Clinicopathological features, treatment approaches, and outcomes in Rosai-Dorfman disease.
Rosai-Dorfman disease is a rare subtype of non-Langerhans cell histiocytosis. With the last major report published in 1990, there is a paucity of contemporary data on this disease. Our objective was to report the clinicopathological features, treatments and outcomes of patients seen at a tertiary referral center. Sixty-four patients with histopathological diagnosis of Rosai-Dorfman disease were identified from 1994 to 2017 (median age 50 years; range, 2-79). The median duration from symptom onset to diagnosis was seven months (range, 0-128), which was also reflected in the number of biopsies required to establish the diagnosis (median 2; range, 1-6). The most common presentation was subcutaneous masses (40%). Of the 64 patients, 8% had classical (nodal only) and 92% had extra-nodal disease (67% extra-nodal only). The most common organs involved were skin and subcutaneous tissue (52%), followed by lymph nodes (33%). Three patients had an overlap with Erdheim-Chester disease, which had not been described before. Two of these were found to have MAP2K1 mutations. Commonly utilized first line treatments were surgical excision (38%) and systemic corticosteroids (27%). Corticosteroids led to a response in 56% of the cases. Of those treated initially, 15 (30%) patients developed recurrent disease. The most commonly used systemic agent was cladribine (n=6), with 67% overall response rate. Our study demonstrates that Rosai-Dorfman disease has diverse clinical manifestations and outcomes. While this disease has been historically considered a benign entity, a subset of patients endures an aggressive course necessitating the use of systemic therapies.


Exploring Selective Exposure and Confirmation Bias as Processes Underlying Employee Work Happiness: An Intervention Study.
Employee psychological capital (PsyCap), perceptions of organizational virtue (OV), and work happiness have been shown to be associated within and over time. This study examines selective exposure and confirmation bias as potential processes underlying PsyCap, OV, and work happiness associations. As part of a quasi-experimental study design, school staff (N = 69) completed surveys at three time points. After the first assessment, some staff (n = 51) completed a positive psychology training intervention. Results of descriptive statistics, correlation, and regression analyses on the intervention group provide some support for selective exposure and confirmation bias as explanatory mechanisms. In focusing on the processes through which employee attitudes may influence work happiness this study advances theoretical understanding, specifically of selective exposure and confirmation bias in a field study context.


PMS2 monoallelic mutation carriers: the known unknown.
Germ-line mutations in MLH1, MSH2, MSH6, and PMS2 have been shown to cause Lynch syndrome. The penetrance of the cancer and tumor spectrum has been repeatedly studied, and multiple professional societies have proposed clinical management guidelines for affected individuals. Several studies have demonstrated a reduced penetrance for monoallelic carriers of PMS2 mutations compared with the other mismatch repair (MMR) genes, but clinical management guidelines have largely proposed the same screening recommendations for all MMR gene carriers. The authors considered whether enough evidence existed to propose new screening guidelines specific to PMS2 mutation carriers with regard to age at onset and frequency of colonic screening. Published reports of PMS2 germ-line mutations were combined with unpublished cases from the authors' research registries and clinical practices, and a discussion of potential modification of cancer screening guidelines was pursued. A total of 234 monoallelic PMS2 mutation carriers from 170 families were included. Approximately 8% of those with colorectal cancer (CRC) were diagnosed before age 30, and each of these tumors presented on the left side of the colon. As it is currently unknown what causes the early onset of CRC in some families with monoallelic PMS2 germline mutations, the authors recommend against reducing cancer surveillance guidelines in families found having monoallelic PMS2 mutations in spite of the reduced penetrance.Genet Med 18 1, 13-19.


Associative memory design for 256 gray-level images using a multilayer neural network.
A design procedure is presented for neural associative memories storing gray-scale images. It is an evolution of a previous work based on the decomposition of the image with 2L gray levels into L binary patterns, stored in L uncoupled neural networks. In this letter, an L-layer neural network is proposed with both intralayer and interlayer connections. The connections between different layers introduce interactions among all the neurons, increasing the recall performance with respect to the uncoupled case. In particular, the proposed network can store images with the commonly used number of 256 gray levels instead of 16, as in the previous approach.


The effects of maternal presence during anesthesia induction on the mother's anxiety and changes in children's behavior.
This study aimed to evaluate whether maternal presence during induction has additional beneficial effects on a mother's anxiety or changes in the child's behavior when an information booklet was given to all mothers and premedication was given to all patients. One hundred children, aged 2-10 years, scheduled for ambulatory surgery were randomly assigned to a mother-present (Group M) or mother-absent group (Group C) after premedication with intranasal midazolam. All mothers were informed about general anesthesia with a detailed information booklet. Preoperatively (pre) and one week after the operation (post), maternal anxiety was assessed using State-Trait Anxiety Inventory (STAI), and Posthospitalization Behavior Questionnaire (PHBQ) was used to measure changes in children's behavior. Anesthesia was induced using sevoflurane-oxygen-nitrous oxide inhalation. The anesthesiologist graded the level of the children's stress at anesthesia induction with a four-point scale. There were no differences between the two groups regarding demographics, anxiety levels of the mothers and postoperative behavioral changes and stress scores of the children (p>0.05 between the groups *p<0.005 within groups). In summary, maternal presence during induction in addition to premedication for children and information booklets for mothers had no additive effects in terms of reducing the mother's or the child's anxiety or postoperative behavioral changes.


Early antiretroviral treatment of infants to attain HIV remission.
Studies in adults and children suggested that starting antiretroviral therapy (ART) soon after infection positively influences early events in HIV infection raising the possibility that remission may be achieved in some. We designed an analytic treatment interruption (ATI) trial to test the hypothesis that a sizable minority of HIV-infected neonates who initiated ART <14 days of birth and maintained on ART would be able to maintain viral suppression when ART was withdrawn. To yield the target cohort for this trial, 73 HIV-infected neonates identified at one hospital in Johannesburg, South Africa, were initiated on ART <14 days of birth and maintained on ART tracking viral load (VL) decline and immune recovery (clinicaltrials.gov # NCT02431975). Three HIV-infected infants (4.1%) died and nine (12.3%) were lost to follow-up before 48 weeks of age. Of those surviving on study, 52.5% attained and sustained VL <50 copies/ml and half of these sustained CD4+ T-cell percentage >30% which were the primary entry criteria for the ATI trial. Proportions achieving ATI eligibility criteria were similar in the 46 infants starting ART <48 h (19.6%) to 27 infants starting 2-14 days (25.9%) (p = 0.567). Very early ART on its own, using regimens available when the trial was designed, is insufficient to attain minimum entry criteria needed to justify our trial of ART interruption. Decisions about how quickly to start ART should be based on optimizing standard clinical outcomes rather than with the expectation that remission can be attained. NICHD/NIAID (U01HD080441), South African Research Chairs Initiative of DST and NRF (South Africa).


NRF2 is essential for adaptative browning of white adipocytes.
White adipose tissue browning, defined by accelerated mitochondrial metabolism and biogenesis, is considered a promising mean to treat or prevent obesity-associated metabolic disturbances. We hypothesize that redox stress acutely leads to increased production of reactive oxygen species (ROS), which activate electrophile sensor nuclear factor erythroid 2-Related Factor 2 (NRF2) that over time results in an adaptive adipose tissue browning process. To test this, we have exploited adipocyte-specific NRF2 knockout mice and cultured adipocytes and analyzed time- and dose-dependent effect of NAC and lactate treatment on antioxidant expression and browning-like processes. We found that short-term antioxidant treatment with N-acetylcysteine (NAC) induced reductive stress as evident from increased intracellular NADH levels, increased ROS-production, reduced oxygen consumption rate (OCR), and increased NRF2 levels in white adipocytes. In contrast, and in line with our hypothesis, longer-term NAC treatment led to a NRF2-dependent browning response. Lactate treatment elicited similar effects as NAC, and mechanistically, these NRF2-dependent adipocyte browning responses in vitro were mediated by increased heme oxygenase-1 (HMOX1) activity. Moreover, this NRF2-HMOX1 axis was also important for β3-adrenergic receptor activation-induced adipose tissue browning in vivo. In conclusion, our findings show that administration of exogenous antioxidants can affect biological function not solely through ROS neutralization, but also through reductive stress. We also demonstrate that NRF2 is essential for white adipose tissue browning processes.


The neurobiology of glucocerebrosidase-associated parkinsonism: a positron emission tomography study of dopamine synthesis and regional cerebral blood flow.
Mutations in GBA, the gene encoding glucocerebrosidase, the enzyme deficient in Gaucher disease, are common risk factors for Parkinson disease, as patients with Parkinson disease are over five times more likely to carry GBA mutations than healthy controls. Patients with GBA mutations generally have an earlier onset of Parkinson disease and more cognitive impairment than those without GBA mutations. We investigated whether GBA mutations alter the neurobiology of Parkinson disease, studying brain dopamine synthesis and resting regional cerebral blood flow in 107 subjects (38 women, 69 men). We measured dopamine synthesis with (18)F-fluorodopa positron emission tomography, and resting regional cerebral blood flow with H(2)(15)O positron emission tomography in the wakeful, resting state in four study groups: (i) patients with Parkinson disease and Gaucher disease (n = 7, average age = 56.6 ± 9.2 years); (ii) patients with Parkinson disease without GBA mutations (n = 11, 62.1 ± 7.1 years); (iii) patients with Gaucher disease without parkinsonism, but with a family history of Parkinson disease (n = 14, 52.6 ± 12.4 years); and (iv) healthy GBA-mutation carriers with a family history of Parkinson disease (n = 7, 50.1 ± 18 years). We compared each study group with a matched control group. Data were analysed with region of interest and voxel-based methods. Disease duration and Parkinson disease functional and staging scores were similar in the two groups with parkinsonism, as was striatal dopamine synthesis: both had greatest loss in the caudal striatum (putamen Ki loss: 44 and 42%, respectively), with less reduction in the caudate (20 and 18% loss). However, the group with both Parkinson and Gaucher diseases showed decreased resting regional cerebral blood flow in the lateral parieto-occipital association cortex and precuneus bilaterally. Furthermore, two subjects with Gaucher disease without parkinsonian manifestations showed diminished striatal dopamine. In conclusion, the pattern of dopamine loss in patients with both Parkinson and Gaucher disease was similar to sporadic Parkinson disease, indicating comparable damage in midbrain neurons. However, H(2)(15)O positron emission tomography studies indicated that these subjects have decreased resting activity in a pattern characteristic of diffuse Lewy body disease. These findings provide insight into the pathophysiology of GBA-associated parkinsonism.


