Bacillus subtilis spore display of laccase for evolution under extreme conditions of high concentrations of organic solvent.
Protein libraries were displayed on the spore coat of Bacillus subtilis, and this method was demonstrated as a tool for directed evolution under extreme conditions. Escherichia coli, yeast, and phage display suffer from protein folding, and viability issues. On the other hand, spores avoid folding concerns by the natural sporulation process, and they remain viable under harsh chemical and physical environments. The naturally occurring B. subtilis spore coat protein, CotA, was evolved for improved activity under conditions of high organic solvent concentrations. CotA is a laccase, which is a copper-containing oxidase enzyme. A CotA library was expressed on the spore coat, and ∼ 3000 clones were screened at 60% dimethyl sulfoxide (DMSO). A Thr480Ala variant (Thr480Ala-CotA) was identified that was 2.38-fold more active than the wild-type CotA. In addition, Thr480Ala-CotA was more active with different concentrations of DMSO ranging from 0 to 70%. The mutant was also found to be more active compared with the wild-type CotA in different concentrations of methanol, ethanol, and acetonitrile.


Whole-genome sequencing shows that patient-to-patient transmission rarely accounts for acquisition of Staphylococcus aureus in an intensive care unit.
Strategies to prevent Staphylococcus aureus infection in hospitals focus on patient-to-patient transmission. We used whole-genome sequencing to investigate the role of colonized patients as the source of new S. aureus acquisitions, and the reliability of identifying patient-to-patient transmission using the conventional approach of spa typing and overlapping patient stay. Over 14 months, all unselected patients admitted to an adult intensive care unit (ICU) were serially screened for S. aureus. All available isolates (n = 275) were spa typed and underwent whole-genome sequencing to investigate their relatedness at high resolution. Staphylococcus aureus was carried by 185 of 1109 patients sampled within 24 hours of ICU admission (16.7%); 59 (5.3%) patients carried methicillin-resistant S. aureus (MRSA). Forty-four S. aureus (22 MRSA) acquisitions while on ICU were detected. Isolates were available for genetic analysis from 37 acquisitions. Whole-genome sequencing indicated that 7 of these 37 (18.9%) were transmissions from other colonized patients. Conventional methods (spa typing combined with overlapping patient stay) falsely identified 3 patient-to-patient transmissions (all MRSA) and failed to detect 2 acquisitions and 4 transmissions (2 MRSA). Only a minority of S. aureus acquisitions can be explained by patient-to-patient transmission. Whole-genome sequencing provides the resolution to disprove transmission events indicated by conventional methods and also to reveal otherwise unsuspected transmission events. Whole-genome sequencing should replace conventional methods for detection of nosocomial S. aureus transmission.


Essential elements of personalized medicine.
Genomic information has been promoted as the basis for "personalized" health care. We considered the benefits provided by genomic testing in context of the concept of personalized medicine. We evaluated current and potential uses of genomic testing in health care, using prostate cancer as an example, and considered their implications for individualizing or otherwise improving health care. Personalized medicine is most accurately seen as a comprehensive effort to tailor health care to the individual, spanning multiple dimensions. While genomic tests will offer many potential opportunities to improve the delivery of care, including the potential for genomic research to offer opportunities to improve prostate cancer screening and treatment, such advances do not in themselves constitute a paradigm shift in the delivery of health care. Rather, personalized medicine is based on a partnership between clinician and patient that utilizes shared decision making to determine the best health care options among the available choices, weighing the patient's personal values and preferences together with clinical findings. This approach is particularly important for difficult clinical decisions involving uncertainty and trade-offs, such as those involved in prostate cancer screening and management. The delivery of personalized medicine also requires adequate health care access and assurance that basic health needs have been met. Substantial research investment will be needed to identify how genomic tests can contribute to this effort.


Incorporating Economic Policy Into A 'Health-In-All-Policies' Agenda.
Recognizing the health effects of nonhealth policies, scholars and others seeking to improve Americans' health have advocated the implementation of a culture of health-which would call attention to and prioritize health as a key outcome of policy making across all levels of government and in the private sector. Adopting this "health-in-all-policies" lens, policy makers are paying increasing attention to health impacts as they debate policies in areas such as urban planning, housing, and transportation. Yet the health impacts of economic policies that shape the distribution of income and wealth are often overlooked. Pooling data from all fifty states for the period 1990-2010, we provide a broad portrait of how economic policies affect health. Overall, we found better health outcomes in states that enacted higher tax credits for the poor or higher minimum wage laws and in states without a right-to-work law that limits union power. Notably, these policies focus on increasing the incomes of low-income and working-class families, instead of on shaping the resources available to wealthier individuals. Incorporating these findings into a health-in-all-policies agenda will require leadership from the health sector, including a willingness to step into core and polarizing debates about redistribution.


Characterization of 28 novel patients expands the mutational and phenotypic spectrum of Lowe syndrome.
The oculocerebrorenal syndrome of Lowe (OCRL) is a rare X-linked multi-systemic disorder, almost always characterized by the triad of congenital cataract, cognitive and behavioral impairment and a proximal tubulopathy. Twenty-eight novel patients with suspected Lowe syndrome were studied. All patients carried OCRL gene defects with mutational hot spots at CpG dinucleotides. Mutations previously unknown in Lowe syndrome were observed in ten of the 28 patients, and carriership was identified in 30.4 % of the mothers investigated. Mapping the exact breakpoints of a complete OCRL gene deletion revealed involvement of several flanking repeat elements. We noted a similar pattern of documented clinically relevant symptoms, and even though the patient cohort comprised relatively young patients, 32 % of these patients already showed advanced chronic kidney disease. Thrombocytopenia was seen in several patients, and hyperosmia and/or hyperacusis were reported recurrently. A p.Asp523Asn mutation in a Polish patient, associated with the typical cerebrorenal spectrum but with late cataract (10 year), was also evident in two milder affected Italian brothers with ocular involvement of similar progression. We have identified clinical features in 28 patients with suspected Lowe syndrome that had not been recognized in Lowe syndrome prior to our study. We also provide further evidence that OCRL mutations cause a phenotypic continuum with selective and/or time-dependent organ involvement. At least some of these mutants might exhibit a genotype-phenotype correlation.


Microstructural differences in the human posterior sclera as a function of age and race.
The purpose of this study was to quantify the age and race-related differences in the microstructural organization of the human posterior sclera. Such differences may contribute to the predisposition of primary open-angle glaucoma in various high-risk populations. Posterior-temporal scleras from 75 right eyes were procured at an average distance of 3.5 mm from the center of the optic nerve head (ONH). A light-scattering device was used to investigate the matrix organization of posterior scleral fibers around the ONH. In addition to the degree of alignment (via eccentricity), the percentage occurrence of fiber angles within equatorial and meridionally aligned bins was quantified as a function of depth, sex, age, and race. There were 20 African Americans, 55 Caucasians, 49 males, 26 females, in this study, all falling within three age groups (<30, n = 8; 30-60, n = 33; and >60 years, n = 34). Three scleral layers, normalized for depth, were examined. For all ages and both races, fibers were preferentially oriented equatorially at all layers (P < 0.001). The African Americans had a significantly higher percentage of occurrence of meridional fibers than did the Caucasians (P < 0.001). The percentage occurrence of meridional fibers decreased significantly from the inner to the outer layers of the posterior sclera (P < 0.001). Statistically significant microstructural differences were found in the posterior sclera between African American and Caucasian donors. Ongoing work is focused on identifying whether such microstructural differences play a role in the higher prevalence of glaucoma in African American populations.


Surgical infection society guidelines for vaccination after traumatic injury.
Recommendations for vaccination of injured patients against infection are evolving. Newly-recognized infections, safety considerations, changing epidemiology, and redefinition of patient groups at risk are factors that may influence vaccine development priorities and recommendations for immunization. However, recommendations must often be formulated based on incomplete data, forcing reliance on expert opinion to address some crucial questions. These guidelines provide evidence-based recommendations for the prevention or treatment of infectious morbidity and mortality after traumatic injury, such as soft tissue wounds, human or animal bites, or after splenectomy. A panel of experts conducted a thorough review of published literature, as well as information posted on the internet at the websites of the U.S. Centers for Disease Control and Prevention, among others. MEDLINE was searched for the period 1966-2004 using relevant terms including "anthrax," "rabies," "tetanus," "tetanus toxoid," and " splenectomy," in combination with "vaccine" and "immunization." The Cochrane database was searched also. Reference lists were cross-referenced for additional relevant citations. All published reports were analyzed for quality and graded, with the strength of the recommendation proportionate to the quality of the supporting evidence. Recommendations are provided for pre- and post-exposure prophylaxis of rabies and anthrax. For tetanus prophylaxis, recommendations are provided for prophylaxis of acute wounds stratified y age and prior immunization status, and for immunization of persons at high risk. After splenectomy, it is recommended that all persons ages 2-64 years receive 23- valent pneumococcal vaccine and meningococcal vaccine, with Haemophilus influenzae type B vaccine administered to high-risk patients as well (all are Grade D recommendations). Vaccination should be given two weeks before elective splenectomy (Grade C), or two weeks after emergency splenectomy (Grade D). A booster dose of pneumococcal vaccine is recommended after five years (Grade D); no re- vaccination recommendation is made for meningococcal or Haemophilus influenzae type B vaccine. Recommendations for prophylaxis of splenectomized children under the age of five years are also provided. There are limited data on the use of vaccines after injury. This document brings together a disparate literature of variable quality into a discussion of the infectious risks after injury relevant to vaccine administration, a summary of safety and adverse effects of vaccines, and evidence-based recommendations for vaccination.


