The tyrosine kinase inhibitors imatinib and dasatinib reduce myeloid suppressor cells and release effector lymphocyte responses.
Immune escape mechanisms promote tumor progression and are hurdles of cancer immunotherapy. Removing immunosuppressive cells before treatment can enhance efficacy. Tyrosine kinase inhibitors (TKI) may be of interest to combine with immunotherapy, as it has been shown that the inhibitor sunitinib reduces myeloid suppressor cells in patients with renal cell carcinoma and dasatinib promotes expansion of natural killer-like lymphocytes in chronic myeloid leukemia (CML). In this study, the capacity of dasatinib and imatinib to reduce myeloid suppressor cells and to induce immunomodulation in vivo was investigated ex vivo. Samples from CML patients treated with imatinib (n = 18) or dasatinib (n = 14) within a Nordic clinical trial (clinicalTrials.gov identifier: NCT00852566) were investigated for the presence of CD11b(+)CD14(-)CD33(+) myeloid cells and inhibitory molecules (arginase I, myeloperoxidase, IL10) as well as the presence of natural killer cells, T cells (naïve/memory), and stimulatory cytokines (IL12, IFNγ, MIG, IP10). Both imatinib and dasatinib decreased the presence of CD11b(+)CD14(-)CD33(+) myeloid cells as well as the inhibitory molecules and the remaining myeloid suppressor cells had an increased CD40 expression. Monocytes also increased CD40 after therapy. Moreover, increased levels of CD40, IL12, natural killer cells, and experienced T cells were noted after TKI initiation. The presence of experienced T cells was correlated to a higher IFNγ and MIG plasma concentration. Taken together, the results demonstrate that both imatinib and dasatinib tilted the immunosuppressive CML tumor milieu towards promoting immune stimulation. Hence, imatinib and dasatinib may be of interest to combine with cancer immunotherapy.


In Arabidopsis thaliana codon volatility scores reflect GC3 composition rather than selective pressure.
Synonymous codon usage bias has typically been correlated with, and attributed to translational efficiency. However, there are other pressures on genomic sequence composition that can affect codon usage patterns such as mutational biases. This study provides an analysis of the codon usage patterns in Arabidopsis thaliana in relation to gene expression levels, codon volatility, mutational biases and selective pressures. We have performed synonymous codon usage and codon volatility analyses for all genes in the A. thaliana genome. In contrast to reports for species from other kingdoms, we find that neither codon usage nor volatility are correlated with selection pressure (as measured by dN/dS), nor with gene expression levels on a genome wide level. Our results show that codon volatility and usage are not synonymous, rather that they are correlated with the abundance of G and C at the third codon position (GC3). Our results indicate that while the A. thaliana genome shows evidence for synonymous codon usage bias, this is not related to the expression levels of its constituent genes. Neither codon volatility nor codon usage are correlated with expression levels or selective pressures but, because they are directly related to the composition of G and C at the third codon position, they are the result of mutational bias. Therefore, in A. thaliana codon volatility and usage do not result from selection for translation efficiency or protein functional shift as measured by positive selection.


Climate change alters leaf anatomy, but has no effects on volatile emissions from Arctic plants.
Biogenic volatile organic compound (BVOC) emissions are expected to change substantially because of the rapid advancement of climate change in the Arctic. BVOC emission changes can feed back both positively and negatively on climate warming. We investigated the effects of elevated temperature and shading on BVOC emissions from arctic plant species Empetrum hermaphroditum, Cassiope tetragona, Betula nana and Salix arctica. Measurements were performed in situ in long-term field experiments in subarctic and high Arctic using a dynamic enclosure system and collection of BVOCs into adsorbent cartridges analysed by gas chromatography-mass spectrometry. In order to assess whether the treatments had resulted in anatomical adaptations, we additionally examined leaf anatomy using light microscopy and scanning electron microscopy. Against expectations based on the known temperature and light-dependency of BVOC emissions, the emissions were barely affected by the treatments. In contrast, leaf anatomy of the studied plants was significantly altered in response to the treatments, and these responses appear to differ from species found at lower latitudes. We suggest that leaf anatomical acclimation may partially explain the lacking treatment effects on BVOC emissions at plant shoot-level. However, more studies are needed to unravel why BVOC emission responses in arctic plants differ from temperate species.


Addiction-like Behavior Associated with Mobile Phone Usage among Medical Students in Delhi.
Mobile phone addiction is a type of technological addiction or nonsubstance addiction. The present study was conducted with the objectives of developing and validating a mobile phone addiction scale in medical students and to assess the burden and factors associated with mobile phone addiction-like behavior. A cross-sectional study was conducted among undergraduate medical students aged ≥18 years studying in a medical college in New Delhi, India from December 2016 to May 2017. A pretested self-administered questionnaire was used for data collection. Mobile phone addiction was assessed using a self-designed 20-item Mobile Phone Addiction Scale (MPAS). Data were analyzed using IBM SPSS Version 17. The study comprised of 233 (60.1%) male and 155 (39.9%) female medical students with a mean age of 20.48 years. MPAS had a high level of internal consistency (Cronbach's alpha 0.90). Bartlett's test of sphericity was statistically significant (P < 0.0001), indicating that the MPAS data were likely factorizable. A principal component analysis found strong loadings on items relating to four components: harmful use, intense desire, impaired control, and tolerance. A subsequent two-stage cluster analysis of all the 20-items of the MPAS classified 155 (39.9%) students with mobile phone addiction-like behavior that was lower in adolescent compared to older students, but there was no significant difference across gender. Mobile phone use with increasing adoption of smartphones promotes an addiction-like behavior that is evolving as a public health problem in a large proportion of Indian youth.


Early, intensive statin treatment reduces 'hard' cardiovascular outcomes after acute coronary syndrome.
Background Early, intensive statin treatment is the standard of care after acute coronary syndrome (ACS). However, the benefit of this approach to prevent major adverse cardiovascular events has been demonstrated in only one randomised, placebo controlled trial. The Myocardial Ischemia Reduction with Aggressive Cholesterol Lowering (MIRACL) trial demonstrated that atorvastatin 80 mg daily, compared with placebo, reduced time to first occurrence of death, non-fatal myocardial infarction, resuscitated cardiac arrest, or hospitalisation for unstable angina (stroke not included) during the 16 week period following ACS. However, there were no significant effects on individual components of the composite endpoint except unstable angina. This led some to question whether early, intensive statin treatment reduces 'hard' events after ACS. Aim The burden of coronary heart disease after ACS, and therefore the efficacy of its treatment, depends not only on the occurrence of one ischaemic event, but rather on cumulative events experienced by patients. Accordingly, we conducted a post-hoc analysis of the MIRACL trial to examine the effect of atorvastatin on first as well as recurrent (i.e. total) hard cardiovascular events after ACS (death, myocardial infarction, stroke, and resuscitated cardiac arrest). Methods and Results In the 3086 patients who comprised the MIRACL trial, atorvastatin 80 mg did not reduce time to first hard event compared with placebo (hazard ratio 0.89, 95% confidence interval 0.72-1.10, P = 0.27). However, atorvastatin significantly reduced total hard events (hazard ratio 0.80, 95% confidence interval 0.66-0.97, P = 0.03). To prevent one hard event during the 16 weeks following ACS, only 11 patient-years of treatment with atorvastatin were required. Conclusion Early, intensive treatment with atorvastatin is an efficient intervention to reduce hard cardiovascular events after ACS.


Nox-4 deletion reduces oxidative stress and injury by PKC-α-associated mechanisms in diabetic nephropathy.
Current treatments for diabetic nephropathy (DN) only result in slowing its progression, thus highlighting a need to identify novel targets. Increased production of reactive oxygen species (ROS) is considered a key downstream pathway of end-organ injury with increasing data implicating both mitochondrial and cytosolic sources of ROS. The enzyme, NADPH oxidase, generates ROS in the kidney and has been implicated in the activation of protein kinase C (PKC), in the pathogenesis of DN, but the link between PKC and Nox-derived ROS has not been evaluated in detail in vivo. In this study, global deletion of a NADPH-oxidase isoform, Nox4, was examined in mice with streptozotocin-induced diabetes (C57Bl6/J) in order to evaluate the effects of Nox4 deletion, not only on renal structure and function but also on the PKC pathway and downstream events. Nox4 deletion attenuated diabetes-associated increases in albuminuria, glomerulosclerosis, and extracellular matrix accumulation. Lack of Nox4 resulted in a decrease in diabetes-induced renal cortical ROS derived from the mitochondria and the cytosol, urinary isoprostanes, and PKC activity. Immunostaining of renal cortex revealed that major isoforms of PKC, PKC-α and PKC-β1, were increased with diabetes and normalized by Nox4 deletion. Downregulation of the PKC pathway was observed in tandem with reduced expression of vascular endothelial growth factor (VEGF), transforming growth factor (TGF)-β1 and restoration of the podocyte slit pore protein nephrin. This study suggests that deletion of Nox4 may alleviate renal injury via PKC-dependent mechanisms, further strengthening the view that Nox4 is a suitable target for renoprotection in diabetes.


Follicle-Stimulating Hormone Glycobiology.
FSH glycosylation varies in two functionally important aspects: microheterogeneity, resulting from oligosaccharide structure variation, and macroheterogeneity, arising from partial FSHβ subunit glycosylation. Although advances in mass spectrometry permit extensive characterization of FSH glycan populations, microheterogeneity remains difficult to illustrate, and comparisons between different studies are challenging because no standard format exists for rendering oligosaccharide structures. FSH microheterogeneity is illustrated using a consistent glycan diagram format to illustrate the large array of structures associated with one hormone. This is extended to commercially available recombinant FSH preparations, which exhibit greatly reduced microheterogeneity at three of four glycosylation sites. Macroheterogeneity is demonstrated by electrophoretic mobility shifts due to the absence of FSHβ glycans that can be assessed by Western blotting of immunopurified FSH. Initially, macroheterogeneity was hoped to matter more than microheterogeneity. However, it now appears that both forms of carbohydrate heterogeneity have to be taken into consideration. FSH glycosylation can reduce its apparent affinity for its cognate receptor by delaying initial interaction with the receptor and limiting access to all of the available binding sites. This is followed by impaired cellular signaling responses that may be related to reduced receptor occupancy or biased signaling. To resolve these alternatives, well-characterized FSH glycoform preparations are necessary.