Two-way antigenic cross-reactivity between severe acute respiratory syndrome coronavirus (SARS-CoV) and group 1 animal CoVs is mediated through an antigenic site in the N-terminal region of the SARS-CoV nucleoprotein.
In 2002, severe acute respiratory syndrome-associated coronavirus (SARS-CoV) emerged in humans, causing a global epidemic. By phylogenetic analysis, SARS-CoV is distinct from known CoVs and most closely related to group 2 CoVs. However, no antigenic cross-reactivity between SARS-CoV and known CoVs was conclusively and consistently demonstrated except for group 1 animal CoVs. We analyzed this cross-reactivity by an enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using specific antisera to animal CoVs and SARS-CoV and SARS patient convalescent-phase or negative sera. Moderate two-way cross-reactivity between SARS-CoV and porcine CoVs (transmissible gastroenteritis CoV [TGEV] and porcine respiratory CoV [PRCV]) was mediated through the N but not the spike protein, whereas weaker cross-reactivity occurred with feline (feline infectious peritonitis virus) and canine CoVs. Using Escherichia coli-expressed recombinant SARS-CoV N protein and fragments, the cross-reactive region was localized between amino acids (aa) 120 to 208. The N-protein fragments comprising aa 360 to 412 and aa 1 to 213 reacted specifically with SARS convalescent-phase sera but not with negative human sera in ELISA; the fragment comprising aa 1 to 213 cross-reacted with antisera to animal CoVs, whereas the fragment comprising aa 360 to 412 did not cross-react and could be a potential candidate for SARS diagnosis. Particularly noteworthy, a single substitution at aa 120 of PRCV N protein diminished the cross-reactivity. We also demonstrated that the cross-reactivity is not universal for all group 1 CoVs, because HCoV-NL63 did not cross-react with SARS-CoV. One-way cross-reactivity of HCoV-NL63 with group 1 CoVs was localized to aa 1 to 39 and at least one other antigenic site in the N-protein C terminus, differing from the cross-reactive region identified in SARS-CoV N protein. The observed cross-reactivity is not a consequence of a higher level of amino acid identity between SARS-CoV and porcine CoV nucleoproteins, because sequence comparisons indicated that SARS-CoV N protein has amino acid identity similar to that of infectious bronchitis virus N protein and shares a higher level of identity with bovine CoV N protein within the cross-reactive region. The TGEV and SARS-CoV N proteins are RNA chaperons with long disordered regions. We speculate that during natural infection, antibodies target similar short antigenic sites within the N proteins of SARS-CoV and porcine group 1 CoVs that are exposed to an immune response. Identification of the cross-reactive and non-cross-reactive N-protein regions allows development of SARS-CoV-specific antibody assays for screening animal and human sera.


Effects of mobile text messaging on breast cancer and breast self-examination (BSE) knowledge, BSE self-efficacy, and BSE frequency: a randomised controlled trial.
Educating women regarding breast cancer and increasing breast self-examination uptake continues to remain a major challenge despite the intensive campaigns. An innovative, cost-effective and easily customised intervention is needed especially in under-resourced and hard to reach communities. This paper is a report examining the effects of mobile text messaging on women's knowledge of breast cancer and BSE, BSE self-efficacy and frequency of BSE. A randomised controlled trial design was used. The study sample consisted of 128 women of reproductive age (WRA) (64 in the experimental group and 64 in the control group) from two communities in the Philippines during the months of August to October 2018. Three to five short messages were sent to the participants in the experimental group daily for one month while the control group did not receive the messages. Women who received mobile text messages had higher knowledge of breast cancer (F = 21.756, p = 0.001) and breast self-examination (F = 6.776, p = 0.010) than the control group. However, no significant improvements were seen in the BSE self-efficacy (F = 1.446, p = 0.232) and frequency of BSE (F = 3.374, p = 0.69). Mobile text messaging significantly improved the knowledge on breast cancer and breast self-examination among women; however, it did not affect their breast self-examination self-efficacy and frequency. Mobile text messaging can be a viable tool that can be used by healthcare workers in order to educate women regarding breast cancer and breast self-examination.


Customized reference ranges for laboratory values decrease false positive alerts in intensive care unit patients.
Traditional electronic medical record (EMR) interfaces mark laboratory tests as abnormal based on standard reference ranges derived from healthy, middle-aged adults. This yields many false positive alerts with subsequent alert-fatigue when applied to complex populations like hospitalized, critically ill patients. Novel EMR interfaces using adjusted reference ranges customized for specific patient populations may ameliorate this problem. To compare accuracy of abnormal laboratory value indicators in a novel vs traditional EMR interface. Laboratory data from intensive care unit (ICU) patients consecutively admitted during a two-day period were recorded. For each patient, available laboratory results and the problem list were sent to two mutually blinded critical care experts, who marked the values about which they would like to be alerted. All disagreements were resolved by an independent super-reviewer. Based on this gold standard, we calculated and compared the sensitivity, specificity, positive and negative predictive values (PPV, NPV) of customized vs traditional abnormal value indicators. Thirty seven patients with a total of 1341 laboratory results were included. Experts' agreement was fair (kappa = 0.39). Compared to the traditional EMR, custom abnormal laboratory value indicators had similar sensitivity (77% vs 85%, P = 0.22) and NPV (97.1% vs 98.6%, P = 0.06) but higher specificity (79% vs 61%, P < 0.001) and PPV (28% vs 11%, P < 0.001). Reference ranges for laboratory values customized for an ICU population decrease false positive alerts. Disagreement among clinicians about which laboratory values should be indicated as abnormal limits the development of customized reference ranges.


Contemporary Management of Achalasia by Pediatric Surgeons: A Survey of the International Pediatric Endosurgical Group.
Achalasia is a rare neurodegenerative disorder of the esophagus. Surgical repair consists of esophagomyotomy, often in conjunction with an antireflux procedure. We sought to determine practice patterns in surgical treatment of pediatric achalasia. Data regarding preferences were collected as part of a comprehensive online-based survey sent to members of the International Pediatric Endosurgery Group (IPEG) completed by 191 surgeons of which 141 performed esophagomyotomies for achalasia. Procedures performed per surgeon were 1-2 (n = 21, 15%); 3-5 (n = 49, 34%); 6-10 (n = 39, 28%); 11-20 (n = 21, 15%); >20 (n = 11, 8%). Most approached the operation laparoscopically (n = 127, 90%). Workup before esophageal myotomy consisted of a diagnostic esophagram (n = 133, 94%) or manometry (n = 102, 73%). Only 60% of surgeons (n = 84) required an EGD. No preference observed in division location of the phrenoesophageal ligament for mobilization of the esophagus. There was a predominant preference for hook cautery (n = 82, 58%) over harmonic shears (n = 30, 21%), heated sealing device LigaSure™ (n = 18, 13%), and other devices (n = 11, 8%) for muscle division. Intraoperatively, 57% (n = 80) had endoscopy and 50% (n = 71) had postoperative esophagram before initiation of enteral feeding. For antireflux procedure, Thal/Dor approach was performed most frequently (n = 111, 79%) followed by the Toupet (n = 18, 13%) and Nissen (n = 4, 3%) and none (n = 7, 5%). Diet restrictions were provided in 76% (n = 107) of postoperative patients. Given the infrequency of achalasia in children, there are a range of treatment plans among pediatric surgeons. We have identified current practices as a first step in developing more standard treatment pathways.


Temporal aspects of Ca(2+) and myosin phosphorylation during myogenic and norepinephrine-induced arteriolar constriction.
Previous studies demonstrated that maintenance of steady-state myogenic tone requires Ca(2+)-dependent myosin phosphorylation. The present studies furthered these observations by examining temporal relationships among Ca(2+), myosin phosphorylation and vessel diameter during acute increases in intraluminal pressure and norepinephrine stimulation. Rat cremaster muscle arterioles were cannulated and loaded with the Ca(2+)-sensitive indicator fura-2. The extent of myosin phosphorylation was measured using two-dimensional gel electrophoresis. Acute increases in intraluminal pressure caused a biphasic increase in intracellular Ca(2+) ([Ca(2+)](i)), characterized by a transient peak followed by a decline to a steady-state level which remained significantly higher than control values. Peak [Ca(2+)](i) was significantly related to vessel distension and increased with the change in wall tension. Increased intraluminal pressure resulted in a monophasic increase in myosin phosphorylation that was significantly correlated with instantaneous wall tension. In general, norepinephrine induced larger [Ca(2+)](i) transients and a biphasic myosin phosphorylation pattern. The results demonstrate: (a) major roles for Ca(2+) and myosin phosphorylation in arteriolar myogenic and norepinephrine-induced responses; (b) that changes in Ca(2+) and phosphorylation during a myogenic response are related to changes in wall tension, and (c) differences in Ca(2+) and phosphorylation patterns between the two modes of contraction reflect possible differences in underlying signaling mechanisms. The data further emphasize that spontaneous arteriolar tone represents a state of maintained smooth muscle activation that requires increases in [Ca(2+)](i) and myosin light-chain phosphorylation.


Resource partitioning among top predators in a Miocene food web.
The exceptional fossil sites of Cerro de los Batallones (Madrid Basin, Spain) contain abundant remains of Late Miocene mammals. From these fossil assemblages, we have inferred diet, resource partitioning and habitat of three sympatric carnivorous mammals based on stable isotopes. The carnivorans include three apex predators: two sabre-toothed cats (Felidae) and a bear dog (Amphicyonidae). Herbivore and carnivore carbon isotope (δ(13)C) values from tooth enamel imply the presence of a woodland ecosystem dominated by C(3) plants. δ(13)C values and mixing-model analyses suggest that the two sabre-toothed cats, one the size of a leopard and the other the size of a tiger, consumed herbivores with similar δ(13)C values from a more wooded portion of the ecosystem. The two sabre-toothed cats probably hunted prey of different body sizes, and the smaller species could have used tree cover to avoid encounters with the larger felid. For the bear dog, δ(13)C values are higher and differ significantly from those of the sabre-toothed cats, suggesting a diet that includes prey from more open woodland. Coexistence of the sabre-toothed cats and the bear dog was likely facilitated by prey capture in different portions of the habitat. This study demonstrates the utility of stable isotope analysis for investigating the behaviour and ecology of members of past carnivoran guilds.