Mammalian numb-interacting protein 1/dual oxidase maturation factor 1 directs neuronal fate in stem cells.
In this study, we describe a role for the mammalian Numb-interacting protein 1 (Nip1) in regulation of neuronal differentiation in stem cells. The expression of Nip1 was detected in the developing mouse brain, embryonic stem cells, primary neuronal stem cells, and retinoic acid-treated P19 embryonal carcinoma cells. The highest expression of Nip1 was observed in undifferentiated neuronal stem cells and was associated with Duox1-mediated reactive oxygen species ROS production. Ectopic nip1 expression in P19 embryonal carcinoma cells induced neuronal differentiation, and this phenotype was also linked to elevated ROS production. The neuronal differentiation in nip1-overexpressing P19 cells was achieved in a retinoic acid-independent manner and was corroborated by an increase in the expression of the neuronal basic helix-loop-helix transcription factors and neural-lineage cell markers. Furthermore, depletion of nip1 by short hairpin RNA led to a decrease in the expression of neuronal basic helix-loop-helix transcription factors and ROS. However, inhibition of ROS production in nip1-overexpressing P19 cells restricted but did not extinguish neuronal differentiation. Microarray and mass spectrometry analysis identified intermediate filaments as the principal cytoskeletal elements affected by up-regulation of nip1. We show here the first evidence for a functional interaction between Nip1 and a component of the nuclear lamina, lamin A/C. associated with a neuronal-specific phenotype. Taken together, our data reveal an important role for Nip1 in the guidance of neuronal differentiation through ROS generation and modulation of intermediate filaments and implicate Nip1 as a novel intrinsic regulator of neuronal cell fate.


The landscape of cardiovascular care in pediatric cancer patients and survivors: a survey by the ACC Pediatric Cardio-Oncology Work Group.
To enhance the understanding of cardiovascular care delivery in childhood cancer patients and survivors. A 20-question survey was created by the Pediatric Cardio-oncology Work Group of the American College of Cardiology (ACC) Cardio-oncology Section to assess the care, management, and surveillance tools utilized to manage pediatric/young adult cardio-oncology patients. The survey distribution was a collaborative effort between Cardio-oncology Section and membership of the Adult Congenital and Pediatric Cardiology Section (ACPC) of the ACC. Sixty-five individuals, all self-identified as physicians, responded to the survey. Most respondents (n = 58,89%) indicated childhood cancer patients are regularly screened prior to and during cancer therapy at their centers, predominantly by electrocardiogram (75%), standard echocardiogram (58%) and advanced echocardiogram (50%) (i.e. strain, stress echo). Evaluation by a cardiologist prior to/during therapy was reported by only 8(12%) respondents, as compared to post-therapy which was reported by 28 (43%, p < 0.01). The most common indications for referral to cardiology at pediatric centers were abnormal test results (n = 31,48%) and history of chemotherapy exposure (n = 27,42%). Of note, during post-treatment counseling, common cardiovascular risk-factors like blood pressure (31,48%), lipid control (22,34%), obesity & smoking (30,46%) and diet/exercise/weight loss (30,46%) were addressed by fewer respondents than was LV function (72%). The survey data demonstrates that pediatric cancer patients are being screened by EKG and/or imaging prior to/during therapy at most centers. Our data, however, highlight the potential for greater involvement of a cardiovascular specialist for pre-treatment evaluation process, and for more systematic cardiac risk factor counseling in posttreatment cancer survivors.


Following polypeptide folding and assembly with conformational switches.
Conformational transitions are a crucial factor in the vast majority of protein misfolding diseases. In most of these cases, the change in conformation is accompanied by the formation of insoluble aggregates, which often precludes a detailed characterization at the molecular level. Therefore, much effort has been put into the development of simplified, easy-to-synthesize peptide models that can be used to elucidate the molecular processes that underlie the conformational switch. For a design to be successful, two, sometimes concomitantly fulfilled, requirements are of importance. First, it is essential to create inherent structural ambiguity. This is usually achieved by combining the most prominent characteristics of different folds within a consensus sequence. Second, a stimulus-sensitive functionality that responds to alterations in the environment, such as pH, ionic strength or the presence of metal ions, is often needed to control structural conversion and to shift the equilibrium in either direction.


Intensive individualized reinforcement education is important for the prevention of hypoglycemia in patients with type 2 diabetes.
We investigated whether an intensive individualized reinforcement education program could influence the prevention of hypoglycemic events in patients with type 2 diabetes. From March 2013 to September 2013, patients aged 35 to 75 years with type 2 diabetes who had not previously participated in diabetes education, and treated with insulin or a sulfonylurea-containing regimen were included in the study. After structured group education, the patients assigned to the intensive individualized education group (IT) were requested to visit for reinforcement. All subjects in the IT were encouraged to self-manage dose adjustments. Participants in both groups (control group [CG, group education only; n=22] and IT [n=24]) attended follow-up visits at 2, 8, 12, and 24 weeks. At each visit, all patients were asked whether they had experienced hypoglycemia. The total study population consisted of 20 men (43.5%; mean age and diabetic duration of 55.9±11.0 and 5.1±7.3 years, respectively). At 24 weeks, there were no significant differences in hemoglobin A1c values between the CG and IT. The total number of hypoglycemic events per patient was 5.26±6.5 in the CG and 2.58±2.3 times in the IT (P=0.004). Adherence to lifestyle modification including frequency of exercise, self-monitoring of blood glucose, or dietary habit was not significantly different between the groups. However, adherence to hypoglycemia management, especially the dose adjustment of medication, was significantly higher in the IT compared with the CG. Compared with the structured group education, additional IT resulted in additional benefits in terms of avoidance of hypoglycemia and treating hypoglycemia in patients with type 2 diabetes.


In ovo electroporation in chick midbrain for studying gene function in dopaminergic neuron development.
Dopaminergic neurons located in the ventral midbrain control movement, emotional behavior, and reward mechanisms. The dysfunction of ventral midbrain dopaminergic neurons is implicated in Parkinson's disease, Schizophrenia, depression, and dementia. Thus, studying the regulation of midbrain dopaminergic neuron differentiation could not only provide important insight into mechanisms regulating midbrain development and neural progenitor fate specification, but also help develop new therapeutic strategies for treating a variety of human neurological disorders. Dopaminergic neurons differentiate from neural progenitors lining the ventricular zone of embryonic ventral midbrain. The development of neural progenitors is controlled by gene expression programs. Here we report techniques utilizing electroporation to express genes specifically in the midbrain of Hamburger Hamilton (HH) stage 11 (thirteen somites, 42 hours) chick embryos. The external development of chick embryos allows for convenient experimental manipulations at specific embryonic stages, with the effects determined at later developmental time points. Chick embryonic neural tubes earlier than HH stage 13 (nineteen somites, 48 hours) consist of multipotent neural progenitors that are capable of differentiating into distinct cell types of the nervous system. The pCAG vector, which contains both a CMV promoter and a chick β-actin enhancer, allows for robust expression of Flag or other epitope-tagged constructs in embryonic chick neural tubes. In this report, we emphasize special measures to achieve regionally restricted gene expression in embryonic midbrain dopaminergic neuron progenitors, including how to inject DNA constructs specifically into the embryonic midbrain region and how to pinpoint electroporation with small custom-made electrodes. Analyzing chick midbrain at later stages provides an excellent in vivo system for plasmid vector-mediated gain-of-function and loss-of-function studies of midbrain development. Modification of the experimental system may extend the assay to other parts of the nervous system for performing fate mapping analysis and for investigating the regulation of gene expression.


Dendritic glycopolymers based on dendritic polyamine scaffolds: view on their synthetic approaches, characteristics and potential for biomedical applications.
In this review we highlight the potential for biomedical applications of dendritic glycopolymers based on polyamine scaffolds. The complex interplay of the molecular characteristics of the dendritic architectures and their specific interactions with various (bio)molecules are elucidated with various examples. A special role of the individual sugar units attached to the dendritic scaffolds and their density is identified, which govern ionic and H-bond interactions, and biological targeting, but to a large extent are also responsible for the significantly reduced toxicity of the dendritic glycopolymers compared to their polyamine scaffolds. Thus, the application of dendritic glycopolymers in drug delivery systems for gene transfection but also as therapeutics in neurodegenerative diseases has great promise.


Expression of truncated PITX3 in the developing lens leads to microphthalmia and aphakia in mice.
Microphthalmia is a severe ocular disorder, and this condition is typically caused by mutations in transcription factors that are involved in eye development. Mice carrying mutations in these transcription factors would be useful tools for defining the mechanisms underlying developmental eye disorders. We discovered a new spontaneous recessive microphthalmos mouse mutant in the Japanese wild-derived inbred strain KOR1/Stm. The homozygous mutant mice were histologically characterized as microphthalmic by the absence of crystallin in the lens, a condition referred to as aphakia. By positional cloning, we identified the nonsense mutation c.444C>A outside the genomic region that encodes the homeodomain of the paired-like homeodomain transcription factor 3 gene (Pitx3) as the mutation responsible for the microphthalmia and aphakia. We examined Pitx3 mRNA expression of mutant mice during embryonic stages using RT-PCR and found that the expression levels are higher than in wild-type mice. Pitx3 over-expression in the lens during developmental stages was also confirmed at the protein level in the microphthalmos mutants via immunohistochemical analyses. Although lens fiber differentiation was not observed in the mutants, strong PITX3 protein signals were observed in the lens vesicles of the mutant lens. Thus, we speculated that abnormal PITX3, which lacks the C-terminus (including the OAR domain) as a result of the nonsense mutation, is expressed in mutant lenses. We showed that the expression of the downstream genes Foxe3, Prox1, and Mip was altered because of the Pitx3 mutation, with large reductions in the lens vesicles in the mutants. Similar profiles were observed by immunohistochemical analysis of these proteins. The expression profiles of crystallins were also altered in the mutants. Therefore, we speculated that the microphthalmos/aphakia in this mutant is caused by the expression of truncated PITX3, resulting in the abnormal expression of downstream targets and lens fiber proteins.


Magnetic nanoparticles as gene delivery agents: enhanced transfection in the presence of oscillating magnet arrays.
Magnetic nanoparticle-based gene transfection has been shown to be effective in combination with both viral vectors and with non-viral agents. In these systems, therapeutic or reporter genes are attached to magnetic nanoparticles which are then focused to the target site/cells via high-field/high-gradient magnets. The technique has been shown to be efficient and rapid for in vitro transfection and compares well with cationic lipid-based reagents, producing good overall transfection levels with lower doses and shorter transfection times. In spite of its potential advantages (particularly for in vivo targeting), the overall transfection levels do not generally exceed those of other non-viral agents. In order to improve the overall transfection levels while maintaining the advantages inherent in this technique, we have developed a novel, oscillating magnet array system which adds lateral motion to the particle/gene complex in order to promote transfection. Experimental results indicate that the system significantly enhances overall in vitro transfection levels in human airway epithelial cells compared to both static field techniques (p<0.005) and the cationic lipids (p<0.001) tested. In addition, it has the previously demonstrated advantages of magnetofection-rapid transfection times and requiring lower levels of DNA than cationic lipid-based transfection agents. This method shows potential for non-viral gene delivery both in vitro and in vivo.