Rate-dependent propagation of cardiac action potentials in a one-dimensional fiber.
Action potential duration (APD) restitution, which relates APD to the preceding diastolic interval (DI), is a useful tool for predicting the onset of abnormal cardiac rhythms. However, it is known that different pacing protocols lead to different APD restitution curves (RCs). This phenomenon, known as APD rate dependence, is a consequence of memory in the tissue. In addition to APD restitution, conduction velocity restitution also plays an important role in the spatiotemporal dynamics of cardiac tissue. We present results concerning rate-dependent restitution in the velocity of propagating action potentials in a one-dimensional fiber. Our numerical simulations show that, independent of the amount of memory in the tissue, the wave-back velocity exhibits pronounced rate dependence and the wave-front velocity does not. Moreover, the discrepancy between wave-back velocity RCs is most significant for a small DI. We provide an analytical explanation of these results, using a system of coupled maps to relate the wave-front and wave-back velocities. Our calculations show that rate-dependent wave-back velocity can be present even if neither APD nor wave-front velocity exhibits rate dependence.


Measurement and significance of sperm morphology.
The measurement or evaluation and clinical significance of human sperm morphology has always been and still is a controversial aspect of the semen analysis for the determination of a male's fertility potential. In this review the background of the development of the evaluation criteria for sperm morphology will be discussed. Aspects of criticism on the strict criteria definition and use of the criteria for sperm morphology evaluation will be discussed as well as possible reasons for the decline in normal sperm morphology values and how we can compromise for this phenomenon resulting in the very low normal reference value as published in the 2010 WHO manual for the Examination and Processing of Human Semen. One of the possible solutions may be to give more attention to a limited number of abnormal sperm morphology categories and the inclusion of sperm morphology patterns. It is concluded in this review that if done correctly and with care and with strict application of existing guidelines as outlined in the 2010 WHO manual, sperm morphology measurement still has a very important role to play in the clinical evaluation of male fertility potential.


Neighborhood poverty and injection cessation in a sample of injection drug users.
Neighborhood socioeconomic environment may be a determinant of injection drug use cessation. The authors used data from a prospective cohort study of Baltimore City, Maryland, injection drug users assessed between 1990 and 2006. The study examined the relation between living in a poorer neighborhood and the probability of injection cessation among active injectors, independent of individual characteristics and while respecting the temporality of potential confounders, exposure, and outcome. Participants' residences were geocoded, and the crude, adjusted, and inverse probability of exposure weighted associations between neighborhood poverty and injection drug use cessation were estimated. Weighted models showed a strong association between neighborhood poverty and injection drug use cessation; living in a neighborhood with fewer than 10%, compared with more than 30%, of residents in poverty was associated with a 44% increased odds of not injecting in the prior 6 months (odds ratio = 1.44, 95% confidence interval: 1.14, 1.82). Results show that neighborhood environment may be an important determinant of drug injection behavior independent of individual-level characteristics.


Key challenges for the creation and maintenance of specialist protein resources.
As the volume of data relating to proteins increases, researchers rely more and more on the analysis of published data, thus increasing the importance of good access to these data that vary from the supplemental material of individual articles, all the way to major reference databases with professional staff and long-term funding. Specialist protein resources fill an important middle ground, providing interactive web interfaces to their databases for a focused topic or family of proteins, using specialized approaches that are not feasible in the major reference databases. Many are labors of love, run by a single lab with little or no dedicated funding and there are many challenges to building and maintaining them. This perspective arose from a meeting of several specialist protein resources and major reference databases held at the Wellcome Trust Genome Campus (Cambridge, UK) on August 11 and 12, 2014. During this meeting some common key challenges involved in creating and maintaining such resources were discussed, along with various approaches to address them. In laying out these challenges, we aim to inform users about how these issues impact our resources and illustrate ways in which our working together could enhance their accuracy, currency, and overall value.


Shared as well as distinct roles of EHD proteins revealed by biochemical and functional comparisons in mammalian cells and C. elegans.
The four highly homologous human EHD proteins (EHD1-4) form a distinct subfamily of the Eps15 homology domain-containing protein family and are thought to regulate endocytic recycling. Certain members of this family have been studied in different cellular contexts; however, a lack of concurrent analyses of all four proteins has impeded an appreciation of their redundant versus distinct functions. Here, we analyzed the four EHD proteins both in mammalian cells and in a cross-species complementation assay using a C. elegans mutant lacking the EHD ortholog RME-1. We show that all human EHD proteins rescue the vacuolated intestinal phenotype of C. elegans rme-1 mutant, are simultaneously expressed in a panel of mammalian cell lines and tissues tested, and variably homo- and hetero-oligomerize and colocalize with each other and Rab11, a recycling endosome marker. Small interfering RNA (siRNA) knock-down of EHD1, 2 and 4, and expression of dominant-negative EH domain deletion mutants showed that loss of EHD1 and 3 (and to a lesser extent EHD4) but not EHD2 function retarded transferrin exit from the endocytic recycling compartment. EH domain deletion mutants of EHD1 and 3 but not 2 or 4, induced a striking perinuclear clustering of co-transfected Rab11. Knock-down analyses indicated that EHD1 and 2 regulate the exit of cargo from the recycling endosome while EHD4, similar to that reported for EHD3 (Naslavsky et al. (2006) Mol. Biol. Cell 17, 163), regulates transport from the early endosome to the recycling endosome. Altogether, our studies suggest that concurrently expressed human EHD proteins perform shared as well as discrete functions in the endocytic recycling pathway and lay a foundation for future studies to identify and characterize the molecular pathways involved.


Relationship of heparan sulfate proteoglycans to the cytoskeleton and extracellular matrix of cultured fibroblasts.
The distribution of heparan sulfate proteoglycans (HSPG) on cultured fibroblasts was monitored using an antiserum raised against cell surface HSPG from rat liver. After seeding, HSPG was detected by immunofluorescence first on cell surfaces and later in fibrillar deposits of an extracellular matrix. Cell surface HSPG aligned with microfilament bundles of rat embryo fibroblasts seen by phase-contrast microscopy but was diffuse on transformed rat dermal fibroblasts (16C cells) which lack obvious stress fibers. Focal adhesions isolated from either cell type and monitored by interference reflection microscopy showed a concentration of HSPG labeling with respect to the rest of the membrane. Increased labeling in these areas was also seen for fibronectin (FN) by using an antiserum that detects both plasma and cell-derived FN. Double immunofluorescent staining of fully adherent rat embryo fibroblast cells showed some co-distribution of HSPG and FN, and this was confirmed by immunoelectron microscopy, which detected HSPG at localized areas of dorsal and ventral cell membranes, overlapping cell margins, and in the extracellular matrix. During cell shape changes on rounding and spreading, HSPG and FN may not co-distribute. Double labeling for actin and either HSPG or FN showed a closer correlation of actin with HSPG than with FN. The studies are consistent with HSPG being closely involved in a transmembrane cytoskeletal-matrix interaction; the possibility that HSPG coordinates the deposition of FN and other matrix components with cytoskeletal organization is discussed.


An international survey in postgraduate training in Oral Medicine.
The aim of this preliminary study was to investigate postgraduate Oral Medicine training worldwide and to begin to identify minimum requirements and/or core content for an International Oral Medicine curriculum. Countries where there was believed to be postgraduate training in Oral Medicine were identified by the working group. Standardized emails were sent inviting participants to complete an online survey regarding the scope of postgraduate training in Oral Medicine in their respective countries. We received 69 total responses from 37 countries. Of these, 22 countries self-identified as having postgraduate Oral Medicine as a distinct field of study, and they served as the study group. While there is currently considerable variation among Oral Medicine postgraduate training parameters, there is considerable congruency in clinical content of the Oral Medicine syllabi. For example, all of the training programs responded that they did evaluate competence in diagnosis and management of oral mucosal disease. This preliminary study provides the first evidence regarding international Oral Medicine postgraduate training, from which recommendations for an international core curriculum could be initiated. It is through such an initiative that a universal clinical core syllabus in postgraduate Oral Medicine training may be more feasible.


Melt-processed all-polymer distributed Bragg reflector laser.
We have assembled and studied melt-processed all-polymer lasers comprising distributed Bragg reflectors that were fabricated in large sheets using a co-extrusion process and define the cavities for dye-doped compression-molded polymer gain core sheets. Distributed Bragg reflector (DBR) resonators consisting of 128 alternating poly(styrene) (PS) and poly(methyl methacrylate) (PMMA) layers were produced by multilayer co-extrusion. Gain media were fabricated by compression-molding thermoplastic host poly notmers doped with organic laser dyes. Both processing methods can be used in high-throughput roll-to-roll manufacturing. Optically pumped DBR lasers assembled from these components display single and multimode lasing in the reflection band of the resonators, with a slope efficiency of nearly 19% and lasing thresholds as low as 90microJ/cm(2). The lasing wavelength can be controlled via the layer thickness of the DBR resonator films, and variation of the laser dye. Studies of threshold and efficiency are in agreement with models for end-pumped lasers.


Automatic bootstrapping and tracking of object contours.
A new fully automatic object tracking and segmentation framework is proposed. The framework consists of a motion-based bootstrapping algorithm concurrent to a shape-based active contour. The shape-based active contour uses finite shape memory that is automatically and continuously built from both the bootstrap process and the active-contour object tracker. A scheme is proposed to ensure that the finite shape memory is continuously updated but forgets unnecessary information. Two new ways of automatically extracting shape information from image data given a region of interest are also proposed. Results demonstrate that the bootstrapping stage provides important motion and shape information to the object tracker. This information is found to be essential for good (fully automatic) initialization of the active contour. Further results also demonstrate convergence properties of the content of the finite shape memory and similar object tracking performance in comparison with an object tracker with unlimited shape memory. Tests with an active contour using a fixed-shape prior also demonstrate superior performance for the proposed bootstrapped finite-shape-memory framework and similar performance when compared with a recently proposed active contour that uses an alternative online learning model.