Mathematical analysis of steady-state solutions in compartment and continuum models of cell polarization.
Cell polarization, in which substances previously uniformly distributed become asymmetric due to external or/and internal stimulation, is a fundamental process underlying cell mobility, cell division, and other polarized functions. The yeast cell S. cerevisiae has been a model system to study cell polarization. During mating, yeast cells sense shallow external spatial gradients and respond by creating steeper internal gradients of protein aligned with the external cue. The complex spatial dynamics during yeast mating polarization consists of positive feedback, degradation, global negative feedback control, and cooperative effects in protein synthesis. Understanding such complex regulations and interactions is critical to studying many important characteristics in cell polarization including signal amplification, tracking dynamic signals, and potential trade-off between achieving both objectives in a robust fashion. In this paper, we study some of these questions by analyzing several models with different spatial complexity: two compartments, three compartments, and continuum in space. The step-wise approach allows detailed characterization of properties of the steady state of the system, providing more insights for biological regulations during cell polarization. For cases without membrane diffusion, our study reveals that increasing the number of spatial compartments results in an increase in the number of steady-state solutions, in particular, the number of stable steady-state solutions, with the continuum models possessing infinitely many steady-state solutions. Through both analysis and simulations, we find that stronger positive feedback, reduced diffusion, and a shallower ligand gradient all result in more steady-state solutions, although most of these are not optimally aligned with the gradient. We explore in the different settings the relationship between the number of steady-state solutions and the extent and accuracy of the polarization. Taken together these results furnish a detailed description of the factors that influence the tradeoff between a single correctly aligned but poorly polarized stable steady-state solution versus multiple more highly polarized stable steady-state solutions that may be incorrectly aligned with the external gradient.


Transplantation of umbilical cord-derived mesenchymal stem cells into the striata of R6/2 mice: behavioral and neuropathological analysis.
Huntington's disease (HD) is an autosomal dominant disorder caused by an expanded CAG repeat on the short arm of chromosome 4 resulting in cognitive decline, motor dysfunction, and death, typically occurring 15 to 20 years after the onset of motor symptoms. Neuropathologically, HD is characterized by a specific loss of medium spiny neurons in the caudate and the putamen, as well as subsequent neuronal loss in the cerebral cortex. The transgenic R6/2 mouse model of HD carries the N-terminal fragment of the human HD gene (145 to 155 repeats) and rapidly develops some of the behavioral characteristics that are analogous to the human form of the disease. Mesenchymal stem cells (MSCs) have shown the ability to slow the onset of behavioral and neuropathological deficits following intrastriatal transplantation in rodent models of HD. Use of MSCs derived from umbilical cord (UC) offers an attractive strategy for transplantation as these cells are isolated from a noncontroversial and inexhaustible source and can be harvested at a low cost. Because UC MSCs represent an intermediate link between adult and embryonic tissue, they may hold more pluripotent properties than adult stem cells derived from other sources. Mesenchymal stem cells, isolated from the UC of day 15 gestation pups, were transplanted intrastriatally into 5-week-old R6/2 mice at either a low-passage (3 to 8) or high-passage (40 to 50). Mice were tested behaviorally for 6 weeks using the rotarod task, the Morris water maze, and the limb-clasping response. Following behavioral testing, tissue sections were analyzed for UC MSC survival, the immune response to the transplanted cells, and neuropathological changes. Following transplantation of UC MSCs, R6/2 mice did not display a reduction in motor deficits but there appeared to be transient sparing in a spatial memory task when compared to untreated R6/2 mice. However, R6/2 mice receiving either low- or high-passage UC MSCs displayed significantly less neuropathological deficits, relative to untreated R6/2 mice. The results from this study demonstrate that UC MSCs hold promise for reducing the neuropathological deficits observed in the R6/2 rodent model of HD.


Heparin alters transdermal transport associated with electroporation.
Short, high-voltage (HV; U(skin,max) approximately 100 V) pulses have been shown to increase rates of transdermal transport by several orders of magnitude via a mechanism hypothesized to involve electroporation. We show that heparin, a linear, highly charged macromolecule, significantly alters the molecular transport capacity and lifetime of aqueous pathways across human stratum corneum (SC) created by such pulses. If co-transported during pulsing, heparin molecules can interact with the SC and other molecules, thereby altering ionic and molecular transport. We also observed an increase in post-pulse skin permeability and persistent lower skin resistance. Because most heparin molecules are long enough to span the five to six lipid bilayer membranes that separate corneocytes within the SC, these results can be explained by the hypothesis that heparin molecules were trapped within the skin, holding open pathway segments connecting adjacent corneocytes. These results support the skin electroporation hypothesis and provide the first demonstration of a chemical enhancer effect for transdermal transport by HV pulsing.


Go big or go home: impact of screening coverage on syphilis infection dynamics.
Syphilis outbreaks in urban men who have sex with men (MSM) are an ongoing public health challenge in many high-income countries, despite intensification of efforts to screen and treat at-risk individuals. We sought to understand how population-level coverage of asymptomatic screening impacts the ability to control syphilis transmission. We developed a risk-structured deterministic compartmental mathematical model of syphilis transmission in a population of sexually active MSM. We assumed a baseline level of treatment of syphilis cases due to seeking medical care in all scenarios. We evaluated the impact of sustained annual population-wide screening coverage ranging from 0% to 90% on syphilis incidence over the short term (20 years) and at endemic equilibrium. The relationship between screening coverage and equilibrium syphilis incidence displayed an inverted U-shape relationship, with peak equilibrium incidence occurring with 20-30% annual screening coverage. Annual screening of 62% of the population was required for local elimination (incidence <1 case per 100 000 population). Results were qualitatively similar in the face of differing programmatic, behavioural and natural history assumptions, although the screening thresholds for local elimination differed. With 6-monthly or 3-monthly screening, the population coverage required to achieve local elimination was reduced to 39% or 23%, respectively. Although screening has the potential to control syphilis outbreaks, suboptimal coverage may paradoxically lead to a higher equilibrium infection incidence than that observed in the absence of intervention. Suboptimal screening programme design should be considered as a possible contributor to unsuccessful syphilis control programmes in the context of the current epidemic.


In situ mapping of the energy flow through the entire photosynthetic apparatus.
Absorption of sunlight is the first step in photosynthesis, which provides energy for the vast majority of organisms on Earth. The primary processes of photosynthesis have been studied extensively in isolated light-harvesting complexes and reaction centres, however, to understand fully the way in which organisms capture light it is crucial to also reveal the functional relationships between the individual complexes. Here we report the use of two-dimensional electronic spectroscopy to track directly the excitation-energy flow through the entire photosynthetic system of green sulfur bacteria. We unravel the functional organization of individual complexes in the photosynthetic unit and show that, whereas energy is transferred within subunits on a timescale of subpicoseconds to a few picoseconds, across the complexes the energy flows at a timescale of tens of picoseconds. Thus, we demonstrate that the bottleneck of energy transfer is between the constituents.


The Direct Assessment of Functional Abilities (DAFA): a comparison to an indirect measure of instrumental activities of daily living.
The Direct Assessment of Functional Abilities (DAFA) was designed as a direct measure of instrumental activities of daily living (IADLs) that could be compared with an indirect assessment of IADLs by the Pfeffer Functional Activities Questionnaire (PFAQ). The DAFA (28 demented and 15 control subjects) and PFAQ (subjects and informants) were administered twice, together with a brief cognitive battery. Demented subjects performed significantly worse on direct assessment (DAFA) than predicted by self-report (PFAQ), and overestimation of abilities increased with severity of dementia. In contrast, informants tended to underestimate abilities of demented subjects, but not to a significant degree. Control (nondemented) subjects had comparable results with the two methods. The DAFA may provide a more objective measure of functional status in demented subjects than do indirect methods of assessment.


The relationship between excessive Internet use and depression: a questionnaire-based study of 1,319 young people and adults.
There is a growing awareness of a psychiatric construct that needs to be better defined and understood: Internet addiction (IA). Recently there has been much public concern over the relationship between Internet use and negative affect. This study explored the concept of IA and examined the relationship between addictive symptoms and depression. An online questionnaire was used to measure participants' Internet use, the functions for which they used the Internet, and their depressive tendencies. Three scales were included: the IA Test, the Internet Function Questionnaire and the Beck Depression Inventory (BDI). 1,319 respondents completed the questionnaires, with 18 (1.2%) identified as falling in the IA category. Correlational analyses were conducted across the whole data sample. In factorial analyses, the 18 IA respondents were compared to a matched group of non-addicted (NA) respondents in terms of their scores on the Function Test and the BDI. Across the whole data sample, there was a close relationship between IA tendencies and depression, such that IA respondents were more depressed; there were also significant differences between the sexes, with men showing more addictive tendencies than women. In addition, young people were significantly more likely to show addictive symptoms than were older people. There was a significant difference between the IA and the NA group in their levels of depressive symptoms, with the NA group firmly in the non-depressed range, and the IA group in the moderately-to-severely depressed range (F(1, 34) = 22.35; p < 0.001). In terms of the function for which they used the Internet, the IA group engaged significantly more than the NA group in sexually gratifying websites, gaming websites and online community/chat websites. The concept of IA is emerging as a construct that must be taken seriously. Moreover, it is linked to depression, such that those who regard themselves as dependent on the Internet report high levels of depressive symptoms. Those who show symptoms of IA are likely to engage proportionately more than the normal population in sites that serve as a replacement for real-life socialising. Further work needs to be done on validating this relationship. Future research is needed to corroborate the existing evidence and address the nature of the relationship between IA and depression: there is comorbidity between these conditions that needs greater investigation.