Long-term efficacy of antitachycardia pacing for supraventricular and ventricular tachycardias.
Over a 14-year period, 53 patients received implanted pacemakers to assist in the control of recurrent tachycardias. Indications were: prevention of tachycardia in 2 patients with supraventricular tachycardia (SVT), and 4 with ventricular tachycardia (VT); termination of tachycardia (15 SVT, 20 VT); and long-term periodic programmed electrical stimulation with potential for tachycardia termination (12 VT). Pacemakers for prevention of VT were implanted in 3 patients with prolonged QT interval syndromes and 1 in whom Holter monitoring showed a significant reduction in ectopic activity during pacing. Pacers were implanted for tachycardia termination only after patients underwent a rigorous protocol aimed at achieving 100 trials of the proposed modality. Patients with tachycardia also requiring antibradycardia pacemakers received pacemakers capable of noninvasive programmed stimulation for use during follow-up. There were no tachycardia recurrences among those patients in whom pacemakers were implanted for prevention. Pacers capable of outpatient programmed stimulation were useful, and it may be desirable to expand their use. The 15 patients with pacers designed for termination of SVT were followed for a mean of 68 months. Among these, actuarial continuation of pacing efficacy was 93% at 1 year, and 78% at 5 years. The 20 patients with pacers for termination of VT were followed for a mean of 37 months. Actuarial efficacy was 78% at 1 year, and 55% at 5 years. Sudden death occurred in 4 of these patients, none clearly pacer related. Pacemakers can play a major therapeutic role in some patients with recurrent tachycardias. The role of such pacemakers in patients with VT may be expanded with the advent of combined pacer-defibrillators.(ABSTRACT TRUNCATED AT 250 WORDS)


Development of the Avian Dorsal Thalamus: Patterns and Gradients of Neurogenesis.
The dorsal thalamus is a region of the diencephalon that relays sensory and motor information between areas of the brain stem and the telencephalon. Although a dorsal thalamic region is recognized in all vertebrates and believed to be homologous, little is known about how the regions within it evolved and whether some or all regions within the dorsal thalamus are homologous among different vertebrate species. To characterize the gradients and patterns of neurogenesis of the avian dorsal thalamus, a single application of a low dose of bromodeoxyuridine (BrdU) was delivered to each chick between embryonic day (E)3 and E8 (stages 21 and 34), and chicks were followed up to E8 or E10 (stage 34 or 36). Comparisons of anti-BrdU labeling patterns across the different injection days suggest that nearly all dorsal thalamic neurons are born early in chick embryogenesis, between E3 and E8. Furthermore, neurons in the lateral, dorsal, and posterior parts of the dorsal thalamus are generally born earlier than those in the medial, ventral, and anterior parts. Analyses of the birth dates for nine regions show that the general pattern of neurogenesis in the avian dorsal thalamus resembles that of homologous regions within the rodent thalamus, with the exception of the auditory region, the nucleus ovoidalis, which is born later than the mammalian auditory medial geniculate nucleus. The similar pattern of neurogenesis in birds and mammals may represent a highly conserved developmental pattern that was present in the common ancestor of living birds and mammals, or may represent independently derived states. Additional studies in reptiles and amphibians are needed to distinguish between these evolutionary histories.


Giant circular polarization conversion in layer-by-layer nonchiral metamaterial.
We studied numerically the transmission properties of a kind of layer-by-layer nonchiral metamaterial. Simulation results show that under certain off-normal incidence, giant circular polarization conversion occurs for both the right and left circularly polarized waves with a roughly 1 GHz operation band. Meanwhile, the copolarization transmissions are almost suppressed to zero, leading to the high purity circular polarization transformation. This phenomenon of giant circular polarization conversion is assumed to suffer from the strong magnetic response, which is illustrated by the surface current distributions of the structure. Compared with chiral structures, this nonchiral structure is easier to design and fabricate and is expected to be used as a promising circular polarization transformer.


Pink frilly dresses and the avoidance of all things "girly": children's appearance rigidity and cognitive theories of gender development.
Many young children pass through a stage of gender appearance rigidity; girls insist on wearing dresses, often pink and frilly, whereas boys refuse to wear anything with a hint of femininity. In 2 studies, we investigated the prevalence of this apparent hallmark of early gender development and its relation to children's growing identification with a gender category. Study 1a examined the prevalence of this behavior and whether it would exhibit a developmental pattern of rigidity followed by flexibility, consistent with past research on identity-related cognitions. Interviews with 76 White, middle-class parents and their 3- to 6-year-old children revealed that about two thirds of parents of 3- and 4-year-old girls and almost half (44%) of parents of 5- and 6-year-old boys reported that their children had exhibited a period of rigidity in their gender-related appearance behavior. Appearance rigidity was not related to parents' preferences for their children's gender-typed clothing. Study 1b examined whether cognitive theories of identity development could shed light on gender appearance rigidity. The more important and positive children considered their gender and the more children understood that gender categories remain stable over time (gender stability), the more likely children were to wear gender-typed outfits. In Study 2, we extended this research to a more diverse population and found that gender appearance rigidity was also prevalent in 267 4-year-old children in the United States from African American, Chinese, Dominican, and Mexican immigrant low-income backgrounds. Results suggest that rigid gender-related appearance behavior can be seen among young children from different backgrounds and might reflect early developing cognitions about gender identity.


A comparison of three methods of pain control for posterior spinal fusions in adolescent idiopathic scoliosis.
Retrospective cohort study. To compare the efficacy of patient-controlled analgesia (PCA) with morphine alone, a single preoperative intrathecal morphine injection and PCA (IT/PCA), and epidural catheter infusion without PCA (EPI) for postoperative pain control after posterior spinal fusion (PSF) and segmental spinal instrumentation (SSI) in adolescent idiopathic scoliosis (AIS). Postoperative pain control after PSF and SSI in AIS can be managed in different ways. EPI provides for a longer period of pain relief but is reliant on the correct placement and maintenance of the catheter in the epidural space. A single preoperative intrathecal morphine injection also provides for long acting analgesia. No direct comparison of these 3 methods of postoperative pain control has been reported. An IRB-approved retrospective chart review was performed at 2 institutions from 1997 to 2005. The medical record was reviewed to determine pain scores after surgery at multiple time periods. The 3 groups were compared using Student t test and chi and significance was defined as P < 0.05. There was no statistical difference in the gender, age, magnitude of curve, or number of levels fused in the IT/PCA (N = 42), PCA (N = 41), or EPI (N = 55) groups. Postoperative pain scores were lowest in the IT/PCA group in the first 8 hours (P < 0.05) but the pain scores in the EPI group were then lower through 24 hours (P < 0.05). Total morphine use (mg/kg) was lower in the IT/PCA group compared with the PCA group at 12 hours and 24 hours (P = 0.0001). Return to solid food ingestion was quickest in the EPI group (2.0 days) followed by the IT/PCA (2.6 days) and PCA alone (3.2 days) (P < 0.002). Respiratory depression and transient neurologic change occurred most frequently in the EPI group (EPI 11/55 pts vs. 1/42 IT/PCA vs. 0/41 PCA P < 0.001). Pruritus was greatest in the epidural group (11/55 P < 0.05). There were no intraoperative somatosensory-evoked potential changes or permanent neurologic injury recorded in any group. An EPI controls postoperative pain for the longest period of time and allows for a quicker return to consumption of solid foods. However, a single preoperative intrathecal morphine injection controls the pain equally for the first 24 hours with less pruritus and with less adverse events thus requiring less nursing and physician intervention after PSF and SSI in AIS. All methods were safe with no neurologic injury recorded.


Optical measurement of picometer displacements of transparent microscopic objects.
An optical laser differential interferometer, based on a modified differential interference contrast microscope, has been developed to measure the thermal motion of microscopic protrusions (stereocilia) that are the sites of mechanoelectrical transduction in auditory hair cells. The measurement sensitivity was limited at high frequencies mainly by shot noise, at intermediate frequencies by acoustic interference, and at low frequencies by thermal drift. The power spectral density of the instrumental noise was found to be as low as 1. pm/ radicalHz in the shot noise regime. We could, thus, measure the Brownian motion of hair bundles over the frequency range from 1 Hz-100 kHz. Experimental data that test and demonstrate the sensitivity and spatial discrimination of the instrument were found to be in agreement with theoretical estimates.


Nuclear overhauser spectroscopy of chiral CHD methylene groups.
Nuclear magnetic resonance spectroscopy (NMR) can provide a great deal of information about structure and dynamics of biomolecules. The quality of an NMR structure strongly depends on the number of experimental observables and on their accurate conversion into geometric restraints. When distance restraints are derived from nuclear Overhauser effect spectroscopy (NOESY), stereo-specific assignments of prochiral atoms can contribute significantly to the accuracy of NMR structures of proteins and nucleic acids. Here we introduce a series of NOESY-based pulse sequences that can assist in the assignment of chiral CHD methylene protons in random fractionally deuterated proteins. Partial deuteration suppresses spin-diffusion between the two protons of CH2 groups that normally impedes the distinction of cross-relaxation networks for these two protons in NOESY spectra. Three and four-dimensional spectra allow one to distinguish cross-relaxation pathways involving either of the two methylene protons so that one can obtain stereospecific assignments. In addition, the analysis provides a large number of stereospecific distance restraints. Non-uniform sampling was used to ensure optimal signal resolution in 4D spectra and reduce ambiguities of the assignments. Automatic assignment procedures were modified for efficient and accurate stereospecific assignments during automated structure calculations based on 3D spectra. The protocol was applied to calcium-loaded calbindin D9k. A large number of stereospecific assignments lead to a significant improvement of the accuracy of the structure.