Facial surface analysis by 3D laser scanning and geometric morphometrics in relation to sexual dimorphism in cerebral--craniofacial morphogenesis and cognitive function.
Over early fetal life the anterior brain, neuroepithelium, neural crest and facial ectoderm constitute a unitary, three-dimensional (3D) developmental process. This intimate embryological relationship between the face and brain means that facial dysmorphogenesis can serve as an accessible and informative index of brain dysmorphogenesis in neurological and psychiatric disorders of early developmental origin. There are three principal challenges in seeking to increase understanding of disorders of early brain dysmorphogenesis through craniofacial dysmorphogenesis: (i) the first, technical, challenge has been to digitize the facial surface in its inherent three-dimensionality; (ii) the second, analytical, challenge has been to develop methodologies for extracting biologically meaningful shape covariance from digitized samples, making statistical comparisons between groups and visualizing in 3D the resultant statistical models on a 'whole face' basis; (iii) the third, biological, challenge is to demonstrate a relationship between facial morphogenesis and brain morphogenesis not only in anatomical-embryological terms but also at the level of brain function. Here we consider each of these challenges in turn and then illustrate the issues by way of our own findings. These use human sexual dimorphism as an exemplar for 3D laser surface scanning of facial shape, analysis using geometric morphometrics and exploration of cognitive correlates of variation in shape of the 'whole face', in the context of studies relating to the early developmental origins of schizophrenia.


Childhood tuberculosis in the Kilimanjaro region: lessons from and for the TB programme.
To determine the magnitude of childhood TB and treatment outcome in Kilimanjaro region. Retrospective review of registration-based data on TB notifications in Kilimanjaro region for the period 2002-2006. Between 2002 and 2006, there were 1615 patients of childhood TB in Kilimanjaro region constituting 13% of total TB burden and the average case detection rate was 147/100 000 for urban and 41.8/100 000 for rural populations. Of them, 54.2% were men and 75.2% had pulmonary TB (PTB); 24.9% were tested for acid-fast bacilli (AFB) by Ziehl-Neelsen staining showing that 5.8% of all patients with TB were AFB smear positive. The remaining 94.2% were presumptively treated for TB. Treatment success rate was 79.9%, mortality 10.9% and default rate was 7%. Unfavourable outcome was more common among unconfirmed TB patients. HIV testing was very rare but increased after 2004 (<2% before 2005, 11-16% afterwards.) The rate of childhood TB in Kilimanjaro region is among the highest in the world. Microbiological diagnosis for TB and AFB smear positivity is very low. Treatment outcome in this region is poor. These findings argue for specific TB control strategies to be designed for children such as more AFB testing using new tools such as induced sputum and laryngeal swabs, active case finding, HIV testing of all suspected TB children, promoting and monitoring adherence. Regular epidemiological studies are also needed.


Acetylcholine negatively regulates development of the neuromuscular junction through distinct cellular mechanisms.
Emerging evidence suggests that the neurotransmitter acetylcholine (ACh) negatively regulates the development of the neuromuscular junction, but it is not clear if ACh exerts its effects exclusively through muscle ACh receptors (AChRs). Here, we used genetic methods to remove AChRs selectively from muscle. Similar to the effects of blocking ACh biosynthesis, eliminating postsynaptic AChRs increased motor axon branching and expanded innervation territory, suggesting that ACh negatively regulates synaptic growth through postsynaptic AChRs. However, in contrast to the effects of blocking ACh biosynthesis, eliminating postsynaptic AChRs in agrin-deficient mice failed to restore deficits in pre- and postsynaptic differentiation, suggesting that ACh negatively regulates synaptic differentiation through nonpostsynaptic receptors. Consistent with this idea, the ACh agonist carbachol inhibited presynaptic specialization of motorneurons in vitro. Together, these data suggest that ACh negatively regulates axon growth and presynaptic specialization at the neuromuscular junction through distinct cellular mechanisms.


Delivery of ligands from sorting endosomes to late endosomes occurs by maturation of sorting endosomes.
After endocytosis, lysosomally targeted ligands pass through a series of endosomal compartments. The endocytic apparatus that accomplishes this passage may be considered to take one of two forms: (a) a system in which lysosomally targeted ligands pass through preexisting, long-lived early sorting endosomes and are then selectively transported to long-lived late endosomes in carrier vesicles, or (b) a system in which lysosomally targeted ligands are delivered to early sorting endosomes which themselves mature into late endosomes. We have previously shown that sorting endosomes in CHO cells fuse with newly formed endocytic vesicles (Dunn, K. W., T. E. McGraw, and F. R. Maxfield. 1989. J. Cell Biol. 109:3303-3314) and that previously endocytosed ligands lose their accessibility to fusion with a half-time of approximately 8 min (Salzman, N. H., and F. R. Maxfield. 1989. J. Cell Biol. 109:2097-2104). Here we have studied the properties of individual endosomes by digital image analysis to distinguish between the two mechanisms for entry of ligands into late endosomes. We incubated TRVb-1 cells (derived from CHO cells) with diO-LDL followed, after a variable chase, by diI-LDL, and measured the diO content of diI-containing endosomes. As the chase period was lengthened, an increasing percentage of the endosomes containing diO-LDL from the initial incubation had no detectable diI-LDL from the second incubation, but those endosomes that contained both probes showed no decrease in the amount of diO-LDL per endosomes. These results indicate that (a) a pulse of fluorescent LDL is retained by individual sorting endosomes, and (b) with time sorting endosomes lose the ability to fuse with primary endocytic vesicles. These data are inconsistent with a preexisting compartment model which predicts that the concentration of ligand in sorting endosomes will decline during a chase interval, but that the ability of the stable sorting endosome to receive newly endocytosed ligands will remain high. These data are consistent with a maturation mechanism in which the sorting endosome retains and accumulates lysosomally directed ligands until it loses its ability to fuse with newly formed endocytic vesicles and matures into a late endosome. We also find that, as expected according to the maturation model, new sorting endosomes are increasingly labeled during the chase period indicating that new sorting endosomes are continuously formed to replace those that have matured into late endosomes.(ABSTRACT TRUNCATED AT 400 WORDS)


A New Method to Analyze the Relative Corneal Refractive Power and Its Association to Myopic Progression Control With Orthokeratology.
We present a new method for analyzing relative corneal refractive power (RCRP) in children undergoing orthokeratology and explore its potential association to effective myopic control. A total of 55 children aged 8 to 12 years participated in the study. Axial growth was calculated as the difference in axial length before and 1 year after orthokeratology. Growth <0.30 mm was considered as effective control. Corneal topography was obtained before and 4 months after lens dispatch. The topography was divided into 36 10° slices and the maximal RCRP (mRCRP) in each was calculated and fitted into a model that integrated the effects of mean refractive power (M), corneal asymmetry (f1), and astigmatism (f2). The relationship between the probability of achieving effective control and the modulation of mRCRP was analyzed with logistic regression. A total of 45 subjects achieved effective control, but for 10 the treatment was ineffective. The M-values were not different between the groups. Modulations of mRCRP were significantly larger in the effective than the ineffective group (1.17 vs. 0.64 diopters [D] for f1, P = 0.02; 0.85 vs. 0.35 D for f2, P = 0.03). The probability to achieve effective control increased with modulation of mRCRP (P = 0.02). With a peak mRCRP > 4.5 D, a subject had an above 80% chance to achieve effective control. The new method reveals that how the combination of spherical equivalent (SE), corneal asymmetry, and astigmatism determines modulation of the mRCRP and a large amplitude of modulation is associated with a higher probability of effective myopic control. Our finding enables clinicians to estimate the outcome early and provides new insights to lens design.


Evaluation of TCR repertoire diversity in patients after hematopoietic stem cell transplantation.
T-cell receptor (TCR) repertoire analyses have been widely used to identify T cell populations of interest in cancer and autoimmunity and for characterizing immune repertoire reconstitution after hematopoietic stem cell transplantation (HSCT). Several decades of development and progress have led to the use of techniques for evaluating TCR repertoires in a more comprehensive, unbiased and fast manner, and the mechanisms of T cell immune reconstitution after HSCT and the new approaches used for recovering T cell repertoire diversity post HSCT have been more exhaustively documented to some degree. To better understand and characterize this progress, here we review recent studies on TCR repertoire diversity recovery in patients with leukemia and autoimmune disease who have received HSCT, impact factors and improvements in approaches for TCR repertoire recovery after HSCT.


Effects of external qigong therapy on osteoarthritis of the knee. A randomized controlled trial.
The objective of our study was to assess the efficacy of external qigong therapy (EQT), a traditional Chinese medicine practice, in reducing pain and improving functionality of patients with knee osteoarthritis (OA). One hundred twelve adults with knee OA were randomized to EQT or sham treatment (control); 106 completed treatment and were analyzed. Two therapists performed EQT individually, five to six sessions in 3 weeks. The sham healer mimicked EQT for the same number of sessions and duration. Patients and examining physician were blinded. Primary outcomes were Western Ontario MacMaster (WOMAC) pain and function; other outcomes included McGill Pain Questionnaire, time to walk 15 m, and range of motion squatting. Results of patients treated by the two healers were analyzed separately. Both treatment groups reported significant reduction in WOMAC scores after intervention. Patients treated by healer 2 reported greater reduction in pain (mean improvement -25.7 +/- 6.6 vs. -13.1 +/- 3.0; p < 0.01) and more improvement in functionality (-28.1 +/- 9.7 vs. -13.2 +/- 3.4; p < 0.01) than those in sham control and reduction in negative mood but not in anxiety or depression. Patients treated by healer 1 experienced improvement similar to control. The results of therapy persisted at 3 months follow-up for all groups. Mixed-effect models confirmed these findings with controlling for possible confounders. EQT might have a role in the treatment of OA, but our data indicate that all EQT healers are not equivalent. The apparent efficacy of EQT appears to be dependent on some quality of the healer. Further study on a larger scale with multiple EQT healers is necessary to determine the role (if any) of EQT in the treatment of OA and to identify differences in EQT techniques.


3D modeling of dual-gate FinFET.
The tendency to have better control of the flow of electrons in a channel of field-effect transistors (FETs) did lead to the design of two gates in junction field-effect transistors, field plates in a variety of metal semiconductor field-effect transistors and high electron mobility transistors, and finally a gate wrapping around three sides of a narrow fin-shaped channel in a FinFET. With the enhanced control, performance trends of all FETs are still challenged by carrier mobility dependence on the strengths of the electrical field along the channel. However, in cases when the ratio of FinFET volume to its surface dramatically decreases, one should carefully consider the surface boundary conditions of the device. Moreover, the inherent non-planar nature of a FinFET demands 3D modeling for accurate analysis of the device performance. Using the Silvaco modeling tool with quantization effects, we modeled a physical FinFET described in the work of Hisamoto et al. (IEEE Tran. Elec. Devices 47:12, 2000) in 3D. We compared it with a 2D model of the same device. We demonstrated that 3D modeling produces more accurate results. As 3D modeling results came close to experimental measurements, we made the next step of the study by designing a dual-gate FinFET biased at Vg1 >Vg2. It is shown that the dual-gate FinFET carries higher transconductance than the single-gate device.


