necessary for IL-1 to stimulate collagenase 1 (MMP-1) in articular chondrocytes and chondrosarcoma cells and found that IL-1 induction of MMP-1 requires different pathways from those required by MMP-13. In chondrosarcoma cells, MMP-1 induction depends on p38 and MEK (an MAPK kinase of the extracellular signal-regulated kinase pathway) and does not require JNK or NF-kappaB. In articular chondrocytes, inhibition of MEK had no effect, while inhibition of p38 gave variable results. These studies demonstrate, for the first time, that p38, JNK, and NF-kappaB are required for IL-1 induction of MMP-13. The results also highlight the differential requirements for signaling pathways in the induction of MMP-1 and MMP-13. Additionally, they demonstrate that induction of MMP-1 by IL-1 in chondrocytic cells depends on unique combinations of signaling pathways that are cell type-specific.


Kinase activation of the non-receptor tyrosine kinase Etk/BMX alone is sufficient to transactivate STAT-mediated gene expression in salivary and lung epithelial cells.
Etk/BMX is a non-receptor protein tyrosine kinase that requires a functional phosphatidylinositol 3-kinase via the pleckstrin homology domain to be activated by cytokine. In the present study, a conditionally active form of Etk was constructed by fusing the hormone-binding domain of estrogen receptor (ER) to an amino terminus truncated form of Etk, PHDelta1-68Etk, to generate DeltaEtk:ER. In stably transfected Pa-4DeltaEtk:ER cells, the activity of DeltaEtk:ER was stimulated within minutes by the treatment of DeltaEtk:ER stimulant, estradiol, and sustained for greater than 24 h. A robust induction in the phosphorylation of signal transducers and activators of transcription (STAT) proteins, including STAT1, STAT3, and STAT5, was accompanied with DeltaEtk:ER activation. Moreover, the conditionally activated Etk stimulated STAT1- and STAT5-dependent reporter activities by approximately 160- and approximately 15-fold, respectively, however, elicited only a modest STAT3-mediated reporter activation. Qualitatively comparable results were obtained in lung A549 cells, indicating that DeltaEtk:ER inducible system could function in an analogous fashion in different epithelial cells. Furthermore, we demonstrated that Etk activation alone augmented cyclin D1 promoter/enhancer activity via its STAT5 response element in both Pa-4DeltaEtk:ER and A549 cells. Altogether, these findings support the notion that the activation of Etk kinase is sufficient to transactivate STAT-mediated gene expression. Hence, our inducible DeltaEtk:ER system represents a novel approach to investigate the biochemical events following Etk activation and to evaluate the contribution by kinase activation of Etk alone or in conjunction with other signaling pathway(s) to the ultimate biological responses.