Birth weight and weight at 1 year are independent determinants of bone mass in the seventh decade: the Hertfordshire cohort study.
Several studies have shown relationships between growth in early life and adult bone mass; in this article, we evaluate the relative contributions of pre- and postnatal factors to bone mass in the seventh decade. A total of 498 eight men and 468 women who were born in Hertfordshire during the period 1931-1939 and still living there were recruited. Detailed birth records were available. Participants attended a clinic where they completed a detailed health questionnaire, before performance of anthropometric measurements and bone densitometry of the proximal femur and lumbar spine (Hologic QDR 4500). Birth weight was associated with bone mineral content in both men (proximal femur: r=0.16, p=0.0003; lumbar spine: r=0.10, p=0.03) and women (proximal femur: r=0.16, p=0.0008; lumbar spine: r=0.11, p=0.03); relationships with bone mineral density were weaker and were significant at the proximal femur in men only (p=0.03). Relationships between weight at 1 y and bone mineral content were even stronger (proximal femur: men r=0.22, p<0.0001; women r=0.14, p=0.002). In men, 18% of the variance in proximal femoral bone area was explained by a model that included birth weight, weight at 1 y, and adult weight, with the relative contributions attributed to each being 2.8, 6.8, and 8.2%, respectively. In women, similar modeling produced figures of 6.7, 4.2, and 3.9% (overall variance of 15% in proximal femoral bone area). Hence, weight at each of these three points in the life course is important in the determination of adult bone mass, with greater contributions of earlier growth to bone size and mineral content than to bone mineral density.


More evidence of cannabis efficacy in restless legs syndrome.
Restless legs syndrome (RLS) is one of the most disabling and sometimes painful sensorimotor ailment of the nervous system that has only in recent years become more widely accepted as a clinical disorder with its own distinct features. Usually, symptoms respond well to dopamine agonists, anticonvulsants, or opiates, but still a subset of patients remains refractory to medical therapy and/or reports serious side effects. Recently, patients' statement of a remarkable and total remission of RLS symptoms following cannabis use has been reported. Here, we confirm and extend these findings to more patients with RLS. The antinociceptive effect of marijuana has been documented in many painful neurological conditions, and the potential benefit of cannabis use in patients with refractory RLS should therefore be questioned by robust clinical trials.


A chimeric mouse model to study human iPSC-derived neurons: the case of a truncating SHANK3 mutation.
Using human induced pluripotent stem cells (iPSC), recent studies have shown that the events underlying autism spectrum disorders (ASD) can occur during neonatal development. We previously analyzed the iPSC-derived pyramidal cortical neurons of a subset of patients with ASD carrying de novo heterozygous mutations in postsynaptic SHANK3 protein, in culture. We reported altered spinogenesis of those neurons. The transplantation of human iPSC-derived neuronal precursors into mouse brain represents a novel option for in vivo analysis of mutations affecting the human brain. In this study, we transplanted the neuronal precursor cells (NPC) into the cortex of newborn mice to analyze their integration and maturation at early stages of development and studied axonal projections of transplanted human neurons into adult mouse brain. We then co-transplanted NPC from a control individual and from a patient carrying a de novo heterozygous SHANK3 mutation. We observed a reduction in cell soma size of selective neuronal categories and in axonal projections at 30 days post-transplantation. In contrast to previous in vitro studies, we did not observe any alteration in spinogenesis at this early age. The humanized chimeric mouse models offer the means to analyze ASD-associated mutations further and provide the opportunity to visualize phenotypes in vivo.


Analysis of a stochastic SIR epidemic on a random network incorporating household structure.
This paper is concerned with a stochastic SIR (susceptible-->infective-->removed) model for the spread of an epidemic amongst a population of individuals, with a random network of social contacts, that is also partitioned into households. The behaviour of the model as the population size tends to infinity in an appropriate fashion is investigated. A threshold parameter which determines whether or not an epidemic with few initial infectives can become established and lead to a major outbreak is obtained, as are the probability that a major outbreak occurs and the expected proportion of the population that are ultimately infected by such an outbreak, together with methods for calculating these quantities. Monte Carlo simulations demonstrate that these asymptotic quantities accurately reflect the behaviour of finite populations, even for only moderately sized finite populations. The model is compared and contrasted with related models previously studied in the literature. The effects of the amount of clustering present in the overall population structure and the infectious period distribution on the outcomes of the model are also explored.


Return to sport after ACL reconstruction.
Objective guidelines permitting safe return to sport following anterior cruciate ligament (ACL) reconstruction are infrequently used. The purpose of this study was to determine the published return to sport guidelines following ACL reconstruction in Level I randomized controlled trials. A systematic review was performed using Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. Level I randomized controlled trials were included if they reported a minimum 2-year follow-up after ACL reconstruction and return to sport criteria. Outcomes analyzed were the timing of initiation of return to sport, follow-up duration, and use of quantitative/qualitative criteria to determine return to sport. Forty-nine studies were included (N=4178; 68% male; mean patient age, 27.5±3.2 years; mean follow-up, 3.0±1.9 years; mean time from injury to reconstruction, 379±321 days). Ninety-six percent of reconstructions used autograft and 87% were single-bundle reconstructions. Lysholm score, single-leg hop, isokinetic strength, and KT-1000 or KT-2000 arthrometer (MEDmetric, San Diego, California) testing were performed in 67%, 31%, 31%, and 82% of studies, respectively. Only 5 studies reported whether patients were able to successfully return to sport. Ninety percent and 65% of studies failed to use objective criteria or any criteria, respectively, to permit return to sport. Description of permission/allowance to return to sport was highly variable and poor. Twenty-four percent of studies failed to report when patients were allowed return to sport without restrictions. Overall, 39%, 45%, and 51% of studies permitted running at 3 months, return to cutting/pivoting sports at 6 months, and return to sport without restrictions at 6 months, respectively. Further research into validated return to sport guidelines is necessary to fill the existing void in contemporary literature and to guide clinical practice.


Clinical outcomes and management associated with major bleeding in patients with atrial fibrillation treated with apixaban or warfarin: insights from the ARISTOTLE trial.
In the Apixaban for Reduction in Stroke and Other Thromboembolic Events in Atrial Fibrillation (ARISTOTLE) trial, apixaban compared with warfarin reduced the risk of stroke, major bleed, and death in patients with atrial fibrillation. In this ancillary study, we evaluated clinical consequences of major bleeds, as well as management and treatment effects of warfarin vs. apixaban. Major International Society on Thrombosis and Haemostasis bleeding was defined as overt bleeding accompanied by a decrease in haemoglobin (Hb) of ≥2 g/dL or transfusion of ≥2 units of packed red cells, occurring at a critical site or resulting in death. Time to event [death, ischaemic stroke, or myocardial infarction (MI)] was evaluated by Cox regression models. The excess risk associated with bleeding was evaluated by separate time-dependent indicators for intracranial (ICH) and non-intracranial haemorrhage. Major bleeding occurred in 848 individuals (4.7%), of whom 126 (14.9%) died within 30 days. Of 176 patients with an ICH, 76 (43.2%) died, and of the 695 patients with major non-ICH, 64 (9.2%) died within 30 days of the bleeding. The risk of death, ischaemic stroke, or MI was increased roughly 12-fold after a major non-ICH bleeding event within 30 days. Corresponding risk of death following an ICH was markedly increased, with HR 121.5 (95% CI 91.3-161.8) as was stroke or MI with HR 21.95 (95% CI 9.88-48.81), respectively. Among patients with major bleeds, 20.8% received vitamin K and/or related medications (fresh frozen plasma, coagulation factors, factor VIIa) to stop bleeding within 3 days, and 37% received blood transfusion. There was no interaction between apixaban and warfarin and major bleeding on the risk of death, stroke, or MI. Major bleeding was associated with substantially increased risk of death, ischaemic stroke, or MI, especially following ICH, and this risk was similarly elevated regardless of treatment with apixaban or warfarin. These results underscore the importance of preventing bleeding in anti-coagulated patients. ClinicalTrials.gov Identifier: NCT00412984.


Selecting, training and supervising nurses to treat depression in the medically ill: experience and recommendations from the SMaRT oncology collaborative care trials.
Collaborative care programs to treat comorbid depression in the medically ill often have general (nonpsychiatric) nurses care managers. In this paper, we aim to provide practical recommendations for their selection, training and supervision. Based on more than 10 years of experience of selecting, training and supervising general nurses to deliver a highly effective collaborative care programme called "Depression Care for People with Cancer," we describe the problems encountered and the solutions adopted to optimize the selection, training and supervision of nurse care managers. To select nurses for the role of care manager, we found that role plays enabled us to assess nurses' ability to interact with distressed patients and their capacity for self-reflection better than simple interviews. To train the nurses, we found that a structured program that mirrored the treatment manual and included simulated practice was best. To achieve effective supervision, we found that having sessions led by senior psychiatrists facilitated both constructive feedback to the nurses and effective review of the management of cases. We recommend that the selection, training and supervision of general nurses use the strategies outlined if they are to maximize the benefit that patients achieve from collaborative care programs.


Incidence and Risk Factors for Dementia in Type 2 Diabetes Mellitus: A Nationwide Population-Based Study in Korea.
Diabetes mellitus is associated with an increased risk of dementia. We aimed to comprehensively analyze the incidence and risk factors for dementia and young-onset dementia (YOD) in diabetic patients in Korea using the National Health Insurance Service data. Between January 1, 2009 and December 31, 2012, a total of 1,917,702 participants with diabetes were included and followed until the date of dementia diagnosis or until December 31, 2015. We evaluated the incidence and risk factors for all dementia, Alzheimer's disease (AD), and vascular dementia (VaD) by Cox proportional hazards analyses. We also compared the impact of risk factors on the occurrence of YOD and late-onset dementia (LOD). During an average of 5.1 years of follow-up, the incidence of all types of dementia, AD, or VaD was 9.5, 6.8, and 1.3/1,000 person-years, respectively, in participants with diabetes. YOD comprised 4.8% of all dementia occurrence, and the ratio of AD/VaD was 2.1 for YOD compared with 5.5 for LOD. Current smokers and subjects with lower income, plasma glucose levels, body mass index (BMI), and subjects with hypertension, dyslipidemia, vascular complications, depression, and insulin treatment developed dementia more frequently. Vascular risk factors such as smoking, hypertension, and previous cardiovascular diseases were more strongly associated with the development of VaD than AD. Low BMI and a history of stroke or depression had a stronger influence on the development of YOD than LOD. The optimal management of modifiable risk factors may be important for preventing dementia in subjects with diabetes mellitus.