Feasibility, acceptability, and cost of tuberculosis testing by whole-blood interferon-gamma assay.
The whole-blood interferon-gamma release assay (IGRA) is recommended in some settings as an alternative to the tuberculin skin test (TST). Outcomes from field implementation of the IGRA for routine tuberculosis (TB) testing have not been reported. We evaluated feasibility, acceptability, and costs after 1.5 years of IGRA use in San Francisco under routine program conditions. Patients seen at six community clinics serving homeless, immigrant, or injection-drug user (IDU) populations were routinely offered IGRA (Quantiferon-TB). Per guidelines, we excluded patients who were <17 years old, HIV-infected, immunocompromised, or pregnant. We reviewed medical records for IGRA results and completion of medical evaluation for TB, and at two clinics reviewed TB screening logs for instances of IGRA refusal or phlebotomy failure. Between November 1, 2003 and February 28, 2005, 4143 persons were evaluated by IGRA. 225(5%) specimens were not tested, and 89 (2%) were IGRA-indeterminate. Positive or negative IGRA results were available for 3829 (92%). Of 819 patients with positive IGRA results, 524 (64%) completed diagnostic evaluation within 30 days of their IGRA test date. Among 503 patients eligible for IGRA testing at two clinics, phlebotomy was refused by 33 (7%) and failed in 40 (8%). Including phlebotomy, laboratory, and personnel costs, IGRA use cost $33.67 per patient tested. IGRA implementation in a routine TB control program setting was feasible and acceptable among homeless, IDU, and immigrant patients in San Francisco, with results more frequently available than the historically described performance of TST. Laboratory-based diagnosis and surveillance for M. tuberculosis infection is now possible.


Altered intrinsic functional connectivity of anterior and posterior insula regions in high-functioning participants with autism spectrum disorder.
Impaired understanding of others' sensations and emotions as well as abnormal experience of their own emotions and sensations is frequently reported in individuals with Autism Spectrum Disorder (ASD). It is hypothesized that these abnormalities are based on altered connectivity within "shared" neural networks involved in emotional awareness of self and others. The insula is considered a central brain region in a network underlying these functions, being located at the transition of information about bodily arousal and the physiological state of the body to subjective feelings. The present study investigated the intrinsic functional connectivity properties of the insula in 14 high-functioning participants with ASD (HF-ASD) and 15 typically developing (TD) participants in the age range between 12 and 20 years by means of "resting state" or "nontask" functional magnetic resonance imaging. Essentially, a distinction was made between anterior and posterior regions of the insular cortex. The results show a reduced functional connectivity in the HF-ASD group, compared with the TD group, between anterior as well as posterior insula and specific brain regions involved in emotional and sensory processing. It is suggested that functional abnormalities in a network involved in emotional and interoceptive awareness might be at the basis of altered emotional experiences and impaired social abilities in ASD, and that these abnormalities are partly based on the intrinsic functional connectivity properties of such a network.


Evaluating the incidence of pathological complete response in current international rectal cancer practice: the barriers to widespread safe deferral of surgery.
The mainstay of management for locally advanced rectal cancer is chemoradiotherapy followed by surgical resection. Following chemoradiotherapy, a complete response may be detected clinically and radiologically (cCR) prior to surgery or pathologically after surgery (pCR). We aim to report the overall complete pathological response (pCR) rate and the reliability of detecting a cCR by conventional pre-operative imaging. A pre-planned analysis of the European Society of Coloproctology (ESCP) 2017 audit was performed. Patients treated by elective rectal resection were included. A pCR was defined as a ypT0 N0 EMVI negative primary tumour; a partial response represented any regression from baseline staging following chemoradiotherapy. The primary endpoint was the pCR rate. The secondary endpoint was agreement between post-treatment MRI restaging (yMRI) and final pathological staging. Of 2572 patients undergoing rectal cancer surgery in 277 participating centres across 44 countries, 673 (26.2%) underwent chemoradiotherapy and surgery. The pCR rate was 10.3% (67/649), with a partial response in 35.9% (233/649) patients. Comparison of AJCC stage determined by post-treatment yMRI with final pathology showed understaging in 13% (55/429) and overstaging in 34% (148/429). Agreement between yMRI and final pathology for T-stage, N-stage, or AJCC status were each graded as 'fair' only (n = 429, Kappa 0.25, 0.26 and 0.35 respectively). The reported pCR rate of 10% highlights the potential for non-operative management in selected cases. The limited strength of agreement between basic conventional post-chemoradiotherapy imaging assessment techniques and pathology suggest alternative markers of response should be considered, in the context of controlled clinical trials.


Comparative Study of the Condensing Effects of Ergosterol and Cholesterol.
Cholesterol, due to its condensing effect, is considered an important regulator of membrane thickness. Other sterols, due to their structural similarities to cholesterol, are often assumed to have a universal effect on membrane properties similar to the condensing effect of cholesterol, albeit possibly to different degrees. We used x-ray diffraction to investigate this assumption. By the combination of lamellar diffraction and grazing-angle scattering, we measured the membrane thickness and the tilt-angle distribution of the lipid's hydrocarbon chains. This method is sensitive to phase separation, which is important for examining the miscibility of sterols and phospholipids. Mixtures of ergosterol or cholesterol with dimyristoylphosphatidylcholine, palmitoyloleoylphosphatidylcholine, and dioleoylphosphatidylcholine were systematically studied. We found that mixing ergosterol with phospholipids into a single phase became increasingly difficult with higher sterol concentrations and also with higher concentrations of unsaturated lipid chains. The only condensing effect of ergosterol was found in dimyristoylphosphatidylcholine, although the effect was less than one-third of the effect of cholesterol. Unlike cholesterol, ergosterol could not maintain a fixed electron density profile of the surrounding lipids independent of hydration. In dioleoylphosphatidylcholine and palmitoyloleoylphosphatidylcholine, ergosterol made the membranes thinner, opposite to the effect of cholesterol. In all cases, the tilt-angle variation of the chain diffraction was consistent with the membrane thickness changes measured by lamellar diffraction, i.e., a thickening was always associated with a reduction of chain tilt angles. Our findings do not support the notion that different sterols have a universal behavior that differs only in degree.


Clients and facilitators' experiences of participating in a Hong Kong self-help group for people recovering from mental illness.
The purpose of this study was to examine the effectiveness of self-help groups in the rehabilitation of people recovering from mental illness. A qualitative approach was adopted, with individual interviews of 12 clients and four focus group interviews of 22 professionals and volunteers, in a Hong Kong self-help organization. Three major categories emerged from the client interviews: the meaning of self-help; experience of self-help group involvement; and changes in life. The major categories which emerged from the focus groups were: the meaning of self-help; therapeutic factors; therapeutic process; facilitators' expectation; and difficulties encountered. Results indicated that self-help group involvement provided positive experiences for the members and led to some changes in their lives which contribute to the rehabilitation of their illness. Meanwhile, the professionals' and volunteers' view of the use of self-help groups was found to coincide with previous literature. Moreover, a spiritual dimension was expressed by all respondents as one of the factors that enhanced the group cohesiveness. It was concluded that self-help group involvement is beneficial to psychiatric rehabilitation. Findings have implications for further utilization of self-help groups for the people recovering from mental illness, as an alternative form of health care to complement the inadequacies in the present health care system.


Graphics in optical coating design.
Graphical displays of complex reflectance for nonabsorbing multilayer optical coatings based upon circle diagrams are presented. Similar displays are then developed for absorbing films. The two types of display are used to analyze several common multilayers, and a repetitive structure with period characteristics is introduced.


Persistence, reticence and the management of multiple time memories by forager honey bees.
Honey bee foragers form time memories that enable them to match their foraging activity to the time of day when a particular food source is most productive. Persistent foragers show food-anticipatory activity by making reconnaissance flights to the previously productive food source and may continue to inspect it for several days. In contrast, reticent foragers do not investigate the source but wait for confirmation from returning persistent foragers. To determine how persistent and reticent foragers might contribute to the colony's ability to rapidly reallocate foragers among sources, we trained foragers to collect sucrose from a feeder at a restricted time of day for several days and then observed their behavior for three consecutive days during which the feeder was empty. In two separate trials, video monitoring of the hive entrance during unrewarded test days in parallel with observing reconnaissance visits to the feeder revealed a high level of activity, in both persistent and reticent foragers, thought to be directed at other food sources. This 'extracurricular' activity showed a high degree of temporal overlap with reconnaissance visits to the feeder. In some cases, inspection flights to the unrewarded feeder were made within the same trip to an extracurricular source, indicating that honey bees have the ability to manage at least two different time memories despite coincidence with respect to time of day. The results have major implications for understanding flower fidelity throughout the day, flower constancy within individual foraging excursions, and the sophisticated cognitive management of spatiotemporal memories in honey bees.


Variants in TTC25 affect autistic trait in patients with autism spectrum disorder and general population.
Autism spectrum disorder (ASD) is a highly heritable neurodevelopmental disorder with a complex genetic architecture. To identify genetic variants underlying ASD, we performed single-variant and gene-based genome-wide association studies using a dense genotyping array containing over 2.3 million single-nucleotide variants in a discovery sample of 160 families with at least one child affected with non-syndromic ASD using a binary (ASD yes/no) phenotype and a quantitative autistic trait. Replication of the top findings was performed in Psychiatric Genomics Consortium and Erasmus Rucphen Family (ERF) cohort study. Significant association of quantitative autistic trait was observed with the TTC25 gene at 17q21.2 (effect size=10.2, P-value=3.4 × 10-7) in the gene-based analysis. The gene also showed nominally significant association in the cohort-based ERF study (effect=1.75, P-value=0.05). Meta-analysis of discovery and replication improved the association signal (P-valuemeta=1.5 × 10-8). No genome-wide significant signal was observed in the single-variant analysis of either the binary ASD phenotype or the quantitative autistic trait. Our study has identified a novel gene TTC25 to be associated with quantitative autistic trait in patients with ASD. The replication of association in a cohort-based study and the effect estimate suggest that variants in TTC25 may also be relevant for broader ASD phenotype in the general population. TTC25 is overexpressed in frontal cortex and testis and is known to be involved in cilium movement and thus an interesting candidate gene for autistic trait.


Can Inner Peace be Improved by Mindfulness Training: A Randomized Controlled Trial.
This article reports a randomized controlled trial to investigate whether mindfulness training can successfully improve inner peace in participants with no known mental disorder. Fifty-seven participants were randomized to either mindfulness training (n = 29) or wait-list control (n = 28). The experience sampling method was used to measure the fleeting momentary experience of inner peace in participants. In addition, we used an experimental approach to assessing ability to focus attention: the Meditation Breath Attention Score, as well as the self-report Five-Facet Mindfulness Questionnaire (FFMQ). Compared with the wait-list control group, mindfulness training led to an increase in scores of inner peace, Meditation Breath Attention Score and FFMQ, using analysis of repeated measures analysis of variance. Change in inner peace was not, however, mediated by changes in self-rated mindfulness (FFMQ) nor by increased attentional focus. The findings provide first evidence suggesting that using mindfulness training improves the participants' inner peace. The focus here was on the immediate effects and future studies need to use follow-up.