Lmo2 and Scl/Tal1 convert non-axial mesoderm into haemangioblasts which differentiate into endothelial cells in the absence of Gata1.
The LIM domain protein Lmo2 and the basic helix-loop-helix transcription factor Scl/Tal1 are expressed in early haematopoietic and endothelial progenitors and interact with each other in haematopoietic cells. While loss-of-function studies have shown that Lmo2 and Scl/Tal1 are essential for haematopoiesis and angiogenic remodelling of the vasculature, gain-of-function studies have suggested an earlier role for Scl/Tal1 in the specification of haemangioblasts, putative bipotential precursors of blood and endothelium. In zebrafish embryos, Scl/Tal1 can induce these progenitors from early mesoderm mainly at the expense of the somitic paraxial mesoderm. We show that this restriction to the somitic paraxial mesoderm correlates well with the ability of Scl/Tal1 to induce ectopic expression of its interaction partner Lmo2. Co-injection of lmo2 mRNA with scl/tal1 dramatically extends its effect to head, heart, pronephros and pronephric duct mesoderm inducing early blood and endothelial genes all along the anteroposterior axis. Erythroid development, however, is expanded only into pronephric mesoderm, remaining excluded from head, heart and somitic paraxial mesoderm territories. This restriction correlates well with activation of gata1 transcription and co-injection of gata1 mRNA along with scl/tal1 and lmo2 induces erythropoiesis more broadly without ventralising or posteriorising the embryo. While no ectopic myeloid development from the Scl/Tal1-Lmo2-induced haemangioblasts was observed, a dramatic increase in the number of endothelial cells was found. These results suggest that, in the absence of inducers of erythroid or myeloid haematopoiesis, Scl/Tal1-Lmo2-induced haemangioblasts differentiate into endothelial cells.


The left atrium, atrial fibrillation, and the risk of stroke in hypertensive patients with left ventricular hypertrophy.
The Losartan Intervention For Endpoint reduction in hypertension (LIFE) study provided extensive data on predisposing factors, consequences, and prevention of atrial fibrillation (AF) in patients with hypertension and left ventricular (LV) hypertrophy. Randomized losartan-based treatment was superior to atenolol-based treatment for reducing new-onset AF and complications, especially stroke, associated with new-onset or pre-existing AF. Potential mechanisms of AF prevention by angiotensin receptor blockade supported by LIFE results include greater reduction in left atrial size and LV hypertrophy. Differential effects of antihypertensive treatment on the left atrium and left ventricle may help prevent AF and reduce risk of stroke associated with hypertensive heart disease.


Communication in a Human biomonitoring study: Focus group work, public engagement and lessons learnt in 17 European countries.
A communication strategy was developed by The Consortium to Perform Human Biomonitoring on a European Scale (COPHES), as part of its objectives to develop a framework and protocols to enable the collection of comparable human biomonitoring data throughout Europe. The framework and protocols were tested in the pilot study DEMOCOPHES (Demonstration of a study to Coordinate and Perform Human biomonitoring on a European Scale). The aims of the communication strategy were to raise awareness of human biomonitoring, encourage participation in the study and to communicate the study results and their public health significance. It identified the audiences and key messages, documented the procedure for dissemination of results and was updated as the project progressed. A communication plan listed the tools and materials such as press releases, flyers, recruitment letters and information leaflets required for each audience with a time frame for releasing them. Public insight research was used to evaluate the recruitment material, and the feedback was used to improve the documents. Dissemination of results was coordinated in a step by step approach by the participating countries within DEMOCOPHES, taking into account specific national messages according to the needs of each country. Participants received individual results, unless they refused to be informed, along with guidance on what the results meant. The aggregate results and policy recommendations were then communicated to the general public and stakeholders, followed by dissemination at European level. Several lessons were learnt that may assist other future human biomonitoring studies. Recruitment took longer than anticipated and so social scientists, to help with community engagement, should be part of the research team from the start. As a European study, involving multiple countries, additional considerations were needed for the numerous organisations, different languages, cultures, policies and priorities. Therefore, communication documents should be seen as templates with essential information clearly indicated and the option for each country to tailor the material to reflect these differences. Future studies should consider setting up multidisciplinary networks of medical professionals and communication experts, and holding training workshops to discuss the interpretation of results and risk communication. Publicity and wide dissemination of the results helped to raise awareness of human biomonitoring to the general public, policy makers and other key stakeholders. Effective and timely communication, at all stages of a study, is essential if the potential of human biomonitoring research to improve public health is to be realised.


Diffusion tensor imaging of peripheral nerves in non-fixed post-mortem subjects.
While standard magnetic resonance imaging (MRI) sequences are increasingly employed in post-mortem (PM) examinations, more advanced techniques such as diffusion tensor imaging (DTI) remain unexplored in forensic sciences. Therefore, we studied the temporal stability and reproducibility of DTI and fiber tractography (FT) in non-fixed PM subjects. In addition, we investigated the lumbosacral nerves with PMDTI and compared their tissue characteristics to in vivo findings. MRI data were acquired on a 1.5T MRI scanner in seven PM subjects, consisting of six non-trauma deaths and one chronic trauma death, and in six living subjects. Inter-scan (within one session) and inter-session (between days) reproducibility of diffusion parameters, fractional anisotropy (FA), and mean diffusivity (MD), were evaluated for the lumbosacral nerves using Bland-Altman and Jones plots. Diffusion parameters in nerves L3-S2 were compared to living subjects using the non-parametric Mann-Whitney U test. Reproducibility of diffusion values of inter-scan 95% limits of agreement ranged from -0.058 to 0.062 for FA, and (-0.037 to 0.052)×10(-3)mm(2)/s for MD. For the inter-session this was -0.0423 to 0.0423, and (-0.0442 to 0.0442)×10(-3)mm(2)/s for FA, and MD, respectively. Although PM subjects showed approximately four-fold lower diffusivity values compared to living subjects, FT results were comparable. The chronic trauma case showed disorganization and asymmetry of the nerves. We demonstrated that DTI was reproducible in characterizing nervous tissue properties and FT in reconstructing the architecture of lumbosacral nerves in PM subjects. We showed differences in diffusion values between PM and in vivo and showed the ability of PMDTI and FT to reconstruct nerve lesions in a chronic trauma case. We expect that PMDTI and FT may become valuable in identification and documentation of PM nerve trauma or pathologies in forensic sciences.


Chimeric origins of ochrophytes and haptophytes revealed through an ancient plastid proteome.
Plastids are supported by a wide range of proteins encoded within the nucleus and imported from the cytoplasm. These plastid-targeted proteins may originate from the endosymbiont, the host, or other sources entirely. Here, we identify and characterise 770 plastid-targeted proteins that are conserved across the ochrophytes, a major group of algae including diatoms, pelagophytes and kelps, that possess plastids derived from red algae. We show that the ancestral ochrophyte plastid proteome was an evolutionary chimera, with 25% of its phylogenetically tractable nucleus-encoded proteins deriving from green algae. We additionally show that functional mixing of host and plastid proteomes, such as through dual-targeting, is an ancestral feature of plastid evolution. Finally, we detect a clear phylogenetic signal from one ochrophyte subgroup, the lineage containing pelagophytes and dictyochophytes, in plastid-targeted proteins from another major algal lineage, the haptophytes. This may represent a possible serial endosymbiosis event deep in eukaryotic evolutionary history.


Characteristics and outcomes of patients with multiple myeloma : Data from a developing country.
Background: Multiple myeloma (MM) is a plasma cell disorder characterized by presence of monoclonal protein in serum or urine or both, increased bone marrow plasma cells, osteolytic lesion, hypercalcemia, and anemia. Several combination regimens are commonly recommended for treatment of multiple myeloma. The present study aimed at determining the characteristics and outcomes of patients with multiple myeloma treated at our centre. Methods: During July 2012 and December 2015, all patients with proven diagnosis of MM were included in this study. Data were collected from hospital information system. The characteristics and outcomes of all patients were analyzed. Progression- free survival and overall survival of patients were also estimated. Kaplan-Meier curves and Log-rank test were applied and SPSS Version19 was used for data analysis. Results: A total of 82 patients, with the median age of 51 years (Range: 23-64 yrs.) were available for final analysis. The number of patients with IgG and IgA type was 48 (58.5%) and 15(18.3%), respectively. There were 7 (8.5%) patients with non-secretory type. Most of the patients (n= 59; 71.9%) were treated with CTD regimen and 13 (15.8%) received bortezomib-based treatment. The median progression-free survival time was 30 months, and overall survival time was 48 months. The cumulative probability of survival at 36 months was 85%. Conclusion: Based on our results, the onset of multiple myeloma occurs in relatively younger age groups. A small number of patients received bortezomib due to cost issues. PFS and OS in our study were comparable with published literature.


Improved Fourier-transform profilometry.
An improved optical geometry of the projected-fringe profilometry technique, in which the exit pupil of the projecting lens and the entrance pupil of the imaging lens are neither at the same height above the reference plane nor coplanar, is discussed and used in Fourier-transform profilometry. Furthermore, an improved fringe-pattern description and phase-height mapping formula based on the improved geometrical generalization is deduced. Employing the new optical geometry, it is easier for us to obtain the full-field fringe by moving either the projector or the imaging device. Therefore the new method offers a flexible way to obtain reliable height distribution of a measured object.