Acetylation of histone H2B mirrors that of H4 and H3 at the chicken beta-globin locus but not at housekeeping genes.
Acetylation of histones H4 and H3 targeted to promoters/enhancers is linked to the activation of transcription, whereas widespread, long range acetylation of the same histones has been linked to the requirement for open chromatin at transcriptionally active loci and regions of V(D)J recombination. Using affinity-purified polyclonal antibodies to tetra/tri-acetylated histone H2B in chromatin immunoprecipitation (ChIP) assays with mononucleosomes from 15-day chicken embryo erythrocytes, a high resolution distribution of H2B acetylation has been determined and compared with that of H4 and H3 at the same genes/loci. At the beta-globin locus, the H2B acetylation is high throughout and in general mirrors that of H3 and H4, consistent with the observation of co-precipitation of hyperacetylated H4 together with the hyperacetylated H2B. In contrast, at the weakly expressed genes glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and Gas41 (housekeeping) and carbonic anhydrase (tissue specific), very little or no hyperacetylated H2B was found despite the presence of acetylated H4 and H3 at their promoters and proximal transcribed sequences. At the inactive lysozyme and ovalbumin genes essentially no acetylation of H2B, H3, or H4 was observed. Acetylation of H2B appears to be principally a feature of only the most actively transcribed genes/loci.


Characterization of insertion mutations in the Saccharomyces cerevisiae MSH1 and MSH2 genes: evidence for separate mitochondrial and nuclear functions.
The MSH1 and MSH2 genes of Saccharomyces cerevisiae are predicted to encode proteins that are homologous to the Escherichia coli MutS and Streptococcus pneumoniae HexA proteins and their homologs. Disruption of the MSH1 gene caused a petite phenotype which was established rapidly. A functional MSH1 gene present on a single-copy centromere plasmid was incapable of rescuing the established msh1 petite phenotype. Analysis of msh1 strains demonstrated that mutagenesis and large-scale rearrangement of mitochondrial DNA had occurred. 4',6-Diamidino-2-phenylindole (DAPI) staining of msh1 yeast revealed an aberrant distribution of mtDNA. Haploid msh2 mutants displayed an increase of 85-fold in the rate of spontaneous mutation to canavanine resistance. Sporulation of homozygous msh2/msh2 diploids gave rise to a high level of lethality which was compounded during increased vegetative growth prior to sporulation. msh2 mutations also affected gene conversion of two HIS4 alleles. The his4x mutation, lying near the 5' end of the gene, was converted with equal frequency in both wild-type and msh2 strains. However, many of the events in the msh2 background were post-meiotic segregation (PMS) events (46.4%) while none (< 0.25%) of the aberrant segregations in wild type were PMS events. The his4b allele, lying 1.6 kb downstream of his4x, was converted at a 10-fold higher frequency in the msh2 background than in the corresponding wild-type strain. Like the his4x allele, his4b showed a high level of PMS (30%) in the msh2 background compared to the corresponding wild-type strain where no (< 0.26%) PMS events were observed. These results indicate that MSH1 plays a role in repair or stability of mtDNA and MSH2 plays a role in repair of 4-bp insertion/deletion mispairs in the nucleus.


Exposure to bloom-like concentrations of two marine Synechococcus cyanobacteria (strains CC9311 and CC9902) differentially alters fish behaviour.
Coastal California experiences large-scale blooms of Synechococcus cyanobacteria, which are predicted to become more prevalent by the end of the 21st century as a result of global climate change. This study investigated whether exposure to bloom-like concentrations of two Synechococcus strains, CC9311 and CC9902, alters fish behaviour. Black perch (Embiotoca jacksoni) were exposed to Synechococcus strain CC9311 or CC9902 (1.5 × 10(6) cells ml(-1)) or to control seawater in experimental aquaria for 3 days. Fish movement inside a testing arena was then recorded and analysed using video camera-based motion-tracking software. Compared with control fish, fish exposed to CC9311 demonstrated a significant preference for the dark zone of the tank in the light-dark test, which is an indication of increased anxiety. Furthermore, fish exposed to CC9311 also had a statistically significant decrease in velocity and increase in immobility and they meandered more in comparison to control fish. There was a similar trend in velocity, immobility and meandering in fish exposed to CC9902, but there were no significant differences in behaviour or locomotion between this group and control fish. Identical results were obtained with a second batch of fish. Additionally, in this second trial we also investigated whether fish would recover after a 3 day period in seawater without cyanobacteria. Indeed, there were no longer any significant differences in behaviour among treatments, demonstrating that the sp. CC9311-induced alteration of behaviour is reversible. These results demonstrate that blooms of specific marine Synechococcus strains can induce differential sublethal effects in fish, namely alterations light-dark preference behaviour and motility.


Evaluation of Two Dosing Regimens for Nivolumab in Combination With Ipilimumab in Patients With Advanced Melanoma: Results From the Phase IIIb/IV CheckMate 511 Trial.
Nivolumab 1 mg/kg plus ipilimumab 3 mg/kg (NIVO1+IPI3) is approved for first-line treatment of patients with advanced melanoma in several countries. We conducted a phase IIIb/IV study (CheckMate 511) to determine if nivolumab 3 mg/kg plus ipilimumab 1 mg/kg (NIVO3+IPI1) improves the safety profile of the combination. Patients (N = 360) age 18 years or older with previously untreated, unresectable stage III or IV melanoma were randomly assigned 1:1 to NIVO3+IPI1 or NIVO1+IPI3 once every 3 weeks for four doses. After 6 weeks, all patients received NIVO 480 mg once every 4 weeks until disease progression or unacceptable toxicity. The primary end point was a comparison of the incidence of treatment-related grade 3 to 5 adverse events (AEs) between groups. Secondary end points included descriptive analyses of objective response rate, progression-free survival, and overall survival. The study was not designed to formally demonstrate noninferiority of NIVO3+IPI1 to NIVO1+IPI3 for efficacy end points. At a minimum follow-up of 12 months, incidence of treatment-related grade 3 to 5 AEs was 34% with NIVO3+IPI1 versus 48% with NIVO1+IPI3 ( P = .006). In descriptive analyses, objective response rate was 45.6% in the NIVO3+IPI1 group and 50.6% in the NIVO1+IPI3 group, with complete responses in 15.0% and 13.5% of patients, respectively. Median progression-free survival was 9.9 months in the NIVO3+IPI1 group and 8.9 months in the NIVO1+IPI3 group. Median overall survival was not reached in either group. The CheckMate 511 study met its primary end point, demonstrating a significantly lower incidence of treatment-related grade 3-5 AEs with NIVO3+IPI1 versus NIVO1+IPI3. Descriptive analyses showed that there were no meaningful differences between the groups for any efficacy end point, although longer follow up may help to better characterize efficacy outcomes.


Modulation of soy isoflavones bioavailability and subsequent effects on bone health in ovariectomized rats: the case for equol.
Soy products are of particular interest because of their potential health benefits in a range of hormonal conditions, such as osteoporosis, due to their high content in phytoestrogens. Because equol, the main metabolite from soy isoflavones, is thought to be powerful, the present study was designated to evaluate the bone-sparing effects of equol by either providing the molecule through the diet or by eliciting its endogenous production by modulating intestinal microflora by short-chain fructooligosaccharides (sc-FOS) or live microbial (Lactobacillus casei) together with daidzein, its precursor. A comparison with daidzein and genistein was also performed. Rats (3 months old) were ovariectomised (OVX) or sham-operated (SH). Ovariectomised rats were randomly assigned to six experimental diets for 3 months: a control diet (OVX), the control diet supplemented with either genistein (G), or daidzein (D), or equol (E) at the level of 10 microg/g body weight/d. The remaining OVX rats were given daidzein at the dose of 10 mug/g body weight/d, simultaneously with short-chain FOS (Actilight, Beghin-Meiji) (D+FOS) or Lactobacillus casei (Actimel, Danone) (D+L). The SH rats were given the same control diet as OVX. Genistein, daidzein or equol exhibited a bone sparing effect. Indeed, total femoral bone mineral density (BMD) was significantly enhanced (compared to that of OVX rats), as was the metaphyseal compartment. Bone strength was improved by E consumption, but not by genistein or daidzein given alone. As far as the FOS diet is concerned, the addition of prebiotics significantly raised efficiency of the daidzein protective effect on both femoral BMD and mechanical properties. The effects of lactobacillus were similar, except that the increase in metaphyseal-BMD was not significant. In conclusion, long-term equol consumption, like genistein and daidzein, in the ovariectomized rat, provides bone sparing effects. Adding indigestible sugars, such as FOS or live microbial as L. casei, in the diet significantly improves daidzein protective effects on the skeleton.


Temperature-Induced uncoupling of cell cycle regulators.
The early stages of development involve complex sequences of morphological changes that are both reproducible from embryo to embryo and often robust to environmental variability. To investigate the relationship between reproducibility and robustness we examined cell cycle progression in early Drosophila embryos at different temperatures. Our experiments show that while the subdivision of cell cycle steps is conserved across a wide range of temperatures (5-35 °C), the relative duration of individual steps varies with temperature. We find that the transition into prometaphase is delayed at lower temperatures relative to other cell cycle events, arguing that it has a different mechanism of regulation. Using an in vivo biosensor, we quantified the ratio of activities of the major mitotic kinase, Cdk1 and one of the major mitotic phosphatases PP1. Comparing activation profile with cell cycle transition times at different temperatures indicates that in early fly embryos activation of Cdk1 drives entry into prometaphase but is not required for earlier cell cycle events. In fact, chromosome condensation can still occur when Cdk1 activity is inhibited pharmacologically. These results demonstrate that different kinases are rate-limiting for different steps of mitosis, arguing that robust inter-regulation may be needed for rapid and ordered mitosis.