Bilirubin conjugates produced by human, dog, and rat hepatocytes transplanted into athymic Gunn rats.
We developed a new hybrid mutant rat (athymic Gunn hybrid strain) that does not form and excrete conjugated bilirubin into bile and accepts xenografts because of T-lymphocyte deficiency. Cryopreserved isolated normal human, dog, or rat liver cells attached to collagen-coated microcarriers were transplanted into the athymic Gunn hybrid rats. Transplantation of human or rat liver cells resulted in biliary excretion of bilirubin conjugates predominantly as glucuronides, whereas isolated dog liver cells likewise transplanted resulted in the excretion of 40% to 50% of the conjugates as a glucoside-glucuronide diconjugate. The biliary bilirubin conjugates of the transplanted athymic Gunn hybrid rats paralleled those of the donor species. The data indicate that the types of bilirubin conjugates excreted by each species are determined by intrinsic properties of the respective hepatocytes rather than nutritional or other environmental factors. The findings also show that conjugation of bilirubin after liver cell transplantation results from functioning of the engrafted cells rather than activation of endogenous uridine diphosphatelucuronosyltransferase activity of the host.


Low prevalence of male microchimerism in women with Mayer-Rokitansky-Küster-Hauser syndrome.
Is there an increased prevalence of male microchimerism in women with Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome, as evidence of fetal exposure to blood and anti-Müllerian hormone (AMH) from a (vanished) male co-twin resulting in regression of the Müllerian duct derivatives? Predominant absence of male microchimerism in adult women with MRKH syndrome does not support our hypothesis that intrauterine blood exchange with a (vanished) male co-twin is the pathophysiological mechanism. The etiology of MRKH is unclear. Research on the phenotype analogous condition in cattle (freemartinism) has yielded the hypothesis that Müllerian duct development is inhibited by exposure to AMH in utero. In cattle, the male co-twin has been identified as the source for AMH, which is transferred via placental blood exchange. In human twins, a similar exchange of cellular material has been documented by detection of chimerism, but it is unknown whether this has clinical consequences. An observational case-control study was performed to compare the presence of male microchimerism in women with MRKH syndrome and control women. Through recruitment via the Dutch patients' association of women with MRKH (comprising 300 members who were informed by email or regular mail), we enrolled 96 patients between January 2017 and July 2017. The control group consisted of 100 women who reported never having been pregnant. After written informed consent, peripheral blood samples were obtained by venipuncture, and genomic DNA was extracted. Male microchimerism was detected by Y-chromosome-specific real-time quantitative PCR, with use of DYS14 marker. Possible other sources for microchimerism, for example older brothers, were evaluated using questionnaire data. The final analysis included 194 women: 95 women with MRKH syndrome with a mean age of 40.9 years and 99 control women with a mean age of 30.2 years. In total, 54 women (56.8%) were identified as having typical MRKH syndrome, and 41 women (43.2%) were identified as having atypical MRKH syndrome (when extra-genital malformations were present). The prevalence of male microchimerism was significantly higher in the control group than in the MRKH group (17.2% versus 5.3%, P = 0.009). After correcting for age, women in the control group were 5.8 times more likely to have male microchimerism (odds ratio 5.84 (CI 1.59-21.47), P = 0.008). The mean concentration of male microchimerism in the positive samples was 56.0 male genome equivalent per 1 000 000 cells. The prevalence of male microchimerism was similar in women with typical MRKH syndrome and atypical MRKH syndrome (5.6% versus 4.9%, P = 0.884). There were no differences between women with or without microchimerism in occurrence of alternative sources of XY cells, such as older brothers, previous blood transfusion, or history of sexual intercourse. We are not able to draw definitive conclusions regarding the occurrence of AMH exchange during embryologic development in women with MRKH syndrome. Our subject population includes all adult women and therefore is reliant on long-term prevalence of microchimerism. Moreover, we have only tested blood, and, theoretically, the cells may have grafted anywhere in the body during development. It must also be considered that the exchange of AMH may occur without the transfusion of XY cells and therefore cannot be discovered by chimerism detection. This is the first study to test the theory that freemartinism causes the MRKH syndrome in humans. The study aimed to test the presence of male microchimerism in women with MRKH syndrome as a reflection of early fetal exposure to blood and AMH from a male (vanished) co-twin. We found that male microchimerism was only present in 5.3% of the women with MRKH syndrome, a significantly lower percentage than in the control group (17.2%). Our results do not provide evidence for an increased male microchimerism in adult women with MRKH as a product of intrauterine blood exchange. However, the significant difference in favor of the control group is of interest to the ongoing discussion on microchimeric cell transfer and the possible sources of XY cells. None. Dutch trial register, NTR5961.


Structure-function analysis of the kinase-CPD domain of yeast tRNA ligase (Trl1) and requirements for complementation of tRNA splicing by a plant Trl1 homolog.
Trl1 is an essential 827 amino acid enzyme that executes the end-healing and end-sealing steps of tRNA splicing in Saccharomyces cerevisiae. Trl1 consists of two domains--an N-terminal ligase component and a C-terminal 5'-kinase/2',3'-cyclic phosphodiesterase (CPD) component--that can function in tRNA splicing in vivo when expressed as separate polypeptides. To understand the structural requirements for the kinase-CPD domain, we performed an alanine scan of 30 amino acids that are conserved in Trl1 homologs from other fungi. We thereby identified four residues (Arg463, His515, Thr675 and Glu741) as essential for activity in vivo. Structure-function relationships at these positions, and at four essential or conditionally essential residues defined previously (Asp425, Arg511, His673 and His777), were clarified by introducing conservative substitutions. Biochemical analysis showed that lethal mutations of Asp425, Arg463, Arg511 and His515 in the kinase module abolished polynucleotide kinase activity in vitro. We report that a recently cloned 1104 amino acid Arabidopsis RNA ligase functions in lieu of yeast Trl1 in vivo and identify essential side chains in the ligase, kinase and CPD modules of the plant enzyme. The plant ligase, like yeast Trl1 but unlike T4 RNA ligase 1, requires a 2'-PO4 end for tRNA splicing in vivo.


Effects of disruption of epiphyseal vasculature on the proximal femoral growth plate.
Proximal femoral growth disturbance is a major complication associated with ischemic osteonecrotic conditions, such as Legg-Calvè-Perthes disease. The extent of ischemic damage and the mechanisms by which ischemic injury to the growing femoral head produces growth disturbance of the proximal femoral growth plate remain unclear. The purpose of this study was to investigate the effects of disruption of the epiphyseal vasculature on the morphology and function of the proximal femoral growth plate in a porcine model. Ischemic osteonecrosis of the femoral head was surgically induced in sixty-five piglets by placing a ligature tightly around the femoral neck. Radiographic, histological, micro-computed tomographic, cellular viability, hypoxia marker, and cellular proliferation studies were performed. Disruption of the epiphyseal vasculature did not lead to diffuse growth plate damage in the majority of the ischemic femoral heads. One of the twelve femoral heads analyzed at four weeks and six of the twenty-six femoral heads analyzed at eight weeks had severe disruption of the growth plate that precluded histological assessment of the growth plate zones. In the remaining animals, the proximal part of the femur continued to elongate following induction of ischemia, albeit at a slower rate than on the normal side. Histologically, normal developmental thinning of the growth plate was seen to be absent on the ischemic side. Severe hypoxia and cellular death were limited to the area of the growth plate bordering on the infarcted osseous epiphysis. Normal chondrocytic organization and continued proliferation were observed in the proliferative zone of the growth plate. In our porcine model, the proximal femoral growth plate was not diffusely damaged following disruption of the epiphyseal vasculature in the majority of the ischemic femoral heads. The majority of the growth plates remained viable and were able to function despite total disruption of the epiphyseal vasculature. These findings suggest that the source of nutrition for the proximal femoral growth plate is not solely the epiphyseal vasculature as has been traditionally believed.


Anti-RDL and Anti-mGlutR1 Receptors Antibody Testing in Honeybee Brain Sections using CRISPR-Cas9.
Cluster Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is a gene editing technique widely used in studies of gene function. We use this method in this study to check for the specificity of antibodies developed against the insect GABAA receptor subunit Resistance to Dieldrin (RDL) and a metabotropic glutamate receptor mGlutR1 (mGluRA). The antibodies were generated in rabbits against the conjugated peptides specific to fruit flies (Drosophila melanogaster) as well to honeybees (Apis mellifera). We used these antibodies in honeybee brain sections to study the distribution of the receptors in honeybee brains. The antibodies were affinity purified against the peptide and tested with immunoblotting and the classical method of preadsorption with peptide conjugates to show that the antibodies are specific to the corresponding peptide conjugates against which they were raised. Here we developed the CRISPR-Cas9 technique to test for the reduction of protein targets in the brain 48 h after CRISPR-Cas9 injection with guide RNAs designed for the corresponding receptor. The CRISPR-Cas9 method can also be used in behavioral analyses in the adult bees when one or multiple genes need to be modified.


The FLC Locus: A Platform for Discoveries in Epigenetics and Adaptation.
Our understanding of the detailed molecular mechanisms underpinning adaptation is still poor. One example for which mechanistic understanding of regulation has converged with studies of life history variation is Arabidopsis thaliana FLOWERING LOCUS C (FLC). FLC determines the need for plants to overwinter and their ability to respond to prolonged cold in a process termed vernalization. This review highlights how molecular analysis of vernalization pathways has revealed important insight into antisense-mediated chromatin silencing mechanisms that regulate FLC. In turn, such insight has enabled molecular dissection of the diversity in vernalization across natural populations of A. thaliana. Changes in both cotranscriptional regulation and epigenetic silencing of FLC are caused by noncoding polymorphisms at FLC. The FLC locus is therefore providing important concepts for how noncoding transcription and chromatin regulation influence gene expression and how these mechanisms can vary to underpin adaptation in natural populations.