SU-E-I-09: Patient Organ Doses from KV-CBCT Acquisitions as Function of Patient Size and Scan Protocols.
To investigate the organ dose dependence on patient size and position resulting from an image guidance procedures. Depending upon the degree of variability between patients, the feasibility tabulating the organ dose based upon patient size and imaging acquisition procedure. Such tables could be useful in estimating and accounting for organ dose resulting from imaging procedures. Monte-Carlo methods were used to calculate radiation dose to the patient resulting from a kV CBCT imaging scan based upon CT image data. A Python program was created to generate DVHs using the calculated 3D dose matrix and the DICOM-RT Structure Set for the patient. A number of head and pelvic CT scans were used in the study, and the resulting DVHs were analyzed to determine the variation of organ dose as a function of patient size. For the head scans in supine position, the average of the mean doses for some organs, along with the standard deviation in those mean doses, are as follows:Brainstem: 0.25cGy +/- 0.02cGyBrain: 0.26cGy +/- 0.01cGyOptic Chiasm: 0.22 +/- 0.03cGyBone: 0.55 +/- 0.04cGyAnterior Skin: 0.13 +/- 0.05cGyPosterior Skin: 0.23 +/- 0.06cGyEye: 0.11cGy +/- 0.05cGyFor Pelvic scans, dose to organs such as rectum, have likewise been shown to have some variation between patients of different sizes. The results show that it is possible to express organ dose as a function of imaging procedure, imaging device, scan location, body size, and orientation. For example, using a standard head scan on an OBI 1.4 system, the dose to the eye ranged from 0.05-0.20cGy, depending upon the patient size. Considering the amount of imaging dose compared to the therapeutic dose, this study indicates that organ dose could be estimated by using tabulated values based upon the scan procedure and patient size.


Comprehensive screen of genetic variation in DNA repair pathway genes and postmenopausal breast cancer risk.
Mistakes in DNA repair can result in sustained damage and genetic instability. We comprehensively evaluated common variants in DNA repair pathway genes for their association with postmenopausal breast cancer risk with and without respect to estrogen receptor (ER) and progesterone receptor (PR) subtypes. In this study of 1,145 prospectively ascertained breast cancer cases and 1,142 matched controls from the Nurses' Health Study Cancer Genetic Markers of Susceptibility project, we evaluated 1,314 common genetic variants in 68 candidate genes. These variants were chosen to represent five DNA repair pathways including base excision repair, nucleotide excision repair, double-strand break repair (homologous recombination and non-homologous end-joining), direct reversal repair, and mismatch repair, along with candidate DNA polymerases, Fanconi Anemia complementation groups, and other genes relevant to DNA damage recognition and response. Main effects, pathway effects, and pair-wise interactions were evaluated using Logistic Regression, and the Admixture Maximum Likelihood (AML) and Kernel Machine tests. Eight linked loci within XRCC4 were associated with susceptibility to PR- breast cancer (main effect p-values corrected for multiple testing at the within-gene level < 0.04). These loci drove the association between the non-homologous end-joining pathway, and PR- breast cancer (AML p-value for the full pathway = 0.002; p-value when the eight loci were removed = 0.86). A Kernel machine test of no linear or quadratic effects, or pairwise interaction, yielded a p-value of 0.85. Common variation alone in DNA repair genes plays at most a small role in determining postmenopausal breast cancer risk among women of European ancestry.


Structural basis for native agonist and synthetic inhibitor recognition by the Pseudomonas aeruginosa quorum sensing regulator PqsR (MvfR).
Bacterial populations co-ordinate gene expression collectively through quorum sensing (QS), a cell-to-cell communication mechanism employing diffusible signal molecules. The LysR-type transcriptional regulator (LTTR) protein PqsR (MvfR) is a key component of alkyl-quinolone (AQ)-dependent QS in Pseudomonas aeruginosa. PqsR is activated by 2-alkyl-4-quinolones including the Pseudomonas quinolone signal (PQS; 2-heptyl-3-hydroxy-4(1H)-quinolone), its precursor 2-heptyl-4-hydroxyquinoline (HHQ) and their C9 congeners, 2-nonyl-3-hydroxy-4(1H)-quinolone (C9-PQS) and 2-nonyl-4-hydroxyquinoline (NHQ). These drive the autoinduction of AQ biosynthesis and the up-regulation of key virulence determinants as a function of bacterial population density. Consequently, PqsR constitutes a potential target for novel antibacterial agents which attenuate infection through the blockade of virulence. Here we present the crystal structures of the PqsR co-inducer binding domain (CBD) and a complex with the native agonist NHQ. We show that the structure of the PqsR CBD has an unusually large ligand-binding pocket in which a native AQ agonist is stabilized entirely by hydrophobic interactions. Through a ligand-based design strategy we synthesized and evaluated a series of 50 AQ and novel quinazolinone (QZN) analogues and measured the impact on AQ biosynthesis, virulence gene expression and biofilm development. The simple exchange of two isosteres (OH for NH₂) switches a QZN agonist to an antagonist with a concomitant impact on the induction of bacterial virulence factor production. We also determined the complex crystal structure of a QZN antagonist bound to PqsR revealing a similar orientation in the ligand binding pocket to the native agonist NHQ. This structure represents the first description of an LTTR-antagonist complex. Overall these studies present novel insights into LTTR ligand binding and ligand-based drug design and provide a chemical scaffold for further anti-P. aeruginosa virulence drug development by targeting the AQ receptor PqsR.


The gap junction protein connexin32 interacts with the Src homology 3/hook domain of discs large homolog 1.
Scaffolding of membrane proteins is a common strategy for forming complexes of proteins, including some connexins, within membrane microdomains. Here we describe studies indicating that Cx32 interacts with a PDZ-containing scaffolding protein, Dlgh1 (Discs Large homolog 1). Initial screens of liver lysates using antibody arrays indicated an interaction between Cx32 and Dlgh1 that was confirmed using coimmunoprecipitation studies. Yeast two-hybrid complementation determined that the Cx32 bound via interaction with the SH3/Hook domain of Dlgh1. Confocal microscopy of liver sections revealed that Cx32 and Dlgh1 could colocalize in hepatocyte membranes in wild type mice. Examination of levels and localization of Dlgh1 in livers from Cx32 null mice indicate that, in the absence of Cx32, Dlgh1 was decreased, and the remainder was translocated from the hepatocyte membrane to the nucleus with some remaining in cytoplasmic compartments. This translocation was confirmed by Western blots comparing Dlgh1 levels in nuclear extracts from wild type and Cx32 null murine livers. Using SKHep cells stably transfected with Cx32 under the control of a tet-off promoter, we found that acute removal of Cx32 led to a decrease of membrane-localized Dlgh1 and an increase in the nuclear localization of this tumor suppressor protein. Together, these results suggest that loss of Cx32 alters the levels, localization, and interactions of the tumor suppressor protein Dlgh1, events known in other systems to alter cell cycle and increase tumorigenicity.


Up-regulation of macrophage migration inhibitory factor induced by endotoxin in experimental otitis media with effusion in mice.
Injection of endotoxin into the middle ear causes production of macrophage migration inhibitory factor (MIF) in an experimental mouse model of otitis media with effusion (OME). Down-regulation of MIF may become a new approach for the management of OME. To determine the role of MIF in OME. Mice were divided into two groups and their middle ears were injected with either endotoxin or phosphate-buffered saline (PBS). Mice were sacrificed at 6 h, 12 h, or 1, 3, 7, or 14 days after injection and concentrations of MIF, interleukin-1 beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in middle ear effusions were measured by enzyme-linked immunosorbent assay. Concentrations of MIF in the endotoxin group at 1 day and 3 days were significantly higher than in the PBS control group. Concentrations of IL-1beta in the endotoxin group at 6 h, 12 h, 1 day, and 3 days were significantly higher than in controls. Concentrations of TNF-alpha in the endotoxin group at 1 day and 3 days were significantly higher than in controls. Concentration of MIF in the endotoxin group was positively correlated with that of IL-1beta and TNF-alpha.


Mainstreamed genetic testing in ovarian cancer: patient experience of the testing process.
Pathogenic BRCA variants account for 5.8-24.8% of ovarian cancers. The identification of such a variant can have a significant impact on the affected individual and their relatives, determining eligibility for targeted therapies, predicting treatment response, and granting access to disease prevention strategies. Cancer services are responding to the increased demand for genetic testing with the introduction of mainstreamed genetic testing via oncology clinics. This study aimed to evaluate patient experience of the mainstreamed genetic testing pathway at a tertiary referral center in London, UK. Study participants were patients diagnosed with high-grade non-mucinous ovarian cancer, tested via a mainstreamed genetic testing pathway at the tertiary referral center between February 2015 and June 2017. Eligible participants were invited to complete the retrospective study questionnaire. Five quantitative measures with additional free-text items were used to evaluate the patient experience of mainstreamed genetic testing. The tertiary referral center tested 170 ovarian cancer patients. Twenty-three pathogenic BRCA mutations were identified (23/170, 13.5%). One-hundred and six patients (106/170, 62.4%) met the study inclusion criteria. Twenty-nine of those invited to participate (29/106, 27.4%) returned the retrospective study questionnaire. Pathogenic BRCA1/2 variants were identified within four respondents (4/29, 13.8%). Motivations for genetic testing related to improved medical management, and the ability to provide relatives with genetic information. Participants did not appear to be adversely affected by result disclosure post-mainstreamed genetic testing. Two individuals with a pathogenic variant reported that the support provided by the tertiary referral center post-result disclosure could have been improved. Results of the current study support further psychosocial research into the expansion of the mainstreamed genetic testing pathway. The results, although promising, have also highlighted the importance of genetic awareness within the multi-disciplinary team and the provision of timely psychological support from genetic specialists.


BRAF mutant non-small cell lung cancer and treatment with BRAF inhibitors.
Inhibitors targeting active protein kinases, such as EGFR or ALK, have demonstrated significant efficacy in the treatment of lung cancer. Activating mutations in the MAPK pathway, which includes the enzymes RAS, RAF, MEK, and ERK, result in constitutive signalling, leading to oncogenic cell proliferation and escape from apoptosis; therefore this pathway is a focus of crucial interest for the development of cancer drugs. In melanoma, the most commonly mutated gene is BRAF, with mutations usually occurring in about 50% of all tumours. The BRAF Val600Glu (V600E) mutation constitutes more than 90% of mutations in melanoma. V600E BRAF mutation shows a great dependency on MEK activity, and offers a rational therapeutic strategy for this genetically defined tumour subtype. The use of vemurafenib and dabrafenib, agents that block MAPK signaling in patients with melanoma and the BRAF V600E mutation, has been associated with prolonged survival and progression-free survival. The frequency of V600E BRAF mutation in lung adenocarcinoma is 1.5% to 2.8%. Treatment of V600E BRAF-mutant lung adenocarcinomas with dabrafenib is under evaluation in a phase 2 trial, and could represent another milestone in individualized therapy for lung cancer patients. The next step will be a combination therapy of BRAF inhibitor dabrafenib and MEK inhibitor trametinib.