Pulsed digital holographic interferometry with 694- and 347-nm wavelengths.
A method for deformation analysis and shape measurement based on digital holography is presented. Two wavelengths, 694 and 347 nm, are used. The object is illuminated with the two wavelengths at the same time, and digital holograms are recorded on a CCD chip. The information corresponding to the two wavelengths is separated in the Fourier domain, and the phase corresponding to the wave fronts is calculated. By recording holograms with two different wavelengths at the same time, we can get measurements of deformations or shape with different sensitivities. Experimental results are presented.


The use of proteomics to identify novel therapeutic targets for the treatment of disease.
The completion of the Human Genome Project has revealed a multitude of potential avenues for the identification of therapeutic targets. Extensive sequence information enables the identification of novel genes but does not facilitate a thorough understanding of how changes in gene expression control the molecular mechanisms underlying the development and regulation of a cell or the progression of disease. Proteomics encompasses the study of proteins expressed by a population of cells, and evaluates changes in protein expression, post-translational modifications, protein interactions, protein structure and splice variants, all of which are imperative for a complete understanding of protein function within the cell. From the outset, proteomics has been used to compare the protein profiles of cells in healthy and diseased states and as such can be used to identify proteins associated with disease development and progression. These candidate proteins might provide novel targets for new therapeutic agents or aid the development of assays for disease biomarkers. This review provides an overview of the current proteomic techniques available and focuses on their application in the search for novel therapeutic targets for the treatment of disease.


Disparities and survival among breast cancer patients.
Although rates of survival for women with breast cancer have improved, the survival disparity between African American and white women in the United States has increased. To determine whether this survival disparity persists in an insured population with access to medical care. In this retrospective cohort study, we extracted data from the tumor registries of six nonprofit, integrated health care delivery systems affiliated with the Cancer Research Network and assessed the survival of African American (n = 2276) and white (n = 18 879) female enrollees who were diagnosed with invasive breast cancer from January 1, 1993, through December 31, 1998. Cox proportional hazards regression was used to estimate the death rate among African American women relative to that of white women after adjustment for potential explanatory factors including stage at diagnosis, tumor characteristics, and treatment. Five-year survival was lower for African American women (73.8%) than for white women (81.6%). African American women were less likely to have tumor characteristics with good prognosis. Controlling for age at diagnosis, stage, grade, tumor size, and estrogen and progesterone receptor status, the adjusted hazard rate ratio for African American women was 1.34 (95% confidence interval = 1.22 to 1.46). Similar risks were found among women ages 20-49 and 50 and older. Controlling for treatment slightly lowered the hazard rate ratio to 1.31 (95% confidence interval = 1.20 to 1.43). Among women with invasive breast cancer, being insured and having access to medical care does not eliminate the survival disparity for African American women.


Cash Transfers, Young Women's Economic Well-Being, and HIV Risk: Evidence from HPTN 068.
Despite the large interest in economic interventions to reduce HIV risk, little research has been done to show whether there are economic gains of these interventions for younger women and what intermediary role economic resources play in changing participants' sexual behavior. This paper contributes to this gap by examining the impacts of a conditional cash transfer (CCT) for young women in South Africa on young women's economic resources and the extent to which they play a role in young women's health and behavior. We used data from HIV Prevention Trials Network 068 study, which provided transfers to young women (in addition to their parents) conditional on the young woman attending at least 80% of school days in the previous month. We found that the CCT increased young women's economic wellbeing in terms of having savings, spending money, being unindebted, and food secure. We also investigated heterogeneous effects of the program by household economic status at baseline because the program was not specifically poverty targeted and found that the results were driven by young women from the poorest families. From these results, we examined heterogeneity by baseline poverty for other outcomes related to HIV risk including sexual behavior and psychosocial well-being. We found psychosocial well-being benefits in young women from the poorest families and that economic wellbeing gains explained much these impacts.


The temporal dynamics of relationships between cannabis, psychosis and depression among young adults with psychotic disorders: findings from a 10-month prospective study.
The aim was to examine the temporal relationships over 10 months between cannabis use and symptoms of psychosis and depression in people with schizophrenia and related disorders. The design was a prospective study of 101 patients with schizophrenia and related disorders who were assessed monthly over 10 months on medication compliance, cannabis and other drug use, symptoms of depression and symptoms of psychosis. Linear regression methods to assess relationships between cannabis use and symptoms of psychosis and depression while adjusting for serial dependence, medication compliance and other demographic and clinical variables. Cannabis use predicted a small but statistically significant increase in symptoms of psychosis, but not depression, after controlling for other differences between cannabis users and non-users. Symptoms of depression and psychosis did not predict cannabis use. Continued cannabis use by persons with schizophrenia predicts a small increase in psychotic symptom severity but not vice versa.


Learning to Anticipate Egocentric Actions by Imagination.
Anticipating actions before they are executed is crucial for a wide range of practical applications, including autonomous driving and robotics. In this paper, we study the egocentric action anticipation task, which predicts future action seconds before it is performed for egocentric videos. Previous approaches focus on summarizing the observed content and directly predicting future action based on past observations. We believe it would benefit the action anticipation if we could mine some cues to compensate for the missing information of the unobserved frames. We then propose to decompose the action anticipation into a series of future feature predictions. We imagine how the visual feature changes in the near future and then predicts future action labels based on these imagined representations. Differently, our ImagineRNN is optimized in a contrastive learning way instead of feature regression. We utilize a proxy task to train the ImagineRNN, i.e., selecting the correct future states from distractors. We further improve ImagineRNN by residual anticipation, i.e., changing its target to predicting the feature difference of adjacent frames instead of the frame content. This promotes the network to focus on our target, i.e., the future action, as the difference between adjacent frame features is more important for forecasting the future. Extensive experiments on two large-scale egocentric action datasets validate the effectiveness of our method. Our method significantly outperforms previous methods on both the seen test set and the unseen test set of the EPIC Kitchens Action Anticipation Challenge.


Delayed enhancement cardiac computed tomography for the assessment of myocardial infarction: from bench to bedside.
A large number of studies support the increasingly relevant prognostic value of the presence and extent of delayed enhancement (DE), a surrogate marker of fibrosis, in diverse etiologies. Gadolinium and iodinated based contrast agents share similar kinetics, thus leading to comparable myocardial characterization with cardiac magnetic resonance (CMR) and cardiac computed tomography (CT) at both first-pass perfusion and DE imaging. We review the available evidence of DE imaging for the assessment of myocardial infarction (MI) using cardiac CT (CTDE), from animal to clinical studies, and from 16-slice CT to dual-energy CT systems (DECT). Although both CMR and gadolinium agents have been originally deemed innocuous, a number of concerns (though inconclusive and very rare) have been recently issued regarding safety issues, including DNA double-strand breaks related to CMR, and gadolinium-associated nephrogenic systemic fibrosis and deposition in the skin and certain brain structures. These concerns have to be considered in the context of non-negligible rates of claustrophobia, increasing rates of patients with implantable cardiac devices, and a number of logistic drawbacks compared with CTDE, such as higher costs, longer scanning times, and difficulties to scan patients with impaired breath-holding capabilities. Overall, these issues might encourage the role of CTDE as an alternative for DE-CMR in selected populations.


Enhanced endocannabinoid tone as a potential target of pharmacotherapy.
The endocannabinoid system is up-regulated in numerous pathophysiological states such as inflammatory, neurodegenerative, gastrointestinal, metabolic and cardiovascular diseases, pain, and cancer. It has been suggested that this phenomenon primarily serves an autoprotective role in inhibiting disease progression and/or diminishing signs and symptoms. Accordingly, enhancement of endogenous endocannabinoid tone by inhibition of endocannabinoid degradation represents a promising therapeutic approach for the treatment of many diseases. Importantly, this allows for the avoidance of unwanted psychotropic side effects that accompany exogenously administered cannabinoids. The effects of endocannabinoid metabolic pathway modulation are complex, as endocannabinoids can exert their actions directly or via numerous metabolites. The two main strategies for blocking endocannabinoid degradation are inhibition of endocannabinoid-degrading enzymes and inhibition of endocannabinoid cellular uptake. To date, the most investigated compounds are inhibitors of fatty acid amide hydrolase (FAAH), an enzyme that degrades the endocannabinoid anandamide. However, application of FAAH inhibitors (and consequently other endocannabinoid degradation inhibitors) in medicine became questionable due to a lack of therapeutic efficacy in clinical trials and serious adverse effects evoked by one specific compound. In this paper, we discuss multiple pathways of endocannabinoid metabolism, changes in endocannabinoid levels across numerous human diseases and corresponding experimental models, pharmacological strategies for enhancing endocannabinoid tone and potential therapeutic applications including multi-target drugs with additional targets outside of the endocannabinoid system (cyclooxygenase-2, cholinesterase, TRPV1, and PGF2α-EA receptors), and currently used medicines or medicinal herbs that additionally enhance endocannabinoid levels. Ultimately, further clinical and preclinical studies are warranted to develop medicines for enhancing endocannabinoid tone.


Predicting the risk of acute kidney injury in primary care: derivation and validation of STRATIFY-AKI.
Antihypertensives reduce the risk of cardiovascular disease but are also associated with harms including acute kidney injury (AKI). Few data exist to guide clinical decision making regarding these risks. To develop a prediction model estimating the risk of AKI in people potentially indicated for antihypertensive treatment. Observational cohort study using routine primary care data from the Clinical Practice Research Datalink (CPRD) in England. People aged ≥40 years, with at least one blood pressure measurement between 130 mmHg and 179 mmHg were included. Outcomes were admission to hospital or death with AKI within 1, 5, and 10 years. The model was derived with data from CPRD GOLD (n = 1 772 618), using a Fine-Gray competing risks approach, with subsequent recalibration using pseudo-values. External validation used data from CPRD Aurum (n = 3 805 322). The mean age of participants was 59.4 years and 52% were female. The final model consisted of 27 predictors and showed good discrimination at 1, 5, and 10 years (C-statistic for 10-year risk 0.821, 95% confidence interval [CI] = 0.818 to 0.823). There was some overprediction at the highest predicted probabilities (ratio of observed to expected event probability for 10-year risk 0.633, 95% CI = 0.621 to 0.645), affecting patients with the highest risk. Most patients (>95%) had a low 1- to 5-year risk of AKI, and at 10 years only 0.1% of the population had a high AKI and low CVD risk. This clinical prediction model enables GPs to accurately identify patients at high risk of AKI, which will aid treatment decisions. As the vast majority of patients were at low risk, such a model may provide useful reassurance that most antihypertensive treatment is safe and appropriate while flagging the few for whom this is not the case.