Relative Capacity and the Spread of Rebellion: Insights from Nepal.
This article explores how insurgencies emerge and spread within a country over time through an analysis of the Maoist insurgency in Nepal. It argues that important processes underpinning the spread of insurgency are likely to change with shifts in the relative military capacity of belligerents. Importantly, insurgents can to a greater extent spread the insurgency by using coercion, material incentives, and movement of forces when they are militarily strong than when they are weak. This in turn leads to changes in the local conditions favorable to insurgency. I hypothesize that inaccessible terrains, preexisting rebel networks, and proximity to insurgent areas are likely to be important determinants of local insurgency onset during rebel weakness, but should decline in importance as the rebels gain strength. I find support for these arguments in a mixed-methods analysis of Nepal's insurgency that combines a qualitative narrative and a quantitative event history analysis.


Reducing demand for antibiotic prescriptions: evidence from an online survey of the general public on the interaction between preferences, beliefs and information, United Kingdom, 2015.
Antimicrobial resistance (AMR), a major public health threat, is strongly associated with human antibiotic consumption. Influenza-like illnesses (ILI) account for substantial inappropriate antibiotic use; patient understanding and expectations probably play an important role. This study aimed to investigate what drives patient expectations of antibiotics for ILI and particularly whether AMR awareness, risk preferences (attitudes to taking risks with health) or time preferences (the extent to which people prioritise good health today over good health in the future) play a role. In 2015, a representative online panel survey of 2,064 adults in the United Kingdom was asked about antibiotic use and effectiveness for ILI. Explanatory variables in multivariable regression included AMR awareness, risk and time preferences and covariates. The tendency not to prioritise immediate gain over later reward was independently strongly associated with greater awareness that antibiotics are inappropriate for ILI. Independently, believing antibiotics were effective for ILI and low AMR awareness significantly predicted reported antibiotic use. However, 272 (39%) of those with low AMR awareness said that the AMR information we provided would lead them to ask a doctor for antibiotics more often, significantly more than would do so less often, and in contrast to those with high AMR awareness (p < 0.0001). Information campaigns to reduce AMR may risk a paradoxical consequence of actually increasing public demand for antibiotics. Public antibiotic stewardship campaigns should be tested on a small scale before wider adoption.


Physico-chemical characteristics of papaya (Carica papaya L.) seed oil of the Hong Kong/Sekaki variety.
A study was carried out to determine the physicochemical characteristics of the oil derived from papaya seeds of the Hong Kong/Sekaki variety. Proximate analysis showed that seeds of the Hong Kong/Sekaki variety contained considerable amount of oil (27.0%). The iodine value, saponification value, unsaponifiable matter and free fatty acid contents of freshly extracted papaya seed oil were 76.9 g I2/100g oil, 193.5 mg KOH/g oil, 1.52% and 0.91%, respectively. The oil had a Lovibond color index of 15.2Y + 5.2B. Papaya seed oil contained ten detectable fatty acids, of which 78.33% were unsaturated. Oleic (73.5%) acid was the dominant fatty acids followed by palmitic acid (15.8%). Based on the high performance liquid chromatography (HPLC) analysis, seven species of triacylglycerols (TAGs) were detected. The predominant TAGs of papaya seed oil were OOO (40.4%), POO (29.1%) and SOO (9.9%) where O, P, and S denote oleic, palmitic and stearic acids, respectively. Thermal analysis by differential scanning calorimetry (DSC) showed that papaya seed oil had its major melting and crystallization transitions at 12.4°C and -48.2°C, respectively. Analysis of the sample by Z-nose (electronic nose) instrument showed that the sample had a high level of volatile compounds.


Phytoplankton assemblage of a small, shallow, tropical African reservoir.
I measured physico-chemical properties and phytoplankton in the small, shallow tropical reservoir of Oyun (Offa, Nigeria) between January 2002 and December 2003. I identified 25 phytoplankton genera in three sampling stations. Bacillariophyceae dominated (75.3%), followed by Chlorophyceae (12.2%), Cyanobacteria (11.1%) and Desmidiaceae (0.73%). The high amount of nutrients (e.g. nitrate, phosphate, sulphate and silica) explain phytoplankton heterogeneity (p<0.05). Phytoplankton was abundant during the rainy season, but the transition period had the richest assemblage and abundance. Fluctuations in phytoplankton density were a result of seasonal changes in concentration of nutrients, grazing pressure and reservoir hydrology. The reservoir is eutrophic with excellent water quality and a diverse phytoplankton assemblage: fish production would be high. These conditions resulted from strategies such as watershed best management practices (BMPs) to control eutrophication and sedimentation, and priorities for water usage established through legislation. Additional measures are recommended to prevent oligotrophy, hypereutrophy, excessive phytoplankton bloom, toxic cyanobacteria, and run-off of organic waste and salts.


Serum and BAL Clara cell 10 kDa protein (CC10) levels and CC10-positive bronchiolar cells are decreased in smokers.
Cigarette smoking has diverse effects on the structure and function of the lung. Smoking appears to reduce the levels of Clara cell 10 kDa protein (CC10) in the alveolar lining fluid, but the influence of smoking serum on CC10 levels is still debated, and it has not been clear whether smoking reduces the number of CC10-producing lung cells. The aims of this study were to clarify the influence of smoking on CC10 levels in the alveolar lining fluid and bloodstream, and on the number of CC10-producing lung cells. CC10 concentrations were measured in sera and bronchoalveolar lavage (BAL) fluids, by means of enzyme-linked immunosorbent assay using monoclonal and polyclonal antibody, and the immunohistochemical expression of CC10 was examined in the lungs of nonsmokers and smokers using the monoclonal antibody, TY-5, against CC10/human urinary protein-1. CC10 concentrations in sera and in BAL fluids from healthy smokers were significantly lower than in healthy nonsmokers. Immunohistochemical expression of CC10 was found exclusively in nonciliated bronchiolar epithelial cells. As compared to that of nonsmokers, the mean percentage of CC10-positive bronchiolar epithelial cells was significantly decreased in lung tissue specimens obtained from smokers who had normal results in pulmonary function tests. It was concluded that smoking reduces the proportion of Clara cell 10 kDa protein-producing bronchiolar epithelial cells, resulting in decreased levels of Clara cell 10 kDa protein in the lower respiratory tract and in the bloodstream. The protein is a new blood biochemical and immunohistochemical marker, reflecting structural changes in peripheral airways induced by cigarette smoking.


Corneal Epithelial Remodeling Induced by Small Incision Lenticule Extraction (SMILE).
To assess the reactive changes of the corneal epithelial thickness (ET) profile induced by small incision lenticule extraction (SMILE) for treatment of myopia. This prospective observational study included 46 eyes of 23 patients that were scheduled for myopic SMILE. High-resolution spectral-domain optical coherence tomography (SD-OCT) and a custom image segmentation algorithm were applied to measure corneal ET at multiple time points within the central 5-mm zone. Postoperative ET alterations were correlated with treatment parameters and refractive outcomes. Mean age was 33 ± 6 years and mean spherical equivalent of surgical refractive correction was -4.78 ± 1.75 diopters. The average ET (averaged over the central 5-mm zone) increased from 52.3 ± 3.6 μm preoperatively to 57.7 ± 5.1 μm 6 months postoperatively (P < 0.01). Average ET was 101.9% of the preoperative thickness at 24 hours postoperatively, 103.2% at 1 week, 106.7% at 6 weeks, 109.3% at 3 months, and 110.4% at 6 months postoperatively. The epithelial thickening response could be modeled by an exponential recovery function and stabilized after 3 months. The extent of epithelial hyperplasia was highly dependent on the magnitude of the induced refractive correction (P = 0.002). In this initial study of corneal ET remodeling after myopic SMILE, significant epithelial thickening was detected as a function of the extent of surgical refractive correction. Moreover, the epithelial remodeling response to the corneal refractive change appeared to decrease with higher age. In our hands, the observed epithelial changes did not appear to affect the refractive outcome of SMILE. (ClinicalTrials.gov number, NCT02614625).


A fresh look at tobacco harm reduction: the case for the electronic cigarette.
Smokers of any age can reap substantial health benefits by quitting. In fact, no other single public health effort is likely to achieve a benefit comparable to large-scale smoking cessation. Surveys document that most smokers would like to quit, and many have made repeated efforts to do so. However, conventional smoking cessation approaches require nicotine addicted smokers to abstain from tobacco and nicotine entirely. Many smokers are unable--or at least unwilling--to achieve this goal, and so they continue smoking in the face of impending adverse health consequences. In effect, the status quo in smoking cessation presents smokers with just two unpleasant alternatives: quit or suffer the harmful effects of continuing smoking. But, there is a third choice for smokers: tobacco harm reduction. It involves the use of alternative sources of nicotine, including modern smokeless tobacco products like snus and the electronic cigarette (E-cig), or even pharmaceutical nicotine products, as a replacement for smoking. E-cigs might be the most promising product for tobacco harm reduction to date, because, besides delivering nicotine vapour without the combustion products that are responsible for nearly all of smoking's damaging effect, they also replace some of the rituals associated with smoking behaviour. Thus it is likely that smokers who switch to E-cigs will achieve large health gains. The focus of this article is on the health effects of using an E-cig, with consideration given to the acceptability, safety and effectiveness of this product as a long-term substitute for smoking.