SMiLE-seq identifies binding motifs of single and dimeric transcription factors.
Resolving the DNA-binding specificities of transcription factors (TFs) is of critical value for understanding gene regulation. Here, we present a novel, semiautomated protein-DNA interaction characterization technology, selective microfluidics-based ligand enrichment followed by sequencing (SMiLE-seq). SMiLE-seq is neither limited by DNA bait length nor biased toward strong affinity binders; it probes the DNA-binding properties of TFs over a wide affinity range in a fast and cost-effective fashion. We validated SMiLE-seq by analyzing 58 full-length human, mouse, and Drosophila TFs from distinct structural classes. All tested TFs yielded DNA-binding models with predictive power comparable to or greater than that of other in vitro assays. De novo motif discovery on all JUN-FOS heterodimers and several nuclear receptor-TF complexes provided novel insights into partner-specific heterodimer DNA-binding preferences. We also successfully analyzed the DNA-binding properties of uncharacterized human C2H2 zinc-finger proteins and validated several using ChIP-exo.


Polarization interferometer for the determination of spectral modulation transfer functions of monochromators.
A polarization interferometer has been constructed to study the spectral modulation transfer function of monochromators in the visible region. The instrument consists of a Soleil compensator and interchangeable fixed compensators mounted between polarizers. As the retardation of the compensator is varied the light reaching the analyzer ranges from plane polarized to circularly polarized. In this way a fringe system is generated, permitting the interferogram of a light distribution to be recorded, just as with a conventional two-beam interferometer. The transfer function of a prism monochromator has been studied under varied conditions of slit width and focusing.


The impact of adjuvant brachytherapy with sublobar resection on pulmonary function and dyspnea in high-risk patients with operable disease: preliminary results from the American College of Surgeons Oncology Group Z4032 trial.
Z4032 was a randomized study conducted by the American College of Surgeons Oncology Group comparing sublobar resection alone versus sublobar resection with brachytherapy for high-risk operable patients with non-small cell lung cancer (NSCLC). This evaluates early impact of adjuvant brachytherapy on pulmonary function tests, dyspnea, and perioperative (30-day) respiratory complications in this impaired patient population. Eligible patients with stage I NSCLC tumors 3 cm or smaller were randomly allocated to undergo sublobar resection with (SRB group) or without (SR group) brachytherapy. Outcomes measured included the percentage predicted forced expiratory volume in 1 second (FEV1%), percentage predicted carbon monoxide diffusion capacity (DLCO%), and dyspnea score per the University of California San Diego Shortness of Breath Questionnaire. Pulmonary morbidity was assessed per the Common Terminology Criteria for Adverse Events version 3.0. Outcomes were measured at baseline and 3 months. A 10% change in pulmonary function test or 10-point change in dyspnea score was deemed clinically meaningful. Z4032 permanently closed to patient accrual in January 2010 at 224 patients. At 3-month follow-up, pulmonary function data are currently available for 148 (74 SR and 74 SRB) patients described in this report. There were no differences in baseline characteristics between arms. In the SR arm, 9 patients (12%) reported grade 3 respiratory adverse events, compared with 12 (16%) in the SRB arm (P = .49). There was no significant change in percentage change in DLCO% or dyspnea score from baseline to 3 months within either arm. In the case of FEV1%, percentage change from baseline to 3 months was significant within the SR arm (P = .03), with patients reporting improvement in FEV1% at month 3. Multivariable regression analysis (adjusted for baseline values) showed no significant impact of treatment arm, tumor location (upper vs other lobe), or surgical approach (video-assisted thoracoscopic surgery vs thoracotomy) on 3-month FEV1%, DLCO%, and dyspnea score. There was no significant difference in incidence of clinically meaningful (10% pulmonary function or 10-point dyspnea score change) change between arms. Twenty-two percent of patients with lower-lobe tumors and 9% with upper-lobe tumors demonstrated 10% decline in FEV1% (odds ratio, 2.79; 95 confidence interval, 1.07-7.25; P = .04). Adjuvant intraoperative brachytherapy in conjunction with sublobar resection did not significantly worsen pulmonary function or dyspnea at 3 months in a high-risk population with NSCLC, nor was it associated with increased perioperative pulmonary adverse events. Lower-lobe resection was the only factor significantly associated with clinically meaningful decline in FEV1%.


Occult serious bacterial infection in infants younger than 60 to 90 days with bronchiolitis: a systematic review.
To summarize the risk of occult serious bacterial infection in the youngest febrile infants presenting with either clinical bronchiolitis or respiratory syncytial virus infection. We performed a systematic search of the Medline database for studies reporting rates of serious bacterial infection in infants younger than 90 days with clinical bronchiolitis and/or respiratory syncytial virus infection. Studies reporting on cultures performed at the time of presentation to care and providing a denominator, ie, total number of each type of culture obtained, were analyzed. Admission for bronchiolitis. Age-specific rates of urinary tract infection, bacteremia, and meningitis were extracted. The weighted rate of urinary tract infections in the youngest infants in the 11 studies analyzed was 3.3% (95% confidence interval, 1.9%-5.7%). No case of bacteremia was reported in 8 of 11 studies. No case of meningitis was reported in any of the studies. Summary statistics for meningitis and bacteremia are not provided because of an excess of zero events in these samples. A screening approach to culturing for serious bacterial infections in febrile infants presenting with bronchiolitis or respiratory syncytial virus infection is very low yield. The rate of urine cultures positive for bacteria remains significant, though asymptomatic bacteriuria may confound these results.


Human non-CpG methylation patterns display both tissue-specific and inter-individual differences suggestive of underlying function.
DNA methylation (DNAm) in mammals is mostly examined within the context of CpG dinucleotides. Non-CpG DNAm is also widespread across the human genome, but the functional relevance, tissue-specific disposition, and inter-individual variability has not been widely studied. Our aim was to examine non-CpG DNAm in the wider methylome across multiple tissues from the same individuals to better understand non-CpG DNAm distribution within different tissues and individuals and in relation to known genomic regulatory features.DNA methylation in umbilical cord and cord blood at birth, and peripheral venous blood at age 12-13 y from 20 individuals from the Southampton Women's Survey cohort was assessed by Agilent SureSelect methyl-seq. Hierarchical cluster analysis (HCA) was performed on CpG and non-CpG sites and stratified by specific cytosine environment. Analysis of tissue and inter-individual variation was then conducted in a second dataset of 12 samples: eight muscle tissues, and four aliquots of cord blood pooled from two individuals.HCA using methylated non-CpG sites showed different clustering patterns specific to the three base-pair triplicate (CNN) sequence. Analysis of CAC sites with non-zero methylation showed that samples clustered first by tissue type, then by individual (as observed for CpG methylation), while analysis using non-zero methylation at CAT sites showed samples grouped predominantly by individual. These clustering patterns were validated in an independent dataset using cord blood and muscle tissue.This research suggests that CAC methylation can have tissue-specific patterns, and that individual effects, either genetic or unmeasured environmental factors, can influence CAT methylation.


Association between methylphenidate treatment and risk of seizure: a population-based, self-controlled case-series study.
Individuals with attention-deficit hyperactivity disorder (ADHD) are at increased risk of seizures. Stimulant medications such as methylphenidate are the most commonly prescribed treatment for ADHD, but the association between their therapeutic use and the risk of seizures is unclear. We aimed to investigate the association between methylphenidate treatment and the risk of seizure. For this population-based observational study, we used the electronic medical record database of the Hong Kong Clinical Data Analysis And Reporting System to identify individuals aged 6-25 years who received at least one methylphenidate prescription during the study period. Individuals with records of seizure or epilepsy before the study period were excluded. Individuals treated with methylphenidate who had seizures during the study period were included in the subsequent analyses, and a self-controlled case-series design was used to control for time-invariant individual characteristics. We did additional analyses using skin infection as a negative control outcome. We compared relative incidence of seizure during periods when individuals were exposed to methylphenidate with that during non-exposed periods. Of 29 604 individuals prescribed methylphenidate between Jan 1, 2001, and Dec 31, 2017, 269 (199 males and 70 females) had incident seizures. The mean age at baseline was 6·66 years (SD 2·01) and the median age at the incident seizure was 9·69 years (IQR 7·62-12·99). The overall incidence of seizure during methylphenidate treatment was 4·4 per 10 000 patient-years. We detected an increased risk of seizure during the first 30 days of methylphenidate treatment compared with that during non-exposed periods, with an incidence rate ratio of 4·01 (95% CI 2·09-7·68). No increase in risk was identified during the following 31-180 days of treatment (1·13, 0·56-2·25) or during subsequent treatment (1·38, 0·92-2·07). We did not identify an increased risk in any risk window for the negative control outcome analysis. No individuals died because of a seizure during the study period. The incidence of seizures was higher in the period immediately after the start of methylphenidate treatment than in the non-exposed period. No increased risk was observed during continuation of methylphenidate treatment. The association between methylphenidate treatment and seizures immediately after initiation of medication can be seen as a potential safety signal. Monitoring of neurological outcomes in individuals with ADHD is recommended when they first start methylphenidate treatment. Hong Kong Research Grants Council.


Molecular identification of Phialophora oxyspora as the cause of mycetoma in a horse.
An 18-year-old mare was evaluated for an oral mass that developed after extraction of a broken incisor. An ulcerated, firm, darkly pigmented, approximately 5-cm-diameter spherical mass involved the gingiva lateral and dorsal to the right first to third maxillary incisors. Osteolysis of the roots of the first and second right maxillary incisors and periosteal proliferation of the adjacent premaxilla margins were apparent on radiographs. Histologic examination of the mass revealed multiple coalescing and ramifying foci of abscess formation, each containing a well-defined, discrete, black mass (2 to 7 mm in diameter). Myriad fungal hyphae enmeshed in a black, granular, cementlike material were within each of the black structures. Mycetoma was the histologic diagnosis. The causative agent could not be identified via culture because of lack of distinguishing characteristics. Fungal DNA was isolated from frozen fungal cultures and paraffin sections. The D1/D2 domains of the large subunit P gene rDNA were amplified and sequenced. The sequences of the D1/D2 domains of both isolates were 96% homologous with those of Phialophora oxyspora. The mass was surgically excised, the local area curetted, and the wound allowed to heal by second intention. Postoperative treatment consisted of administration of phenylbutazone and IV administration of sodium iodide followed by oral administration of potassium iodide. There was no evidence of recurrence 1 year later. Mycetomata should be a differential diagnosis for equine gingival masses. Identification of the fungal agent can be critical for selection of optimal treatments. Molecular methods may permit definitive identification when standard phenotypic-based identification criteria are inconclusive.