Nutritional screening in community-dwelling older adults: a systematic literature review.
Nutrition screening is a process used to quickly identify those who may be at risk of malnutrition so that a full nutrition assessment and appropriate nutrition intervention can be provided. While many nutrition screening tools have been developed, few have been evaluated for use in older adults in the community setting. The aim of this paper is to determine the most appropriate nutrition screening tool/s, in terms of validity and reliability, for identifying malnutrition risk in older adults living in the community. Electronic databases MEDLINE, PUBMED, CINAHL and the Cochrane Library were searched for nutrition screening tools to identify malnutrition or under-nutrition for adults greater than 65 years living in the community. Ten screening tools were found for use in community-dwelling older adults and subjected to validity and/or reliability testing: Mini Nutritional Assessment-Short Form (MNA-SF), Malnutrition Universal Screening Tool (MUST), Nutrition Screening Initiative (NSI), which includes the DETERMINE Checklist and Level I and II Screen, Australian Nutritional Screening Initiative (ANSI), Seniors in the Community: Risk Evaluation for Eating and Nutrition (SCREEN I and SCREEN II), Short Nutritional Assessment Questionnaire (SNAQ), Simplified Nutritional Appetite Questionnaire (SNAQ), and two unnamed tools. MNA-SF appears to be the most appropriate nutrition screening tool for use in community-dwelling older adults although MUST and SCREEN II also have evidence to support their use. Further research into the acceptability of screening tools focusing on the outcomes of nutrition screening and appropriate nutrition intervention are required.


Socio-economic inequalities in childhood mortality in low- and middle-income countries: a review of the international evidence.
In low- and middle-income countries (LMICs), the probability of dying in childhood is strongly related to the socio-economic position of the parents or household in which the child is born. This article reviews the evidence on the magnitude of socio-economic inequalities in childhood mortality within LMICs, discusses possible causes and highlights entry points for intervention. Sources of data Evidence on socio-economic inequalities in childhood mortality in LMICs is mostly based on data from household surveys and demographic surveillance sites. Childhood mortality is systematically and considerably higher among lower socio-economic groups within countries. Also most proximate mortality determinants, including malnutrition, exposure to infections, maternal characteristics and health care use show worse levels among more deprived groups. The magnitude of inequality varies between countries and over time, suggesting its amenability to intervention. Reducing inequalities in childhood mortality would substantially contribute to improving population health and reaching the Millennium Development Goals (MDGs). The contribution of specific determinants, including national policies, to childhood mortality inequalities remains uncertain. What works to reduce these inequalities, in particular whether policies should be universal or targeted to the poor, is much debated. The increasing political attention for addressing health inequalities needs to be accompanied by more evidence on the contribution of specific determinants, and on ways to ensure that interventions reach lower socio-economic groups.


Intravenous insulin infusion in diabetic emergencies.
Continuous intravenous insulin and dextrose infusions were used in managing various diabetic emergencies. Standard and constant rates of insulin and dextrose infusion resulted in satisfactory control of blood glucose concentrations during labour, after major surgery, and in patients recovering from ketoacidosis (average insulin infusion rates 1, 2, and 3 U/h respectively). Higher infusion rates were used to correct or prevent ketoacidosis in pregnant diabetic women who had received steroids and sympathomimetic agents. The infusion method is simple, reliable, and flexible, and may help to simplify management of diverse types of diabetic emergencies.


The Evasive Enemy: Insights into the Virulence and Epidemiology of the Emerging Attaching and Effacing Pathogen Escherichia albertii.
The diarrheic attaching and effacing (A/E) pathogen Escherichia albertii was first isolated from infants in Bangladesh in 1991, although the bacterium was initially classified as Hafnia alvei Subsequent genetic and biochemical interrogation of these isolates raised concerns about their initial taxonomic placement. It was not until 2003 that these isolates were reassigned to the novel taxon Escherichia albertii because they were genetically more closely related to E. coli, although they had diverged sufficiently to warrant a novel species name. Unfortunately, new isolates continue to be mistyped as enteropathogenic E. coli (EPEC) or enterohemorrhagic E. coli (EHEC) owing to shared traits, most notably the ability to form A/E lesions. Consequently, E. albertii remains an underappreciated A/E pathogen, despite multiple reports demonstrating that many provisional EPEC and EHEC isolates incriminated in disease outbreaks are actually E. albertii Metagenomic studies on dozens of E. albertii isolates reveal a genetic architecture that boasts an arsenal of candidate virulence factors to rival that of its better-characterized cousins, EPEC and EHEC. Beyond these computational comparisons, studies addressing the regulation, structure, function, and mechanism of action of its repertoire of virulence factors are lacking. Thus, the paucity of knowledge about the epidemiology, virulence, and antibiotic resistance of E. albertii, coupled with its misclassification and its ability to develop multidrug resistance in a single step, highlights the challenges in combating this emerging pathogen. This review seeks to synthesize our current but incomplete understanding of the biology of E. albertii.


Rock-shells (Thais clavigera) as an indicator of As, Cu, and Zn contamination on the Putai Coast of the black-foot disease area in Taiwan.
This study presents the distribution of arsenic (As), copper (Cu), and zinc (Zn) in various seafoods-oysters (Crassostrea gigas), false fusus (Hemifuscus tuba), venus clams (Cyclina sineasis), green mussels (Perna viridis), blood clams (Arca granosa), flounders (Psettodes erumei), and rock-shells (Thais clavigera) collected from the Putai coast of the black-foot disease (BFD) area in Taiwan. Special attention is paid to evaluate the relationships among As, Cu, and Zn and effect of body size on metal concentration in Thais clavigera. Maximum Zn and Cu geometric mean (GM) concentrations (GM = 615 and 376 microg/g, dry wt, respectively) are found in oysters (Crassostrea gigas), and the values are much higher than those of the other organisms by about 1.7-208 and 1.8-375 times, respectively. Similarly, Thais clavigera has a high capacity for accumulating Cu and Zn collected from the same location. One interesting point is that relatively high As concentrations (GM = 65.7 microg/g, dry wt) in Thais clavigera are found as compared with those in other organisms (range from GM = 2.37 to 40.2 microg/g, dry wt). The As concentrations are significantly higher in Thais clavigera (1.62-27.7 times) than those in other organisms (p < 0.05), except for the false fusus (Hamifuscus tuba). A linear regression analysis shows a significant increase in Zn concentration with increasing Cu concentration in Thais clavigera. On the other hand, the As concentration is correlated with Cu and Zn concentrations (r = 0.77 and 0.77, respectively; p < 0.05) in Thais clavigera. Double logarithmic plots of metal content and concentration against dry-body weight and shell length show linear relationships. The result indicates that large individuals have higher contents of Cu, Zn and As, and have slopes of 1.58, 1.38, and 1.34, respectively. In addition, metal concentrations against shell length for all animal sizes also indicate that Cu, Zn and As have slopes of 1.92, 1.18, and 1.11, respectively. In conclusion, Thais clavigera has a high capactiy for accumulating As, Cu, and Zn and is a potential bioindicator for monitoring As, Cu and Zn.


Amniotic fluid phospholipid profile determined by two-dimensional thin-layer chromatography as index of fetal lung maturation.
A phospholipid profile, the main features of which were the lecithin/sphingomyelin (L/S) ratio and the presence or absence of phosphatidylglycerol (PG), was determined in amniotic fluid from 188 patients. There was a mature profile (L/S ratio of at least 2 . 0 and detectable PG) in 145 patients, including seven insulin-dependent diabetics, and noe of their babies developed respiratory distress syndrome (RDS). The L/S ratio was less than 2 . 0 and PG absent in 12 patients, nine of whose babies developed RDS, whereas only three small babies (delivered between 28 and 35 weeks because of fulminant pre-eclampsia or severe abruptio placentae) out of 31 developed RDS when the L/S ratio was less than 2 . 0 but PG was present. When amniotic fluid was collected from the vagina only one out of 69 babies developed RDS when PG was present (regardless of the L/S ratio), while all of seven babies developed RDS when PG was absent. It is concluded that the amniotic fluid phospholipid profile, particularly the presence or absence of PG, gives an accurate assessment of fetal lung maturation. The profile may prove a useful adjunct to the management of high-risk pregnancies, especially after premature membrane rupture and perhaps also when the mother is diabetic.


Germline excision of transgenes in Aedes aegypti by homing endonucleases.
Aedes (Ae.) aegypti is the primary vector for dengue viruses (serotypes1-4) and chikungunya virus. Homing endonucleases (HEs) are ancient selfish elements that catalyze double-stranded DNA breaks (DSB) in a highly specific manner. In this report, we show that the HEs Y2-I-AniI, I-CreI and I-SceI are all capable of catalyzing the excision of genomic segments from the Ae. aegypti genome in a heritable manner. Y2-I-AniI demonstrated the highest efficiency at two independent genomic targets, with 20-40% of Y2-I-AniI-treated individuals producing offspring that had lost the target transgene. HE-induced DSBs were found to be repaired via the single-strand annealing (SSA) and non-homologous end-joining (NHEJ) pathways in a manner dependent on the availability of direct repeat sequences in the transgene. These results support the development of HE-based gene editing and gene drive strategies in Ae. aegypti, and confirm the utility of HEs in the manipulation and modification of transgenes in this important vector.