A proposed radiosurgery-based grading system for arteriovenous malformations.
Radiosurgery is an effective treatment strategy for properly selected patients harboring arteriovenous malformations (AVMs). Grading scales that are currently used to predict patient outcomes after AVM resection are unreliable tools for the prediction of the results of AVM radiosurgery. A grading system was developed to predict outcomes following AVM radiosurgery, based on the multivariate analysis of data obtained in 220 patients treated between 1987 and 1991 (Group 1). The dependent variable in all analyses was excellent patient outcome (complete AVM obliteration without any new neurological deficit). The grading scale was tested on a separate set of 136 patients with AVMs treated between 1990 and 1996 at a different center (Group 2). One hundred twenty-one (55%) of 220 Group 1 patients had excellent outcomes. Multivariate analysis identified five variables related to excellent patient outcomes: AVM volume (p = 0.001), patient age (p < 0.001), AVM location (p < 0.001), previous embolization (p = 0.02), and number of draining veins (p < 0.001). Regression analysis modeling permitted removal of two significant variables (previous embolization and number of draining veins) and resulted in the following equation to predict patient outcomes after AVM radiosurgery: AVM score = (0.1)(AVM volume in cm3) + (0.02)(patient age in years) + (0.3)(location of lesion: frontal or temporal) = 0; parietal, occipital, intraventricular, corpus callosum, cerebellar = 1; or basal ganglia, thalamic, or brainstem = 2). Seventy-nine (58%) of 136 Group 2 patients had excellent outcomes. All variables in the model remained significant for the Group 2 patients: AVM volume (p = 0.01), patient age (p = 0.01), and AVM location (p < 0.001). Testing of the entire model on the Group 2 patients demonstrated that the AVM score could be used to predict patient outcomes after radiosurgery (p < 0.0001). All patients with an AVM score of 1 or lower had an excellent outcome compared with only 39% of patients with an AVM score higher than 2. The Spetzler-Martin grade (p = 0.13), the K index (p = 0.26), and the obliteration prediction index (p = 0.21) did not correlate with excellent patient outcomes. Despite significant differences in preoperative patient characteristics and dose prescription guidelines at the two centers, the proposed AVM grading system strongly correlated with patient outcomes after single-session radiosurgery for both patient groups. Although further testing of this model by independent centers using prospective methodology is still required, this system allows a more accurate prediction of outcomes from radiosurgery to guide choices between surgical and radiosurgical management for individual patients with AVMs.


Harnessing global fisheries to tackle micronutrient deficiencies.
Micronutrient deficiencies account for an estimated one million premature deaths annually, and for some nations can reduce gross domestic product1,2 by up to 11%, highlighting the need for food policies that focus on improving nutrition rather than simply increasing the volume of food produced3. People gain nutrients from a varied diet, although fish-which are a rich source of bioavailable micronutrients that are essential to human health4-are often overlooked. A lack of understanding of the nutrient composition of most fish5 and how nutrient yields vary among fisheries has hindered the policy shifts that are needed to effectively harness the potential of fisheries for food and nutrition security6. Here, using the concentration of 7 nutrients in more than 350 species of marine fish, we estimate how environmental and ecological traits predict nutrient content of marine finfish species. We use this predictive model to quantify the global spatial patterns of the concentrations of nutrients in marine fisheries and compare nutrient yields to the prevalence of micronutrient deficiencies in human populations. We find that species from tropical thermal regimes contain higher concentrations of calcium, iron and zinc; smaller species contain higher concentrations of calcium, iron and omega-3 fatty acids; and species from cold thermal regimes or those with a pelagic feeding pathway contain higher concentrations of omega-3 fatty acids. There is no relationship between nutrient concentrations and total fishery yield, highlighting that the nutrient quality of a fishery is determined by the species composition. For a number of countries in which nutrient intakes are inadequate, nutrients available in marine finfish catches exceed the dietary requirements for populations that live within 100 km of the coast, and a fraction of current landings could be particularly impactful for children under 5 years of age. Our analyses suggest that fish-based food strategies have the potential to substantially contribute to global food and nutrition security.


Supine body position is an important factor influencing postprandial ambulatory blood pressure.
Although body position is known to be an important factor influencing the results of ambulatory blood pressure monitoring (ABPM), even very recent studies assessing postprandial blood pressure (BP) have not taken into account the possibility that the examined subjects were lying down after a meal. We addressed this issue by analysing diurnal BP profiles together with subject's reports on supine rest periods and meal consumption during ABPM. The ABPM results of 109 non-medicated subjects were analysed (age 40+/-12 years, daytime BP 132/84+/-15/11 mmHg). BP and heart rate (HR) changes were assessed on the basis of 2-hour means. Subjects who lay down within 2 hours after a meal showed significantly greater decreases in systolic and diastolic BP and HR than individuals who did not: -10+/-10 vs. 1+/-10 mmHg, -11+/-10 vs. -3+/-7 mmHg, and -7+/-9 vs. 0+/-11 beats/min, respectively. The BP decrease was greater when the duration of supine rest was longer. There were no significant differences between BP/HR changes during those periods when the subjects lay down within 2 hours after the meal and BP/HR changes during those periods when supine rest was not preceded by a meal. The mean changes for the latter group were -9+/-10 mmHg, -11+/-8 mmHg, and -9+/-9 beats/min. for SBP, DBP, and HR, respectively. Our results indicate that supine body position is an important factor, which may influence postprandial BP investigated by ABPM. Body position should always be taken into consideration when ABPM results are analysed.


Genotoxicity and carcinogenicity of cobalt-, nickel- and copper-based nanoparticles.
The nanotechnology industry has matured and expanded at a rapid pace in the last decade, leading to the research and development of nanomaterials with enormous potential. The largest source of these nanomaterials is the transitional metals. It has been revealed that numerous properties of these nano-sized elements are not present in their bulk states. The nano size of these particles means they are easily transported into biological systems, thus, raising the question of their effects on the susceptible systems. Although advances have been made and insights have been gained on the effect of transitional metals on susceptible biological systems, there still is much ground to be covered, particularly with respect to our knowledge on the genotoxic and carcinogenic effects. Therefore, this review intends to summarize the current knowledge on the genotoxic and carcinogenic potential of cobalt-, nickel- and copper-based nanoparticles indicated in in vitro and in vivo mammalian studies. In the present review, we briefly state the sources, use and exposure routes of these nanoparticles and summarize the current literature findings on their in vivo and in vitro genotoxic and carcinogenic effects. Due to the increasing evidence of their role in carcinogenicity, we have also included studies that have reported epigenetic factors, such as abnormal apoptosis, enhanced oxidative stress and pro-inflammatory effects involving these nanoparticles.


The role of p53 in myelodysplastic syndromes and acute myeloid leukemia: molecular aspects and clinical implications.
TP53 gene mutations occurring in patients with myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) are associated with high-risk karyotypes including 17p abnormalities, monosomal and complex cytogenetics. TP53 mutations in these disorders portend rapid disease progression and resistance to conventional therapeutics. Notably, the size of the TP53 mutant clone as measured by mutation allele burden is directly linked to overall survival (OS) confirming the importance of p53 as a negative prognostic variable. In nucleolar stress-induced ribosomopathies, such as del(5q) MDS, disassociation of MDM2 and p53 results in p53 accumulation in erythroid precursors manifested as erythroid hypoplasia. P53 antagonism by lenalidomide or other therapeutics such as antisense oligonucleotides, repopulates erythroid precursors and enhances effective erythropoiesis. These findings demonstrate that p53 is an intriguing therapeutic target that is currently under investigation in MDS and AML. This study reviews molecular advances in understanding the role of p53 in MDS and AML, and explores potential therapeutic strategies in this era of personalized medicine.


Factors Affecting Perceptions of Open, Mini-Open, and Arthroscopic Rotator Cuff Repair Techniques Among Medical Professionals.
Although no long-term difference between arthroscopic and mini-open rotator cuff repairs has been documented, use of arthroscopic repair has exploded. We conducted a study to determine which repair technique medical professionals preferred for their own surgery and to analyze the perceptions shaping those opinions. A survey was emailed to selected professionals at our institution: attendings, residents, and allied health professionals; 84 (41, 20, and 23, respectively) responded. Irrespective of specialty or career length, almost half (39, 46%) preferred deferring the repair choice to their surgeon; the other 45 preferred arthroscopic (22, 26%), mini-open (19, 23%), open (2, 2%), or no (2, 2%) repair. Most agreed repairs were safe and fast but had no opinion about cost-effectiveness or which technique provided the best outcome. Significantly (P < .05) more respondents thought arthroscopic and mini-open repairs promoted quick healing, good cosmetic results, and patient satisfaction compared with open repair, but these repairs were also perceived as significantly (P < .05) harder to learn and more challenging than open repair. It is important for medical professionals to recognize these biases, especially given that many defer to the judgment of their medical peers.


Selectivity of nitrate and chloride ions in microporous carbons: the role of anisotropic hydration and applied potentials.
Understanding ion transport in porous carbons is critical for a wide range of technologies, including supercapacitors and capacitive deionization for water desalination, yet many details remain poorly understood. For instance, an atomistic understanding of how ion selectivity is influenced by the molecular shape of ions, morphology of the micropores and applied voltages is largely lacking. In this work, we combined molecular dynamics simulations with enhanced sampling methods to elucidate the mechanism of nitrate and chloride selectivity in subnanometer graphene slit-pores. We show that nitrate is preferentially adsorbed over chloride in the slit-like micropores. This preferential adsorption was found to stem from the weaker hydration energy and unique anisotropy of the ion solvation of nitrate. Beside the effects of ion dehydration, we found that applied potential plays an important role in determining the ion selectivity, leading to a lower selectivity of nitrate over chloride at a high applied potential. We conclude that the measured ion selectivity results from a complex interplay between voltage, confinement, and specific ion effects-including ion shape and local hydration structure.


Associations of general and central adiposity with hypertension and cardiovascular disease among South Asian populations: a systematic review and meta-analysis.
The relevance of measures of general and central adiposity for cardiovascular disease (CVD) risks in populations of European descent is well established. However, it is less well characterised in South Asian populations, who characteristically manifest larger waist circumferences (WC) for equivalent body mass index (BMI). This systematic review and meta-analysis provide an overview of the literature on the association of different anthropometric measures with CVD risk among South Asians. MEDLINE and Embase were searched from 1990 to the present for studies in South Asian populations investigating associations of two or more adiposity measures with CVD. Random-effects meta-analyses were conducted on the associations of BMI, WC and waist-to-hip ratio (WHR) with blood pressure, hypertension and CVD. Quality assessment was performed using the Newcastle-Ottawa scale. Titles and abstracts were screened for 7327 studies, yielding 147 full-text reviews. The final sample (n=30) included 2 prospective, 5 case-control and 23 cross-sectional studies. Studies reported generally higher risks of hypertension and CVD at higher adiposity levels. The pooled mean difference in systolic blood pressure (SBP) per 5 kg/m2 higher BMI was 3 mmHg (2.90 (95% CI 1.30 to 4.50)) and 6 mmHg (6.31 (95% CI 4.81 to 7.81) per 13 cm larger WC. The odds ratio (OR) of hypertension per 5 kg/m2 higher BMI was 1.33 (95% CI 1.18 to 1.51), 1.45 (95% CI 1.05 to 1.98) per 13 cm larger WC and 1.22 (95% CI 1.04 to 1.41) per 0.1-unit larger WHR. Pooled risk of CVD for BMI-defined overweight versus healthy-weight was 1.65 (95% CI 1.55 to 1.75) and 1.48 (95% CI 1.21 to 1.80) and 2.51 (95% CI 0.94 to 6.69) for normal versus large WC and WHR, respectively. Study quality was average with significant heterogeneity. Measures of both general and central adiposity had similar, strong positive associations with the risk of CVD in South Asians. Larger prospective studies are required to clarify which measures of body composition are more informative for targeted CVD primary prevention in this population.