Inorganic polymerization: an attractive route to biocompatible hybrid hydrogels.
As an intermediate state between liquid and solid materials, hydrogels display unique properties, opening a wide scope of applications, especially in the biomedical field. Organic hydrogels are composed of an organic network cross-linked via chemical or physical reticulation nodes. In contrast, hybrid hydrogels are defined by the coexistence of organic and inorganic moieties in water. Inorganic polymerization, i.e. the sol-gel process, is one of the main techniques leading to hybrid hydrogels. The chemoselectivity of this method proceeds through hydrolysis and condensation reactions of metal oxide moieties. In addition, the mild reaction conditions make this process very promising for the preparation of water-containing materials and their bio-applications.


Genome evolutionary dynamics followed by diversifying selection explains the complexity of the Sesamum indicum genome.
Whole genome duplication (WGD) and tandem duplication (TD) provide two critical sources of raw genetic material for genome complexity and evolutionary novelty. Little is known about the complexity of the Sesamum indicum genome after it diverged from a common ancestor with the paleodiploid Vitis vinifera and further experienced WGD and TD events. Here, we analyzed the functional divergence of different classes of inter- and intra-genome gene pairs from ancestral events to uncover multiple-layers of evolutionary dynamics acting during the process of forming the modern S. indicum genome. Comprehensive inter-genome analyses revealed that 60% and 70% of syntenic orthologous gene pairs were retained among the two subgenomes in S. indicum compared to V. vinifera, although there was no evidence of significant differences under selection pressure. For the intra-genomic analyses, 5,932 duplicated gene pairs experienced fractionation, with the remaining 1,236 duplicated gene pairs having undergone functional divergence under diversifying selection. Analysis of the TD events indicated that 2,945 paralogous gene pairs, from 1,089 tandem arrays of 2-16 genes, experienced functional divergence under diversifying selection. Sequence diversification of different classes of gene pairs revealed that most of TD events occurred after the WGD event, with others following the ancestral gene order indicating ancient TD events at some time prior to the WGD event. Our comparison-of-function analyses for different classes of gene pairs indicated that the WGD and TD evolutionary events were both responsible for introducing genes that enabled exploration of novel and complementary functionalities, whilst maintaining individual plant ruggedness. In this study, we first investigated functional divergence of different classes of gene pairs to characterize the dynamic processes associated with each evolutionary event in S. indicum. The data demonstrated massive and distinct functional divergence among different classes of gene pairs, and provided a genome-scale view of gene function diversification explaining the complexity of the S. indicum genome. We hope this provides a biological model to study the mechanism of plant species formation, particularly in the context of the evolutionary history of flowering plants, and offers novel insights for the study of angiosperm genomes.


Estimating the Absorbed Dose of Organs in Pediatric Imaging of 99mTc-DTPATc-DTPA Radiopharmaceutical using MIRDOSE Software.
In this study, organ radiation doses were calculated for the renal agent 99mTc-DTPATc-DTPA in children. Bio-kinetic energy of 99mTc-DTPATc-DTPA was evaluated by scintigraphy and estimates for absorbed radiation dose were provided using standard medical internal radiation dosimetry (MIRD) techniques. In this applied research, fourteen children patients (6 males and 8 females) were imaged using a series of planar and SPECT images after injecting with technetium-99mTc-DTPA diethylenetriaminepentaacetic acid (99mTc-DTPATc-DTPA). A hybrid planar/SPECT method was used to plot time-activity curves to obtain the residence time of the source organs and also MIRDOSE software was used to calculate the absorbed dose of every organ. P-values were calculated using t-tests in order to make a comparison between the adsorbed doses of male and female groups. Mean absorbed doses (µGy/MBq) for urinary bladder wall, kidneys, gonads, liver and adrenals were 213.5±47.8, 12.97±6.23, 12.0±2.5, 4.29±1.47, and 3.31±0.96, respectively. Furthermore, the mean effective dose was 17.5±3.7 µSv/MBq. There was not any significant difference in the mean absorbed dose of the two groups. Bladder cumulated activity was the most contribution in the effective dose of patients scanned with 99mTc-DTPATc-DTPA. Using a hybrid planar/SPECT method can cause an increase in accumulated activity accuracy for the region of interest. Moreover, patient-specified internal dosimetry is recommended.


Fahr's disease: a differential diagnosis of frontal lobe syndrome.
Fahr's disease refers to a rare syndrome characterised by symmetrical and bilateral intracranial calcification. The basal ganglia are the most common site of involvement and most cases present with extra-pyramidal symptoms. We describe two men with Fahr's diseases who presented with prominent frontal lobe symptoms. The first man presented with frequent uncontrollable bursts of laughter and crying spells. He later developed mild dysarthric speech and choreoathetoid movement. The second man presented with progressive changes in personality and behaviour. In both cases, there were no parkinsonian features. Computed tomographic scans of both patients demonstrated extensive symmetrical calcification over the basal ganglia and dentate nuclei. A repeated imaging scan in the second patient revealed progressive cerebral atrophy but reduction in the calcification. No underlying cause for the bilateral calcification was found. As frontal lobe symptoms are usually inconspicuous in the early stage, the presence of these symptoms might be overlooked in clinical practice when compared with those suffering from prominent movement disorders.


Pharmacological Management of Anxiety on Pain Occurrence During Root Canal Treatment: A Systematic Review.
To answer the question: 'Does the pharmacological management of dental anxiety influence pain occurrence during root canal treatment?' Searches on MEDLINE/PubMed, Cochrane Library, Web of Science, Scopus, EMBASE and Open Grey were conducted until September 02, 2022. Only randomised clinical trials were included. The Cochrane risk of bias tool for randomized trials (RoB 2) was used. The overall quality of evidence was assessed through the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) tool. Initial screening resulted in 811 studies. Three hundred seventy-three were excluded for being duplicates. Of 438 eligible papers, ten studies met the inclusion criteria and were selected for full-text reading. Four studies were included in the final analysis. Three studies had a low risk of bias, and one was a high risk. GRADE demonstrated a low quality of evidence. There is insufficient evidence to determine whether the pharmacological control of anxiety can influence intraoperative pain occurrence. (EEJ-2022-08-096).


Atmospheric mercury deposition to forests in the eastern USA.
Atmospheric mercury (Hg) deposition to forests is important because half of the land cover in the eastern USA is forest. Mercury was measured in autumn litterfall and weekly precipitation samples at a total of 27 National Atmospheric Deposition Program (NADP) monitoring sites in deciduous and mixed deciduous-coniferous forests in 16 states in the eastern USA during 2007-2014. These simultaneous, uniform, repeated, annual measurements of forest Hg include the broadest area and longest time frame to date. The autumn litterfall-Hg concentrations and litterfall mass at the study sites each year were combined with annual precipitation-Hg data. Rates of litterfall-Hg deposition were higher than or equal to precipitation-Hg deposition rates in 70% of the annual data, which indicates a substantial contribution from litterfall to total atmospheric-Hg deposition. Annual litterfall-Hg deposition in this study had a median of 11.7 μg per square meter per year (μg/m2/yr) and ranged from 2.2 to 23.4 μg/m2/yr. It closely matched modeled dry-Hg deposition, based on land cover at selected NADP Hg-monitoring sites. Mean annual atmospheric-Hg deposition at forest study sites exhibited a spatial pattern partly explained by statistical differences among five forest-cover types and related to the mapped density of Hg emissions. Forest canopies apparently recorded changes in atmospheric-Hg concentrations over time because litterfall-Hg concentrations decreased year to year and litterfall-Hg concentrations were significantly higher in 2007-2009 than in 2012-2014. These findings reinforce reported decreases in Hg emissions and atmospheric elemental-Hg concentrations during this same time period. Methylmercury (MeHg) was detected in all litterfall samples at all sites, compared with MeHg detections in less than half the precipitation samples at selected sites during the study. These results indicate MeHg in litterfall is a pathway into the terrestrial food web where it can accumulate in the prey of songbirds, bats, and raptors.


Corticosteroids and pentoxifylline for the treatment of alcoholic hepatitis: Current status.
The treatment of choice for patients with severe alcoholic hepatitis (AH) is use of corticosteroids. Many randomized well designed studies have been reported from all over the world on the use of corticosteroids in the treatment of AH. However, the data on the efficacy of corticosteroids in these patients have been conflicting. Initial meta-analyses also failed to show beneficial effects of corticosteroids. Based on individual data meta-analysis showing clear benefit of corticosteroids amongst patients with severe AH (modified discriminant function of 32 or more), led American College of Gastroenterology to recommend use of corticosteroids as the first line treatment option amongst patients with severe AH. However, corticosteroids are relatively contraindicated amongst patients with severe AH and coexistent sepsis, gastrointestinal bleeding, and acute pancreatitis. These patients may be candidates for second line treatment with pentoxifylline. Further, specific treatment of AH with corticosteroids far from satisfactory with as many as 40%-50% of patients failing to respond to steroids, thus classified as non-responsive to steroids. The management of these patients is a continuing challenge for physicians. Better treatment modalities need to be developed for this group of patients in order to improve the outcome of patients with severe AH. This article describes at length the available trials on use of corticosteroids and pentoxifylline with their current status. Route of administration, dosage, adverse effects, and mechanisms of action of these two drugs are also discussed. Finally, an algorithm with clinical approach to management of patients who present with clinical syndrome of AH is described.