A Robust Incubator to Improve Access to Microbiological Culture in Low Resource Environments.
To help address the limitations of operating conventional microbiological culture incubators in low resource environments, a new incubator design was developed and tested to meet the requirements of operation in laboratories without reliable power (power outages up to 12 contiguous hours) or climate control (ambient indoor temperatures from 5 °C to 45 °C). The device is designed to enable adherence to incubation temperatures recommended for growth detection, identification, and drug susceptibility testing (DST) of human pathogenic bacteria. During power outages, stable temperatures are maintained in the device's internal sample compartment by employing phase change material (PCM) as a bi-directional thermal battery to maintain incubation temperature. Five prototypes were tested in a laboratory setting using environmental test chambers and programmable power supplies, and three were field tested in the Lao PDR in situations of intended use. The prototypes successfully held their temperature to within ±1 °C in both laboratory environmental chamber testing as well as during the field test. The results indicate that the device will maintain stable culture temperatures across periods of intermittent power supply, while enabling normal workflow of this could greatly increase the availability of microbiological culture for diagnosis and antimicrobial resistance (AMR) monitoring.


Surgical management of hidradenitis suppurativa.
Hidradenitis suppurativa (HS) is a chronic, relapsing inflammatory disease of skin, characterized by recurrent draining sinuses and abscesses, predominantly in skin folds carrying terminal hairs and apocrine glands. This study reviewed 54 sites in 27 patients with moderate to extensive chronic inflammatory skin lesions treated surgically in our hospital from 2004 through 2009, with a follow-up of at least 6 months. A total number of 54 operative procedures were performed during the study period with 42% (23 sites) involving the axilla, 20% (11 sites) involving the gluteal area, %24 (13 sites) involving the perineal area and 12% (7 sites) involving the inguinal region. Conservative treatment methods have little or no effects especially on gluteal, perineal/perianal, axillary hidradenitis suppurativa. The morbidity associated with the established form of this disease is significant, and the only successful treatment is wide surgical excision.


Nematosomes in the human placenta.
In this study, the ultrastructure of the human placental 'nematosome' or 'glomerular body' is described. It is usually found as electron-dense aggregates forming an inclusion 0.3 to 0.5 micron in diameter and of variable length, and occurs within cytotrophoblast cells in different configurations which include spirals, concentric rings, parallel arrays, random coils and paracrystalline structures. Analysis of random electron micrographs of placentae from normal and complicated pregnancies showed that the nematosome is present in low frequencies in the first and second trimesters but is more commonly found at full term; their incidence did not differ markedly from normal in any of the pathological conditions studied, ranging from 10.4 to 21.9 per cent, and their frequency suggests that there may be several nematosomes per cell. The relationship of the nematosome to the centriole is discussed, and it is suggested that morphometric analysis may elucidate the three-dimensional structure of nematosomes, their distribution within cells and their relationship to the plasma and other membranes.


Virus surveys of Capsicum spp. in the Republic of Benin reveal the prevalence of pepper vein yellows virus and the identification of a previously uncharacterised polerovirus species.
Surveys were conducted in 2014 and 2015 in Southern and Northern Benin, respectively, to identify the viruses infecting peppers (Capsicum spp.). The samples were screened by ELISA for cucumber mosaic virus (CMV), pepper veinal mottle virus (PVMV), potato virus Y (PVY) and tomato yellow leaf curl virus (TYLCV). A generic reverse transcription PCR (RT-PCR) was used to test for the presence of poleroviruses. ELISA tests confirmed the prevalence of all viruses, while the RT-PCR detected pepper vein yellows virus (PeVYV) which is reported for the first time in Benin. A further, divergent polerovirus isolate was detected from a single pepper sample originating from southern Benin. Screening of samples collected from solanaceous plants during virus surveys in Mali (conducted in 2009) also detected this divergent polerovirus isolate in two samples from African eggplants. The complete genome sequence was obtained from the Mali isolate using transcriptome sequencing and by conventional Sanger sequencing of overlapping RT-PCR products. Based on the sequence characteristics of this isolate we propose a new polerovirus species, African eggplant yellowing virus (AeYV).


Technical and implementation issues in using next-generation sequencing of cancers in clinical practice.
Next-generation sequencing (NGS) of cancer genomes promises to revolutionise oncology, with the ability to design and use targeted drugs, to predict outcome and response, and to classify tumours. It is continually becoming cheaper, faster and more reliable, with the capability to identify rare yet clinically important somatic mutations. Technical challenges include sequencing samples of low quality and/or quantity, reliable identification of structural and copy number variation, and assessment of intratumour heterogeneity. Once these problems are overcome, the use of the data to guide clinical decision making is not straightforward, and there is a risk of premature use of molecular changes to guide patient management in the absence of supporting evidence. Paradoxically, NGS may simply move the bottleneck of personalised medicine from data acquisition to the identification of reliable biomarkers. Standardised cancer NGS data collection on an international scale would be a significant step towards optimising patient care.


DNA barcode data reveal biogeographic trends in Arctic non-biting midges.
Chironomid flies (non-biting midges) are among the most abundant and diverse animals in Arctic regions, but detailed analyses of species distributions and biogeographical patterns are hampered by challenging taxonomy and reliance on morphology for species-level identification. Here we take advantage of available DNA barcode data of Arctic Chironomidae in BOLD to analyse similarities in species distributions across a northern Nearctic - West Palearctic gradient. Using more than 260 000 barcodes representing 4666 BINs (Barcode Index Numbers) and 826 named species (some with interim names) from a combination of public and novel data, we show that the Greenland chironomid fauna shows affinities to both the Nearctic and the West Palearctic regions. While raw taxon counts indicate a strong Greenland - North American affinity, comparisons using Chao's dissimilarity metric support a slightly higher similarity between Greenland and West Palearctic chironomid communities. Results were relatively consistent across different definitions of species taxonomic units, including morphologically determined species, BINs, and superBINs based on a ∼4.5% threshold. While most taxa found in Greenland are shared with at least one other region, reflecting circum-Arctic dispersal, our results also reveal that Greenland harbours a small endemic biodiversity. Our exploratory study showcases how DNA barcoding efforts using standardized gene regions contribute to an understanding of broad-scale patterns in biogeography by enabling joint analysis of public DNA sequence data derived from diverse prior studies.


Effect of nanohypericum (Hypericum perforatum gold nanoparticles) treatment on restraint stressinduced behavioral and biochemical alteration in male albino mice.
Hypericum perforatum extract (HPE), is known for its antidepressant effect. In the present study we investigated the effect of H. perforatum gold nanoparticles (Nanohypericum-HPGNPs) protective role against restraint stress-induced behavioral and biochemical alterations in mice. Animals were immobilized for a period of 6 hrs/day. HPE (200 mg/kg) and nanohypericum (20 mg/kg) were administered 30 minutes before the animals were subjected to acute immobilized stress. Behavioral test parameters for anxiety and spatial memory were assessed followed by biochemical parmeters (lipid peroxidation, super oxide dismutase, catalase, glutathione peroxidase, reduced glutathione, etc.) subsequently. The behavior study showed severe anxiety and memory loss compared to unstressed animals. Biochemical analyses revealed an increase in lipid per oxidation, depletion of super oxide dismutase, reduced glutathione, catalase activity and glutathione per oxidase as compared to unstressed animal. Twenty one days of H. perforatum and nanohypericum treatment in a dose of 200 mg/kg and 20 mg/kg, respectively, significantly attenuated restraint stress-induced behavioral and oxidative damage. In conclusion nanohypericum prove the modest activity than the HPE.


Human mucosal-associated invariant T cells respond to Mucorales species in a MR1-dependent manner.
Activation of mucosal-associated invariant T cells (MAIT cells) by certain bacteria, viruses, and yeast is well studied, but the activation potential of filamentous moulds from the order Mucorales is not known. Here, we show a rapid response of human MAIT cells against the Mucorales species Mucor circinelloides, Rhizopus arrhizus, and Rhizopus microsporus. This activation included upregulation of CD69 and degranulation marked by increased CD107a expression, while intracellular perforin and granzyme A expression were reduced. Furthermore, blocking of the antigen-presenting molecule major histocompatibility complex class I-related abrogated MAIT cell activation demonstrating a T cell receptor-dependent stimulation by Mucorales.


Dynamic norm-based encoding for unfamiliar shapes in human visual cortex.
Previous studies have argued that faces and other objects are encoded in terms of their deviation from a class prototype or norm. This prototype is associated with a smaller neural population response compared with nonprototype objects. However, it is still unclear (1) whether a norm-based representation can emerge for unfamiliar or novel object classes through visual experience at the time scale of an experiment and (2) whether the results from previous studies are caused by the prototypicality of a stimulus, by the physical properties of individual stimuli independent from the stimulus distribution, and/or by the trial-to-trial adaptation. Here we show with a combined behavioral and event-related fMRI study in humans that a short amount of visual experience with exemplars from novel object classes determines which stimulus is represented as the norm. Prototypicality effects were observed at the behavioral level by behavioral asymmetries during a stimulus comparison task. The fMRI data revealed that class exemplars closest to the prototypes--the perceived average of each class--were associated with a smaller response in the anterior part of the visual object-selective cortex compared with other class exemplars. By dissociating between the physical characteristics and the prototypicality status of the stimuli and by controlling for trial-to-trial adaptation, we can firmly conclude for the first time that high-level visual areas represent the identity of exemplars using a dynamic, norm-based encoding principle.


Mechanisms of photodynamic inactivation of herpes simplex viruses: comparison between methylene blue, light plus electricity, and hematoporhyrin plus light.
Herpes simplex virus (HSV) types 1 and 2 have been inactivated in vitro using low concentrations of methylene blue (MB), light (lambda) plus electricity (E), or hematoporphyrin derivative (HPD) plus lambda. Both techniques introduce single strand interruptions into viral DNA, but do not make double strand ruptions into viral DNA, but do not make double strand breaks. MB, lambda plus E-treated virions adsorb normally to and penetrate susceptible cells, whereas HSV inactivated with HPC and light does not. This difference is emphasized by the induction of new viral and cell DNA synthesis after infection with MB, lambda plus E-treated virions, whereas only cell, DNA but no HSV DNA, is made subsequent to HPD and lambda exposure. These observations reflect disparate mechanisms of viral inactivation. A block(s) in viral maturation, subsequent to viral DNA synthesis, occurs as a result of treatment with MB, lambda and E, whereas HPD plus lambda-treated particles fail to enter a susceptible cell, and therefore do not initiate an infection.