Neuropsychology of acute stroke.
Neuropsychology includes both the psychiatric manifestations of neurological illness (primary brain-based disorders) and neurobiology of "idiopathic" psychiatric disorders. Neurological primary brain disorders provoke broad spectrum of brain pathophysiology that cause deficit sin human behaviour, and the magnitude of neurobehavioral-related problems is a world wide health concern. Speech disorders of aphasic type, unilateral neglect, anosognosia (deficit disorders), delirium and mood disorders (productive disorders) in urgent neurology, first of all in acute phase of stroke are more frequent disorders then it verified in routine exam, not only in the developed and large neurological departments. Aphasia is common consequence of left hemispheric lesion and most common neuropsychological consequence of stroke, with prevalence of one third of all stroke patients in acute phase although exist reports on greater frequency. Unilateral neglect is a disorder that mostly effects the patient after the lesion of the right hemisphere, mostly caused by a cerebrovascular insult (infarct or haemorrhage affecting a large area - up to two thirds of the right hemisphere), and in general the left-side neglect is the most widespread neuropsychological deficit after the lesion of the right cerebral hemisphere. Reports on the incidence of visual neglect vary and they range from 13 to 85%. Anosognosia is on the second place as neuropsychological syndrome of stroke in right hemisphere, characterized by the denial of the motor, visual or cognitive deficit. This syndrome, defined as denial of hemiparesis or hemianopsia, is a common disorder verified in 17-28% of all patents with acute brain stoke. There are different reports on frequency of delirium in acute stroke, from 24 to 48%, and it is more frequent in hemorrhagic then ischemic stoke. Post stroke depression (PSD) is one of the more frequent consequences on the stroke, and the prevalence of PSD has ranged from 5 to 63% of patients in several cross-sectional studies, peaking three to six months after a stroke.


Ocular cosmesis in retinopathy of prematurity. The Cryotherapy for Retinopathy of Prematurity Cooperative Group.
We report the externally apparent outcome in the natural history cohort (n = 4099) that was followed up prospectively in the Multicenter Trial of Cryotherapy for Retinopathy of Prematurity. The overall incidence of an adverse cosmetic outcome in the survivors who were examined 12 months post term (n = 2759) was 15.1%. Adverse cosmetic outcomes included strabismus (12.8%), nystagmus (3.3%), total retrolental membrane (1%), epiphora (0.6%), corneal opacity (0.6%), cataract (0.3%), and episcleral hyperemia (0.3%). A comparable subgroup examined 24 months post term showed strabismus (14.4%), nystagmus (2.2%), epiphora (0.5%), corneal opacity (0.7%), cataract (0.5%), episcleral hyperemia (0.5%), lid fissure asymmetry (2.4%), and corneal diameter asymmetry (2.0%). The rate of adverse aesthetic outcome was greatest in eyes that had developed more severe acute retinopathy of prematurity and an unfavorable structural outcome. In patients with bilateral threshold retinopathy of prematurity who underwent no therapeutic ocular procedures, other than randomized assignment to undergo cryotherapy in one eye, more frequent adverse cosmetic outcomes were found in the untreated eyes.


Estimating the Effects of Habitat and Biological Interactions in an Avian Community.
We used repeated sightings of individual birds encountered in community-level surveys to investigate the relative roles of habitat and biological interactions in determining the distribution and abundance of each species. To analyze these data, we developed a multispecies N-mixture model that allowed estimation of both positive and negative correlations between abundances of different species while also estimating the effects of habitat and the effects of errors in detection of each species. Using a combination of single- and multispecies N-mixture modeling, we examined for each species whether our measures of habitat were sufficient to account for the variation in encounter histories of individual birds or whether other habitat variables or interactions with other species needed to be considered. In the community that we studied, habitat appeared to be more influential than biological interactions in determining the distribution and abundance of most avian species. Our results lend support to the hypothesis that abundances of forest specialists are negatively affected by forest fragmentation. Our results also suggest that many species were associated with particular types of vegetation as measured by structural attributes of the forests. The abundances of 6 of the 73 species observed in our study were strongly correlated. These species included large birds (American Crow (Corvus brachyrhynchos) and Red-winged Blackbird (Agelaius phoeniceus)) that forage on the ground in open habitats and small birds (Red-eyed Vireo (Vireo olivaceus), House Wren (Troglodytes aedon), Hooded Warbler (Setophaga citrina), and Prairie Warbler (Setophaga discolor)) that are associated with dense shrub cover. Species abundances were positively correlated within each size group and negatively correlated between groups. Except for the American Crow, which preys on eggs and nestlings of small song birds, none of the other 5 species is known to display direct interactions, so we suspect that the correlations may have been associated with species-specific responses to habitat components not adequately measured by our covariates.


Monochloramine induced DNA fragmentation in gastric cell line MKN45.
Monochloramine (NH2Cl) is known to be one of the virulence factors in Helicobacter pylori-associated gastric mucosal injury. The present study was designed to examine NH2Cl-evoked DNA fragmentation in the gastric epithelial cell line MKN45. NH2Cl was produced by mixing NH3 with sodium hypochlorite (NaClO). MKN45 cells were exposed to NH2Cl, NH3, or NaClO in Hanks' balanced salt solution. DNA cleavage was evaluated quantitatively by photometeric enzyme immunoassay for the in vitro determination of cytoplasmic mono- and oligonucleosomes. Damage to the plasma membrane was assessed by measuring the activity of lactate dehydrogenase in the supernatants. Separately, DNA ladder formation was performed to confirm the incidence of DNA fragmentation. NH2Cl (0.001-0.01 mM) significantly increased the cytoplasmic mono- and oligonucleosomes, suggesting the incidence of DNA cleavage. The DNA ladder was clearly evoked by NH2Cl. NH2Cl induced a DNA fragmentation, one of the important aspects in apoptosis, in the gastric cell line MKN45.


Neuroendocrine neoplasms of the genitourinary tract in adults: cross-sectional imaging spectrum.
Neuroendocrine (NE) neoplasms of the genitourinary (GU) tract in adults are rare tumors with distinct histopathology and variable biological behavior and imaging findings. They may be primary or metastatic in origin. The spectrum of primary GU tract NE neoplasms includes carcinoid, small cell carcinoma, large cell NE carcinoma, and paraganglioma. The tumors commonly show positivity to specific immunohistochemical markers and characteristic dense-core granules at the ultra-structural level. Although imaging findings are nonspecific and accurate differentiation from the more common malignancies of the individual organs is not possible, cross-sectional imaging modalities play an important role in the diagnosis, staging, and surveillance of these tumors. Somatostatin receptor scintigraphy (octreotide scan) may be useful in the detection and treatment of metastatic disease in select patients. Knowledge of the various NE tumors of the adult GU tract and familiarity with their pathological and imaging findings permit optimal patient management.


Wearable technology: using Google Glass as a teaching tool.
Wearable technology holds great promise in revolutionising healthcare delivery. The benefits can also be seen in medical education and delivering healthcare in remote places. We report the use of Google Glass technology as a teaching tool in broadcasting a procedure onto a mobile phone as a viewer, replacing expensive and often cumbersome existing equipment.


Variations in protein-flavin hydrogen bonding in a light, oxygen, voltage domain produce non-Arrhenius kinetics of adduct decay.
Light, oxygen, voltage (LOV) domains utilize a conserved blue light-dependent mechanism to control a diverse array of effector domains in biological and engineered proteins. Variations in the kinetics and efficiency of LOV photochemistry fine-tune various aspects of the photic response. Characterization of the kinetics of a key aspect of this photochemical mechanism in EL222, a blue light responsive DNA binding protein from Erythrobacter litoralis HTCC2594, reveals unique non-Arrhenius behavior in the rate of dark-state cleavage of the photochemically generated adduct. Sequence analysis and mutagenesis studies establish that this effect stems from a Gln to Ala mutation unique to EL222 and homologous proteins from marine bacteria. Kinetic and spectroscopic analyses reveal that hydrogen bonding interactions between the FMN N1, O2, and ribityl hydroxyls and the surrounding protein regulate photocycle kinetics and stabilize the LOV active site from temperature-induced alteration in local structure. Substitution of residues interacting with the N1-O2 locus modulates adduct stability, structural flexibility, and sequestration of the active site from bulk solvent without perturbation of light-activated DNA binding. Together, these variants link non-Arrhenius behavior to specific alteration of an H-bonding network, while affording tunability of photocycle kinetics.


Determination of DNA methylation levels using Illumina HumanMethylation450 BeadChips.
DNA methylation is a modifiable epigenetic phenomenon that has a strong influence over transcriptional regulation and as such has been consistently implicated in development and disease. Several platforms are targeted toward the identification of DNA methylation changes that might be pertinent to the disease process and include regional analysis (e.g., pyrosequencing) as well as genome-wide analysis (e.g., next-generation sequencing and microarray). The Illumina HumanMethylation450 BeadChip is one of the most comprehensive microarray platforms available, and due to the high costs associated with next-generation sequencing, it is becoming a widely used tool for the analysis of genome-wide DNA methylation levels. Providing quantitative DNA methylation levels at 482,421 CpG sites within CpG islands, shores, and shelves, as well as intergenic regions, the HumanMethylation450 BeadChip can allow accurate assessment of differential methylation across large studies. This chapter outlines the laboratory methodologies associated with performing the Illumina Infinium Methylation Assay, including bisulfite conversion, whole-genome amplification, BeadChip hybridization, XStain procedures, and imaging systems. Furthermore, this chapter provides an outline of data analysis tools, including the GenomeStudio pipeline, quality control measures, and additional statistical considerations. This comprehensive overview can aid not only in performing the Illumina Infinium Methylation Assay but also in the interpretation of data derived from this platform.