Reduced oligodendrocyte exosome secretion in multiple system atrophy involves SNARE dysfunction.
Transportation of key proteins via extracellular vesicles has been recently implicated in various neurodegenerative disorders, including Parkinson's disease, as a new mechanism of disease spreading and a new source of biomarkers. Extracellular vesicles likely to be derived from the brain can be isolated from peripheral blood and have been reported to contain higher levels of α-synuclein (α-syn) in Parkinson's disease patients. However, very little is known about extracellular vesicles in multiple system atrophy, a disease that, like Parkinson's disease, involves pathological α-syn aggregation, though the process is centred around oligodendrocytes in multiple system atrophy. In this study, a novel immunocapture technology was developed to isolate blood CNPase-positive, oligodendrocyte-derived enriched microvesicles (OEMVs), followed by fluorescent nanoparticle tracking analysis and assessment of α-syn levels contained within the OEMVs. The results demonstrated that the concentrations of OEMVs were significantly lower in multiple system atrophy patients, compared to Parkinson's disease patients and healthy control subjects. It is also noted that the population of OEMVs involved was mainly in the size range closer to that of exosomes, and that the average α-syn concentrations (per vesicle) contained in these OEMVs were not significantly different among the three groups. The phenomenon of reduced OEMVs was again observed in a transgenic mouse model of multiple system atrophy and in primary oligodendrocyte cultures, and the mechanism involved was likely related, at least in part, to an α-syn-mediated interference in the interaction between syntaxin 4 and VAMP2, leading to the dysfunction of the SNARE complex. These results suggest that reduced OEMVs could be an important mechanism related to pathological α-syn aggregation in oligodendrocytes, and the OEMVs found in peripheral blood could be further explored for their potential as multiple system atrophy biomarkers.


Farm-based day care services - a prospective study protocol on health benefits for people with dementia and next of kin.
The increasing number of people with dementia will place a considerable challenge on the health care system and will necessitate innovation and new solutions. Day care services aim to provide meaningful activities and coping experiences for the target group. The aim of the present study is to explore the experience and potential health benefits for people with dementia attending farm-based day care services in Norway and their next of kin. The present study is a prospective study organized into several qualitative and quantitative sub-studies. Study period will last from 2016-2020. We use a multi-method approach and both an empirical and a constructivist view on knowledge to explore this multi-facetted phenomenon. Several challenges are expected, for instance, the ability to include participants with dementia and keep them within the study for the whole study period. The functioning level of participants regarding language and understanding could challenge their ability to answer. Despite having several limitations, we believe that this study with its different approaches and methods will add important knowledge to the field. Knowledge from our study could be essential for creating day care services of good quality for people with dementia and such services could reduce the burden of care for their next of kin.


Antiviral Activities of Honey, Royal Jelly, and Acyclovir Against HSV-1.
Herpes simplex virus type 1 (HSV-1) belongs to the Herpesviridae family and genus simplex virus. This virus is usually acquired during childhood and is transmitted through direct mucocutaneous contact or droplet infection from infected secretions. The aim of the present study was to compare antiviral effects of honey, royal jelly, and acyclovir on herpes simplex virus-1 in an extra-somatic environment. Vero cells were cultured in the Dulbecco's Modified Eagle's Medium (DMEM) along with 10% fetal bovine serum (FBS) in 12-welled microplates. Various dilutions of honey, royal jelly, and acyclovir (5, 10, 50, 100, 2500, 500, and 800 μg/mL) were added to the Vero cells along with a 100-virus concentration of TCID50. The plaque assay technique was used to evaluate the antiviral activities. The results showed that honey, royal jelly, and acyclovir have the highest inhibitory effects on HSV-1 at concentrations of 500, 250, and 100 μg/mL, respectively. In addition, honey, royal jelly, and acyclovir decreased the viral load from 70 795 to 43.3, 30, and 0 PFU/mL at a concentration of 100 μg/mL, respectively. The results of the present study showed that honey and royal jelly, which are natural products with no reports about their deleterious effect at least in laboratory conditions, can be considered alternatives to acyclovir in the treatment of herpetic lesions. However, it should be pointed out that further studies are necessary to substantiate their efficacy because hard evidence on their effectiveness is not available at present.


Adriamycin-induced DNA damage mediated by mammalian DNA topoisomerase II.
Adriamycin (doxorubicin), a potent antitumor drug in clinical use, interacts with nucleic acids and cell membranes, but the molecular basis for its antitumor activity is unknown. Similar to a number of intercalative antitumor drugs and nonintercalative epipodophyllotoxins (VP-16 and VM-26), adriamycin has been shown to induce single- and double-strand breaks in DNA. These strand breaks are unusual because a covalently bound protein appears to be associated with each broken phosphodiester bond. In studies in vitro, mammalian DNA topoisomerase II mediates DNA damage by adriamycin and other related antitumor drugs.


High SVR12 with 8-week and 12-week glecaprevir/pibrentasvir therapy: An integrated analysis of HCV genotype 1-6 patients without cirrhosis.
Glecaprevir plus pibrentasvir (G/P) is a pangenotypic, once-daily, ribavirin-free direct-acting antiviral (DAA) treatment for hepatitis C virus (HCV) infection. In nine phase II or III clinical trials, G/P therapy achieved rates of sustained virologic response 12 weeks after treatment (SVR12) of 93-100% across all six major HCV genotypes (GTs). An integrated efficacy analysis of 8- and 12-week G/P therapy in patients without cirrhosis with HCV GT 1-6 infection was performed. Data were pooled from nine phase II and III trials including patients with chronic HCV GT 1-6 infection without cirrhosis who received G/P (300 mg/120 mg) for either 8 or 12 weeks. Patients were treatment naïve or treatment experienced with peginterferon, ribavirin, and/or sofosbuvir; all patients infected with HCV GT 3 were treatment naïve. Efficacy was evaluated as the SVR12 rate. The analysis included 2,041 patients without cirrhosis. In the intent-to-treat population, 943/965 patients (98%) achieved SVR12 when treated for eight weeks, and 1,060/1,076 patients (99%) achieved SVR12 when treated for 12 weeks; the difference in rates was not significant (p = 0.2). A subgroup analysis demonstrated SVR12 rates > 95% across baseline factors traditionally associated with lower efficacy. G/P was well tolerated, with one DAA-related serious adverse event (<0.1%); grade 3 laboratory abnormalities were rare. G/P therapy for eight weeks in patients with chronic HCV GT 1-6 infection without cirrhosis achieved an overall SVR12 rate of 98% irrespective of baseline patient or viral characteristics; four additional weeks of treatment did not significantly increase the SVR12 rate, demonstrating that the optimal treatment duration in this population is eight weeks. In this integrated analysis of nine clinical trials, patients with chronic HCV genotype 1-6 infection without cirrhosis were treated for either 8 or 12 weeks with the direct-acting antiviral regimen glecaprevir/pibrentasvir (G/P). The cure rate was 98% and 99% following 8 and 12 weeks of treatment, respectively; the difference in rates was not significant (p = 0.2), nor was there a significant difference in the cure rates across the two treatment durations on the basis of baseline patient or viral characteristics. These results, along with a favourable safety profile, indicate that G/P is a highly efficacious and well-tolerated pangenotypic eight-week therapy for most patients with chronic HCV infection.


Serial measurements of higher-order aberrations after blinking in normal subjects.
To investigate sequential changes in the optical quality of normal eyes associated with blinking. Ocular higher-order aberrations (HOAs) were measured sequentially by using a wavefront sensor for 30 seconds in 20 eyes of 20 normal subjects. During the measurement, subjects were forced to blink every 10 seconds. The obtained aberration data were analyzed in the central 4-mm diameter for coma-like, spherical-like, and total HOAs up to the sixth-order Zernike polynomials. The serial changes in the HOAs with blinking were classified into four groups by pattern: stable (25%), small-fluctuation (45%), sawtooth (20%), and others (10%). In the subjects with the sawtooth pattern, the total HOAs increased significantly (P < 0.001, one-way repeated-measures ANOVA) with time between blinks. Increased total HOAs and coma-like aberrations in the subjects with the sawtooth pattern suggested that the inferosuperior asymmetric change in tear film thickness is responsible. Dynamic changes in HOAs after blinking showed variations even in clinically normal subjects. Serial measurements of HOAs may be useful in evaluating the dynamic changes in tear film and the effects on the quality of vision after blinking.


The antioxidative activity of the mucoregulatory agents: ambroxol, bromhexine and N-acetyl-L-cysteine. A pulse radiolysis study.
Ambroxol and bromhexine are shown to be scavengers of both superoxide and hydroxyl radicals as determined by pulse radiolysis experiments. The dismutation of superoxide was accelerated 3-fold by bromhexine and 2.5-fold by ambroxol over the rate of spontaneous dismutation. The reaction constants of hydroxyl radicals with bromhexine and ambroxol were determined by competition kinetics to be 1.58 +/- 0.15 x 10(10) M-1S-1 and 1.04 +/- 0.1 x 10(10) M-1S-1, respectively. N-acetyl-L-cysteine also reacted with hydroxyl radicals (1.28 +/- 0.14 x 10(10) M-1S-1) but not with superoxide radical. These effects may be clinically relevant in the treatment of oxidant-associated lung damage induced by inflammatory agents and/or environmental pollutants.


Clinical assessment of ankle joint dorsiflexion: a review of measurement techniques.
Ankle dorsiflexion measurement is important for clinical and research use. With so much evidence on the unreliability of goniometric measurements, a systematic review was performed to investigate various alternative techniques for measuring ankle dorsiflexion in the nonneurologic patient. All of the major databases were queried electronically to identify studies that used any method of ankle dorsiflexion measurement in the nonneurologic subject. Keywords included ankle dorsiflexion NOT cerebral palsy NOT stroke, the latter to exclude neurologic conditions. In 755 studies that used some form of ankle joint dorsiflexion measurement, ten different techniques were identified that included various apparatuses designed specifically for this purpose. Reliability testing of these techniques involved test-retest trials with small student populations as subjects, which returned high intraclass correlation coefficient scores. However, their methodological quality would have benefitted from the use of an actual patient population and comparison with a reference standard. When validating ankle dorsiflexion measurement techniques, actual patient populations should be used, otherwise papers would score poorly on methodological quality assessment. Standardizing patient position, foot posture, amount of moment applied, and reference landmarks will ensure that various trial results can be compared directly.


Wildlife and wildlife management in Tanzania.
Tanzania, arguably mainland Africa's most important nation for conservation, is losing habitat and natural resources rapidly. Moving away from a charcoal energy base and developing sustainable finance mechanisms for natural forests are critical to slowing persistent deforestation. Addressing governance and capacity deficits, including law enforcement, technical skills, and funding, across parts of the wildlife sector are key to effective wildlife protection. These changes could occur in tandem with bringing new models of natural resource management into play that include capacity building, corporate payment for ecosystem services, empowering nongovernmental organizations in law enforcement, greater private-sector involvement, and novel community conservation strategies. The future of Tanzania's wildlife looks uncertain-as epitomized by the current elephant crisis-unless the country confronts issues of governance, embraces innovation, and fosters greater collaboration with the international community.